Neste trabalho avaliamos os efeitos da exposição de embriões bovinos produzidos in vitro à hipotermia moderada (33,0 ºC), visando o estudo futuro das proteínas de choque frio e a congelabilidade destes embriões. Para isso complexos cumulus-oócitos (CCOs) foram maturados e fecundamos in vitro. Então os embriões foram cultivados em normotermia (38,5 ºC) até o 6º dia de cultivo (FIV dia zero) quando então foram expostos a hipotermia moderada (estufa de CO2 ajustada para 33,0 ºC) por 6 e 12h (grupos cold-6 e cold-12, respectivamente) e então retornaram para o cultivo em normotermia. Um grupo de embriões foi mantido o tempo todo em condições normotérmicas (Controle) e todos os grupos foram avaliados no 7º e 8º dia quanto a taxa de formação de blastocistos e quanto à cinética de desenvolvimento destes embriões formados. Observamos que exposição a hipotermia no 6º dia de cultivo por 6h e 12h não afetou significativamente a sobrevivência dos embriões (p > 0,05). Contudo no 7º dia de análise foi possível notar a redução numérica na taxa de desenvolvimento dos blastocistos (Cold 6=32.0, Cold 12=32.5 em relação ao Controle= 34.8 p > 0,05) . Sendo que no 8º dia de análise houve uma recuperação na taxa em todos os grupos experimentais (Cold 6= 34.9, Cold12= 42.2 e Controle 37.2%; p > 0,05). Com relação a cinética observamos que a exposição a hipotermia por 6h (grupo cold-6) gerou uma maior quantidade de blastocistos iniciais em comparação aos grupos cold-12 e controle (58.5% vs. 51.2% e 47.4%, respectivamente; p<0,05) e consequente mente uma menor taxa de blastocistos expandidos (38.1% vs. 45.6% e 50.4%) no 7º dia de cultivo. Além disso, a análise cinética no 8º dia revelou que o grupo cold-6 gerou uma menor proporção (p<0,05) de blastocistos eclodidos (22.4%) em comparação aos grupos cold-12 (23.0%) e controle (38.6%).Entretanto, com relação a cinética observamos que a exposição a hipotermia por 12h (grupo cold-12) gerou uma maior quantidade de blastocistos iniciais em comparação aos grupos cold-6 e Controle (78.0% vs. 63.7% e 59.4%, respectivamente; p<0,05) no 7º dia de cultivo. Conjuntamente, nossos resultados indicam que a exposição de embriões D6 por 12h a hipotermia moderada não afetou a taxa de formação de blastocistos no 7º dia de cultivo mas retardou a evolução do desenvolvimento embrionário analisado até o 8º dia de cultivo. Sendo assim, esses resultados preliminares indicam que o metabolismo das proteínas de choque-frio embrionárias pode ter sido ativado e portanto a análise desse mecanismo precisa ser realizada para melhor compreender os mecanismos de resposta ao frio e o papel das proteínas de choque frio neste processo. 

Breast tumors are the most common neoplasm in unneutered female dogs, with unilateral radical mastectomy considered the main treatment for these conditions. As it is a long-lasting procedure, with a high nociceptive stimulus and risk of post-surgical complications, an adequate anesthetic protocol, focused on multimodal analgesia, which includes administration of opioids in the perioperative period, leads to a shorter recovery time. However, gastroesophageal reflux is related to the action of anesthetic drugs during general anesthesia, caused by a reduction in the tone of the lower esophageal sphincter. This reflux is associated with consequences such as esophagitis, esophageal stenosis and even aspiration pneumonia. Thus, this study sought to determine the incidence of gastric reflux and the behavior of the lower esophageal sphincter in bitches undergoing mastectomy surgery, normally using premedication with midazolam and morphine or tramadol as part of the anesthetic protocol. 12 bitches undergoing total unilateral mastectomy surgery were evaluated as part of a routine routine at the Veterinary Hospital of the Federal University of Pará. During the operation, endoscopic evaluations were carried out to analyze the behavior of the esophagogastric sphincter and confirm gastric reflux. 100% of the dogs presented some degree of gastroesophageal reflux and relaxation of the lower esophageal sphincter during the operation. The use of the evaluated anesthetic protocols resulted in a high number of intraoperative gastric reflux, probably caused by the relaxing effect of the lower esophageal sphincter. This study found a high number of gastroesophageal reflux by intraoperative endoscopy in dogs undergoing mastectomy surgery using a combination of midazolam and morphine or tramadol as part of the anesthetic protocol, triggered by relaxation of the lower esophageal sphincter. To the detriment of the risk of postoperative complications, especially in geriatric and oncology patients, future prospective studies are necessary to evaluate and propose possible ways to minimize this effect.

This thesis was divided into two moments. In the first (Experiment 1) we studied the technique of videolaparoscopic ovarian follicular aspiration (LOPU) in buffalo calves, describing in detail its particularities, intercurrences and the result of oocyte aspiration. Six lactating Murrah heifers aged between 3 and 5 months, raised on pasture in a milking regime. Three LOPUs were performed on each animal, with intervals of 15 days between surgeries, totaling 18 procedures. After being sedated, the calves were placed in the Trendelenburg position at 45°. Two 10 mm and one 5 mm trocars were used to establish the three laparoscopic portals. The abdomen was inflated with CO₂ at a speed of 5L/min, stabilizing intra-abdominal pressure between 5-8 mmHg. For ovarian manipulation, two atraumatic tweezers (5 mm and 10 mm) were used and for aspiration, a 20G transabdominal needle connected to a vacuum system calibrated for 50 mmHg. The oocytes were recovered in a 50 mL tube containing saline solution at a concentration of 1% fetal bovine serum and 10 IU/mL of Heparin, kept at 38ºC and then they were selected and classified, still on the farm, according to the cumulus cells and the homogeneous aspect of the cytoplasm. For statistical analysis, GraphPad Prism 9^® Software was used. Data were presented as mean ± standard deviation (SD) in experiment 1 and as mean ± standard error of the mean (SEM) in experiment 2. The normal distribution of data was tested using the Kolmogorov-Smirnov test in the first experiment and the test of Shapiro-Wilk in the second, then underwent analysis of variance (ANOVA) and Tukey's post-hoc test. The average time of complete surgery from anesthesia to removal of the animal from the stretcher was 49.8 ± 10.1 min, and the step that took the most time was manipulation and follicular aspiration of both ovaries (20.6 ± 9 .7 min) (p≤0.05), with 22.2% of cases of pre-peritoneal insufflation. The total number of aspirated follicles, retrieved oocytes and viable oocytes, at first experiment, were 126, 76 and 31, respectively. The same parameters per procedure were 7.17 ± 7.17; 4.22 ± 7.1 and 1.72 ± 4.2; respectively. The recovery rate was 60.3%. In the second experiment, the use of equine chorionic gonadotropin (eCG) in different concentrations in ovarian stimulation in pre-LOPU buffalo calves was studied. For this, 20 lactating Murrah heifers with similar age and management to the first experiment were used. A 3 x 3 Latin square design was applied, with LOPUs being performed every 15 days. All animals received on day 0 (D0) a P4 progesterone device (Primer Pr^® - 0.36g; Agener, Brazil), on day 3 (D3) eCG was administered according to the experimental groups, 500 IU of eCG (Ecegon^® - Biogénesis Bagó/Brazil) (Group 500), 1000 IU of eCG (Group 1000) or did not receive gonadotropin (Control Group), on day 5 (D5) LOPU was performed in all animals. The LOPU procedure followed the technique described in experiment 1, with the oocytes sent to the laboratory for the parthenogenetic activation procedure. The dependent variables were presented as mean and standard error of mean (mean ± SEM) considering a significance level of 5%. In this experiment, 430 follicles were aspirated, 248 oocytes were recovered, of which 73 were classified as viable. It was observed that the use of eCG increased the number of medium (6 – 10mm) and large (>10mm) follicles (Medium: Control = 5.19%; G500 = 38.36% and G1000 = 25.69% and Large: Control = 3.90%; G500 = 13.70% and G1000 = 25). There was no influence of eCG on the number of aspirated follicles (Control = 9.00 ± 1.40; G500 = 5.00 ± 0.67 and G1000 = 10.65 ± 1.83; P=0.1744); oocyte recovery (Control = 6.94 ± 1.73; G500 = 2.70 ± 0.67 and G1000 = 5.33 ± 1.23; P = 0.0736) and recovery rate (Control = 59.14% ± 7.9; G500 = 61.69% ± 9.9 and G1000 = 55.30% ± 8.2; P = 0.8052). The dose of 1000UI of eCG significantly influenced oocyte viability per treatment (Control = 0.64 ± 0.2; G500 = 0.55 ± 0.2 and G1000 = 3.05 ± 0.6; P = 0.0086). With the results obtained, we can observe that this is a safe technique to be performed even in a farm environment, presenting an excellent postoperative response, allowing it to be performed several times, without harming the female and that the application of eCG before LOPU, is an economical alternative available on the market to stimulate the follicular development of buffalo calves, creating alternatives for the best use of the ovarian potential of females and thus obtaining increasingly better results with PIVE.

Global warming with the consequent rise in water temperature has been shown to considerably alter aspects inherent in the life cycle of certain fish species, such as reproduction. Thus, aiming at minimizing the impacts caused to fish by the rise in environmental temperature, germ cell transplantation technology and fish sterilization have been widely used as biotechniques capable of contributing to the reproduction, maintenance and preservation of different fish species, either in risk of extinction or economically important. Thus, this work aimed to evaluate the effect of high temperature on embryonic development, as well as hybridization in the adult phase of two Amazonian fish species. The species chosen were Astyanax bimaculatus (Linnaeus, 1758) and Moenkhausia oligolepis, which belong to the Characidae family, which is composed largely of freshwater fish from Brazil, and both are valuable objects of study because they present unique characteristics for the development of aquaculture and scientific production. The results showed that high temperature had relevant effects on the embryonic development of the animals studied, and as a possible tool for sterilization in fish, hybridization between the two species proved to be very promising.

An investigative and epidemiological study was carried out for equine herpesvirus type 1 (EHV-1) in 10 outbreaks of neurological disease, with 25 horses, 22 with nervous symptoms and three apparently healthy, from 10 different rural properties located in the state of Pará, Brazil. In these outbreaks of neurological disease, six male and 19 female horses, aged between one and 13 years. Of these, six horses underwent necropsy, and the others recovered with or without the use of drug therapy. The main clinical signs observed were motor incoordination, thoracic and/or pelvic limbs abducted after induction to the clinical examination, flexion of the fetlocks, pendular movements of the hindquarters (horses standing still or when walking) and paresis, which was sometimes so severe that the animal almost dragged its hindquarters on the ground, on a steep slope, giving the appearance of a “crouched” animal. Other signs such as tachycardia, tachypnea, decreased tongue tone, difficulties in getting up, localized sweating, bed sores, pedaling movements, muscle fasciculations in the face, frequent urination, priapism, spasticity of the limbs and wear of tweezers were also observed, however, less frequently. The animals that received the treatment protocols recovered after eight days. In the laboratory diagnosis via serum neutralization, 72% (18/25) of the animals showed a positive reaction for EHV-1. The nested PCR for EHV-1, from whole blood, revealed 88% (22/25) of positive animals, including three apparently healthy animals, and the three negative animals in the molecular tool were positive in serology. At necropsy, foci of hemorrhage in the gray matter were evidenced in the spinal cord, which slightly extended into the white matter. On histopathological analysis, there was perivasculitis and neutrophilic vasculitis in the gray matter of the spinal cord and brain. Thus, this study proves that EHV-1 is circulating in the Amazon Biome, mainly in the state of Pará.

O presente estudo teve como objetivo avaliar a influência da cinética espermática na fertilidade de búfalas submetidas à IATF. Foram utilizadas 332 vacas bubalinas multíparas com 30 a 60 dias pós- parto, predominantemente da raça Murrah e escore corporal de 3,5 ± 0,5. O protocolo de sincronização se deu no d 0, com implantação do dispositivo intravaginal contendo 1,0 grama de progesterona (Sincrogest/Ourofino) mais a aplicação de 2,0 mg de benzoato de estradiol e administração de 10 ml de suplementação mineral injetável via intramuscular as 16:00 h. No d 9, também as 16h, foi realizado remoção do dispositivo de progesterona vaginal e realizado a aplicação de 0,265 mg de pgf2α + 300 u.i de gonadotrofina coriônica equina. Ao d 11, às 16:00 h, foi administrado 0,1 mg de GnRH. No d12, 16 horas após a aplicação de gnrh (às 08:00 h) foi realizada a inseminação artificial. Doze amostras de sêmen de seis touros bubalinos da raça Murrah foram analisadas, sendo que todas as amostras foram criopreservadas em centros de reprodução animal a nível comercial. As análises da cinética espermática movimento espermático foram realizadas pelo programa Sperm Class Analyser® Mot. Foram obtidas as seguintes médias de parâmetros cinéticos: MOT (80,38% ± 4,20%), BCF (10,94 Hz ± 0,09 Hz), PRO (48,35% ± 3,57%) e PRA (23,80% ± 0,09%), VCL (68,06μm/s ± 3,58μm/s), VAP (32,65 μm/s ± 3,58 μm/s) e VSL (29,27μm/s ± 3,58 μm/s), WOB (59,25% ± 3,58%), STR (50,00% ± 3,58%) e (49,03 ± 3,8%) e ALH (1,58 μ ± 0,09 μ). Foi observado no estudo que os parâmetros de correlação que mais influenciaram na taxa de prenhez foram velocidade progressiva rápida, motilidade progressiva, velocidade progressiva, movimento progressivo rápido e a frequência de batimento cruzado. O trabalho foi concluído considerando que houve influência dos diversos parâmetros cinéticos espermáticos analisados no CASA com a taxa de prenhez a campo, e evidenciando que há uma alta correlação com os parâmetros de PRO, VAP, PRA, VSL e BCF.

Puberty is a time process characterized by the transition to adulthood and sexual development of the individual to be able to reproduce. Reproductive disorders during the pubertal stage impact the fertility of the animal in its adult life. Multiple infectious agents are noted as a threat to male fertility, one of these being Zika virus. Studies in adult specimens point to the Zika virus' ability to damage sperm lineage cells, as well as Sertoli and Leydig cells. However, studies addressing the effects of the virus in immature animals are still scarce. Neotropical primates are mentioned as possible reservoirs in the wild cycle of Zika virus transmission. The squirrel monkey (Saimiri collinsi) is a neotropical primate recently presented as a candidate model to study the reproductive pathogenesis of this virus. Thus, in this work we aimed to investigate the effects of Zika virus (ZIKV) on reproductive parameters (fecal testosterone levels, testicular biometry, testicular echogenicity and histomorphometric evaluation with Johnsen's score) of pubertal squirrel monkey (Saimiri collinsi). Experiment period was categorized according to the stages of ZIKV infection: pre-inoculation, acute phase, and convalescent phase. Infected animals had reduced fecal testosterone levels in the acute phase when compared to pre-inoculation and convalescent phases. Within groups, testicular volume and gonadosomatic index of infected animals were lower than non-infected animals in both acute and convalescent phases. Infected animals also showed reduced testicular volume, gonadosomatic index, and testicular echogenicity in the convalescence phase. Furthermore, the infected group showed significantly lower values than the non-infected group in all parameters evaluated in testicular histomorphometry (tubular area, seminiferous epithelium, major and minor diameter, epithelium height, and Johnsen score). In light of the above-mentioned factors, we can conclude that the experimental infection by ZIKV causes multiple reproductive alterations in these animals. In addition, the parameters adopted in this study allowed the quantitative evaluation of the changes caused by ZIKV in male pubertal squirrel monkeys (Saimiri collinsi).

Pyometra is the endometrial disorder of greatest clinical importance in small animals. Therefore, studies with the purpose of obtaining epidemiological data related to the occurrence of pyometra in bitches are fundamental. The objective of the study was to determine epidemiological indicators of canine piometra, using the statistical method of main component analysis, evaluating the casuistry attended to university veterinary hospitals in the northeast region of the state of Pará. Thus, draw the set of parameters that facilitate diagnosis and a possible clinical-epidemiological profile. For this purpose, 142 medical records of cases seen between the years 2017 to 2019 were consulted. A total of 18 variables related to the disease were evaluated. Data were analyzed using principal component analysis (PCA) and descriptive statistics. The PCA identified three groups of epidemiological indicators that together explained 71.05% of the variability of the data. The first group (abdominal pain, abdominal distension, critical condition, and leukocytosis), explained 29.2%, the second (apathy, anorexia, pyometra type, and vaginal discharge) explained 24.05%, and the third (age, race, parity, and pseudocyesis) explained 17.75% of the variability in the data. It was observed that 47.7% of the bitches were five to 10 years old, 63.8% were of undefined breed, 62.4% were nulliparous (p<0.05), 82.4% (p<0.05) had open pyometra and 46.8% of the bitches had already received contraceptive administration (p>0.05). The most frequent clinical signs were apathy (94.4%), anorexia/hyporexia (90.1%), vaginal discharge (83%), abdominal pain and distension (67.9%) and leukocytosis (63.7%). From the results it was identified that the variables abdominal pain and distension and leukocytosis are epidemiological indicators for the occurrence of the disease in severe stage. That the variables apathy, anorexia/hyporexia and vaginal secretion are epidemiological indicators for the occurrence of the disease in mild stage and that the variables age and race act as epidemiological indicators of predisposition. Moreover, it was possible to identify that the clinical-epidemiological profile of affected animals is that of middle-aged, nonbreed, nulliparous bitches with open pyometra. And that the presence of leukocytosis, apathy, anorexia/hyporexia and vaginal secretion are common findings in patients diagnosed with this disease.

A produção in vitro de embriões (PIVE) bovinos é uma biotécnica mundialmente difundida para o aumento do ganho genético da espécie, mas também pode ser usada como modelo para pesquisas de toxicologia na reprodução. Em 2019, com o aparecimento do SARS-CoV-2, a hidroxicloroquina (HCQ) foi amplamente utilizada como tratamento off label para a COVID-19 mesmo sob grande controvérsia em relação à sua eficácia e feitos colaterais. Diante disso, este trabalho teve como objetivo utilizar a técnica de PIVE para avaliar a citotoxicidade da HCQ em oócitos bovinos. Deste modo, óocitos bovinos foram maturados in vitro com HCQ (6, 12. 24 µM) e sem HCQ (Controle) por 24h. Após a maturação os oócitos foram ativados partenogeneticamente e os embriões cultivados in vitro por 7 dias. Os resultados indicaram que o tratamento com a HCQ não afetou a taxa de extrusão do primeiro corpúsculo polar e nem taxa de metáfase II (p>0,05). Quanto ao desenvolvimento, não foi observado efeito do tratamento sobre a clivagem (p>0,05), porém ocorreu uma redução significativa (p<0,05) na taxa (%) de blastocistos proporcionalmente ao aumento das concentrações de HQC utilizadas na maturação (Crtl=48,8; 6µM=43,0; 12µM=37,7 e 24µM=30,0). Finalmente, analisamos o processo de autofagia em oócitos e suas respectivas células do cumulus oophorus após a maturação, mediante a coloração com o corante Laranja de Acridina. O tratamento com 24 µM de HCQ promoveu aumento significativo (p<0,05) do bloqueio da autofagia (aumento da fluorescência em relação ao Ctrl) tanto nos oócitos (1,7X maior) quanto nas células do cumulus (2,8X maior). Concluímos que o tratamento com HCQ apesar de não ter afetado a maturação nuclear dos oócitos, afetou a posterior capacidade de desenvolvimento embrionário dos mesmos sendo que este efeito negativo pode estar relacionado ao distúrbio do mecanismo de autofagia na maturação causado pelo tratamento com a droga.

The International Union for the Conservation of Nature, estimated that 49% of the world's primates are in some category of extinction risk. In Brazil, the genus Saimiri has endangered species, the conservation of the genetic variability of these animals can be guaranteed through in situ (in the natural environment) and ex situ (in captivity or through the formation of banks) protection strategies. It is known that for the planning of conservation programs, it is important to know the reproductive aspects of the species or genus in focus. To elucidate some aspects of Saimiri reproduction, the work aims to verify the hormonal influence (testosterone and estradiol) on seminal quality as a function of age, in male individuals of the species S. collinsi kept in captivity.The experimental group consisted of 8 adult male squirrel monkeys (Saimiri collinsi), 4 animals with 5-6 years old and 4 animals aged 12-16 years. Collections were carried out for 18 months (January 2015 to July 2016) The animals were previously sedated by a hydrochloride association of ketamine (20 mg/kg; intramuscular) and xylazine hydrochloride (1 mg/kg; intramuscular). Subsequently, blood samples of ~1ml were collected monthly by venipuncture, centrifuged (2100 g /15 min) for plasma separation and stored (-20 º C). After chemical restraint, the animals were weighed, and the measurements of width, height and length of the testes were obtained to obtain the volume of the testes. Semen collection was performed by electroejaculation, and the basic parameters of seminal quality were: seminal volume, color, pH, degree of coagulation, motility, vigor, integrity and functionality of the plasma membrane. Testosterone and estradiol plasma levels were measured in duplicate by the amplified electrochemiluminescence method. For statistical analysis, the animals were divided into two groups according to age: animals aged 5-6 years and animals aged 12-16 years. All data were expressed as Mean ± SEM (standard error of the mean) and analyzed using the Minitab program. Data were checked for normality using the Kolmogorov-Smirnov test. To verify the differences between the means of body mass, testicular volume, hormonal dosage and seminal quality, in the parametric data, it was performed by Analysis of Variance (ANOVA), followed by the significant difference by Fisher's test, while for the data there was no The Wilcoxon test was used for parametric parameters. The correlation between the hormonal levels of testosterone and estradiol with parameters of body mass, testicular biometry and seminal parameters was performed by Pearson correlation for parametric data and Spearman correlation for non-parametric data. Results were considered significant when P < 0.05.