Effect of the addition of antifreeze protein type I on the quality of post-thawed domestic cat epididymal sperm.

Cryopreservation of domestic cat semen is mainly performed as a model for the establishment of endangered wild feline protocols. The supplementation of antifreeze protein type I (AFP I) to cryopreservation medium has shown improvement in frozen-thawed sperm quality in other species, but its effect on cat semen has not yet been tested. This study aimed to assess the addition of AFP I to cryopreservation medium in domestic cats. Sperm was obtained from the cauda epididymis of orchiectomized cats; sperm was then pooled in Tris buffer and allocated into three treatments, according to AFP I final concentration: 0 (control), 0.1, and 0.5 µg/ml. Nine replicates were cryopreserved in a two-step protocol and subsequently thawed at 37°C for 30 s. There was no difference (P > 0.05) among the control, 0.1 and 0.5 µg/ml groups for parameters such as motility, vitality, functional membrane integrity, mature chromatin, normal morphology, and sperm binding to egg perivitelline membrane. In the 0.5 µg/ml group only, percentages of live sperm with intact acrosome and of sperm with most inactive mitochondria (DAB III) showed a significant reduction, along with a tendency (P = 0.053) to an increase in the percentage of sperm with most active mitochondria (DAB II). In conclusion, the supplementation of 0.1 and 0.5 µg/ml of AFP I did not promote consistent beneficial effects on the overall sperm cryotolerance in domestic cats.

Effects of d-cloprostenol administrations with 7.5 and 11.5-day intervals between administrations on pregnancy rates after artificial insemination in estrous cyclic dairy goats.

This study was conducted to evaluate effects of two administrations of d-cloprostenol at different intervals to synchronize the time of estrus and ovulation among estrous cyclic goats. In Experiment 1, 32 does were treated with 30 µg d-cloprostenol at 7.5 (T7.5, n = 16) or 11.5-day (T11.5, n = 16) intervals. In Experiment 2, the same treatments were administered and there was AI of the does (T7.5, n = 40 and T11.5, n = 38). In Experiment 1, ultrasonic assessments of ovaries were conducted at the time of the second administration of d-cloprostenol, every 12 h until detection of ovulation, and 7 days after estrous onset to detect the corpora lutea, as well as for pregnancy diagnosis 40 days after AI. In Experiment 1, the estrous response (90.6%, 29/32) was similar (P > 0.05) in both groups. Diameter of the largest follicle at the time of administration of the second dose was larger (P = 0.01) in the T7.5 than T11.5 group (7.0 compared with 5.7 mm), while the values for ovarian variables were similar (P > 0.05). In Experiment 2, the greatest (P < 0.001) synchrony in timing of initiation of estrus in does (T7.5 = 83.3% and T11.5 = 50.0%) occurred after the second day (36-48 h). The pregnancy rate tended (P = 0.0836) to be greater for does in the T7.5 (71.4%, 40/56) than T11.5 (55.6%, 30/54) group. With use of both protocols, there were acceptable estrous synchronization and pregnancy rates in estrous cyclic dairy goats.

Why do children and adolescents neglect dental flossing?

AIM: To identify the variables and actual difficulties related to children and adolescents' non-compliance with dental flossing. METHODS: This cross-sectional study with 36 children and 59 adolescents were selected from dental clinics at the Dental School, University of São Paulo. The percentage of surfaces with disclosed biofilm was used to evaluate general oral hygiene. Participants answered questions concerning dental flossing (difficulties, self-reported motivation, and previous instruction). An examiner observed how the participants flossed their teeth and their possible faults. Univariate and multiple logistic regression analyses were performed and odds ratio values were calculated in order to verify the association between non-compliance (or difficulties) with flossing and possible reasons for that. RESULTS: Similar non-compliance with daily flossing was observed among children and adolescents (p = 0.95). Children's flossing negligence was strongly associated with self-reported laziness in flossing (p = 0.02), and negatively associated with their previous practice by some dentists (p = 0.009). Self-described difficulties in flossing also showed an association with laziness in flossing (p = 0.03). No association was found between negligence of flossing and all variables tested among adolescents (p ≥ 0.05). CONCLUSIONS: Low compliance and difficulties in flossing among children and adolescents seemed to be more related to lack of motivation, although problems concerning manual skills were also observed.

Hybrid Self-Adaptive Evolution Strategies Guided by Neighborhood Structures for Combinatorial Optimization Problems.

This article presents an Evolution Strategy (ES)--based algorithm, designed to self-adapt its mutation operators, guiding the search into the solution space using a Self-Adaptive Reduced Variable Neighborhood Search procedure. In view of the specific local search operators for each individual, the proposed population-based approach also fits into the context of the Memetic Algorithms. The proposed variant uses the Greedy Randomized Adaptive Search Procedure with different greedy parameters for generating its initial population, providing an interesting exploration-exploitation balance. To validate the proposal, this framework is applied to solve three different [Formula: see text]-Hard combinatorial optimization problems: an Open-Pit-Mining Operational Planning Problem with dynamic allocation of trucks, an Unrelated Parallel Machine Scheduling Problem with Setup Times, and the calibration of a hybrid fuzzy model for Short-Term Load Forecasting. Computational results point out the convergence of the proposed model and highlight its ability in combining the application of move operations from distinct neighborhood structures along the optimization. The results gathered and reported in this article represent a collective evidence of the performance of the method in challenging combinatorial optimization problems from different application domains. The proposed evolution strategy demonstrates an ability of adapting the strength of the mutation disturbance during the generations of its evolution process. The effectiveness of the proposal motivates the application of this novel evolutionary framework for solving other combinatorial optimization problems.

Goats reinfected with toxoplasma gondii: loss of viable prolificacy and gross revenue / Cabras reinfectadas com toxoplasma gondii: perdas de prolificidade viável e receita bruta

We determined the reproductive parameters and clinical disorders in pregnant goats infected and reinfected with Toxoplasma gondii, and posteriorly the loss of gross revenue due to congenital toxoplasmosis was estimated. Of the 25 non-pregnant females negative for T. gondii, 20 were orally inoculated (ME 49 strain) and of these, 15 pregnant females chronically infected were orally reinoculated (VEG strain) with T. gondii oocysts. Five groups were formed (n=5): GI, GII and GIII (reinoculations at 40, 80 and 120 days of gestation, respectively), GIV (inoculation) and GV (no inoculation). Clinical and serological exams were performed on days 0 (prior to inoculation), 3, 6 9, 15 and 21 and every 7 days post-inoculation. Exams were also performed on day 3 and every 7 days post-reinoculation. Reproductive management was performed on all females and initiated when the females infected displayed IgG titers IFAT<1,024. From the average prolificacy indexes of each experimental group were estimated: total production of kilograms of live weight (total kg LW) of goats for slaughter, gross revenue and loss of gross revenue in U.S. dollars (US$), designed for a herd of 1,000 matrices. The unviable prolificacy indexes were 0.8 (GI), 1.2 (GII) and 0.2 (GIII). Clinical disorders affected 57.1% (GI), 75.0% (GII) and 16.7% (GIII) of the offspring of goats reinfected with T. gondii. Congenital toxoplasmosis in goats reinfected resulted in the loss of 26.5% of gross revenues, being GI (US$ 10,577.60 or 57.1%) and GII (US$ 12,693.12 or 60%) holders of the highest values and percentages of economic losses. It was found that congenital toxoplasmosis reinfection cause clinical disorders in goats chronically infected with T. gondii and their offspring with birth of unviable animals and loss of gross revenue, at different stages of pregnancy (40, 80 and 120 days of gestation)...

Nós determinamos os parâmetros reprodutivos e distúrbios clínicos em cabras gestantes infectadas e reinfectados com Toxoplasma gondii, e posteriormente, foi estimada a perda de receita bruta devido à toxoplasmose congênita. Das 25 fêmeas não prenhes negativas para T. gondii, 20 foram inoculadas oralmente (cepa ME 49) e, destas, 15 fêmeas gestantes infectadas cronicamente foram reinoculadas (cepa VEG), via oral, com oocistos de T. gondii. Cinco grupos foram formados (n = 5): GI, GII e GIII (reinoculações aos 40, 80 e 120 dias de gestação, respectivamente), GIV (inoculação) e GV (não inoculação). Exames clínicos e sorológicos foram realizados nos dias 0 (antes da inoculação), 3, 6 9, 15 e 21 e a cada sete dias após a inoculação. Os exames também foram realizados nos dias 3 e a cada sete dias de pós-reinoculação. Manejo reprodutivo foi realizado em todas as fêmeas e iniciou-se quando as fêmeas infectadas exibiram títulos de anticorpos IgG<1.024. A partir dos índices médios de prolificidade de cada grupo experimental foram estimados: a produção total de kg de peso vivo (total kg PV) de cabritos para o abate, receita bruta e perda de receita bruta em dólares norte-americanos (US$), projetadas para um rebanho de 1000 matrizes. Os índices de prolificidade inviáveis foram de 0,8 (GI), 1.2 (GII) e 0,2 (GIII). Distúrbios clínicos afetaram 57,1% (GI), 75,0% (GII) e 16,7% (GIII) das crias de cabras reinfectados com T. gondii. A toxoplasmose congênita em crias das cabras reinfectadas com T. gondii resultou na perda de 26,5% da receita bruta, sendo GI (US $ 10,577.60 e 57,1%) e GII (US $ 12,693.12 e 60.0%) os detentores dos mais altos valores e porcentagens de perdas econômicas. Verificou-se que a reinfecção toxoplásmica congênita causa distúrbios clínicos em cabras cronicamente infectadas com T. gondii e sua prole com o nascimento de animais inviáveis e perda de receita bruta, em diferentes fases da gestação (40, 80 e 120 dias de gestação)...

Effects of anesthesia with the essential oil of Ocimum gratissimum L. in parameters of fish stress / Efeitos da anestesia com o óleo essencial de Ocimum gratissimum L. em parâmetros de estresse de peixes

RESUMO: The effects of anesthesia with the essential oil of Ocimum gratissimum (EOO) in parameters of stress after handling were investigated in silver catfish (Rhamdia quelen). EOO was obtained from the aerial parts by hydrodistillation. Juveniles were anesthetized with 70 or 300 mg L-1 EOO and submitted to air exposure for 1 minute. The fishes were sampled immediately or transferred to anesthetic-free aquaria until sampling. In the first experiment, juveniles had their blood collected at 0, 1, 4, and 8 h after handling to assay plasma cortisol and blood glucose levels. The unanesthetized animals were restrained manually for blood collection. In the second experiment, water samples of the recovery aquaria were collected to evaluate net ion fluxes at 0 - 4 h and 4 - 8 h. Water and ethanol controls were also performed under the same conditions. The results showed that the cortisol levels did not differ among the treatments. Hyperglycemia was verified in fish exposed to 70 and 300 mg L-1 EOO at 1 h and 4 h after handling. After 8 h, cortisol and glucose concentrations were lower or similar than those from immediately after handling for all treatments. EOO anesthesia prevented Na+ efflux observed in the control groups in both flux periods. There were net Cl- and K+ effluxes at 0 - 4 h and influxes at 4 - 8 h after handling in most treatments, and these fluxes did not differ among the treatments. The results suggest that EOO did not impair stress recovery and did not act as an additional handling stressor in silver catfish.

RESUMO: Os efeitos da anestesia com o óleo essencial de Ocimum gratissimum (EOO) em parâmetros de estresse após manuseio foram investigados em jundiás (Rhamdia quelen). EOO foi obtido a partir das partes aéreas por hidrodestilação. Os juvenis foram anestesiados com 70 ou 300 mg L-1 de EOO e expostos ao ar por 1 minuto. Os peixes foram amostrados imediatamente ou transferidos para aquários sem anestésico até amostragem. No primeiro experimento, os juvenis tiveram seu sangue coletado em 0, 1, 4, e 8 h após manuseio para avaliar os níveis de cortisol e glicemia. Os animais não anestesiados foram contidos manualmente para coleta sanguínea. No segundo experimento, amostras de água foram coletadas do aquário de recuperação dos animais para avaliação do fluxo iônico entre 0 - 4 h e 4 - 8 h. Grupos controles em água e etanol também foram realizados sobre as mesmas condições. Os resultados demonstraram que os níveis de cortisol não diferiram entre os tratamentos. Hiperglicemia foi detectada em peixes expostos a 70 e 300 mg L-1 de EOO em 1 h e 4 h após o manuseio. Após 8 h, os teores de cortisol e glicose foram menores ou similares aqueles imediatamente após o manuseio para todos os tratamentos. A anestesia com EOO preveniu o efluxo de Na+ observado para os grupos controle em ambos os períodos avaliados. Ocorreram efluxos de Cl- and K+ entre 0 - 4 h e influxos entre 4 - 8 h após o manuseio para a maioria dos tratamentos, e estes eventos não diferiram entre os tratamentos. Os resultados sugerem que o EOO não prejudica a recuperação do animal frente ao evento estressor ou atua como estressor adicional ao manuseio em jundiás.

Bioprospection of bacteria and yeasts from Atlantic Rainforest soil capable of growing in crude-glycerol residues.

The increasing world production of biodiesel has resulted in an accumulation of crude glycerol as the major byproduct. This could be used as carbon source for industrial microbiology, with economic and environmental advantages for the biodiesel industry. We explored an Atlantic Rainforest soil sample to search for crude glycerol-degrading microorganisms. Microcosms of this soil were established containing minimal medium + 8% crude glycerol (w/w); the biological activity was measured by respirometry. High CO2 levels were found in some of the crude glycerol microcosms, suggesting the activity of microorganisms capable of degrading this residue. In an attempt to isolate and cultivate these microorganisms in vitro, aliquots of the soil suspension were plated on minimal medium containing 10% crude glycerol (v/v). Out of 19 morphologically distinct isolates, 12 bacteria and 6 yeasts were identified by PCR from universal primers 16S and 26S rDNA, respectively. Optical density readings revealed growth differences among cultures. Two yeasts and three bacteria with distinct growth profiles stood out and appeared to have potential for liquid fermentation of crude glycerol. The yeasts adapted rapidly, but produced relatively little biomass. Opposite tendencies were found in the bacteria. Amplicon sequencing placed the bacterial isolates as close to Staphylococcus arlettae, Pseudomonas citronellolis, and Bacillus megaterium, and the yeasts to Trichosporon moniliiforme and Meyerozyma guilliermondii. We concluded that these species have potential for use in crude glycerol bioreactors and for bioremediation processes.

Detection of fetal fibronectin in twin pregnancies in relation to gestational age.

CONTEXT: The presence of fetal fibronectin in the cervix or vagina has been investigated as a possible marker for the risk of preterm birth. Fetal fibronectin in cervical fluid can provide direct evidence of pathologic changes at the interface of fetal and maternal tissues. OBJECTIVE: To evaluate the presence of fetal fibronectin as a predictor of premature delivery in twin pregnancies in relation to gestational age. DESIGN: Accuracy study. SETTING: University referral unit. PARTICIPANTS: 52 pregnant women with twin pregnancies and gestational age of between 24 and 34 weeks. MAIN MEASUREMENTS: Sensitivity, specificity, predictive values and relative risk ratios of the correlation between fetal fibronectin and preterm birth before 34 and 37 weeks using an immediate-reading membrane test on cervicovaginal secretions obtained from participants. RESULT: The sensitivity varied between 66.7% and 85.7%, whereas the specificity was from 58.3% to 81.8% according to gestational age at the time of sampling. The relative risk of spontaneous preterm birth after a positive fetal fibronectin test, as compared with a negative fetal fibronectin test, rose from 2.8 at 24-26 weeks to 4.1 at 27-30 weeks. Analyses of the risk of delivery before 34 weeks were not statistically significant. CONCLUSION: Fetal fibronectin in the cervicovaginal secretions of patients with twin pregnancies is a useful tool for the early identification of twin pregnancies likely to deliver before 37 weeks. However, the clinical value of the fibronectin test is limited because of low indices for prediction of delivery before 34 weeks. The best period for performing the fetal fibronectin test in twin pregnancies to predict preterm delivery is between 27 and 30 weeks.

Obstetric and perinatal aspects in patients with congenital heart diseases.

The benefits of surgical treatment for patients with congenital heart disease in relation to pregnancy are still controversial. We studied 48 pregnant women (mean age = 25 years) with surgically-corrected congenital heart diseases (Group 1). This included 15 cyanotic diseases: Fallot's tetralogy (11 cases); Ebstein's anomaly (2 cases); transposition of the great arteries (1 case); and double outlet of the right ventricle (1 case). We compared them to 52 pregnant women (mean age = 26 years) with untreated congenital heart diseases, which included 11 cases of Eisenmenger's syndrome (Group 2). Group 2 showed a higher incidence of maternal death (12 vs. 0 percent; p = 0.01), perinatal mortality (15 vs. 0 percent; p = 0.01) and prematurity (32 vs. 7 percent; p = 0.01). Spontaneous abortion (4 vs. 10 percent), Caesarean deliveries (48 vs. 66 percent) or growth retardation (13 vs. 28 percent) did not present any significant differences between these groups. Surgical treatment in patients with heart diseases is associated with a better maternal and fetal prognosis. Therefore, surgery must be considered when counseling patients with congenital heart diseases.

PIP: In a prospective study of 100 pregnant Brazilian women with congenital heart diseases, the 48 patients who had undergone corrective surgery an average of 10 years before the index pregnancy (group 1) had substantially better maternal and fetal/neonatal outcomes than the 52 women with no history of surgery (group 2). Group 1 included 16 women with cyanotic diseases and 9 with residual heart lesions. In Group 2, 11 women had Eisenmenger's syndrome. There were 6 maternal deaths (12%) in Group 2: 3 related to Eisenmenger's syndrome and 3 in women with aortic or subaortic stenosis, but none in Group 1. There were 2 fetal and 5 neonatal deaths in Group 2, but, again, no such deaths in Group 1. Most maternal and fetal/neonatal deaths were associated with left obstructive involvement and cyanosis. Group 2 also included significantly more cases than Group 1 of low birth weight (38% versus 13%), premature birth (32% versus 7%), and intrauterine growth retardation (28% versus 13%). The highest rates of these adverse outcomes were recorded among infants of mothers with Eisenmenger's syndrome, obstructive involvement, and cyanosis. Residual surgical lesions had no adverse effects on maternal or infant outcomes. Surgery should be considered in patients with correctable heart disease who desire pregnancy. However, pregnancy is not recommended for women with Eisenmenger's syndrome and aortic stenosis.