Impact of ethanol and heat stress-dependent effect of ultra-diluted Arnica montana 6 cH on in vitro embryo production in cattle.

This study evaluated the effect of adding ultra-diluted and dynamized Arnica montana 6 cH, and its vehicle (0.3% ethanol) to the in vitro maturation (IVM) medium, in the absence (experiment 1) or presence (experiment 2) of heat stress (HS), on bovine oocyte maturation and in vitro embryo production (IVEP). In experiment 1 (n = 902 cumulus oocyte complexes, COCs), the treatments were 1) IVM medium (Control treatment), 2) IVM medium + 0.3% ethanol, and 3) IVM medium + Arnica montana 6 cH. In experiment 2 (n = 1064 COCs), the treatments were 1) IVM medium without HS, 2) IVM medium under HS, 3) IVM medium + ethanol under HS, and 4) IVM medium + Arnica montana under HS. In the absence of HS (experiment 1), the addition of Arnica montana to the IVM medium had a deleterious effect on the IVEP (cleavage and blastocyst rates) and the total cell number/blastocysts. On the other hand, ethanol (0.3%) increased IVEP in relation to the Control and Arnica montana treatments. However, in the presence of HS during IVM (experiment 2), the addition of ethanol or Arnica montana increased IVEP when compared to the HS treatment alone, and the Arnica montana treatment resulted in greater total cell number/blastocysts compared to the other treatments. In conclusion, this study showed for the first time that the negative or positive effect of Arnica montana 6 cH on IVEP depends on the culture condition (i.e., absence or presence of HS during IVM). On the other hand, ethanol showed beneficial and consistent results on IVEP regardless of exposure to HS.

Cholelithiasis and associated complications in pediatric patients. / Colelitiasis y complicaciones asociadas en pacientes pediátricos.

OBJECTIVE: To review the clinical characteristics and complications associated with cholelithiasis in pediatric patients by identifying risk factors and assessing surgical management and results. METHODS: Retrospective study in patients under 18 years of age undergoing cholelithiasis surgery. The following data were analyzed: age, sex, body mass index (BMI), associated comorbidities, clinical presentation, symptom duration, surgical treatment, pathological report, postoperative complications, and hospital stay. RESULTS: 135 cholelithiasis patients underwent surgery from 2013 to 2018, with an increasing trend in the annual number of cholecystectomies. Most patients were adolescents (86.7%) and female (72.6%), and they had cholesterol gallstones (86.6%) and >85 BMI (33%). Mean symptom duration was 85 days (SD: 148). Symptomatic cholelithiasis was present in 131 cases (97%). CL associated complications were recorded in 64.4% of patients, with cholecystitis, pancreatitis, and choledocholithiasis being the most frequent ones. All patients underwent laparoscopic cholecystectomy; 4 (2.9%) required conversion to open surgery, and 6 (4.4%) had postoperative complications. Mean hospital stay and postoperative follow-up were 5.7 days (SD: 4) and 2.3 months (SD: 1.9), respectively. CONCLUSIONS: Non-hemolytic cholelithiasis in the pediatric population is more frequent in female overweight and obese adolescents. Symptomatic cholelithiasis is associated with higher risk of complications such as cholecystitis, pancreatitis, or choledocholithiasis, which supports early surgical management.

OBJETIVO: Revisión de las características clínicas y complicaciones asociadas de los pacientes pediátricos con colelitiasis, mediante la identificación de factores de riesgo, la evaluación del manejo quirúrgico y sus resultados. METODOS: Estudio retrospectivo en pacientes menores de 18 años, intervenidos quirúrgicamente por colelitiasis. Se analizaron datos como edad, género, índice de masa corporal (IMC), comorbilidades asociadas, presentación clínica, tiempo de los síntomas, tratamiento quirúrgico, reporte de patología, complicaciones posoperatorias y estancia hospitalaria. RESULTADOS: Se intervinieron quirúrgicamente 135 pacientes con colelitiasis entre los años 2013 a 2018, con una tendencia ascendente en el número de colecistectomías realizadas anualmente. La mayoría fueron pacientes adolescentes (86,7%), con cálculos de colesterol (86,6%), de género femenino (72,6%) y con IMC > 85 (33%). El tiempo de los síntomas en promedio fue de 85 días (DE 148). Colelitiasis sintomática se presentó en 131 casos (97%). Complicaciones asociadas a CL se registraron en 64,4% pacientes, siendo las más frecuentes colecistitis, pancreatitis y coledocolitiasis. En todos los pacientes se realizó colecistectomía laparoscópica, 4 (2,9%) requirieron conversión a cirugía abierta y 6 (4,4%) presentaron complicaciones postoperatorias. La estancia hospitalaria y el seguimiento posoperatorio fueron en promedio 5,7 días (DE 4) y 2,3 meses (DE 1,9), respectivamentea. CONCLUSIONES: La colelitiasis no hemolítica en la población pediátrica, es más frecuente en pacientes adolescentes de género femenino con sobrepeso u obesidad. La colelitiasis sintomática está asociada con mayor riesgo de complicaciones como colecistitis, pancreatitis o coledocolitiasis, en consecuencia, se justifica una intervención quirúrgica temprana.

Influences of trace mineral nutrition and maternal flock age on broiler embryo bone development.

At hatch, the chick skeleton is a miniature of that of the adult bird. The hen deposits calcium, phosphorus, and trace minerals (copper, zinc, and manganese) along with vitamin D into the egg to allow development of the embryonic skeleton. The main source of calcium is the eggshell, whereas phosphorus, trace minerals, and vitamin D are mainly derived from the yolk. Calcium is absorbed from the eggshell and transferred to the embryo and yolk through the chorioallantoic membrane, whereas phosphorus and trace minerals are simultaneously mobilized by the yolk sac membrane. These processes start at day 12 of incubation and peak at around day 17. While the eggshell provides a steady supply of calcium until 19 d of incubation, phosphorus and trace mineral reserves decrease considerably and minimal skeletal development occurs in the last 3 d of incubation. Whether the low levels of phosphorus and trace minerals at late incubation prevent further bone growth, or some other biological control exists preventing further mineralization towards hatching is unknown. Maternal transfer of minerals and the influence of trace mineral form in the hen diet to advance the state of skeletal development at hatch have received increased research attention. Minimal effects on yolk mineral composition and bone growth were observed in the offspring of hens fed different forms of trace minerals. Embryos from young hens had inferior bone development towards the end of incubation and at hatch relative to chicks from older hens. This effect is likely a consequence of limited egg nutrient resources in eggs from young hens. The influence of maternal nutrient transfer on embryonic bone development has been clearly established. However, attempts to increase the state of skeletal development at hatch through increasing egg mineral content have met with limited success. The focus of this paper is the relationship between skeletal mineralization of the chicken embryo throughout incubation and egg mineral supply.

Physiological response of broiler embryos to different incubator temperature profiles and maternal flock age during incubation. 1. Embryonic metabolism and day-old chick quality.

Broiler strain, maternal age, and incubation temperature influence embryo metabolism. Hatching eggs were obtained from young (Y; 28 to 34 wk, $\bar{\rm x}$ = 31.2 wk), mid (M; 36 to 45 wk, $\bar{\rm x}$ = 40.5 wk) and old (O; 49 to 54 wk, $\bar{\rm x}$ = 51.4 wk) Ross 708 (n = 88; Experiment 1) and Ross 308 [(n = 45; Experiment 2: (Y; 25 to 34 wk, $\bar{\rm x}$ = 30.5 wk), (M; 35 to 44 wk, $\bar{\rm x}$ = 40.2 wk), and (O; 49 to 54 wk, $\bar{\rm x}$ = 51.6 wk)] breeders. Eggs were stored for 2 to 4 d (18°C, 73% RH), and incubated for 14 d at 37.5°C and 56% RH. At 15 d (E15), 8 fertile eggs per flock age were incubated in individual metabolic chambers at 36.0, 36.5, 37.0, or 37.5°C until E21.5. Each temperature was repeated one additional time. O2 consumption and CO2 production were used to calculate embryonic heat production (EHP). Embryo temperature was measured as eggshell temperature (EST). Initial egg weight was used as a covariate; significance was assessed at P < 0.05. In Ross 708, daily EHP tended to be higher in M and O than Y treatments at E16; EHP of M was higher than Y and O eggs at E18; M and O were higher than O eggs at E19. Incubation at 37.0°C resulted in the highest EHP from E15 to E21, except at E17. Embryos at 37.5°C had reduced EHP beyond E17. Daily EST from E15 to E21 was higher at 37.5 and 37.0°C than at 36.0 and 36.5°C. In Ross 308, daily EST was highest at 37.5°C except at E20. Incubation temperature and EST were highly correlated (R2 = 0.90 to 0.89; P < 0.001). Ross 708 chicks were longer and hatched earlier at 37.0°C than at 36.0 and 37.5°C. EST and EHP increased with incubation temperature in Ross 708. In Ross 308, maternal flock age and incubation temperature did not impact EHP. However, EST was highest at 37.5°C except at E20. Ross 708 was more sensitive to incubation temperature than Ross 308.

Ovarian follicular dynamic and plasma progesterone concentration in Alpine goats on the breeding season / Dinâmica folicular ovariana e concentração plasmática de progesterona em cabras Alpinas durante a estação reprodutiva

Os objetivos deste estudo foram relatar a dinâmica folicular ovariana e a concentração plasmática de progesterona durante ciclos estrais de duração irregular em cabras da raça Alpina. Vinte e nove cabras tiveram sua atividade ovariana monitorada diariamente por ultrassonografia transretal durante um, dois ou três períodos interovulatórios consecutivos. Amostras de sangue foram coletadas para análise da concentração plasmática de progesterona. Dos 47 ciclos avaliados, 28 (59,57%) foram de duração normal (17 a 25 dias), sete (14,89%) de curta duração (<17 dias) e 12 (25,53%) de longa duração (>25 dias). Os ciclos estrais de curta duração tiveram 6,14±0,69 dias e uma onda folicular que emergiu no dia 0,43±1,13. Os ciclos estrais de longa duração tiveram entre 26 e 79 dias, com duas a nove ondas foliculares. Os ciclos estrais de curta duração ocorreram durante a estação reprodutiva e foram associados à regressão prematura do corpo lúteo (CL). Os ciclos estrais de longa duração ocorreram na transição entre o anestro e a estação reprodutiva e foram associados principalmente à persistência do CL ou à fase folicular prolongada.(AU)

Gamma and Deltacoronaviruses in quail and pheasants from Northern Italy1.

In view of the restricted knowledge on the diversity of coronaviruses in poultry other than chicken, this study aimed to investigate the genetic diversity of coronaviruses in quail, pheasant, and partridge from two regions of Northern Italy. To this end, pools of tracheal and cloacal swabs from European quail (Coturnix Coturnix) and intestinal tract from pheasants (Phasianus Colchicus) and partridge (Perdix Perdix) flocks, with or without enteric signs, were collected during 2015. Avian coronavirus (Gammacoronavirus) was detected in quail not vaccinated against Infectious Bronchitis Virus (IBV) and in pheasants vaccinated with an IBV Massachusetts serotype. Based on DNA sequences for the gene encoding the S protein, the avian coronaviruses detected in the quail and pheasant are related to the IBV 793B and Massachusetts types, respectively. However, RNA-dependent RNA polymerase (RdRp) analyses showed the susceptibility of quail also to Deltacoronaviruses, suggesting that quail and pheasant avian coronaviruses share spike genes identical to chicken IBV spike genes and quail might host Deltacoronavirus.

Influência da condição corporal ao parto no balanço energético e desempenho reprodutivo de cabras leiteiras no pós-parto / Influence of body condition at calving on energy balance and reproductive performance of dairy goats in the postpartum

O estudo teve como objetivo avaliar a influência da condição corporal (CC) ao parto no balanço energético (BE) e o desempenho reprodutivo de cabras Alpinas no pós-parto. Foram utilizadas 68 cabras distribuídas em três grupos (G), sendo: G1- cabras com baixa CC (CC entre 1,5 e 2,5); G2: cabras com moderada CC (entre 2,75 e 3,5) e G3: cabras com alta CC (entre 3,75 e 5,0). Os animais receberam dieta (silagem de milho e concentrado, com 18% de proteína bruta e 1,7Mcal/kg de matéria seca de energia líquida) ad libitum. As avaliações foram feitas nas oito primeiras semanas de lactação, para determinação do BE, do intervalo do parto à primeira ovulação e ao primeiro estro e da concentração plasmática de progesterona. Os dados foram analisados por análise de variância a 5% de probabilidade. Não houve efeito da CC ao parto sobre o BE dos animais. Todas as cabras apresentaram BE negativo ao parto, com mudança para BE positivo aos 51, 58 e 64 dias pós-parto, respectivamente. Não houve diferença na concentração plasmática de progesterona nos três grupos, sendo caracterizada função luteal (concentração ≥1ng/dL) em apenas 5,9% dos animais avaliados. O intervalo do parto à primeira ovulação foi de 46 dias. Apenas 4,4% das cabras exibiram estro. Cabras leiteiras de média produção, entre 2,5 e 3,0kg de leite/dia, entram em BE negativo ao parto, independentemente da CC apresentada, influenciando negativamente o reinício da atividade ovariana no pós-parto.(AU)

The concentration of progesterone in the blood was used to study the effect of body condition score (BCS) on the post-partum ovarian activity of dairy goats. Sixty-eight female goats were distributed between three treatments. Treatment 1: goats with BCS between 1.00 and 2.75; Treatment 2: goats with BCS between 2.75 and 3.50; and Treatment 3: goats with BCS between 3.50 and 5.00. The goats had the blood collected immediately after birth, and from that time up to the eighth week post-partum at three day intervals. The progesterone analysis on the blood was used to determine the interval between the birth and the first ovulation. The females were teasing two times per day for estrus detection and determination of the time interval from birth to first estrus. No significant difference was found on the progesterone concentration between treatments. The luteal functionality was characterized in only 5.9% of goats, with a concentration higher than 1ng/mL. The period from birth to first ovulation was 46 days. The detectable estrus was found in only 4.4% of goats. The BCS at birth did not influence the recovery of ovarian activity of dairy goats.(AU)

Gammacoronavirus and Deltacoronavirus in Quail.

This paper expands on a previous report about coronaviruses in quail. After surveillance carried out in 2009 and 2010, some farmers started vaccinating quail with the Massachusetts avian infectious bronchitis virus serotype. The samples for this study were collected in 2013 from São Paulo state in southeastern Brazil. Pools of trachea, lungs, reproductive tract, kidneys, and enteric contents from quail and laying hens kept in the same farms and from quail-only farms as well as from both healthy birds and those showing infectious bronchitis-like symptoms were sampled in this study. The samples were screened using nested RT-PCR targeting the 3'-untranslated region of the Gammacoronavirus genus. Based on the DNA sequence for the RNA-dependent RNA polymerase (RdRp) gene, the strains isolated from quail clustered within either the Gammacoronavirus or Deltacoronavirus genus, and sequences from both genera were found in one quail sample. The phylogeny based on the partial S1 subunit sequence showed that the gammacoronaviruses detected in quail and layers belonged to the Brazil type. These results suggest that quail are susceptible to Gammacoronavirus and Deltacoronavirus viruses and indicate that the Massachusetts vaccination was not controlling IBV in quail or chickens.

Increased level of protection of respiratory tract and kidney by combining different infectious bronchitis virus vaccines against challenge with nephropathogenic Brazilian genotype subcluster 4 strains.

Genotyping of seven infectious bronchitis virus (IBV) strains isolated in Brazil showed that all belonged to the common Brazilian genotype and that these strains were closest to the subcluster of strain IBV/Brazil/2007/USP-19. Pathotyping of four selected Brazilian strains showed that they all caused a considerable level of ciliostasis in the trachea but at a somewhat lower level than did M41 and Brazilian strains 50/96, 57/96, 62/96 and 64/96 representing four different serotypes that had been reported earlier. In contrast to the M41 challenge strain, all Brazilian isolates replicated in kidney tissue in a high percentage of non-vaccinated challenged birds, clearly showing that they are nephropathogenic. As for the tracheal protection, the results using Massachusetts (Mass) vaccination against the recent strains seemed to show protection higher on average than for the strains reported earlier. A single or twofold vaccination with a Mass vaccine resulted in a mean tracheal protection level against the four challenge strains of 92% and 90%, respectively, whereas a single and twofold vaccination with a Mass vaccine halved the percentage of infected kidneys (14% and 13%, respectively, P < .05) compared to that of the unvaccinated birds (27%). The combination of the Mass and the 793B vaccine provided on average a tracheal protection of 99% and a reduction of the percentage of infected kidneys to a mean of 2%. This was a significantly (P < .05) higher protection than that achieved by a single or twofold Mass vaccination, showing the added value of the 793B vaccination following priming with a vaccine of the Mass type.

Influência do Hormônio do Crescimento na concentração de testosterona plasmática e nas características seminais de touros jovens e adultos da raça Nelore / Influence of Growth Hormone on plasma testosterone concentration and seminal characteristics of young and adult Nellore bulls

Estudou-se o efeito do Hormônio do Crescimento bovino (bGH) sobre os parâmetros seminais e a concentração sérica de testosterona de touros da raça Nelore. Dezesseis touros foram distribuídos em um delineamento fatorial 2 x 2 (duas idades e dois níveis de r-bST), com quatro animais em cada tratamento. As aplicações de r-bST foram realizadas a cada 14 dias, totalizando nove aplicações por animal, em um período experimental de 120 dias. As coletas de sêmen foram realizadas a cada 15 dias e, para determinar as concentrações de testosterona, foram realizadas coletas de sangue a cada quatro horas, num total de 24 horas, e a cada 30 dias a partir da primeira aplicação de r-bST. O sêmen dos touros adultos tratados com r-bST apresentaram maior motilidade e vigor (P<0,05) em relação aos animais controles (79,70±11,56% vs 64,06±23,65%) e (3,80±0,88 vs 2,92±1,25), respectivamente; entretanto, as demais características seminais não foram influenciadas (P>0,05) pelo tratamento com r-bST. Os aspectos físicos do sêmen dos touros jovens não foram afetados (P>0,05) pelo tratamento com r-bST. Entretanto o tratamento com r-bST diminuiu os defeitos morfológicos menores (delgado, gigante normal, pequeno normal, globoso, cabeça isolada normal, abaxial, cauda dobrada e enrolada levemente e gota distal) em relação aos jovens não tratados (P<0,05). As concentrações séricas de testosterona foram influenciadas (P<0,05) pelo tratamento, aos 120 dias após a primeira aplicação do r-bST, e apresentaram tendência de três picos no período de 24 horas.

The aim of this trial was to study the effect of Growth Hormone (GH) on the seminal parameters and testosterone profile from Nellore bulls. Sixteen bulls were allocated in a 2 x 2 factorial arrangement (two ages and two r-bST levels), with four animals per treatment. The r-bST was injected every 14 days, with a total of nine injections per animal, during the 120 days of the experimental period. The semen collections were done every 15 days and blood samples were collected in four hour intervals for 24 hours, and in 30 day intervals from the first r-bST injection for testosterone profile. The semen of adult bulls treated with r-bST showed greater MOT and VIG (P<0.05) in relation to the control (79.70±11.56% vs 64.06±23.65%) and (3.80±0.88 vs 2.92±1.25) respectively; however, the other evaluated seminal parameters were not affected (P>0.05) by the r-bST treatment. The physical measurements of the semen of young animals was also unaffected (P>0.05) by r-bST injection. However, the treatment with r-bST decreased the number of minor morphologic defects (narrow head, giant head, small normal head, round head, free normal head, abaxial implantation, simple bent tail and distal droplet) compared to untreated young bulls (P<0.05). The serum testosterone concentration was influenced (P<0.05) by treatment, at 120 days after the first injection of r-bST and they showed a tendency of three picks in a 24 hour period of sampling.

Assessing the economic burden of avian infectious bronchitis on poultry farms in Brazil.

Avian infectious bronchitis (IB), caused by avian infectious bronchitis virus (IBV), is a worldwide endemic disease of chickens that affects all branches of the poultry industry. Multiple geno/serotypes occur, and lowvaccine cross-protection results from the highly divergent IBV types. In view of the lack of consistent data on the economic losses caused by IB and the poor protection resulting from the use of the Massachusetts type as a live vaccine in Brazil, this survey aimed to estimate the losses per 1000 birds in broiler and breeder flocks positive for IBV. Thirty-two different IBV genetic types were found. In breeders, the total loss per 1,000 birds was US $3567.4 and US $4210.8 at 25-26 and 42 weeks old, respectively, whereas in broilers (48 days old), the estimated loss was US $266.3 per 1,000 birds. Taken together, the results show a significant and measurable economic impact on the broiler and breeder industries, with an age-dependent increasing trend and an association with multiple genetic types of the virus.

Using virtual human technology to capture dentists' decision policies about pain.

Healthcare professionals use race, gender, and age cues when making pain management decisions. Use of these demographic cues, therefore, is an important topic in the study of healthcare disparities. This study used virtual human (VH) technology to investigate the effects of VH patients' demographic cues on dentists' pain management decisions. Eighty-nine dentists viewed patients with different demographic cues. Analyses revealed that dentists rated pain intensity higher and were more willing to prescribe opioids to female, African-American, and younger patients than to their demographic counterparts. Results also found significant 2-way interactions between race and age for both pain assessment and treatment decisions. The interaction results suggest that the race difference (Caucasian < African American) was more pronounced for younger than for older patients. This is the first study to examine demographic cue use in dentists' decision-making for pain. The study found that dentists used demographic cues when making pain management decisions. Currently, there are no guidelines for decision- making practices for gender-, race-, or age-related pain. Since dentists see thousands of patients during their careers, the use of demographic cues could affect a substantial portion of the population. The findings could improve future training programs for dentists and dental students.

GHEP-ISFG proficiency test 2011: Paper challenge on evaluation of mitochondrial DNA results.

The GHEP-ISFG Working Group performed a collaborative exercise to monitor the current practice of mitochondrial (mt)DNA reporting. The participating laboratories were invited to evaluate a hypothetical case example and assess the statistical significance of a match between the haplotypes of a case (hair) sample and a suspect. A total of 31 forensic laboratories participated of which all but one used the EMPOP database. Nevertheless, we observed a tenfold range of reported LR values (32-333.4), which was mainly due to the selection of different reference datasets in EMPOP but also due to different applied formulae. The results suggest the need for more standardization as well as additional research to harmonize the reporting of mtDNA evidence.

An Avian coronavirus in quail with respiratory and reproductive signs.

An Avian coronavirus was detected in pools of lungs, tracheas, female reproductive tracts, kidneys, and enteric contents from quail (Coturnix coturnix japonica) and laying hen flocks, with and without infectious bronchitis (IB)-like signs, cohoused in farms located in two states of southeastern Brazil during 2009-2010. Although Avian metapneumovirus subtype B was found in two layers samples, Newcastle disease virus was not found in quail or in hens. Based on DNA sequences for the 3'-untranslated region and the gene encoding the RNA-dependent RNA polymerase, this avian coronaviruses in quail is an IB virus-like gammacoronavirus.

Effects of prostaglandin administration 10 days apart on reproductive parameters of cyclic dairy nulliparous goats / Efeitos da aplicação de prostaglandinas intervaladas de 10 dias sobre características reprodutivas de cabras leiteiras nulíparas cíclicas

This study reported the effects of prostaglandin (PGF2a) administration 10 days apart on reproductive parameters of cyclic artificial inseminated (AI) nulliparous Alpine (n=9) and Saanen (n=9) goats. Animals received two doses of 22.5mg PGF2a 10 days apart. After 1st and 2nd PGF2a administrations, estrus was monitored at 12 h intervals, with a buck teaser. Plasma progesterone concentration (ng/mL) was determined from blood sampled on day 0 (1st PGF2a) and the following 5, 10 (2nd PGF2a), 15, 20, 25 and 30 days. After the onset of the second estrus, females were transrectally (5 MHz probe) scanned at 4 hour intervals until at least 8h after ovulation. Pregnancy was checked through transrectal ultrasound on days 20, 25, 30, 35 and 90 after insemination. All parameters studied did not differ between breeds (P>0.05). Estrous response and interval to estrus, respectively, after 1st (78.9% and 50.6±17.2h) and 2nd PGF2a (88.9% and 50.0±14.8h) administration did not differ (P>0.05). Overall animals ovulating (100.0%), interval to ovulation after 2nd PGF2a (64.5±19.5h) and after estrous onset (18.0±9.1h), ovulation rate (1.3±0.5), diameter of ovulatory follicle (8.1±1.1mm) were recorded. Embryo loss occurred before day 30 of pregnancy. Estrus can be efficiently synchronized in nulliparous Alpine and Saanen goats with two doses of prostaglandin 10 days apart.

Relataram-se os efeitos da aplicação de prostaglandina sobre características reprodutivas de cabras leiteiras nulíparas cíclicas. Cabras Alpinas (n=9) e Saanen (n=9) receberam duas doses de 22,5mg PGF2a com 10 dias de intervalo. A progesterona plasmática (ng/mL) foi determinada a partir de amostras de sangue coletadas nos dias 0 (primeira dose), 5, 10 (segunda dose), 15, 20, 25 e 30. Após início do segundo estro, as fêmeas foram monitoradas por ultrassonografia transretal a cada quatro horas até oito horas após a ovulação. A gestação foi verificada por ultrassonografia transretal nos dias 20, 25, 30, 35 e 90 após a segunda dose. As características estudadas foram semelhantes entre as raças (P>0,05). Animais em estro e o intervalo parto-estro de, respectivamente, 78,9% e 50,6±17,2h e 88,9% e 50,0±14,8h após a primeira e segunda administrações de prostaglandina, não diferiram (P>0,05). Todas as cabras ovularam e registraram-se valores do intervalo parto-ovulação após a segunda aplicação de prostaglandina de 64,5±19,5h e após início do estro de 18,0±9,1h, a taxa de ovulação de 1,3±0,5 e diâmetro do folículo ovulatório de 8,1±1,1mm. Perda embrionária ocorreu antes de 30 dias de gestação. O estro pode ser eficientemente sincronizado em cabras leiteiras núliparas com duas doses de prostaglandina intervaladas de 10 dias.

Autoclaved, previously used intravaginal progesterone devices induces estrus and ovulation in anestrous Toggenburg goats.

Intravaginal progesterone devices are used worldwide for estrus induction in goats. Reused devices are able to induce estrus; however, this can be a health risk within a flock. The objective was to compare new and previously used (and autoclaved) progesterone-releasing intravaginal devices for induction of estrus and ovulation in seasonally anestrous Toggenburg goats. Anestrous goats (n=42) received new intravaginal devices containing 0.3g progesterone (CONTROL), or similar devices previously used for either 6 (USED6) or 12d (USED12) and subsequently autoclaved. All goats received 5mg dinoprost at device insertion and 200 IU eCG 5d later, and all devices were removed after 6d. After device removal, estrus was monitored and females displaying signs of estrus were mated by fertile bucks. Transrectal ovarian ultrasonography was performed after device removal until detection of ovulation. Blood samples were collected for determination of plasma progesterone concentration at different times. There was no difference (P>0.05) among groups CONTROL, USED6 or USED12 for: estrus response (87, 100 or 100%, respectively); duration of estrus (32.3±2.3, 25.2±3.4 or 27.3±4.1h); ovulation rate (100, 88 or 100%); number of ovulations (1.5±0.2, 1.9±0.3 or 1.7±0.3); and pregnancy rate (60, 58 or 67%). Plasma progesterone (P4) concentrations were greater (P<0.05) in CONTROL than in USED6-treated and USED12-treated goats (7.2±1.2, 4.7±0.7 and 4.3±0.6 ng/mL, respectively) at 6h after device insertion; these differences were maintained until 4d after device insertion (3.4±0.4, 2.3±0.2, and 2.5±0.2 ng/mL). Overall, plasma progesterone concentrations were greater (P<0.05) in nulliparous than in lactating goats (3.1±0.8 compared to 2.4±0.6 ng/mL, respectively). In conclusion, autoclaved, previously used intravaginal progesterone-releasing devices resulted in significant lesser plasma progesterone concentrations than new devices, but were similarly effective in inducing estrus and ovulation in anestrous goats.

Reproductive parameters of Mangalarga Marchador mares in a commercial embryo transfer programme.

The objective of this study is to evaluate the reproductive efficiency in donors and recipient Mangalarga Marchador mares in commercial programmes of embryo transfer (ET) and the effects of some reproductive characteristics and ET methodology on conception rates in the recipient mares. A total of 1140 flushing procedures were performed and 830 embryos (72.8%) were recovered. There were no differences between the rates of embryonic recovery in the different breeding seasons (p > 0.05) and 92.8% of the recovered embryos were 8-9 days old. There was no difference in the embryonic recovery regarding the collection order from the first to the ninth embryo collection along the breeding season, as well as among mares inseminated during the foal heat or subsequent cycles (p > 0.05). Pregnancy rates observed in the total period of all reproductive seasons at 15, 30, 45 and 60 days of pregnancy were 73.4, 69.9, 66.7 and 64.5%, respectively. Differences in pregnancy rate and early embryonic loss rates were not observed between embryos transferred immediately after collection (66.8% and 13.5%) and embryos transported at room temperature for periods of < 1 h (62.9% and 14.4%; p > 0.05). Pregnancy rates were higher when the interval between ovulations of donor and recipient mares remained between -3 and -2 days (p < 0.05), and the lowest rates were observed for intervals of -6 days (p < 0.05) with intermediary values for intervals of -1, 0 and +1 (p > 0.05). Embryonic loss rates, however, did not differ between intervals of ovulation's synchronism between donor and recipient mares (p > 0.05). This flexibilization in the ovulatory synchronism between donor and recipient mares optimizes the use of recipient mares, thus reducing costs and facilitating management of horse breeding farms.

Efeito de fontes de óleo e níveis de suplementação de vitamina E na ração sobre as características físicas e morfológicas do sêmen in natura de suínos / Effect of oil sources and vitamin E supplementation in the diet on physical and morphological characteristics of fresh boar semen

Avaliou-se o efeito da adição de diferentes fontes de óleo e níveis de suplementação de vitamina E na ração sobre as características do sêmen in natura de suínos reprodutores. Foram utilizados 24 reprodutores Dalboar 85, com idades entre 12 e 18 meses, distribuídos em delineamento inteiramente ao acaso, em arranjo fatorial 2 x 3, com duas fontes de óleo, soja e salmão, e três níveis de antioxidantes, 150, 300 e 450mg de vitamina E/kg. Volume, motilidade espermática total, teste hiposmótico, porcentagem de espermatozoides vivos e morfologia espermática não diferiram (P>0,05) entre os tratamentos. Óleo de salmão (P<0,05) melhorou o vigor espermático. A inclusão de vitamina E na ração melhorou (P<0,05) a concentração espermática, e não foi observada diferença entre as fontes de óleo (P>0,05). Os animais tratados com óleo de salmão apresentaram menor (P<0,05) concentração de antioxidantes totais no sêmen do que os tratados com óleo de soja. Observou-se efeito linear da vitamina E sobre a concentração de antioxidantes totais (P<0,05). A fonte de óleo de salmão da ração melhora o vigor espermático e a concentração de antioxidantes totais no sêmen.

The addition of oil sources and dietary vitamin E supplementation was evaluated on the characteristics of fresh boar sperm. Twenty-four mature Dalboar 85 boars of proven fertility and in routine semen production for artificial insemination were randomly distributed in a factorial arrangement 2 X 3, with two oil sources, soy and salmon, and three antioxidant levels: 150, 300, and 450mg/kg of vitamin E. Volume, total motile sperm, hyposmotic swelling test, percentage of live cell, and morphology did not differ (P<0.05) between the animals from the treatments. Salmon oil improved (P<0.05) sperm vigor. The vitamin E added to the diet improved (P<0.05) the sperm concentration, and there was no difference between the oil sources (P>0.05). Salmon oil treatment showed the lowest (P<0.05) concentration of total antioxidants in the semen. Vitamin E and concentration of total antioxidant showed a linear effect (P<0.05). The semen vigor and the concentration of total antioxidants were improved by oil sources, but the sperm concentration was dependent on the levels of vitamin E. Therefore, salmon oil can be used in the diet of male pigs.

Polyhydroxyalkanoate (PHA) production by a mixed microbial culture using sugar molasses: effect of the influent substrate concentration on culture selection.

In Polyhydroxyalkanoate (PHA) production processes using Mixed Microbial Culture (MMC), the success of the culture selection step determines, to a great extent, the PHA accumulation performance obtained in the final PHA production stage. In this study, the effect of the influent substrate concentration (30-60Cmmol VFA/L) on the selection of a PHA-storing culture using a complex feedstock, fermented sugar molasses, was assessed. At 30 and 45Cmmol VFA/L, substrate concentration impacted on the process kinetics through a substrate dependent kinetic limitation effect. However, further increasing the carbon substrate concentration to 60Cmmol VFA/L, resulted in an unforeseen growth limitation effect associated with a micronutrient deficiency of the fermented feedstock (magnesium) and high operating pH. Struvite precipitation caused a nutrient limitation which prevented biomass concentration increase, thus causing the feast to famine length ratio to vary in the selection reactor, with subsequent impact on the selective pressure for PHA-storing organisms. A highly dynamic response of the selected population to transient conditions of feast to famine ratio, in the range of 0.21-1.1, was observed. Kinetic (limiting concentration of carbon source) and physiological (loss of internal growth limitation due to the shorter length of famine phase) effects, resulting from variation of the influent substrate concentration, were subsequently demonstrated in batch studies. The culture selected at an influent substrate concentration of 45Cmmol VFA/L showed the best PHA-storing capacity since neither substrate concentration nor feast to famine ratio were limiting factors. This culture, highly enriched in PHA-storing organisms (88%), reached a maximum PHA content of 74.6%.

Effect of serial subculturing on the genetic composition and cytotoxic activity of Mycobacterium tuberculosis.

Continuous subculture has been observed to produce changes in the virulence of micro-organisms, e.g. rabies virus, poliovirus and Mycobacterium bovis BCG. The latter has been used as a vaccine for tuberculosis for the last 100 years; however, in some instances its efficacy has been observed to be very low. In order to determine whether similar changes can be produced in Mycobacterium tuberculosis, we selected four isolates, M. tuberculosis H37Rv, a Beijing strain (DR-689), and two more isolates with deletion of the phospholipase C locus (plcA-plcB-plcC ), and subjected them to serial culturing on Middlebrook 7H9 medium, with or without ox bile. After 100 passages, we performed RFLP-IS6110 analysis to determine whether genomic changes were produced. We also checked their genomic composition by microarray analysis. Changes in virulence were studied by measuring the cytotoxic effect of parental and subcultured isolates on a THP-1 macrophage monolayer. The most visible change was the change of position of an IS6110 band of approximately 1400 bp to approximately 1600 bp in the Beijing isolate subcultured in the ox bile medium. Analysis by microarray and PCR confirmation did not reveal any genomic changes. Cytotoxic activity was decreased in the isolates at levels close to that of BCG, and more consistently in those subcultured in the presence of ox bile.