Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802)

Abstract This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Resumo Este estudo objetivou avaliar os efeitos da temperatura e motilidade embrionária sobre o desenvolvimento esquelético de jacaré-do-pantanal (Caiman yacare). Os ovos foram incubados com 90% de umidade e empregou-se a temperatura de 29°C por 45 dias. Após, para a incubação do Grupo I a temperatura continuou em 29°C, mas associou-se à injeção de 4-aminopiridina (29°C-4AP, n = 15) aplicada nos dias 46, 47, 48 e 49, do Grupo II permaneceu em 29°C (n = 14) e do Grupo III elevou-se para 33°C (n = 14). A movimentação foi mensurada através do monitor digital Egg Buddy® nos dias 30, 35, 42, 49, 56 e 60 dias. Aos 60 dias, os embriões foram eutanasiados e coletadas amostras embrionárias. Na análise estatística não foram observadas diferenças entre os grupos para o fator temperatura sobre a motilidade embrionária no desenvolvimento esquelético. Em contraste, a motilidade evidenciou diferença estatística no dia 49 para o Grupo I (P 0,001) e apresentou maiores proporções de nariz e mão. Esses dados demonstraram pela primeira vez que o aumento na motilidade, induzidos farmacologicamente resultam em divergências fenotípicas na proporção de segmentos anatômicos durante a ontogenia pré-natal, podendo alterar efetivamente a adaptação dos animais em ambientes específicos.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802) / Influência da temperatura de incubação e da motilidade embrionária sobre o crescimento dos membros de Caiman yacare (Daudin, 1802)

Abstract This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P < 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Resumo Este estudo objetivou avaliar os efeitos da temperatura e motilidade embrionária sobre o desenvolvimento esquelético de jacaré-do-pantanal (Caiman yacare). Os ovos foram incubados com 90% de umidade e empregou-se a temperatura de 29°C por 45 dias. Após, para a incubação do Grupo I a temperatura continuou em 29°C, mas associou-se à injeção de 4-aminopiridina (29°C-4AP, n = 15) aplicada nos dias 46, 47, 48 e 49, do Grupo II permaneceu em 29°C (n = 14) e do Grupo III elevou-se para 33°C (n = 14). A movimentação foi mensurada através do monitor digital Egg Buddy® nos dias 30, 35, 42, 49, 56 e 60 dias. Aos 60 dias, os embriões foram eutanasiados e coletadas amostras embrionárias. Na análise estatística não foram observadas diferenças entre os grupos para o fator temperatura sobre a motilidade embrionária no desenvolvimento esquelético. Em contraste, a motilidade evidenciou diferença estatística no dia 49 para o Grupo I (P < 0,001) e apresentou maiores proporções de nariz e mão. Esses dados demonstraram pela primeira vez que o aumento na motilidade, induzidos farmacologicamente resultam em divergências fenotípicas na proporção de segmentos anatômicos durante a ontogenia pré-natal, podendo alterar efetivamente a adaptação dos animais em ambientes específicos.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802).

This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P < 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Recuperação anestésica e analgesia residual da infusão continua intravenosa de fantanil, lidocaína, cetamina e fentanyl-lidocaína-cetamina associados à anesthesia total intravenosa com Propofol em cadelas submetidas à ovariossalpingo-histerectomia eletiva / Anesthetic recovery and residual analgesia of constant rate infusion intraoperative of fentanyl, lidocaine, ketamine or fentanyl-lidocaine-ketamine associated with total intravenous anesthesia with propofol in bitches submitted to elective ovariohysterectomy

Avaliou-se a recuperação anestésica e a analgesia residual da infusão contínua (IC) de fentanil (F), lidocaína (L), cetamina (K) e fentanil-lidocaína-cetamina (FLK), associados à anestesia total intravenosa com o propofol, em cadelas submetidas à ovariossalpingo-histerectomia. Foram utilizados 32 animais pré-medicados com acepromazina, distribuídos em quatro grupos de acordo com o tratamento analgésico: F: bolus de 0,0036mg/kg de fentanil e IC de 0,0036mg mg/kg/h; L: bolus de 3mg/kg de lidocaína e IC de 3mg/kg/h; K: bolus de 0,6mg/kg de cetamina e IC de 0,6mg/kg/h; e FLK: bolus e IC dos três fármacos nas doses supracitadas. Após o bolus do tratamento analgésico, foi realizada a indução e o início da IC do tratamento analgésico e do propofol. Para avaliação da recuperação anestésica, foram considerados os tempos de extubação, decúbito esternal, posição quadrupedal e os efeitos adversos. A avaliação da analgesia foi realizada por meio da escala visual analógica e modificada de Glasgow, durante seis horas. Os efeitos adversos observados foram vômito, sialorreia e tremor muscular. Receberam analgesia de resgate 100% dos animais do grupo F, 87,5% do K, 50% do L e 12,5% do FLK. O FLK demonstrou maior analgesia, e a recuperação anestésica foi semelhante em todos os grupos.(AU)

The anesthetic recovery and residual analgesia of continuous rate infusion (CRI) of fentanyl (F), lidocaine (L), ketamine (K) and fentanyl-lidocaine-ketamine (FLK) associated with total intravenous anesthesia with propofol in bitches submitted to ovariohysterectomy were evaluated. 32 animals were used, pre-medicated with acepromazine and distributed into four groups according to analgesic treatment: F loading dose (LD) of 0.0036mg/kg fentanyl, and CRI of 0.0036mg/kg/h, L: LD of 3mg/kg lidocaine, and CRI of 3mg/kg/h; K: LD of 0.6mg/kg ketamine, and CRI of 0.6mg/kg/h and FLK: LD and CRI of the three drugs in the above mentioned doses. After the LD of analgesic treatment, the induction was performed and the CRI of the analgesic treatment and propofol started. To evaluate the anesthetic recovery, the time of extubation, sternal decubitus, quadrupedal position and adverse effects were considered. The analgesia evaluation was performed using the visual scale and modified Glasgow for six hours. The adverse effects observed were vomiting, sialorrhea and muscle tremor. 100% of the animals in group F, 87.5% of K, 50% of L and 12.5% of FLK received rescue analgesia. FLK demonstrated greater analgesia, and anesthesia recovery was similar in all groups.(AU)

Autotransfusão imediata em cão - relato de caso / Instant autotransfusion in dog - case report

As hemorragias podem levar à morte, em caso de não interrupção e recuperação da volemia. A substituição do sangue pode ser realizada por transfusão homóloga ou autóloga. Na transfusão homóloga, o sangue é obtido de um doador, na autotransfusão ou transfusão autóloga, o sangue é coletado do próprio paciente. Objetivou-se relatar a autotransfusão em um cão como um método simples, rápido e barato, e discutir esse procedimento como uma alternativa emergencial para clínicas e hospitais que não possuem bancos de sangue ou animais doadores. Foi atendido um cão Rottweiler fêmea, 42,8kg, 10 anos de idade, com queixa principal de tumor mamário e para realização de castração. Após o procedimento, a paciente apresentou hemoperitônio, sendo encaminhada para novo procedimento cirúrgico, durante o qual se observou grande quantidade de sangue livre na cavidade abdominal. Foi realizada a autotransfusão de emergência, pela técnica das duas seringas, devido à indi1111sponibilidade de sangue ou animal doador no momento do procedimento cirúrgico emergencial, demonstrando ser uma opção eficiente, econômica, de fácil acesso e segura, por ser uma transfusão normotérmica, apresentar diminuição do risco de sobrecarga circulatória e por ser o sangue compatível, devendo ser mais explorada, pois existem poucos dados descritos na literatura.(AU)

Hemorrhages can lead to death in case of non-interruption and recovery of blood volume. Blood replacement may be performed by homologous or autologous transfusion. In homologous transfusion, blood is obtained from a donor, in autotransfusion or autologous transfusion, blood is collected from the patient himself. We aimed to report autotransfusion in a dog as a simple, quick, and inexpensive method, and discuss it as an emergency alternative for clinics and hospitals that do not have blood banks or animal donors. A female Rottweiler dog, 42.8kg, 10 years of age, was treated with a primary complaint of mammary tumor and for castration. After the procedure the patient presented hemoperitoneum, being referred to a new surgical procedure, where a large amount of free blood was observed in the abdominal cavity. The emergency autotransfusion was performed by the two syringes technique, due to the unavailability of blood or donor animal at the time of the emergency surgical procedure, proving to be an efficient, economical, easily accessible and safe option because it is a normothermic transfusion, risk of circulatory overload and blood is compatible. It should be more exploited because there are few data described in the literature.(AU)

Método não invasivo para avaliação da movimentação embrionária de Caiman yacare (Daudin, 1802) / Non-invasive method for evaluation of embrionic moviment of Caiman yacare (Daudin, 1802)

This study aimed to evaluate the viability of using a non-invasive digital monitor to monitor heart rate (HR) and motility during the embryonic development of Pantanal alligator (Caiman yacare) using Egg Buddy ® , at different incubation temperatures. The collection of the eggs followed the Ranching system and egg identification, transportation, and incubation were performed with the required care; all eggs were incubated with 90% humidity at 29°C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29°C, increased to 33°C or maintained at 29°C and embryos simultaneously treated with 4-aminopyridine on days 46, 47, 48, and 49 (29°C-4AP). Embryo movement was measured with a digital non-invasive monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were sacrificed. In the statistical analysis no differences were observed between the groups for the temperature (33°C and 29°C); for motility, a difference was observed at day 49 for the 29°C-4AP group. This revealed that the non-invasive evaluation method can be used to verify embryonic motility and HR effectively in Caiman yacare embryos.(AU)

Clinical, laboratory and epidemiological aspects of HPV infection in a low-income population from South Bahia, Brazil.

The aim of this study was to determine the prevalence and risk factors for human papillomavirus (HPV) infection in the Southern region of the State of Bahia, evaluating the performance of alternative complementary methods for cervical lesion detection. Cervical samples from women who attended healthcare units were collected and diagnosed by visual inspection, cervical cytology and nested polymerase chain reaction (PCR). Moreover, hemi-nested PCR was performed to detect different HPV genotypes. The prevalence of HPV infection was 47·7%, with genotype 16 detected in most cases. Infection was associated with dyspareunia and bleeding (P < 0·001, odds ratio (OR) 5·6, 95% confidence interval (CI) 2·815-11·14) and hormonal contraceptive use (P = 0·007, OR 2·33, 95% CI 1·25-4·34). There was a positive correlation between positive PCR and positive visual inspection, cervical cytology and symptoms reported. Furthermore, visual inspection was twice as specific, and had a greater positive predictive value than cytology. We showed a high prevalence of HPV infection in Southern Bahia, with HPV 16 being the most common type, and visual inspection being most effective at detecting HPV lesions, corroborating the suggestion that it can be applied in routine gynecologic examinations for low-income populations.

Aspectos clínicos, cirúrgicos, histológicos e pós-operatórios de oito cadelas com leiomioma vaginal / Clinical, surgical, histological and postoperative aspects in the case of eight bitches with vaginal leiomyoma

O presente trabalho reporta as características clínicas e cirúrgicas de oito casos de cadelas portadoras de leiomioma vaginal, com histórico de aumento de volume na região perineal, aparecimento de massa pela vulva, disquezia, tenesmo e disúria. As pacientes foram submetidas à cirurgia de episiotomia para ressecção da massa neoplásica, e em quatro casos foi necessária a ovariossalpingo-histerectomia. Porém, uma delas apresentou recidiva, e, no segundo procedimento cirúrgico de exérese de neoformação, o histopatológico diagnosticou leiomiossarcoma. A paciente ainda apresentou mais duas recidivas, até o proprietário optar pela eutanásia. Decorridos quatro meses, as sete cadelas diagnosticadas com leiomioma não apresentaram recidiva, e os proprietários relataram ausência de sinais relacionados com a doença descrita. Conclui-se que a episiotomia para ressecção da massa vaginal e a ovariossalpingo-histerectomia nas cadelas inteiras nos casos de tumores vaginais benignos são tratamentos satisfatórios, com bom prognóstico, ao contrário do leiomiossarcoma.

This study reports the clinical and surgical procedures of eight cases of bitches with vaginal leiomyoma. All animals presented increased volume in the perineal region, and exteriorization of the vulva, dyschezia, tenesmus and dysuria. The patients underwent episiotomy surgery for resection of the neoplastic mass, and ovariosalpingohisterectomy in four cases. However, one had recurrence on the second surgical procedure for removal of neoformation, histopathological diagnosed leiomyosarcoma, the patient also had two more relapses, until the owner chose euthanasia. After four months, the seven dogs diagnosed with leiomyoma showed no recurrence and the owners reported no signs of the disease described. We concluded that episiotomy for vaginal mass resection combined with ovariohysterectomy in cases of benign vaginal tumors have shown satisfactory treatment with good prognostic.

CYP21A2 genotypes do not predict the severity of hyperandrogenic manifestations in the nonclassical form of congenital adrenal hyperplasia.

There is a strong correlation between the severity of genotypes and 17OH-progesterone levels in patients with the nonclassical form of 21-hydroxylase deficiency (NC-CAH); however, there are few studies regarding the correlation with clinical signs. The aim of the study was to evaluate whether genotypes correlate with the severity of the hyperandrogenic phenotype. A cohort of 114 NC-CAH patients were diagnosed by stimulated-17OHP ≥10 ng/ml. CYP21A2 genotypes were divided into 2 groups according to the severity of enzymatic impairment; mild and severe. Clinical data and hormonal profiles were compared between the 2 groups. Age at onset of manifestations did not differ between children or adults carrying both mild and severe genotypes. Frequencies of precocious pubarche and hirsutism, with or without menstrual abnormalities, were similar between the 2 groups. There were no differences in basal testosterone levels of adult symptomatic females carrying both genotypes, but there were differences between adult females with (92.9±49.5 ng/dl) and without hirsutism (43.8±38 ng/dl) (p=0.0002). Similar frequencies of both genotypes were observed in asymptomatic females and in those with clitoromegaly. Nonclassical genotypes do not predict the severity of phenotype. Asymptomatic and virilized females carrying the same genotype suggest that there is a modulatory effect of genes involved in the androgen pathway on the phenotype.