Ethanol, Carnoy, and paraformaldehyde as fixative solutions for histological evaluation of preantral follicles in equine ovarian tissue.

The most adequate fixative solution for equine ovarian tissue is still to be determined as a tool to evaluate the improvement of methodological studies in assisted reproductive techniques and fertility preservation. This study aimed to evaluate a short-time ethanol 70% (ST-EtOH, 45 min) exposure as an alternative fixative compared with two classically fixatives [Carnoy's (CAR) solution and paraformaldehyde 4% (PFA)] at different fixation times (6 h, 12 h). The end points evaluated were morphology and classes of preantral follicles, follicular and stromal cell densities, and follicular and oocyte nuclear diameters in equine ovarian tissue. Ovaries (n = 6) from ovariectomized young mares were fragmented (3 × 3 × 1 mm; 20 fragments/ovary) and fixed in the tested treatments. Overall, a total of 11,661 preantral follicles were evaluated in 1444 histological slides. The ST-EtOH similarly preserved the preantral follicle morphometry and stromal cell density compared to the PFA fixative, regardless of the exposure time. Nonetheless, the CAR fixative solution had the greatest percentage of normal preantral follicles and the highest stromal cell density among all treatments. In conclusion, Carnoy's solution must be preferred compared with ST-EtOH and PFA fixatives for studies concerning the cellular morphology of equine ovarian tissue. Moreover, ST-EtOH fixative is a good alternative for equine ovarian tissue when a quick histological evaluation is required instead of more time-consuming and expensive techniques. Additional studies concerning the impact of different fixatives on the ultrastructure of cellular populations and their compatibility with IHC and molecular techniques in equine ovarian tissue are warranted.

[Descriptive comparison of serum progesterone levels during intravaginal device insertion in mares] / Comparação descritiva de dosagens séricas de progesterona durante inserção de dispositivo intravaginal em éguas

RESUMO O objetivo foi descrever os níveis séricos de progesterona (P4) de éguas tratadas com um ou dois dispositivos intravaginais de primeiro uso e com um dispositivo de segundo uso de P4, em um período de 10 dias e comparar a concentração de P4. Três éguas foram tratadas com implante intravaginal de P4 (Reprosync®; 2g de P4) por 10 dias (D0 a D10). A égua 1 recebeu dois implantes novos, a égua 2 recebeu um implante novo e a égua 3 recebeu um implante usado de P4. Coletas de sangue foram realizadas diariamente (a.m./p.m.) do D0 ao D10 e concentrações de P4 foram analisadas por quimioluminescência. A égua que recebeu dois dispositivos apresentou numericamente maior concentração sérica média de P4 (5,11±2,24ng/mL) do que a égua que recebeu um dispositivo novo (3,27±0,94) e do que a égua que recebeu um dispositivo usado (2,78±0,72). As éguas que receberam um dispositivo (novo ou usado) tiveram médias de P4 relativamente semelhantes numericamente. Concluiu-se que a concentração sérica de P4 de éguas recebendo diferentes quantidades de dispositivos intravaginais de P4 parece diferir de acordo com o número de implantes inseridos. Ainda assim, todos os tratamentos atingiram concentrações séricas de P4 capazes de bloquear a ovulação.

The Effect of Treatment with Oestradiol Benzoate on Oestrus Expression and Endometrial Oedema in Anovulatory and Cyclic Mares.

Oestrogens treatment is often used to induce oestrus behaviour in anoestrous mares to aid in the collection of stallion semen and as recipient mares to receive embryos when combined with progesterone. However, there are no studies to describe the effect of dose and individual mare on the intensity and duration of the response, in both anoestrous and cyclic mares. In Experiment 1, 13 anoestrous mares were treated with one of five doses of oestradiol benzoate (OB) (1, 1.5, 2, 3 and 4 mg) per mare in five consecutive treatment periods (n = 65), to determine the response in terms of endometrial oedema and oestrous behaviour. Experiment 2 and 3 used 3 mg of OB in cyclic mares to confirm or deny the presence of an active corpus luteum (CL). There was a dose rate of OB and individual mare effect (p < 0.05) on the intensity and persistence of endometrial oedema and oestrous behaviour. A total of 2 mg OB was enough to induce endometrial oedema and oestrous behaviour within 48 h in most mares. Mares with an active CL did not show endometrial oedema following treatment of 3 mg OB.

Luz artificial na reprodução equina ­ Revisão / Artificial lighting in the equine reproduction ­ A review

Os equinos são animais poliéstricos sazonais, fotoperiódico positivos, apresentando ciclos reprodutivos em estações definidas do ano, primavera e verão, e estações com ausência ou baixa atividade reprodutiva, outono e inverno. A necessidade da maximização do período reprodutivo, bem como a definição de um ano hípico iniciando em julho, no hemisfério sul (para algumas raças como o Puro Sangue de Corrida e o Quarto de milha) levou ao desenvolvimento de técnicas para diminuir o tempo que essas éguas permanecem em anestro reprodutivo. Uma das técnicas mais utilizadas destaca-se o uso de iluminação artificial. Consistindo no fornecimento de luz branca incandescente ou led azul ou branco em baias, piquetes ou por meio de máscaras, a fim de simular os efeitos de dias longos no organismo assim liberar o eixo hipotalâmico-hipofisário-gonadal. Para estimular o retorno antecipado à ciclicidade, a égua deve ser exposta a um estímulo de 14 a 15 horas de luz diárias, totalizando 9 a 10 horas de escuridão, ou receber um estímulo de 1 hora de luz, aproximadamente 9 horas após o anoitecer. É fundamental que a égua esteja com escore corporal adequado e alguns cuidados com o fornecimento da luz artificial sejam atendidos, como a continuidade no fornecimento para o sucesso da técnica.(AU)

Horses are seasonally breeding animals which means that they natural breed in specific seasons (Spring, and Summer), with non or low reproductive activity on the other seasons (Winter and Fall). The development of techniques to shorten the anoestrus can be necessary to anticipate the breeding season, starting in July in the Southern Hemisphere, for some breeds such as Thoroughbred and Quarter Horses. The most common technique is the use of artificial lighting with incandescent, or blue/white led light, applied in stalls, paddocks or with a light mask to mimic the effects of a long day and release the hipotalamic-hipophysarie-gonadal axis. To stimulate an early return to cyclicity, the mare should be exposed to a 14-to-15-hour light stimulus daily, totalling 9 to 10 hours of darkness, or receive a 1-hour light stimulus, approximately 9 hours after dusk. It is fundamental that the mare has an adequate body condition score and some awareness with the provision of artificial light are considered, such as the continuity of the light program for the success of the technique.(AU)

Biópsia endometrial na égua: importância no diagnóstico da subfertilidade e principais achados / Endometrial biopsy in the mare: importance in the diagnosis of subfertility and main findings

A biópsia endometrial na égua é uma técnica simples e bastante antiga. É uma importante ferramenta para o diagnóstico de patologia no endométrio nesta espécie utilizada para determinar o estágio do ciclo estral, para detectar a presença de inflamações agudas ou crônicas e para determina a possibilidade do útero levar a gestação a termo. Mais recentemente outras alterações tais como a má-diferenciação e as angiopatias, também estão sendo avaliadas em amostras de biópsia endometrial em éguas, uma vez que também podem interferir no prognóstico reprodutivo destas fêmeas. Esta revisão tem por objetivo descrever a avaliação da biópsia endometrial e as principais alterações que ocorrem no endométrio das éguas.(AU)

Endometrial biopsy in the mare is a simple and quite old technique. It is an important tool for the diagnosis of pathology in the endometrium in this species, used to determine the stage of the estrous cycle, to detect the presence of acute or chronic inflammation and to determine the possibility of the uterus carrying the pregnancy to term. More recently, other alterations, such as poor differentiation and angiopathies, are also being evaluated in endometrial biopsy samples in mares, since they may also interfere with the reproductive prognosis of these females. This review aims to describe the evaluation of the endometrial biopsy and the main changes that occur in the endometrium of mares.(AU)

The equine endometrium and its multiple connections, interrelationships and clinical effects in aged mares

Mares, which make up 90% of animals requiring reproductive assistance in commercial equine production systems, are subjected to strong breeding and reproductive efficiency pressure throughout their lives, but the effects of aging on overall and reproductive systems in particular; it often means that we are forcing the natural homeostasis mechanisms under pressure over time. The objective of this brief communication is to summarize some concepts related to the role and importance of equine endometrium, particularly in old mares, the most challenged category to achieve productive and efficiency goals established by humans. Endometritis is one of the most frequent and recurrent pathologies in equine gynecology and is responsible for enormous economic losses in the industry. Alterations of the microbiota, molecular signals and uterine endocrine microenvironment can lead to infertility, but degenerative phenomena associated with age and parity can lead to endometriosis altering maternal recognition of pregnancy and pregnancy maintenance. Maintaining horses in an adequate nutrition status and environment is essential to achieve pregnancy but also to obtain healthy offspring.(AU)

Uma história em construção: Manguinhos-Maré no tempo presente / A history under construction: Manguinhos-Maré in the present time

Resumo As duas primeiras décadas do século XXI foram representativas para a história contemporânea da Fundação Oswaldo Cruz e demonstraram seu protagonismo no cenário nacional da saúde. O artigo apresenta transformações espaciais ocorridas no território ocupado pela instituição a partir de 2000. Relacionam-se as transformações no uso dos territórios com sua política institucional no contexto nacional e são ressaltadas as demandas sociais que impactaram a instituição - transformações urbanas de grande porte e pandemias. O trabalho averigua, sob os pressupostos da história do tempo presente, as mudanças ocorridas no campus Manguinhos-Maré e no discurso de sujeitos institucionais gestores e projetistas. Para isso, conjuga análise dos relatórios institucionais, da base digital cartográfica e de imagens aéreas.

Abstract The two first decades of the twenty-first century were representative of the contemporary history of the Oswaldo Cruz Foundation (Fundação Oswaldo Cruz) and demonstrated its active role in the national health scenario. This article discusses the spatial transformations that took place in the territory occupied by the foundation from the year 2000 onward. The transformations in the use of this territory and it's institutional policy within the national context are described, along with the social demands that have impacted the institution, namely large-scale urban transformations and pandemics. This research uses the premises of the history of the present to investigate the changes that took place on the Manguinhos-Maré campus and in the discourse of the institutions involved in managing and planning these changes through analysis of institutional reports, digital mapping, and aerial images.

The effect of priming and duration of oestradiol benzoate treatment before progesterone administration on embryo development and survival in anestrous recipient mares.

The effect of three different hormonal protocols to prepare anestrous recipient mares on embryo survival was evaluated. The first group consisted of only progesterone administration (NE) 4 days before embryo transfer, while the recipients from the other two groups received a single administration of 2.5 mg of oestradiol benzoate (SE) 2 days earlier or 8 mg of oestradiol split in increasing doses for 5 consecutive days (LE) ending 3 days before progesterone treatment. The likelihood of recovering an embryo 2 days after transfer was 46.1% (6/13), 62.5% (5/8) and 85.7% (6/7) for recipient mares from the no oestrus, short and long oestrous groups respectively (p = .09). In conclusion, the presence and duration of oestradiol treatment before progesterone administration tended to influence the embryo survival in anestrous recipients 2 days after transfer. The surviving embryos recovered from the three different groups of recipients did not show any difference in size and morphology.

Hematological, serum biochemical results, and treatment approach of an Arabian mare with squamous cell carcinoma of the vulva - case report / Resultados hematológicos, bioquímicos séricos e abordagem de tratamento de uma égua Árabe com carcinoma escamoso de células da vulva - relato de caso

A 20-year-old Arabian mare, was brought to the obstetrics and gynecology clinic with the complaint of an unknown wound in the genital area. Although the mare's general condition was good, examination of the genital area revealed a wide-open wound in the vulva and posterior vagina, characterized by fibrotic growth. The biopsy sample was sent to the pathology laboratory. A diagnosis of squamous cell carcinoma (SCC) was made after histopathological examination. Blood samples for serum biochemistry and hematological tests were collected from the jugular vein. Hematological and serum biochemical findings are generally used to diagnose diseases in animals and humans. Hematological and serum biochemical findings are important with genital SCC. Therefore, the findings of these values are given in this case report.

Uma égua de 20 anos (raça árabe), foi levada à clínica de obstetrícia e ginecologia com a queixa de uma ferida desconhecida na área genital. Embora o estado geral da égua fosse bom, o exame da área genital revelou uma ferida amplamente aberta na vulva e na vagina posterior, caracterizada pelo crescimento fibrótico. A amostra da biópsia foi enviada para o laboratório de patologia. Um diagnóstico de carcinoma espinocelular (CEC) foi feito como resultado do exame histopatológico. Amostras de sangue para bioquímica sérica e testes hematológicos foram coletadas da veia jugular. Os resultados hematológicos e bioquímicos séricos são geralmente usados para diagnosticar doenças em animais e seres humanos. Os achados hematológicos e bioquímicos séricos são importantes com o SCC genital. Portanto, as descobertas destes valores são dadas neste relatório de caso.

Fluconazol intra-estromal - eficácia na cirurgia de abscesso estromal em equino / Intrastromal fluconazole - effectiveness in the surgery of stromal abscess in a horse

Background: Due to its ocular microflora, the equine species is predisposed to develop mycotic ulcers which, when notproperly treated, can lead to the formation of a stromal abscess. A stromal abscess occurs through the introduction ofmicroorganisms into the corneal stroma. During re-epithelialization, the foreign body is encapsulated, thus creating abarrier that protects bacteria or fungi from treatment with antimicrobial medication. This framework can end up resultingin blindness due to chronic iridocyclitis, putting the animal’s vision at risk. The current work aims to report a case of corrective surgery for stromal abscess in a mare with the administration of intraoperative intrastromal fluconazole, in orderto corroborate the effectiveness of the technique.Case: A 9-year-old mare was evaluated, with the complaint that her right eye was closed and “yellowish” and that she hadalready been treated with intramuscular injectable anti-inflammatory drugs based on flunexin meglumine (Banamine® -50 mg) for 15 days, referring to a possible ulcer in the right eye. Ophthalmic screening resulted in a negative direct reflexand no threat response in the right eye. Examination of the conjunctiva showed congestion and chemosis. Examination ofthe cornea of the right eye was negative for Fluorescein and Green Lissamine tests, and opacity and corneal neovascularization were noted. The final diagnosis was a corneal abscess of probable fungal origin secondary to a keratomycosis. Afterthe consultation, complementary blood and biochemical tests were performed, which showed normal results for the speciesin question, and treatment was started with eye drops based on atropine 1% (Fagra® - 20 mL), ciprofloxacin antimicrobialeye drops (Ciprovet Colirio® - 5 mL), and antifungal eye drops based on ketoconazole...

Characterization of microbial communities and predicted metabolic pathways in the uterus of healthy mares.

Background: Culture-independent techniques have made it possible to expand the knowledge about the composition of bacterial communities present in the healthy uterus and their role in health and disease, mainly in humans. However, in animals like mares, there is a dearth of information regarding this area. Aim: To narrow this knowledge gap, the objective of this study was to identify and characterize the composition and function of the uterine microbiome of a group of Chilean purebred mares (CPM), an equine breed with the oldest genealogical record in South America and an economical important reproductive industry. Methods: From uterine biopsy samples obtained during estrus, DNA extraction and targeted sequencing were performed to investigate the bacterial diversity and its probable metabolic function. Results: CPM biopsy samples were characterized by having a varied microbial composition, where the four most relatively abundant phyla were Proteobacteria (69.6%), Firmicutes (21.1%), Bacteroidetes (7.8%), and Actinobacteria (1.06%); which made up 99.6% of the total identified phyla. In contrast, Actinobacteria and Fusobacteria were the phyla not identified in all samples. Of a total of 59 genera identified across all samples, Staphylococcus was the most abundant genus with an average relative abundance of 18.88%, followed by Pseudomonas (17.9%), Escherichia/Shigella (10.42%), and Klebsiella (9.92%). Conclusion: These findings contribute to the knowledge of microbes' presence in the uterus, while future studies are required to demonstrate the role of these microorganisms in health and disease.

Rainfall effects on Anomalocardia flexuosa densities on the northeastern Brazilian coast using distributed lag models / Efeitos da chuva na densidade de Anomalocardia flexuosa na costa nordeste do Brasil usando modelos de defasagem distribuída

In this study, the effects of precipitation on Anomalocardia flexuosa densities were analyzed from the perspective of temporal delays between the variables. The collections occurred bi-monthly between April 2016 and February 2018 at Mangue Seco beach, Pernambuco, Brazil. Total densities and densities according to the category of size (small, medium and large), and precipitation were analyzed with autocorrelation and cross-correlation functions, with a retrospective analysis of up to 12 months. Distributed lag models were used among the variables. The maximum density was found in June 2017 (378 ind.m2) for the medium-size category of individuals. Significant autocorrelations occurred for densities (total and average size) and precipitation. In cross-correlation functions, high precipitation for a given month was followed by high densities of total and average individuals for up to two months. Distributed lag models also showed significant values between densities (total and medium-sized) and precipitation, which explained more than 50% of the variability of these two groups. The effect of precipitation was responsible for the increase in the density of A. flexuosa for up to two months, mainly by increasing individuals with medium shell lengths.

Neste estudo, foram analisados os efeitos da precipitação nas densidades de Anomalocardia flexuosa sob a perspectiva de atrasos temporais entre as variáveis. As coletas foram bimensais, entre abril de 2016 e fevereiro de 2018, na praia de Mangue Seco, Pernambuco, Brasil. As densidades totais e por categoria de tamanho (pequeno, médio e grande) e a precipitação foram analisadas com as funções de autocorrelação e correlação cruzada, com análise retrospectiva de até 12 meses. Modelos de defasagem distribuída foram empregados entre as variáveis. A densidade máxima foi encontrada em junho de 2017 (378 ind.m²) na categoria de tamanho médio dos indivíduos. Autocorrelações significativas ocorreram nas densidades (total e tamanho médio) e precipitação. Nas funções de correlação cruzada, a alta precipitação de determinado mês foi seguida por alta nas densidades total e de indivíduos médios por até dois meses. Os Modelos de defasagem distribuída também apresentaram valores significativos entre as densidades (total e tamanho médio) e a precipitação, explicando mais de 50% da variabilidade desses dois grupos. O efeito da precipitação foi responsável pelo aumento da densidade de A. flexuosa por até dois meses, principalmente pelo incremento de indivíduos com comprimentos de concha medianos.

Clinical-biochemical and Fibroelastometric Results of the Mare's Milk Application in Non-Alcoholic Steathepatitis / Resultados clínico-bioquímicos y fibroelastométricos de la aplicación de la leche de yegua en la esteatohepatitis no alcohólica

Introduction: Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the world, representing the hepatic component of metabolic syndrome, and currently affects an average of 20-33 percent of the adult population. Non-alcoholic steatohepatitis (NASH) is a severe form of NAFLD and is a consequence of overweight, obesity, and metabolic syndrome. Objective: The study aimed to evaluate the effectiveness of the use of freeze-dried mare's milk in NASH. Methods: Clinical-biochemical, ultrasound, and fibroelastometric research methods were used in NASH patients receiving mare's milk in comparison with the control groups. Results: The result of the study demonstrates noticeable gain not only in the clinical symptoms of the disease but also in the laboratory and instrumental indicators, as well as health improvement and a decrease in symptoms of concomitant pathology. Taking mare's milk not only normalized liver biochemical parameters but also decreased cholesterol metabolism (total cholesterol, LDL, TG), the degree of liver steatosis, and existing hepatomegaly declined too. This pleiotropic effect of mare's milk points to the pathogenetic feasibility for the use of Saumal in NAFLD, including NASH. Conclusions: Addressing the problems of healthy nutrition in NASH, mare's milk can be considered as a pathogenetically justified, highly effective, and affordable natural therapeutic and prophylactic agent. The value of the product is determined by its multicomponent balanced qualitative composition. Hepatoprotective, hypocholesterolemic, and lipotropic properties of mare's milk in this pathology are marked(AU)

Introducción: La enfermedad del hígado graso no alcohólico es la enfermedad hepática más común en el mundo, representa el componente hepático del síndrome metabólico y actualmente afecta a un promedio de 20 a 33 por ciento de la población adulta. La esteatohepatitis no alcohólica es una forma grave de hígado graso no alcohólico y es una consecuencia del sobrepeso, la obesidad y el síndrome metabólico. Objetivo: Evaluar la efectividad del uso de leche de yegua liofilizada en hígado graso no alcohólico. Métodos: Se utilizaron métodos de investigación clínico-bioquímicos, ecográficos y fibroelastométricos en pacientes con hígado graso no alcohólico que recibieron leche de yegua en comparación con los grupos de control. Resultados: El resultado del estudio demuestra una ganancia notable no solo en los síntomas clínicos de la enfermedad, sino también en los indicadores de laboratorio e instrumentales, así como una mejora de la salud y una disminución de los síntomas de la enfermedad concomitante. La ingesta de leche de yegua no solo normalizó los parámetros bioquímicos del hígado, sino que también disminuyó el metabolismo del colesterol (colesterol total, LDL, TG), el grado de esteatosis hepática y la hepatomegalia existente también disminuyeron. Este efecto pleiotrópico de la leche de yegua apunta a la viabilidad patogénica del uso de Saumal en hígado graso no alcohólico, incluida la esteatohepatitis no alcohólica. Conclusión: Al abordar los problemas de una nutrición saludable en la enfermedad hígado graso no alcohólico, la leche de yegua se puede considerar como un agente terapéutico y profiláctico natural patogénicamente justificado, altamente efectivo y asequible. El valor del producto está determinado por su composición cualitativa equilibrada multicomponente. Las propiedades hepatoprotectoras, hipocolesterolémicas y lipotrópicas de la leche de yegua en esta enfermedad son marcadas(AU)

Parto gemelar de 310 días de gestación en yegua con resultado de un potro vivo y otro en estado de autolisis / Twin Delivery of 310 Days of Gestation in a Mare Resulting in One Live Foal and One Autolysed Foal

Resumen Las preñeces gemelares son gestaciones de alto riesgo, tanto para la yegua como para ambos fetos. Hoy en día existen diferentes técnicas para evitar el avance de este tipo de eventos, tanto en sus inicios como en gestaciones avanzadas. Aun así, existen múltiples casos de gestaciones dobles que presentan diferentes resultados ya evidenciados. En este caso se reportó una yegua de raza polo argentino de ocho años, que llegó al Centro de Perinatología Equina Foal Care por presentar gestación gemelar de 292 días. Se prolongó la gestación durante dieciocho días más, con ayuda de terapia farmacológica. Finalmente, nació un potro vivo con múltiples complicaciones, incluyendo el hecho de ser un potro prematuro. El segundo potro nació muerto con características de momificación y autolisis. En conclusión, se requirió de un adecuado monitoreo reproductivo por parte del veterinario para prevenir el avance de este tipo de gestación.

Abstract Nowadays, there are different techniques to avoid the progression of this type of events, both in early and advanced gestations. Even so, there are multiple cases of double gestations that present different results already evidenced. In this case, an eight-year-old Argentine polo mare was reported, who arrived at the Foal Care Equine Perinatology Center because she presented a twin gestation of 292 days. The gestation was prolonged for eighteen more days, with the help of pharmacological therapy. Finally, a live foal was born with multiple complications, including being a premature foal. The second foal was stillborn with mummification and autolysis characteristics. In conclusion, adequate reproductive monitoring by the veterinarian was required to prevent the progression of this type of gestation.

Segmental Cervical Aplasia in a Colombian Creole Mare with Mosaic X-Chromosome Aneuploidy.

A 4-year-old Colombian Creole mare was presented for diagnosis because the external orifice of her cervix was not detectable when a uterine lavage as therapy for uterine fluid accumulation was attempted. Clinical and ultrasonographic evaluation of the genital tract revealed that ovaries were of normal size and showed structures suggestive of regular ovarian activity. However, granular free-floating fluid material distending the uterus was detected by ultrasound. Upon vaginal examination, the normal external cervical morphology was not evident. The vagina ended in a blind bag with a small papilla with no evident external cervical os. Cytology of uterine fluid obtained by transvaginal aspiration showed findings compatible with mucometra. Cytogenetic analysis revealed an abnormal karyotype (63,X and 64,XX both 45% and 65,XXX 10%). A diagnosis of congenital segmental cervical aplasia was proposed possibly related to the mosaicism detected. To our knowledge, this is the first case of this reproductive pathology in a mare with regular ovarian activity and confirmed aneuploidy in mosaic form of the X sex chromosome.

Use of Intravaginal Progesterone-Releasing Device Results in Similar Pregnancy Rates and Losses to Long-Acting Progesterone to Synchronize Acyclic Embryo Recipient Mares.

The objectives of this study were: (1) to assess uterine features and serum progesterone concentrations of acyclic mares synchronized and resynchronized with intravaginal progesterone release device (IPRD), and (2) to compare pregnancy rates and losses of cyclic and acyclic embryo recipient mares treated with different synchronization protocols. In Experiment 1, mares (n = 12) received estradiol for 3 days (E2-3d), and then 24 h after the last injection, an IPRD was inserted and kept in place for 9 days. Three days after IPRD removal, mares were treated with E2-3d, and then a new IPRD was inserted and maintained for three days. Serum progesterone concentrations were assessed 2, 6, and 12 h after insertion and removal of IPRD, and then daily from the insertion of the first IPRD to one day after removal of the second IPRD. Experiment 2 was conducted with embryo recipient mares randomly assigned to four groups: (1) Cyclic: mares (n = 75) had ovulation confirmed after receiving a single dose of histrelin when a periovulatory follicle was first detected, (2) LAP4: acyclic mares (n = 92) were treated with E2-3d and then administered a single dose of LAP4 24 h after the last estradiol injection, (3) IPRD: acyclic mares (n = 130) were treated with E2-3d and an IPRD for 4-8 days, and (4) RE-IPRD: acyclic mares (n = 32) were synchronized as in the IPRD group but not used for embryo transfer (ET), then 8 to 15 days later, the mares were resynchronized with E2-3d and an IPRD for 4-8 days. In vivo-produced Day-8 embryos were collected and transferred 4-8 days after ovulation or progesterone treatments. Mares in IPRD and RE-IPRD groups had the intravaginal device removed immediately before ET, and then a new IPRD was inserted right after ET. Pregnancy diagnosis was performed at 5, 30, and 60 days after ET. Once pregnancy was confirmed, mares in the three acyclic groups received weekly injections of LAP4 (1.5 g) until 120 days of pregnancy. Mares in IPRD and RE-IPRD groups had the device removed three days after the first pregnancy diagnosis. In Experiment 1, progesterone concentrations increased rapidly starting 2 h after insertion of IPRD (p < 0.05); then, concentrations plateaued well above pregnancy maintenance until removal on days 9 and 3, respectively. Progesterone concentrations were reduced to baseline 24 h after IPRD removal (p < 0.05). For experiment 2, there was no difference in pregnancy rates across groups (65-74%) or pregnancy losses by 60 days of gestation (7-12%) (p > 0.05). In conclusion, the IPRD used herein resulted in a rapid increase and a sharp decline in progesterone concentrations upon its insertion and removal, respectively. Finally, our results demonstrated that IPRD could be a compatible alternative to LAP4 to synchronize and resynchronize acyclic embryo recipient mares.

Modelación hidrodinámica y de calidad del agua en un ecosistema estuarino urbano con incidencia maregráfica y vertimientos de aguas residuales / Hydrodynamic and water quality modeling in an urban estuarine ecosystem with tidal incidence and sewage discharges / Modelagem hidrodinâmica e de qualidade da água em um ecossistema estuarino urbano com incidência da maré e descarga de esgoto

RESUMEN El efecto del régimen de mareas y la descarga directa de aguas residuales en áreas estuarinas pueden incrementar los problemas de contaminación debido a la retención y transporte de cargas contaminantes. El uso de modelos matemáticos se ha convertido en una herramienta esencial para la gestión del recurso hídrico en estos ambientes. En esta investigación se modeló la incidencia del régimen de mareas y los vertimientos de aguas residuales en la calidad de agua en un ambiente estuarino urbano utilizando el modelo HEC-RAS. El modelo fue calibrado/validado utilizando el caudal medio de largo periodo (QMLT) y parámetros de calidad de agua medidos en campo, tales como: DBO, OD, N-O2 y N-O3. Los resultados evidenciaron que los niveles de agua, caudales y parámetros de calidad de agua medidos en campo fueron bien representados por el modelo. Las simulaciones mostraron que la calidad del agua en todos los trechos aguas abajo de la PTAR se encuentran eutrofizados. Adicionalmente, el régimen de marea provoca retención y transporte de contaminantes en áreas de baja pendiente. El abordaje propuesto se constituye en una valiosa herramienta para evaluar la calidad de agua en ambientes estuarinos urbanos y áreas circundantes con incidencia de régimen maregráfico.

ABSTRACT The effect of the tidal regime and sewage discharge on estuarine areas can cause pollution problems, due to the retention and transport of pollutants. The use of mathematical models has become an essential tool for the water resources management in these environments. In this paper, the incidence of tidal regime and sewage discharge on the water quality of an urban estuarine ecosystem was modeled using the HEC-RAS model. The model was calibrated / validated using the medium long-term streamflow (QMLT) and water quality parameters measured in field, such as: BOD, DO, N-O2 N-O3. The results showed that the water level, streamflow and the water quality parameters measured in field were well represented from the model. The simulations showed that the water quality for all the sections downstream of the Wastewater Treatment Plant are eutrophied. Additionally the tidal regime causes retention and transport of pollutants in areas of low slope. The proposed approach is a valuable tool for assessing the water quality in urban estuarine environments and surrounding areas with incidence of tidal regime.

RESUMO O efeito do regime de marés e a descarga directa de esgoto em zonas estuarinas pode aumentar os problemas de poluição devido à retenção e transporte de cargas poluentes. A utilização de modelos matemáticos tornou-se uma ferramenta essencial para a gestão dos recursos hídricos nestes ambientes. Nesta investigação, a incidência do regime de marés e descargas de esgoto na qualidade da água em um ambiente estuarino urbano foi simulada utilizando o modelo HEC-RAS. O modelo foi calibrado/validado utilizando o caudal médio de longo período (QMLT) e parâmetros de qualidade da água medidos no campo, tais como DBO, OD, N-O2 e N-O3. Os resultados mostraram que os níveis da água, vazões e parâmetros de qualidade da água medidos no campo foram bem representados no modelo. As simulações mostraram que a qualidade da água em todos os trechos a jusante da ETE estão eutrofizados. Além disso, o regime de maré causa retenção e transporte de poluentes em áreas de baixa declividade. A abordagem proposta é uma ferramenta valiosa para avaliar a qualidade da água em ambientes estuarinos urbanos e áreas circundantes com incidência de regime de marés.

Integração do SWMM e ferramentas SIG para modelagem hidrológico-hidráulica de bacia complexa / SWMM and GIS tools integration for hydrological-hydraulic modeling of a complex basin

RESUMO O presente trabalho aplicou o Storm Water Management Model (SWMM) para simular e avaliar a resposta hidrológica do sistema de macrodrenagem de um rio costeiro, cuja bacia situa-se em área de transição urbano-rural e apresenta influência marítima em seu exutório. Assim, como condição de contorno de jusante, reconstruiu-se a curva de maré existente no exutório da bacia por meio do método harmônico. Ademais, realizou-se a delimitação dos trechos de escoamento e a discretização das sub-bacias automaticamente e desenvolveu-se um mapa de uso e ocupação do solo para obtenção dos parâmetros físicos e espaciais dos elementos da drenagem, etapas estas auxiliadas por ferramentas de Sistemas de Informações Geográficas (SIG). Procedeu-se à simulação das vazões no sistema para o período de um mês sob condição de precipitação moderada. A validação do modelo resultou em um coeficiente de Nash-Sutcliffe de 0,57. Os resultados mostraram que o SWMM simulou as vazões nos trechos de drenagem das sub-bacias rurais com desvio médio de 8,6% das vazões medidas em campo. Para os trechos sob influência marinha, o SWMM conseguiu reproduzir o efeito da maré nos trechos, apresentando uma oscilação completa. Ademais, obteve-se a descarga fluvial média da bacia ao estuário de 2,35 m3.s-1 e a vazão média mensal afluente ao maior reservatório da bacia de 0,33 m3.s-1, resultados estes em consonância com trabalhos anteriores realizados na bacia em estudo. Em síntese, os resultados obtidos e o modelo proposto têm potencial para auxiliar na gestão integrada de águas rurais, urbanas e costeiras.

ABSTRACT The present work applied the Storm Water Management Model (SWMM) to simulate and evaluate the hydrological response of a coastal river macrodrainage system whose basin is located in an urban-rural transition area and has a tidal influence in its outflow. Thus, as a downstream contour condition, the existing tidal curve in the basin's outlet was reconstructed using the harmonic method. In addition, drainage stretches and sub-basins were automatically delimited, and a land use and occupation maps were developed to obtain the physical and spatial parameters of the drainage elements, these steps being aided by Geographic Information Systems (GIS) tools. The flow was simulated in the system for a period of one month under a moderate precipitation condition. The validation of the model resulted in a Nash-Sutcliffe coefficient of 0.57. The results showed that the SWMM simulated the flows in the drainage stretches of the rural sub-basins with an average deviation of 8.6% of the flows measured in the field. For stretches under tidal influence, SWMM was able to reproduce the effect of the tide in the stretches with a complete oscillation. In addition, the river discharge from the basin to the estuary was 2.35 m3.s-1, on average, and the average monthly flow affluent to the largest reservoir in the basin was 0.33 m3.s-1, consistently with previous work carried out in the study basin. In summary, the results obtained and the proposed model have the potential to assist in the integrated management of rural, urban, and coastal waters.

Heat shock protein HSP90 immunoexpression in equine endometrium during oestrus, dioestrus and anoestrus.

Heat shock proteins play a crucial role in cellular development, proliferation, differentiation and apoptosis. Heat shock protein 90 (HSP90) has been localised in the human endometrium, where its immunoexpression changes during the menstrual cycle. Similar studies have not been done for the equid species, so the present study aimed to describe endometrial HSP90 immunoexpression in mare endometrium. Endometrial biopsies were formalin-fixed and paraffin-embedded, and sections were stained with haematoxylin-eosin in preparation for HSP90 immunohistochemistry. Immunostaining and morphometric analyses were performed on the epithelial lining, endometrial glands and connective stroma during oestrus, dioestrus phase and anoestrus period (n = 7 per phase or period). Immunoexpression was localised in the basal region of the epithelial cells lining the lumen. Immunoexpression was greater during oestrus than during either dioestrus or anoestrus. During anoestrus, there was little immunostaining in the endometrium, suggesting that HSP90 is involved in the functional modulation of sex steroid receptors in cyclic mares. Indeed, the function of HSP90 as a chaperone in the folding of proteins, such as steroid receptors, might explain the greater intensity of immunostaining during the oestrus and dioestrus phases, compared the anoestrus period. We conclude that, in the mare, HSP90 plays a role in endometrial function and that further studies are needed to test whether it is important in pathological conditions as endometritis.

Equine ovarian tissue xenografting: impacts of cooling, vitrification, and VEGF.

Ovarian tissue transplantation methods using cooled and cryopreserved samples have been attractive options for fertility preservation in animal models and humans. The aim of this study was to evaluate the impact of previous exposure to cooling, cryopreservation, and VEGF on the overall efficiency of equine ovarian tissue after heterotopic xenotransplantation in mice. The end points evaluated were follicular morphology and development, follicular and stromal cell densities, angiogenesis (i.e. the density of new and mature blood vessels), collagen types I and III fiber densities, and total fibrosis. Ovaries of adult mares were harvested after ovariectomy, and ovarian fragments were xenografted in the i.p. wall of BALB nude mice. Ten types of treatments involving different combinations of cooling, cryopreservation, xenografting procedures, and VEGF exposure were compared. The novel aspect of this study was the use of equine ovarian tissue xenotransplantation in mice, challenging the fragments with different combinations of treatments. The main findings were (i) cooling but not cryopreservation was effective in preserving the follicular morphology, (ii) a greater percentage of developing follicles but lower follicular and stromal cell densities were observed after ovarian tissue engraftment, (iii) exposure to VEGF increased new and mature vessels in cryopreserved-transplanted tissue, and (iv) an appropriate balance in the collagen types I and III fiber ratio in cooling-transplanted tissue was observed after exposure to VEGF. This study contributes to advancing knowledge in the preservation of ovarian tissue after cooling-cryopreservation and transplantation aiming to be applied to genetically superior/valuable horses, livestock, endangered animals, and, possibly, humans. LAY SUMMARY: Due to ethical limitations involving humans, the female horse (mare) has recently emerged as an alternative model for reproductive comparisons with women to optimize fertility restoration using ovarian tissue transplantation techniques. This study determined if ovarian tissue from donor mares (n = 3), exposed or not to vascular endothelial growth factor (VEGF) before transplantation, better survives for 7 days after transplantation into mouse hosts (n = 12). Tissues submitted to different combinations of cooling, freezing, and transplanting treatments, along with control groups, were evaluated using the parameters morphology, development, the density of immature eggs (follicles), the density of supportive (stromal) cells, collagen protein proportions, and density of blood vessels. Frozen-thawed treatments had lower percentages of normal follicles. Exposure to VEGF increased blood vessel densities in frozen tissue and favored adequate collagen levels in cooled-transplanted treatments. In conclusion, VEGF exposure seems to be beneficial for mare ovarian tissue transplantation and warrants further investigation.