Juvenile panhypopituitarism in a dog - what are the therapeutic challenges?

Background: Juvenile panhypopituitarism is an endocrinopathy that affects young dogs and must be differentiated from other causes of height disorders, as one could have a deficiency of one or more adenohypophysis hormones, besides growth hormone. Diagnosis consists of excluding endocrine and non-endocrine causes. Treatment requires administration of heterologous porcine growth hormone or progestins, as well as supplementing other hormones that are deficient in the circulation. The prognosis for these patients tends to be unfavorable and they have a shorter life expectancy. The present case aimed to report the therapeutic challenges in a dog diagnosed with juvenile panhypopituitarism. Case: This study presents the case of a 6-month-old crossbred Labrador/Brazilian Fila dog, not neutered, 6.7 kg of body weight (body condition score 5/9), with a growth disorder and persistence of the puppy coat, when compared to other members of the same litter. During inspection, it was possible to observe an undersized dog and soft puppy coat, besides no bone irregularities or joint tenderness was noted. The other physical examination parameters were within the normal range for the species. No changes in complete blood count and only increases in urea, cholesterol and alkaline phosphatase activity were observed. Thyroid and abdominal ultrasound (US) examination did not reveal any remarkable changes. After serum dosage of insulin-like growth factor, thyroxine, thyrostimulating hormone, and cortisol, the patient was diagnosed with juvenile panhypopituitarism and underwent therapy with medroxyprogesterone and thyroid hormone supplementation. Monitoring was instituted at intervals of 3, 6, and 12 weeks, and currently every 3 or 4 months and the IGF-1 values normalized after 6 months with. After 8 months of therapy, the patient had good body growth and bone mineralization compared to the time of diagnosis. However, skeletal development was completed only 12 months after hormone replacement, accompanied by the presence of vertebral osteophytes and coxofemoral osteoarticular alterations. Considering chronic use of progestins, ovariosalpingohysterectomy (OSH) was recommended, but for personal reasons, the owners chose to do not submit her to surgery. After 18 months of treatment, the dog starts to exhibit prostration, selective appetite, and increased abdominal volume. After imaging exam, she was forwarded for OSH, due to consistent findings of pyometra. Subsequently, even the supervision of possible comorbidities involved in the chronic use of progestins was maintained at half-yearly intervals, the bitch returned to the service with skin thickening, increased limb volume, and macroglossia after 60 months of therapy. At this point IGF-1 values were higher when compared to the previous measurements and the application of medroxyprogesterone was suspended. Its supplementation was reintroduced only after 8 months when IGF-1 was significantly reduced. To date, the patient is close to 6 years of age and with a good quality of life. Discussion: The greatest therapeutic challenge for these patients involves dealing with the adverse effects of progestins, mainly related to reproductive disorders and alopecia at the application site, as well as maintaining adequate hormone replacement in order to avoid hypersomatotropism. Although longevity is lower in these individuals, the patient in this report has achieved 6 years of age and has had an excellent quality of life so far.

Pituitary carcinoma in a bitch: clinical, tomographic, histopathological and immunohistochemistry findings

Background: Primary pituitary carcinoma is rarely reported in dogs and only few reports describe its malignancy. In veterinary literature, primary pituitary carcinomas correspond up to 2.4% to 3.4% of intracranial neoplasms found in dogs andinformation regarding its biological behavior is quite limited. In humans, primary pituitary carcinomas represent less than1.0% of all tumors found in the pituitary gland. The proposed classification for pituitary carcinoma in humans and dogsdetermines that the tumor must have its origin in adenohypophyseal region and disseminated metastasis by cerebrospinalfluid or systemically to other organs must be observed. In dogs, a few reports have described primary pituitary carcinoma.The goal of this report was to describe clinical, tomographic, histopathological and immunohistochemistry features of abitch with primary pituitary carcinoma with adjacent invasion.Case: A 7-year-old female spayed Golden Retriever dog was assessed by general practice due progressive weight loss,muscular atrophy, lethargy, blindness, head pressing, and hyporexia for 21 days. Computed tomography (CT) showeda cerebral parenchyma with expansive extra-axial base formation, originating from sella turcica topography, measuringabout 2.0 cm dorsally, displacing the third ventricle, suggesting the diagnosis of pituitary neoplasia. The hormones thyroidstimulating hormone (TSH) and total thyroxine (T4) as well as stimulation ACTH test were unremarkable. After 7 days,neurological clinical signs progressed and unfortunately the patient died ten days later after hospitalization. A necropsyexam revealed pituitary gland with increased dimensions (2.5 x 2.0 cm). Histopathological fi ndings revealed tumor proliferation in pituitary gland. The neoplasm showed invasion to the nervous parenchyma and metastatic foci between the brainlobes. Immunohistochemistry was positive for keratin and neuron-specifi c enolase...

Investigation of stress in primates maintained in zoo and its main implications on behavior and well-being

Brazil is considered the country which holds the largest number of known primate species. At captives without environmental enrichment program, especially in zoos, it is common for the animals to be easily discouraged from captive life, being bound to develop chronic stress condition. This review aimed to provide veterinary professionals and students with updated bibliography on stress in zoo captive primates, focusing on reactions resulting from these changes, as well as their implications for behavior and well-being. Integrative literature review using descriptors "Captivity", "Stress", "Primates", "Zoos" and "Wellbeing", on the main scientific research bases. The present work highlighted stress responses and its consequences on primate’s health and behavior captive in zoo through bibliographic survey, performing selected articles analysis, results interpretation, and discussion. It is concluded behavioral profile individual variations expose primates differently to stressful conditions. Public visitation, lack of structure, reduced enclosure, improper nutritional management, and unstimulated environment may be potential stress sources for some housed primates in zoo.

Investigation of stress in primates maintained in zoo and its main implications on behavior and well-being

Brazil is considered the country which holds the largest number of known primate species. At captives without environmental enrichment program, especially in zoos, it is common for the animals to be easily discouraged from captive life, being bound to develop chronic stress condition. This review aimed to provide veterinary professionals and students with updated bibliography on stress in zoo captive primates, focusing on reactions resulting from these changes, as well as their implications for behavior and well-being. Integrative literature review using descriptors "Captivity", "Stress", "Primates", "Zoos" and "Wellbeing", on the main scientific research bases. The present work highlighted stress responses and its consequences on primates health and behavior captive in zoo through bibliographic survey, performing selected articles analysis, results interpretation, and discussion. It is concluded behavioral profile individual variations expose primates differently to stressful conditions. Public visitation, lack of structure, reduced enclosure, improper nutritional management, and unstimulated environment may be potential stress sources for some housed primates in zoo.(AU)

The role of prostaglandin F2α on ovulation and LH release in cows / A função da prostaglandina F2α na ovulação e na liberação de LH em vacas

This study aimed to evaluate the role of prostaglandin F2α (PGF) on ovulation. In Experiment 1, cows were randomly allocated to two treatments to receive 150 µg of d-Cloprostenol (PGF Group, n = 12) or 2 mL of NaCl 0.9% (Control Group, n = 11) and CIDRs, were removed 4 days later. No cow ovulated in Control and PGF groups. In Experiment 2, cows were randomly separated into two experimental groups to receive 4 injections of 150 µg of d-Cloprostenol (n = 9) or 2 mL of NaCL 0.9% (n = 9). In this experiment, ovulation was not observed in any cows. In Experiment 3, ovariectomized cows receive three injections of 300µg of PGF analog (PGF Group, n = 5), 100µg of Lecirelin (GnRH Group, n = 5) or 2 mL of PBS (Control Group, n = 4). The LH concentration was higher (P <0.0001) in cows from the GnRH group than in the PGF and Control groups. In experiment 4, cows with preovulatory follicles (>11.5 mm) were treated with Saline (Control Group, n = 6); Lecirelin (GnRH Group, n = 7) or Cloprostenol Sodium (PGF Group, n = 6). There was a significant increase in the vascular area of follicles from 0 to 24 h in GnRH and PGF treatments. In conclusion, PGF was not able to induce ovulation in cows with high or low plasma progesterone concentration. Additionally, PGF alone was not able to induce LH release and follicle luteinization, but increased follicular vascularization.(AU)

O objetivo deste estudo foi avaliar o papel da prostaglandina F2α (PGF) na ovulação. No Experimento 1, as vacas foram alocadas aleatoriamente em dois tratamentos para receber 150 µg de d-Cloprostenol (Grupo PGF, n = 12) ou 2 mL de NaCl 0,9% (Grupo Controle, n = 11) e os CIDR, foram removidos 4 dias depois. Nenhuma vaca ovulou nos grupos Controle e PGF. No Experimento 2, as vacas foram separadas aleatoriamente em dois grupos experimentais para receber 4 injeções de 150 µg de d-Cloprostenol (n = 9) ou 2 mL de NaCL 0,9% (n = 9). Não foi observada ovulação em nenhum dos animais deste experimento. No Experimento 3, vacas ovariectomizadas receberam três injeções de 300µg de análogo de PGF (Grupo PGF, n = 5), 100µg de Lecirelina (Grupo GnRH, n = 5) ou 2 mL de PBS (Grupo Controle, n = 4). A concentração de LH foi maior (P <0,0001) nas vacas do grupo GnRH do que nos grupos PGF e Controle. No Experimento 4, vacas com folículos pré-ovulatórios (> 11,5 mm) foram tratadas com solução salina (Grupo Controle, n = 6), Lecirelina (Grupo GnRH, n = 7) ou Cloprostenol Sódico (Grupo PGF, n = 6). Houve um aumento significativo na área vascular dos folículos de 0 a 24h nos tratamentos com GnRH e PGF. Em conclusão, a PGF não foi capaz de induzir ovulação em vacas com alta ou baixa concentração plasmática de progesterona. Além disso, a PGF sozinha não foi capaz de induzir a liberação de LH e a luteinização do folículo, mas aumentou a vascularização folicular.(AU)

Conduta diagnóstica em equino da raça Crioula com síndrome de Cushing / Diagnostic process in a crioulo horse with Cushing’s syndrome

Background: Equine pituitary pars intermedia dysfunction, also known as equine Cushing’s syndrome, is a neurodegenerative disease. An important risk factor for Cushing’s is advanced aging and it is the most common endocrine disorder inolder horses. The prevalence in horses aged over 10 and 15 years is reported as 9.3% and 21%, respectively. Due to the slowprogressive nature of the disease, seasonal variation in hormone output and overlapping endocrine response to other events,accurate diagnosis is challenging. The diagnosis requires the combination of anamnesis, clinical signs, in addition to laboratory tests results. This study aimed to report Cushing’s syndrome in a Crioulo breed horse focusing on diagnostic methods.Case: A 13-year-old male Crioulo breed, orchiectomized, was attended at the Universidade de Passo Fundo (UPF), in PassoFundo, RS, Brazil. The owner reported that the animal had progressive weight loss and coat abnormal growth, with curlyappearance. From visual inspection, body condition score was 4 (1-9) bulging abdomen was noticed, hirsutism, depressionand lethargy. Also, there was a large neoplastic mass on the left side of gluteal region. Later, this mass was classified inhistopathological examination as a fibroblastic sarcoid and was treated. The animal presented physical parameters withinthe physiological limits of the specie. Normochromic normocytic anemia and neutrophilic leukocytosis were reported in thehematologic evaluation. In coproparasitological examination, there were 300 eggs per gram of feaces. Hyperadrenocorticismwas suspected in the clinical examination and dexamethasone suppression test was performed to confirm the fact. Basal serumwas collected at 17 h (M0) and subsequently 40 µg/kg of dexamethasone was administered intramuscular...

The role of prostaglandin F2α on ovulation and LH release in cows / A função da prostaglandina F2α na ovulação e na liberação de LH em vacas

This study aimed to evaluate the role of prostaglandin F2α (PGF) on ovulation. In Experiment 1, cows were randomly allocated to two treatments to receive 150 µg of d-Cloprostenol (PGF Group, n = 12) or 2 mL of NaCl 0.9% (Control Group, n = 11) and CIDRs, were removed 4 days later. No cow ovulated in Control and PGF groups. In Experiment 2, cows were randomly separated into two experimental groups to receive 4 injections of 150 µg of d-Cloprostenol (n = 9) or 2 mL of NaCL 0.9% (n = 9). In this experiment, ovulation was not observed in any cows. In Experiment 3, ovariectomized cows receive three injections of 300µg of PGF analog (PGF Group, n = 5), 100µg of Lecirelin (GnRH Group, n = 5) or 2 mL of PBS (Control Group, n = 4). The LH concentration was higher (P <0.0001) in cows from the GnRH group than in the PGF and Control groups. In experiment 4, cows with preovulatory follicles (>11.5 mm) were treated with Saline (Control Group, n = 6); Lecirelin (GnRH Group, n = 7) or Cloprostenol Sodium (PGF Group, n = 6). There was a significant increase in the vascular area of follicles from 0 to 24 h in GnRH and PGF treatments. In conclusion, PGF was not able to induce ovulation in cows with high or low plasma progesterone concentration. Additionally, PGF alone was not able to induce LH release and follicle luteinization, but increased follicular vascularization.(AU)

O objetivo deste estudo foi avaliar o papel da prostaglandina F2α (PGF) na ovulação. No Experimento 1, as vacas foram alocadas aleatoriamente em dois tratamentos para receber 150 µg de d-Cloprostenol (Grupo PGF, n = 12) ou 2 mL de NaCl 0,9% (Grupo Controle, n = 11) e os CIDR, foram removidos 4 dias depois. Nenhuma vaca ovulou nos grupos Controle e PGF. No Experimento 2, as vacas foram separadas aleatoriamente em dois grupos experimentais para receber 4 injeções de 150 µg de d-Cloprostenol (n = 9) ou 2 mL de NaCL 0,9% (n = 9). Não foi observada ovulação em nenhum dos animais deste experimento. No Experimento 3, vacas ovariectomizadas receberam três injeções de 300µg de análogo de PGF (Grupo PGF, n = 5), 100µg de Lecirelina (Grupo GnRH, n = 5) ou 2 mL de PBS (Grupo Controle, n = 4). A concentração de LH foi maior (P <0,0001) nas vacas do grupo GnRH do que nos grupos PGF e Controle. No Experimento 4, vacas com folículos pré-ovulatórios (> 11,5 mm) foram tratadas com solução salina (Grupo Controle, n = 6), Lecirelina (Grupo GnRH, n = 7) ou Cloprostenol Sódico (Grupo PGF, n = 6). Houve um aumento significativo na área vascular dos folículos de 0 a 24h nos tratamentos com GnRH e PGF. Em conclusão, a PGF não foi capaz de induzir ovulação em vacas com alta ou baixa concentração plasmática de progesterona. Além disso, a PGF sozinha não foi capaz de induzir a liberação de LH e a luteinização do folículo, mas aumentou a vascularização folicular.(AU)

Conduta diagnóstica em equino da raça Crioula com síndrome de Cushing / Diagnostic process in a crioulo horse with Cushings syndrome

Background: Equine pituitary pars intermedia dysfunction, also known as equine Cushings syndrome, is a neurodegenerative disease. An important risk factor for Cushings is advanced aging and it is the most common endocrine disorder inolder horses. The prevalence in horses aged over 10 and 15 years is reported as 9.3% and 21%, respectively. Due to the slowprogressive nature of the disease, seasonal variation in hormone output and overlapping endocrine response to other events,accurate diagnosis is challenging. The diagnosis requires the combination of anamnesis, clinical signs, in addition to laboratory tests results. This study aimed to report Cushings syndrome in a Crioulo breed horse focusing on diagnostic methods.Case: A 13-year-old male Crioulo breed, orchiectomized, was attended at the Universidade de Passo Fundo (UPF), in PassoFundo, RS, Brazil. The owner reported that the animal had progressive weight loss and coat abnormal growth, with curlyappearance. From visual inspection, body condition score was 4 (1-9) bulging abdomen was noticed, hirsutism, depressionand lethargy. Also, there was a large neoplastic mass on the left side of gluteal region. Later, this mass was classified inhistopathological examination as a fibroblastic sarcoid and was treated. The animal presented physical parameters withinthe physiological limits of the specie. Normochromic normocytic anemia and neutrophilic leukocytosis were reported in thehematologic evaluation. In coproparasitological examination, there were 300 eggs per gram of feaces. Hyperadrenocorticismwas suspected in the clinical examination and dexamethasone suppression test was performed to confirm the fact. Basal serumwas collected at 17 h (M0) and subsequently 40 µg/kg of dexamethasone was administered intramuscular...(AU)

Effect of Heat Stress on Expression of Main Reproductive Hormone in Hypothalamic-Pituitary-Gonadal Axis of Wenchang Chicks

ABSTRACT The hypothalamic-pituitary-gonadal (HPG) axis is a key neuroen-docrine regulation system involved in the growth and reproduction of poultry. High-temperature conditions lead to the physiological dysfunction of target organs of the HPG axis of poultry, ultimately affecting the animals growth and development. In this study, we evaluated the effect of heat stress (HS) on the development of cells secreting major reproductive hormones of the HPG axis (i.e., hypothalamus, pituitary gland, ovary, and testis) of Wenchang chicks. Seventy-two one-day-old healthy Wenchang chicks were randomly divided into control (CK) and HS groups. The HS group was placed in a 40 ± 0.5°C artificial climate chamber for heat-stress treatment from 13:00 to 15:00 daily for six consecutive weeks. As development progressed, compared with the CK group, the gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels in the hypothalamus and testosterone (T) levers in the testes of male chicks in the HS group were significantly decreased at three weeks of age (p 0.05). However, GnRH levels in the hypothalamus and estradiol (E2) levels in the ovaries of female chicks in the HS group were variable and significantly lower than those of the CK group at four and five weeks of age (p 0.05). In addition, the LH and FSH levels in the pituitary gland were significantly lower than those in the CK group at two and four weeks of age and at four and six weeks of age, respectively (p 0.05). In brief, HS caused dysfunction of the corresponding target organs of the HPG axis in Wenchang chicks, and this affected the normal growth and development of the cells HPG axis.

Evidence of pituitary hypoplasia associated with partial central diabetes insipidus in a young Persian cat

Background: Congenital anomalies are an uncommon pituitary hypofunction cause associated to multiple hormone deficiencies. Congenital hyposomatotropism is often related to an inherited anomaly, characterized mainly by delayed growth. It is not uncommon to find associated thyroid-stimulating hormone and gonadotropin deficiencies. Pituitary malformation may be associated to progressive cystic lesion expansion. Central diabetes insipidus (CDI) is another rare disease associated to polyuria (PU) and polydipsia (PD) secondary to antidiuretic hormone (ADH) deficient secretion. The aim of this report is to describe a likely case of pituitary hypoplasia, associated with partial CDI in a cat. Case: A 9-month-old unneutered male Persian cat weighing 2 kg was presented due to severe polyuria and polydipsia associated with growth deficit when compared with its sibling. After clinical and laboratory evaluations during the months in which the patient was monitored, reduced serum concentration of insulin-like growth factor-1 (IGF-1), thyroid-stimulating hormone (TSH), thyroid hormones, and testosterone were documented, confirming the diagnosis of hyposomatotropism, hypogonadism, and secondary hypothyroidism. Furthermore, therapeutic diagnosis with desmopressin revealed partial central diabetes insipidus (CDI). As the sibling showed normal development aging 13-months, a radiographic examination of the forelimb (carpus) was performed on both cats. There was lack of growth plate fusion in the patient, without any other evidence of dysgenesis, whereas complete epiphyseal closure was observed in the sibling. Despite therapeutic prescriptions of desmopressin and levothyroxine, the owners refused further follow-up to the...

Evidence of pituitary hypoplasia associated with partial central diabetes insipidus in a young Persian cat

Background: Congenital anomalies are an uncommon pituitary hypofunction cause associated to multiple hormone deficiencies. Congenital hyposomatotropism is often related to an inherited anomaly, characterized mainly by delayed growth. It is not uncommon to find associated thyroid-stimulating hormone and gonadotropin deficiencies. Pituitary malformation may be associated to progressive cystic lesion expansion. Central diabetes insipidus (CDI) is another rare disease associated to polyuria (PU) and polydipsia (PD) secondary to antidiuretic hormone (ADH) deficient secretion. The aim of this report is to describe a likely case of pituitary hypoplasia, associated with partial CDI in a cat. Case: A 9-month-old unneutered male Persian cat weighing 2 kg was presented due to severe polyuria and polydipsia associated with growth deficit when compared with its sibling. After clinical and laboratory evaluations during the months in which the patient was monitored, reduced serum concentration of insulin-like growth factor-1 (IGF-1), thyroid-stimulating hormone (TSH), thyroid hormones, and testosterone were documented, confirming the diagnosis of hyposomatotropism, hypogonadism, and secondary hypothyroidism. Furthermore, therapeutic diagnosis with desmopressin revealed partial central diabetes insipidus (CDI). As the sibling showed normal development aging 13-months, a radiographic examination of the forelimb (carpus) was performed on both cats. There was lack of growth plate fusion in the patient, without any other evidence of dysgenesis, whereas complete epiphyseal closure was observed in the sibling. Despite therapeutic prescriptions of desmopressin and levothyroxine, the owners refused further follow-up to the...(AU)

Central blindness associated with a pituitary adenoma in a mare / Cegueira de origem central associado a adenoma de pars intermedia de hipófise em uma égua

Background: Pituitary pars intermedia (PI) adenoma is a benign adenohypophysis neoplasm, rare in Brazil, which maycompress adjacent structures and lead to dysfunctions of the endocrine organs. The most affected species are equines,particularly aged animals. This neoplastic disease is often associated with Cushing’s syndrome, when the pituitaryadrenocortical axis is affected. However, this neoplasm is seldom associated with clinical blindness. This paper describesa case of pituitary pars intermedia (PI) adenoma which caused blindness in a mare.Case: An emaciated, blind mare that had difficulty finding water and food was referred to “Hospital Veterinário de GrandesAnimais” of “Universidade Federal Rural do Rio de Janeiro”, in Seropédica, state of Rio de Janeiro, Brazil, and later tothe “Setor de Anatomia Patológica” (SAP/UFRRJ) for necropsy. During necropsy it was found that the animal’s pituitarygland consisted of a circumscribed globous mass measuring 3.5x2.5x2.5 cm, symmetrical and well delimited at the sellaturcica. Several organs were collected, fixed in 10% formalin and then processed for histological examination. The brainwas subjected to serial cleavage along the path of the vision organs. All collected tissues were stained with hematoxylinand eosin; the pituitary was stained with Schiff Periodic Acid (PAS) to differentiate neoplastic cells and with Luxol FastBlue to evidence demyelination. Histopathology found that the pars intermedia (PI) was thickened and compressed theneurohypophysis and adenohypophysis. Cells were arranged in a disorganized manner or formed follicles or cysts containing eosinophilic material (colloid), with granules strongly positive after staining with PAS. Proliferated cells werelarge, polyhedral to ovoid, and contained clear basophilic granular material. The nuclei were spherical to ovoid and therewere mild anisocytosis and anisocariasis. Cysts were frequently found in the pars intermedia (PI)...

Central blindness associated with a pituitary adenoma in a mare / Cegueira de origem central associado a adenoma de pars intermedia de hipófise em uma égua

Background: Pituitary pars intermedia (PI) adenoma is a benign adenohypophysis neoplasm, rare in Brazil, which maycompress adjacent structures and lead to dysfunctions of the endocrine organs. The most affected species are equines,particularly aged animals. This neoplastic disease is often associated with Cushings syndrome, when the pituitaryadrenocortical axis is affected. However, this neoplasm is seldom associated with clinical blindness. This paper describesa case of pituitary pars intermedia (PI) adenoma which caused blindness in a mare.Case: An emaciated, blind mare that had difficulty finding water and food was referred to “Hospital Veterinário de GrandesAnimais” of “Universidade Federal Rural do Rio de Janeiro”, in Seropédica, state of Rio de Janeiro, Brazil, and later tothe “Setor de Anatomia Patológica” (SAP/UFRRJ) for necropsy. During necropsy it was found that the animals pituitarygland consisted of a circumscribed globous mass measuring 3.5x2.5x2.5 cm, symmetrical and well delimited at the sellaturcica. Several organs were collected, fixed in 10% formalin and then processed for histological examination. The brainwas subjected to serial cleavage along the path of the vision organs. All collected tissues were stained with hematoxylinand eosin; the pituitary was stained with Schiff Periodic Acid (PAS) to differentiate neoplastic cells and with Luxol FastBlue to evidence demyelination. Histopathology found that the pars intermedia (PI) was thickened and compressed theneurohypophysis and adenohypophysis. Cells were arranged in a disorganized manner or formed follicles or cysts containing eosinophilic material (colloid), with granules strongly positive after staining with PAS. Proliferated cells werelarge, polyhedral to ovoid, and contained clear basophilic granular material. The nuclei were spherical to ovoid and therewere mild anisocytosis and anisocariasis. Cysts were frequently found in the pars intermedia (PI)...(AU)

REPRODUÇÃO DE LAMBARI-DO-RABO-AMARELO (Astyanax lacustris) UTILIZANDO DIFERENTES INDUTORES

O lambari é um peixe com pequeno porte, que apresentou um aumento em sua produção nacional de 37% de 2015 a 2019 com um total de 661,019 Kg produzidos em 2019. O objetivo desse estudo foi avaliar a desova e características reprodutivas de diferentes indutores hormonais em adultos de lambari-do-rabo-amarelo (Astyanax lacustris). Foram utilizados 135 fêmeas (12,54 g ± 2,33 e 7,66 cm ± 0,63 cm) e 135 machos (5,83 g ± 0,39 g e 6,14 cm ± 0,64 cm), todos em idade reprodutiva. Foram avaliados três diferentes indutores: (i) 0,4 pellets de Ovopel®/kg de peso vivo; (ii) 0,5 mL de acetato de buserelina/kg de peso vivo; (iii) Extrato de Hipófise de Carpa (EHC) (5,5 mg de EHC/kg de peso vivo para fêmeas e 2,5 mg de EHC/kg de peso vivo para machos). A variável horas-grau foi maior (P<0,05) para o tratamento com Ovopel© e Acetato de buserelina em relação ao tratamento com EHC. O número de larvas por peso das fêmeas foi maior com a utilização do Ovopel®. Fecundidade absoluta e fecundidade relativa foram maiores para os tratamentos com Ovopel® e EHC. A maior taxa de fertilização ocorreu no tratamento com acetato de buserelina. A taxa de eclosão foi maior para o acetato de buserelina e Ovopel®. A utilização de Ovopel® proporcionou melhores resultados para a reprodução de A. lacustris quando comparados ao EHC e acetato de buserelina.

Lambari is a small fish, which had an increase of 37% in its production from 2015 to 2019 with a total amount of 661.019 Kg produced in the year of 2019. The objective of this study was to evaluate the spawning and reproductive characteristics of different spawning inducers in adults of yellow-tailed lambari (Astyanax lacustris). A total of 135 females (12.54 g ± 2.33 and 7.66 cm ± 0.63 cm) and 135 males (5.83 g ± 0.39 g and 6.14 cm ± 0.64 cm), all of reproductive age, were used. Three different inducers were evaluated: (i) 0.4 pellets of Ovopel®/kg of body weight; (ii) 0.5 ml of buserelin acetate/kg of body weight; (iii) Carp's Pituitary Extract (CPE) (5.5 mg CPE/kg body weight for females and 2.5 mg of CPE/kg of body weight for males). The variable hour-degree was higher (P <0.05) for the treatment with Ovopel © and buserelin acetate compared to the treatment with CPE. There number of larvae per female body weight was greater when using Ovopel®. Absolute fecundity (AF) and relative fecundity (RF) were higher for treatments with Ovopel® and CPE. The higher fertilization rate occurred in the treatment with buserelin acetate. The hatching rate was higher for buserelin acetate and Ovopel®. The use of Ovopel® provided greater results in the reproduction of A. lacustris when compared to CPE and buserelin acetate.

Seminal characteristics of piabanha before and after induction with different hormones / Características seminais de piabanha antes e após a indução com diferentes hormônios

The migratory species piabanha does not reproduce in lentic environments since it requires environmental stimuli for the maturation and extrusion of gametes, and therefore hormonal induction is mandatory. Current study compares the seminal characteristics of Brycon insignis without any hormonal induction (Control - Ctrl) and with two types of hormonal inductors, or rather, carp pituitary extract (T1 - 2.5 mg kg-1 body weight) and GnRH analogues, the latter applied in two different concentrations (T2 - 0.7 mg kg-1 body weight and T3 - 1.4 mg kg-1 body weight). Post-induction analyses showed that the hormones increased the motility rate - Ctrl (95%), T1 (100%), T2 (100%) and T3 (98%), although sperm concentration - Ctrl (11.52 x 109); T1 (4.37 x 109); T2 (4.34 x 109); T3 (4.01 x 109) decreased. Assessments for sperm vigor, motility time and spermatic morphology did not vary with hormonal induction. Hormonal inducer does not alter negatively the seminal characteristics of the piabanha, and the choice for the proper hormone depends on the preference of the dispenser. (AU)

A espécie migradora piabanha não possui a capacidade de reproduzir em ambientes lênticos devido à necessidade de estímulos ambientais para a maturação e extrusão dos gametas, por isso a necessidade da indução hormonal. No presente estudo, as características seminais do Brycon insignis foram comparadas sem indução hormonal (Ctrl) e utilizando dois tipos de indutores hormonais - Extrato de Hipófise de Carpa (T1 - 2,5 mg kg-1 de peso vivo) e Análogos de GnRH, sendo este último aplicado em duas concentrações distintas (T2 - 0,7 mg kg-1 de peso vivo e T3 - 1,4 mg kg-1 de peso vivo). As análises realizadas após a indução mostraram que os hormônios utilizados produziram um aumento da taxa de motilidade - Ctrl (95%), T1 (100%), T2 (100%) e T3 (98%), porém houve uma diminuição na concentração espermática - Ctrl (11,52 x 109 ), T1 (4,37 x 109 ), T2 (4,34 x 109 ) e T3 (4,01 x 109 ). Os restantes das avaliações, vigor espermático, tempo de motilidade e morfologia espermática não apresentaram variações com a indução hormonal. Portanto, a utilização do indutor hormonal não altera negativamente as características seminais de piabanha, e a escolha do mesmo se deve à preferência do manipulador.(AU)

Histopathological hoof laminar changes in horses with Pituitary Pars Intermedia Adenoma: cases report / Alterações histopatológicas do tecido laminar do casco em cavalos com adenoma da Pars Intermedia da Hipófise: relato de casos

Laminitis in horses is often associated with endocrine disorders, especially the pituitary pars intermedia dysfunction (PPID) in older animals. Morphologic exams of the laminar tissue of the hoof were performed in two horses with suspected PPID, with no clinical signs of laminitis. Changes compatible with laminitis of endocrine origin were observed, such as rounding of the nuclei of the basal cells, thinning and stretching of the secondary epidermal laminae and tissue proliferation. PPID horses with no clinical signs of laminitis may be affected by lesions of the laminar tissue of the hoof that compromise the integrity of the dermal-epidermal junction and may develop clinical symptoms of the disease. It has been suggested that the development stage of endocrine laminitis is longer, but further studies should be conducted to confirm it.(AU)

A laminite em cavalos está frequentemente associada a distúrbios endócrinos, como a disfunção da pars intermedia da pituitária (PPID) em animais mais velhos. Exames morfológicos do tecido laminar do casco foram realizados em dois cavalos com suspeita de PPID após o óbito, os quais não apresentaram sinais clínicos de laminite. Alterações compatíveis com a laminite de origem endócrina foram observadas, como arredondamento da núcleo das células basais, alongamento e afilamento das lâminas epidérmicas secundárias e proliferação tecidual. Cavalos com PPID sem sinais clínicos de laminite podem estar acometidos por lesões do tecido laminar do casco que comprometam a integridade das interdigitações dérmico-epidérmicas e podem desenvolver a sintomatologia clínica da doença. Sugere-se que o período de desenvolvimento da laminite endócrina seja mais longo, porém estudos adicionais devem ser realizados para confirmar essa hipótese.(AU)

ANÁLISE COMPORTAMENTAL, HEMATOLÓGICA E HORMONAL DE CÃES EM ABRIGO RECEBENDO SUPLEMENTAÇÃO COM NO STRESS

O estresse é um fenômeno onipresente e fundamental para a expressão de um comportamento adaptativo. Contudo, quando exacerbado ou existente de forma contínua, este perde sua função adaptativa e passa a prejudicar o indivíduo. Avalia-lo é uma tentativa de medir experiências subjetivas e individuais de cada indivíduo, sendo a atividade do eixo hipotálamo-pituitária-adrenal (HPA) o principal mecanismo de resposta fisiológica frente a diferentes fatores estressores, utilizando como variável métrica a mensuração de cortisol. Vários estudos já demonstraram a importância da terapia farmacológica e a aceleração do efeito terapêutico sobre a modificação comportamental, com destaque aos princípios naturais e com menos efeitos colaterais. O objetivo deste estudo é avaliar de maneira multifatorial, sob os aspectos comportamental, hematológico e hormonal, a resposta ao estresse de cães mantidos em canil e tratados com o complemento NO STRESS® em comparação à um grupo placebo, em um estudo de padrão duplo cego. Participaram da pesquisa 20 cães de abrigo diagnosticados com comportamentos estereotipados e distribuídos entre os grupos placebo e tratado, recebendo comprimidos diariamente durante 90 dias. Os cães permaneceram em baias individuais e foram realizadas coletas sanguíneas a cada 30 dias para realização de exames hematológicos e mensuração de cortisol. Foi empregado um etograma de comportamento estereotipado no dia 0 e no dia 90, o qual demonstrou diminuição significativa dos comportamentos estereotipados em ambos os grupos e diminuição significativa dos comportamentos locomotores no grupo tratado. A dinâmica de flutuação do cortisol demonstrou uma queda considerável dos níveis de cortisol até o dia 30 em ambos os grupos, com queda contínua apenas do grupo tratado até o fim do estudo. Enquanto o grupo placebo apresentou inversão desta curva, com aumento da frequência de comportamentos estereotipados a partir do dia 30. Os valores hematológicos não apresentaram alterações relevantes e acompanham o bom estado geral de saúde dos cães. O medicamento fitoterápico demonstrou eficácia e seguridade pela somatória das análises.

Stress is an ubiquitous and fundamental phenomenon for the expression of adaptive behavior. However, when exacerbated or existing continuously, it loses its adaptive function and starts to harm the individual. Evaluating it is an attempt to measure subjective and individual experiences of each individual, with the activity of the hypothalamic-pituitary-adrenal axis (HPA) being the main mechanism of physiological response to different stressors, using the measurement of cortisol as a metric variable. Several studies have already demonstrated the importance of pharmacological therapy and the acceleration of the therapeutic effect on behavioral modification, with emphasis on natural principles and with less side effects. The objective of this study is to evaluate in a multifactorial way, under the behavioral, hematological and hormonal aspects, the stress response of dogs kept in kennel and treated with the NO STRESS® complement in comparison to a placebo group, in a double blind standard study. . The study included 20 shelter dogs diagnosed with stereotyped behavior and distributed between the placebo and treated groups, receiving pills daily for 90 days. The dogs remained in individual stalls and blood samples were taken every 30 days for hematological exams and measurement of cortisol. A stereotyped behavior ethogram was used on day 0 and day 90, which demonstrated a significant decrease in stereotyped behaviors in both groups and a significant decrease in locomotor behaviors in the treated group. The dynamics of cortisol fluctuation demonstrated a considerable drop in cortisol levels up to the 30th day in both groups, with continuous drop only in the treated group until the end of the study. While the placebo group presented an inversion of this curve with an increase in the frequency of stereotyped behaviors starting on the 30th. The hematological values did not show relevant changes and follow the good general health of dogs. The herbal medicine demonstrated efficacy and safety due to the sum of the analyzes.

EFEITO DA ADMINISTRAÇÃO DE ANTICOAGULANTES SOBRE A RECUPERAÇÃO EMBRIONÁRIA DE ÉGUAS SUPEROVULADAS

A superovulação é uma biotécnica utilizada nos programas de transferência de embrião e proporciona diversos benefícios devido ao maior número de ovulações por ciclo. Entretanto, os protocolos superovulatórios na espécie equina ainda não são utilizados rotineiramente, pois apresentam resultados inconsistentes relacionados a baixa taxa de recuperação embrionária frente ao número de ovulações, estando muito aquém do esperado quando comparado a espécie bovina. Atualmente, sabe-se que uma das possíveis causas responsáveis por este fato pode estar relacionada pela formação de grandes coágulos de sangue obstruindo a fossa da ovulação, dificultando assim a captação do oócito para o interior da tuba uterina. Desta forma, o objetivo desse estudo foi avaliar o efeito da utilização de dois tipos de anticoagulantes parenterais (heparina não fracionada - HNF e heparina de baixo peso molecular - HBPM) em éguas superovuladas com extrato de pituitária equina (EPE), sobre a posterior taxa de recuperação embrionária. Foram utilizados quatro ciclos estrais de 11 éguas, sendo subdivididos em quatro grupos: Grupo 1 (G1) utilizado como controle; Grupo 2 (G2) receberam 25 mg de EPE intramuscular (IM) a cada 12 horas sendo a primeira e a terceira administração associada a 5 mg de dinoprost-trometamina (IM) e solução fisiológica 0,9% única aplicação 35 horas após indução da ovulação; Grupo 3 (G3) foi utilizado mesmo protocolo superovulatório, sendo substituído a solução fisiológica por um dos anticoagulantes, Heparina Sódica 450 UI/Kg/IV, e Grupo 4 (G4) onde as éguas foram superovuladas pelo mesmo protocolo mencionada acima e receberam 1 mg/Kg/IV de Enoxaparina Sódica. Os animais foram aleatoriamente designados aos quatros grupos em cross-over design, nos quais todos foram submetidos a todos os tratamentos. O intervalo de um ciclo estral para cada égua foi utilizado entre os tratamentos. Os tratamentos foram iniciados no sétimo dia pós-ovulação. A duração do tratamento com EPE não excedeu oito dias, sendo interrompido quando a maioria dos folículos atingissem diâmetro adequado para indução da ovulação. A indução da ovulação foi realizada utilizando 1 ml (250µg) de GnRH (Histrelina Acetato) intramuscular e 2500 UI de hCG (Gonadotrofina Coriônica Humana) intravenoso quando o(s) folículo(s) atingisse um diâmetro médio entre eles de 35 mm em ambos os grupos. A partir da indução da ovulação, 24 horas após as éguas foram inseminadas com sêmen a fresco de um garanhão de fertilidade comprovada. A colheita de embriões foi realizada no nono dia após a primeira ovulação detectada. Para cada animal submetido ao tratamento com anticoagulante foram realizadas avaliações laboratoriais (hemograma e coagulograma). Para isso foram padronizados os seguintes momentos de avaliação; para hemograma as amostras de sangue foram coletadas nos momentos prévio a administração do anticoagulante (T0 hora) servindo como controle, e três horas após a aplicação do anticoagulante (T3 horas), já para o coagulograma as amostras foram coletadas nos momentos T0, T1, T2, T3, T6, T9 e T12 horas após aplicação do anticoagulante. Os resultados demostraram maior (p<0,05) número de ovulações nos grupos de éguas superovuladas (G2 4,6±0,4; G3 5,2±0,5; G4 4,9±0,6) em relação ao G1 (1±0). Já o número de embriões recuperados por égua não diferiu (p>0,05) entre o G1 (0,6±0,2) e G2 (1,6±0,5), no entanto, foi maior para os grupos que receberam anticoagulantes G3 (2,5±0,3) e G4 (2,6±0,6) em relação ao G1. Quando a taxa de recuperação embrionária por folículo ovulado foi avaliado, os resultados demostraram que os grupos tratados com anticoagulantes apresentaram taxas similares (G3 50%; G4 52%) em comparação ao grupo controle (G1 64%), já o grupo superovulado controle (G2 36%) apresentou tendência de redução (P = 0,8) em relação ao G1. Em relação a avaliação laboratorial, os valores hematológicos dos animais em ambos os grupos que receberam anticoagulante (G3 e G4) no momento T3 horas não diferiu (p>0,05) em relação ao momento T0 hora, mantendo-se dentro dos valores de referência. Quanto ao coagulograma, para o G3 os valores de tempo de tromboplastina parcial ativado (TTPA) a partir de uma hora após aplicação (T1 hora) apresentou aumento significativo se manteve até doze horas após aplicação (T12 horas), onde todos os momentos diferiram-se (p<0,05) em relação ao momento (T0 hora), já para o tempo de protrombina (TP) houve um aumento (p<0,05) após aplicação do anticoagulante (T1-T2 horas) em relação ao tempo de avaliação pré aplicação (T0 hora), reduzindo-se gradativamente a partir de três horas até doze horas após aplicação (T3-T12 horas), não diferindo (p>0,05) em relação do T0 hora. Para o G4, uma hora após a aplicação (T1 hora) houve um aumento (p<0,05) da atividade plasmática fator anti-Xa, apresentando um constante declínio até doze horas (T12), não apresentando diferença (p>0,05) apenas neste momento em relação ao T9 horas e T0 hora. Desta forma, pode-se concluir que o protocolo proposto no presente estudo é uma alternativa para melhoria das taxas de recuperação embrionária em éguas superovuladas com EPE, mostrando-se seguro para o uso de ambos os tipos de anticoagulantes sem apresentar alterações hematológicas ou efeitos adversos que possam comprometer ou colocar em risco a condição clínica do animal.

Superovulation is a biotechnic from embryo transfer programs providing several benefits due to the higher number of ovulations per cycle. However, superovulation protocols in the equine species are not yet routinely used, as they present inconsistent results related to the low embryonic recovery rate concerning the number of ovulations, being far below expectations when compared to the bovine species. Currently, one of the possible causes responsible for this fact may be related to the development of large blood clots obstructing the ovulation fossa, making it difficult to capture the oocyte into the fallopian tube. Therefore, this study aimed to evaluate the effect of two types of parenteral anticoagulants (unfractionated heparin - HNF and low molecular weight heparin - LMWH) on equine pituitary extract (EPE) superovulated mares of embryonic recovery. Four estrous cycles of 11 mares were used and subdivided into four groups: Group 1 (G1) as a control; Group 2 (G2) received 25 mg intramuscular EPE (MI) every 12 hours being the first and third administration associated with 5 mg dinoprost-tromethamine (IM) and 0.9% saline only application 35 hours after ovulation induction; Group 3 (G3) in the same superovulation protocol, replacing the physiological solution with one of the anticoagulants, Sodium Heparin 450 IU / kg / IV, and Group 4 (G4) where the mares were superovulated by the same protocol mentioned above and received 1 mg. / Kg / IV of Enoxaparin Sodium. All animals were randomly assigned to the four cross-over design groups, in which all underwent all treatments. The interval of one estrous cycle for each mare was used between treatments. Treatments started on the seventh day after ovulation. EPE treatment did not exceed eight days being interrupted when most follicles reached the appropriate diameter for ovulation induction. Ovulation induction was performed using 1 ml (250µg) of intramuscular GnRH (Histrelin Acetate) and 2500 IU of intravenous hCG (Human Chorionic Gonadotropin) when the follicle (s) reached a mean diameter of 35 mm in both groups. Twenty-four hours afterward ovulation induction the mares were inseminated with fresh semen from a proven fertility stallion. Embryo collection was performed on the ninth day after the first ovulation detected. Each animal at anticoagulant treatment was submitted to laboratory evaluations (blood count and coagulogram). For blood count blood samples were collected before anticoagulant administration (T0 hour) serving as a control, and three hours after applying anticoagulant (T3 hours), while for the coagulogram the samples were collected at T0, T1, T2, T3, T6, T9 and T12 hours after anticoagulant application. Groups of superovulated mares showed higher (p <0.05) number of ovulations (G2 - 4.6 ± 0.4; G3 - 5.2 ± 0.5; G4 - 4.9 ± 0.6) in relation to G1 (1 ± 0). The number of embryos recovered by mare did not differ (p> 0.05) between G1 (0.6 ± 0.2) and G2 (1.6 ± 0.5), however, it was higher for the groups that received anticoagulants G3 (2.5 ± 0.3) and G4 (2.6 ± 0.6) concerning G1. Evaluating the embryo recovery rate by ovulated follicle, anticoagulant-treated groups had similar rates (G3 - 50%; G4 - 52%) compared to the control group (G1 - 64%). Superovulated control (G2 - 36%) showed a tendency to decrease (P = 0.8) concerning G1. Hematological values in both groups receiving anticoagulant (G3 and G4) at T3 hours did not differ (p> 0.05) from T0 hour, remaining the reference values. As for the coagulogram, for G3 the activated partial thromboplastin time (TTPA) values from one hour after application (T1 hour) showed a significant increase until twelve hours after application (T12 hours), where all moments differed (p <0.05) to (T0 hour). Prothrombin time (PT) had an increase (p <0.05) after anticoagulant application (T1-T2 hours) about pre-application evaluation (T0 hour), gradually decreasing from three hours to twelve hours after application (T3-T12 hours), not differing (p> 0.05) from T0 hour. For G4, one hour after application (T1 hour) there was an increase (p <0.05) of anti-Xa factor plasma activity, showing a constant decline up to twelve hours (T12), with no difference (p> 0, 05) only at this moment in relation to T9 hours and T0 hour. The protocol proposed in this study is an alternative for improving embryo recovery rates in EPE superovulated mares, proving safe use of both types of anticoagulants without presenting hematological alterations or adverse effects compromising or endangering the clinical condition of the animal.

INFLUÊNCIA DA ORIGEM E DA CONCENTRAÇÃO DE FSH SOBRE O CULTIVO IN VITRO DE FOLÍCULOS OVARIANOS ISOLADOS NA ESPÉCIE CAPRINA

Os objetivos deste estudo foram: 1) investigar uma curva concentração-resposta do FSH recombinante humano (hrFSH) no cultivo in vitro de folículos pré-antrais (FOPA) e antrais precoces (FOA) isolados caprinos (Fase 1); 2) comparar três diferentes origens de FSH (hipofisário porcino e recombinantes bovino e humano) durante o cultivo in vitro de FOPA e FOA caprinos isolados (Fase 2); 3) investigar se a extensão do período de cultivo, juntamente com mudanças dinâmicas na composição do meio de cultivo para cada estádio folicular, poderia melhorar a eficiência do sistema de cultivo in vitro para FOPA caprinos a taxas equivalentes aos FOA (Fase 3). Na fase 1, folículos isolados foram cultivados por 18 dias nos seguintes tratamentos: meio básico de cultivo (controle); ou meio controle suplementado com 10, 50 ou 100 mUI/mL de rhFSH. Na fase 2, FOPA e FOA foram cultivados por 18 dias e os tratamentos foram: meio base suplementado sem FSH (controle), 10, 50, ou 100 mUI/mL de pFSH (pFSH10, pFSH50 e pFSH100, respectivamente), 100 ng/mL de rbFSH (rbFSH) e 50 mUI/mL de rhFSH (rhFSH). Na fase 3, foram utilizados meios específicos para o cultivo in vitro de FOPA e FOA durante 24 ou 18 dias, respectivamente. A composição do meio de cultivo foi fixa (FOPA e FOA) ou dinâmica (FOPA). Os FOPA foram cultivados em meio fixo com alta concentração de insulina ou em meio dinâmico (insulina alta nos primeiros 6 dias, seguida de baixa insulina + GH até o final do período de cultivo) com ou sem GDF-9. Foram avaliadas a morfologia folicular, a formação de antro, a taxa de crescimento e a produção de estradiol, a viabilidade oocitária e a configuração da cromatina, bem como a expressão gênica relativa do mRNA da parede folicular, nas três fases. Na fase 1, nos FOA, a adição de FSH50 aumentou o diâmetro folicular e a taxa de crescimento, o percentual de oócitos totalmente crescidos e o diâmetro oocitário (P < 0,05) e tendeu a aumentar o percentual de oócitos em MII quando comparados ao controle (P = 0,07). Na fase 2, os tratamentos contendo pFSH apresentaram um percentual menor de folículos intactos e produção de estradiol e maior taxa de extrusão (P < 0,05). Para FOPA, a adição de pFSH10, pFSH100 e rhFSH diminuiu (P < 0,05) a retomada meiótica de oócitos em comparação aos tratamentos controle e rbFSH. Nos folículos antrais, rhFSH e pFSH10 aumentaram (P < 0,05, P = 0,08) a maturação oocitária. Em conclusão, a fonte de FSH afetou diferentemente a expressão gênica e a dinâmica folicular dos folículos pré-antrais e antrais in vitro. Na fase 3, o sistema de cultivo (fixo vs dinâmico) afetou diferentemente o resultado de todos os parâmetros avaliados, ou seja, morfologia e crescimento folicular, produção de estradiol, expressão gênica e maturação oocitária. Dependendo do sistema de cultivo, FOPA e FOA apresentaram taxas de crescimento e de maturação oocitária semelhantes quando cultivados in vitro sob as mesmas condições experimentais.

The aims of this study were: 1) investigate a concentration-response curve of human recombinant FSH (rhFSH) for in vitro culture of isolated caprine preantral and early antral follicles. (Phase 1); 2) compare three different sources of FSH (recombinant bovine, recombinant human, and porcine pituitary) during the in vitro culture of isolated caprine preantral and early antral follicles under the same experimental conditions (Phase 2); 3) investigate whether the extension of the culture period together with dynamic changes in the culture medium composition for each follicular stage could improve the efficiency of the in vitro culture system for late caprine preantral follicles (LPF) to rates equivalent to early antral follicles (EAF) (Phase 3). In phase 1, isolated follicles were cultured for 18 days in the following treatments: basic culture medium (control); or control medium supplemented with 10, 50, and 100 mIU/mL of hrFSH.In phase 2, preantral and early antral follicles were cultured for 18 days and the treatments were: base medium supplemented with no FSH (control), 10, 50, or 100 mIU/mL pFSH (pFSH10, pFSH50, and pFSH100, respectively), 100 ng/mL rbFSH (rbFSH), and 50 mIU/mL rhFSH (rhFSH). In phase 3, specific media were used for the in vitro culture of LPF and EAF during 24 or 18 days, respectively. Culture medium composition was either fixed (LPF and EAF) or dynamic (LPF). LPF were cultured either in a fixed medium with high insulin concentration, or in a dynamic medium (high insulin in the first 6 days followed by low insulin + GH until the end of the culture period) with or without GDF-9. EAF were cultured in a fixed medium containing low insulin, GH, and with or without GDF-9. Follicle morphology, antrum formation, growth rate, and estradiol production, and oocyte viability and chromatin configuration, and follicle wall mRNA relative gene expression were evaluated in the three phases. In phase 1, in antral follicles, the FSH addition at 50 mIU/mL increased follicular diameter and growth rate, the percentage of fully grown oocytes, and oocyte diameter (P < 0.05), and tended to increase the percentage of MII oocytes when compared to the control (P = 0.07). For preantral follicles, the addition of pFSH10, pFSH100, and rhFSH decreased (P < 0.05) oocyte meiotic resumption compared to control and rbFSH treatments. For antral follicles, rhFSH and pFSH10 increased (P = 0.08 P < 0.05) oocyte maturation. In conclusion, the source of FSH differently affected gene expression and follicular dynamics of preantral and antral follicles in vitro. In phase 3, the culture system (fixed vs. dynamic) differently affected the outcome of all parameters evaluated, i.e., follicle morphology and growth, estradiol production, gene expression, and oocyte maturation. Depending on the culture system, LPF and EAF had similar growth rates and oocyte maturation rates when cultured in vitro under the same experimental conditions.

Abordagem de biologia de sistemas para a determinação de mecanismos moleculares associados à eficiência alimentar de bovinos Nelore

ALEXANDRE, P. A. Abordagem de biologia de sistemas para a determinação de mecanismos moleculares associados à eficiência alimentar de bovinos Nelore. XXX f. Tese (Doutorado) Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, 2018. A eficiência alimentar (EA) é um fenótipo complexo, controlado por diversos processos biológicos. Determinar e entender esses processos é fundamental para selecionar animais superiores ou mesmo orientar decisões de manejo com o objetivo de aumentar a produtividade e diminuir o impacto ambiental da pecuária. Neste trabalho, propusemos analisar a EA através de uma abordagem de biologia de sistemas, baseada em transcriptômica multitecidual, a fim de gerar um entendimento sistêmico dessa característica. Para isso, 18 animais extremos para consumo alimentar residual foram selecionados a partir de um grupo de 98 bovinos Nelore machos inteiros e tiveram seu transcriptoma de hipotálamo, pituitária, adrenal, músculo e fígado sequenciado (RNAseq). Os reads gerados foram alinhados com o genoma de referência bovino (UMD3.1), filtrados e a expressão de cada gene foi estimada. A partir desses dados três abordagens de análises de dados foram desenvolvidas. Na primeira, cinco critérios de inclusão foram definidos para selecionar genes e construir uma rede de co-expressão para os cinco tecidos, de forma que além de indicarmos diversos genes e processos associados à EA, também fomos capazes de determinar dois genes reguladores, o NR2F6 e o TGFB1. Na segunda abordagem focamos no eixo hipotálamo-pituitária-adrenal, também utilizando análises de co-expressão, mas dessa vez sem partir de prévia seleção de genes e concluímos que o sistema de recompensa do cérebro pode estar envolvido no estímulo para maior consumo de alimentos observado no grupo de baixa EA. Finalmente, com a terceira abordagem, identificamos RNAs longos não codificadores (lncRNAs) expressos nos cinco tecidos e encontramos 30 transcritos expressos diferencialmente entre a alta e baixa EA na pituitária, músculo e adrenal, sendo que alguns deles se mostraram relacionados a processos já previamente demostrados como sendo associados a essa característica. Concluímos que, apesar de não conseguirmos determinar nesse momento o papel da maior susceptibilidade ao estresse, reportado na literatura para animais de baixa EA, no estímulo para maior ingestão de alimentos desse grupo, o sistema de recompensa hipotalâmico parece estar envolvido nesse processo. A maior ingestão pode ser a causa da resposta inflamatória observada no fígado, sendo ela de origem bacteriana, indicada pela maior concentração de endotoxina sérica nos animais menos eficientes. O maior turnover de proteínas no músculo de animais de baixa EA já havia sido indicado como um dos fatores que levam ao maior gasto energético nesses indivíduos e foi confirmado nesse trabalho. Além de alguns fatores de transcrição serem indicados como reguladores centrais desse fenótipo, lncRNAs também parecem ter função regulatória importante na EA.

ALEXANDRE, P. A. Systems biology approach for determination of molecular mechanisms associated with feed efficiency in Nellore cattle. 143 f. PhD Thesis Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, 2018. Feed efficiency (FE) is a complex phenotype, controlled by several biological processes. Determining and understanding these processes is fundamental to select superior animals or even guide management decisions, aiming to increase productivity and reduce the environmental impact of livestock. In this work, we propose to analyze FE through a systems biology approach, based on multi-tissue transcriptomics, in order to generate a systemic understanding of this trait. For this purpose, 18 extreme animals for residual feed intake were selected from a group of 98 male Nellore cattle and had their hypothalamus, pituitary, adrenal gland, muscle and liver transcriptome sequenced (RNAseq). Reads generated were aligned with the bovine reference genome (UMD3.1), filtered and the expression of each gene was estimated. From these data three experiments were developed. In the first one, five inclusion criteria were defined to select genes and to construct a network of coexpression for the five tissues, so that besides indicating several genes and processes associated with EA, we were also able to determine two regulatory genes, NR2F6 and TGFB1. In the second experiment, we focused on the hypothalamic-pituitary-adrenal axis, also using co-expression analysis, but this time without starting from previous selected genes. We conclude that the reward system of the brain might be involved in the stimulus for higher feed intake observed in the low EA group. Finally, in the third experiment, we identified long noncoding RNAs (lncRNAs) expressed in the five tissues and found 30 transcripts differentially expressed between the high and low FE in the pituitary, muscle and adrenal, and some of them were related to previously demonstrated processes associated to this trait. We conclude that although we cannot determine at this time the role of higher susceptibility to stress, reported in the literature for animals of low FE, in the stimulus for higher feed intake of this group, the hypothalamic reward system seems to be involved in this process. The higher ingestion might be the cause of the inflammatory response observed in the liver of, being of bacterial origin, indicated by the higher concentration of serum endotoxin in less efficient animals. The higher turnover of proteins in the muscle of low FE animals had already been indicated as one of the factors that lead to higher energy expenditure in these individuals and it was confirmed in this study. In addition to some transcription factors being indicated as central regulators of this phenotype, lncRNAs also appear to play an important regulatory role in FE.