Investigation of the influence of different moisture levels on acrylamide formation/elimination reactions using multiresponse analysis

The influence of water activity on the formation and elimination reactions of acrylamide was examined by means of multiresponse modeling on two different levels of complexity: basic equimolar asparagine-glucose systems and equimolar potato-based asparagine-glucose systems. To this end, model systems were first equilibrated to initial water activities in the range of 0.88-0.99 (corresponding roughly to the moisture gradient observed in French fries) and then heated at temperatures between 120 and 200 degrees C during different reaction times. For each sample, the concentration of acrylamide, glucose, asparagine, and aspartic acid was measured, as well as the extent of browning. A mechanistic model was proposed to model the five measured responses simultaneously. For both types of model systems, the model prediction was quite adequate, with the exception of the extent of browning, especially in the case of the potato-based model system. Moreover, the corresponding estimated kinetic parameters for acrylamide formation and elimination did not change significantly (based on a 95% confidence level) within the range of water activities tested, nor between the systems in the absence or presence of the potato matrix. The only remarkable difference was observed for the activation energy of acrylamide elimination, which was lower in the presence of the potato matrix, although not always significant. In general, these results confirm the generic nature of the model proposed and show that the influence of different moisture levels on acrylamide formation and elimination is minimal and that the addition of a potato matrix has little or no influence on the kinetic model and corresponding kinetic parameters.     

Bovine thyroglobulin gene polymorphisms and their association with sexual precocity in Guzerat bulls

Puberty is a stage of sexual development determined by the interaction of environmental factors and genetic mechanisms. Among them, thyroid function plays a key role in sexual development and spermatogenic function and is under the control of several genes, including the well‐described thyroglobulin gene (TG). Previous reports have shown genetic association between thyroid function and selected single nucleotide polymorphisms (SNPs) in taurine cattle. Therefore, the identification of genetic mechanisms involved in the regulation of this trait can assist with the selection for early pubertal bulls, thus improving genetic progress in livestock breeding. The aim of this study was to validate the association between TG SNPs and age at puberty in zebuine bulls. Three SNPs (rs110406764, rs109662686, rs109057985) were genotyped in 159 Guzerat animals using SEQUENOM technology. Results showed a significant association (p < .05) between the studied SNPs and puberty age, in agreement with our previous reports in a taurine breed. Interestingly, allele frequencies were different from those already reported, being GAT the most favourable allele for age at puberty in Guzerat (94.4 days lower). Overall, our findings corroborate previous reports and reinforce the importance of genetic influence in the regulation of sexual development and puberty through a thyroid pathway in zebuine cattle.     

Effect of sperm pretreatment with glutathione and membrane destabilizing agents lysolecithin and Triton X‐100, on the efficiency of bovine intracytoplasmic sperm injection

Intracytoplasmic sperm injection (ICSI) is an assisted reproduction tool with several applications. Its effectiveness in bovines is lower than that in other species, mainly because of difficulties in the decondensation of the sperm nucleus after injection, and the presence of the acrosome and the plasma membrane which remain intact in this procedure. In this study, we assessed the effect of lysolecithin (LL) and Triton X‐100 (TX), in combination with glutathione (GSH) as sperm pretreatments prior to ICSI. The GSH‐LL and GSH‐TX groups showed 0% of spermatozoa with intact membrane (SYBR 14+/PI), in comparison with the control (63.3%) and GSH (65.7%) groups. The proportions of spermatozoa with damaged acrosome membrane in the GSH‐LL, GSH‐TX, GSH and control groups were 46%, 35.9%, 10.5% and 7.5%, respectively. Sperm chromatin decondensation analysis showed that the groups incubated for 3 hr with GSH presented greater decondensation (p < .05). Although fertilization was improved in all treatment groups evaluated, no differences were observed in the cleavage rate 72 hr after activation in the GSH (73.7%), GSH‐LL (80.2%) and GSH‐TX (77.8%) groups compared to the control (66.3%), neither in the blastocyst rate on day 8 (24.0%, 26.2%, 27.1% and 28.4% for the control, GSH, GSH‐LL and GSH‐TX groups, respectively). No differences were also observed in the total number of cells in all groups. In conclusion, although these sperm treatments promoted nuclear decondensation and induced plasma membrane disruption, these effects were not sufficient to improve bovine embryonic development after ICSI.     

Different Doses of Porcine Luteinizing Hormone in Precocious Puberty Induction in Gilts

The use of hormonal protocols in puberty induction and synchronization of oestrus has lead to an increase in the efficiency of replacement gilts. The aim of this study was to evaluate different doses of porcine LH in precocious puberty induction and oestrus synchronization in a homogeneous group of gilts. Sixty-seven homogeneous prepubertal gilts (Camborough 22) at 137 +/- 4 days of age and 87 +/- 7 kg were treated with three different hormonal protocols: T1--600 UI of equine chorionic gonadotrophin (eCG; Novormon) and after a 72-h period 5 mg of LH (Lutropin); T2--600 UI of eCG and a 72-h period 2.5 mg of LH; T3--600 UI of eCG and a 72-h period 1.25 mg of LH. The ovaries were examined at slaughter, on day 6 after the hormonal treatment. There were no statistical differences (p > 0.05) between the different LH doses in the percentage of the detected oestrus (T1 = 42.85%; T2 = 60.87%, T3 = 52.18%), oestrus duration (T1 = 41.44 +/- 16.30 h; T2 = 48.57 +/- 16.29 h, T3 = 39.33 +/- 11.42 h), number of corpora lutea (T1 = 9.61 +/- 5.43; T2 = 9.86 +/- 3.32, T3 = 8.13 +/- 5.52) and percentage of animals presenting ovarian cystic degeneration (T1 = 33.33%; T2 = 39.13%, T3 = 39.13%). The T2 (2.5 mg of LH) presented the lowest dispersion (p < 0.05) of the LH-ovulation interval (T1 = 37.17 +/- 4.07 h; T2 = 38.26 +/- 2.84 h; T3 = 36.25 +/- 5.69 h). The LH dose reduction to 2.50 and 1.25 mg presented equal results with the recommended dose of 5.0 mg, and could be used in the precocious induction of oestrus in gilts. The 2.5-mg LH dose showed the lowest dispersion of ovulation and it can be used in fixed-time artificial insemination programmes.     

Evaluation of a Burdizzo Castrator for Neutering of Dogs

A Burdizzo castrator was evaluated for the neutering of dogs. Histological and morphological changes of spermatic cells and peripheral serum testosterone after challenge with a GnRH-analogue (gonadorelin) were assessed. There was a control group (G1), a surgically castrated group (G2) and a Burdizzo group (G3) divided in two, G3a receiving two crunches in each spermatic cord and G3b receiving one crunch in each spermatic cord. Sixteen days after application of the Burdizzo blood samples were taken from the dogs at 30 min interval during 2 h; after the second sample the dogs were treated with 1 mug/kg body weight of gonadorelin i.v. The same protocol of gonadorelin challenge was performed in G1 and G2 dogs. The G2 dogs were surgically castrated after the second blood sample, before the gonadorelin treatment, and the G1 dogs after the last blood sample. The excised gonads were examined histologically, and sperm smears were prepared from the caudae epididymidis. The testes and plexus pampiniformis of the G1 and G2 dogs had a normal histological appearance, and they had morphologically normal epididymal sperm cells. In all G3 dogs, there was an acute fibrosis with an inflammatory reaction in the plexus pampiniformis. The testes from the G3a dogs showed diffuse areas of infarction and degeneration of the parenchyma. Similar but less diffuse lesions were seen in group 3b dogs. The deferent ducts from all G3 dogs showed vasitis and/or sperm granulomas. Azoospermia or sperm malformations were observed in the epididymal smears from the G3 dogs. Testosterone concentration in the G1 dogs increased after gonadorelin application (p < 0.0001). The G2 dogs had basal testosterone levels after castration (p < 0.001) and did not respond to gonadorelin. Groups 3a and b showed a slight but non-significant increase in testosterone concentration after gonadorelin challenge, supposedly due to the reduction of testicular blood flow and loss of testicular interstitial tissue.     

Inactivation of orange pectinesterase by combined high-pressure and -temperature treatments: a kinetic study

Pressure and/or temperature inactivation of orange pectinesterase (PE) was investigated. Thermal inactivation showed a biphasic behavior, indicating the presence of labile and stable fractions of the enzyme. In a first part, the inactivation of the labile fraction was studied in detail. The combined pressure-temperature inactivation of the labile fraction was studied in the pressure range 0.1-900 MPa combined with temperatures from 15 to 65 degrees C. Inactivation in the pressure-temperature domain specified could be accurately described by a first-order fractional conversion model, estimating the inactivation rate constant of the labile fraction and the remaining activity of the stable fraction. Pressure and temperature dependence of the inactivation rate constants of the labile fraction was quantified using the Eyring and Arrhenius relations, respectively. By replacing in the latter equation the pressure-dependent parameters (E(a), k(ref)(T)()) by mathematical expressions, a global model was formulated. This mathematical model could accurately predict the inactivation rate constant of the labile fraction of orange PE as a function of pressure and temperature. In a second part, the stable fraction was studied in more detail. The stable fraction inactivated at temperatures exceeding 75 degrees C. Acidification (pH 3.7) enhanced thermal inactivation of the stable fraction, whereas addition of Ca(2+) ions (1 M) suppressed inactivation. At elevated pressure (up to 900 MPa), an antagonistic effect of pressure and temperature on the inactivation of the stable fraction was observed. The antagonistic effect was more pronounced in the presence of a 1 M CaCl(2) solution as compared to the inactivation in water, whereas it was less pronounced for the inactivation in acid medium.     

Biomechanical Properties of Skin and Wounds in Ehlers-Danlos Syndrome

The biomechanical properties of wounded and nonwounded skin were studied in three dogs and three cats affected with type I Ehlers-Danlos syndrome. Three nonaffected dogs and one nonaffected cat served as controls. Samples of wounded skin and adjacent normal skin were harvested at days 75, 138, 141, 144, 147, and 150. Samples were subjected to uniaxial tensile strength testing. Tensile strength, energy absorbed, and site of failure were recorded. In the dogs with Ehlers-Danlos syndrome, there was an increase in tensile strength in samples containing a scar over adjacent intact skin. In nonaffected dogs, affected cats and the nonaffected cat, the nonwounded skin samples had greater tensile strength. The energy absorbed by the skin samples during testing was highly correlated with tensile strength.     

Haematological and biochemical reference values for the koala (Phascolarctos cinereus)

Haematological and biochemical reference values were established from 45 clinically healthy koalas. Statistical analysis revealed no significant differences for sex and season of sampling. Immature koalas had significantly higher alkaline phosphatase and inorganic phosphate values, and significantly lower total protein concentrations due to low globulins values. Enzyme reference values tended to be wide and could limit their usefulness in detecting disease. In the reference values for leukocytes, neutrophils and lymphocytes, the inclusion of low values which were not actually seen may interfere with the detection of reduced levels due to disease.     

Influence de la densité et de la période de concurrence de Solanum nigrum L. sur la tomate de semis direct, en relation avec le désherbage

Effects of density and period of competition by Solanum nigrum L. on direct seeded tomatoes in relation to weed control The effects of density and period of competition from Solanum nigrum L. were measured in direct seeded tomatoes given weed control treatments currently used in south-east France. S. nigrum emerging after a diquat treatment at the 2–3 leaf stage of the crop and thinned to low densities (<12.8 plants ha^?1) at the 5–6 leaf stage of the crop caused significant yield loss if left to compete with the crop until harvest. Yield reduction was smaller if the same weed densities were present only until the onset of flowering. The regression curves of yield on weed density differed as annual climatic variations affected sowing date and plant growth; a comparison between years was made using the relation ‘crop yield × weed biomass/crop biomass’. Significant interactions between weed density and period of competition were found with yield of both green and red fruit. For late sown crops with low densities of S. nigrum two weed control treatments at the 5–6 leaf stage and at the onset of flowering were sufficient to prevent yield loss.