Detection and differentiation of Leptospira spp. serovars in bovine semen by polymerase chain reaction and restriction fragment length polymorphism

In view of the importance of venereal transmission of bovine leptospirosis, the objective of the present study was to apply the polymerase chain reaction (PCR) to 26 serovars of Leptospira interrogans, L. borgpetersenii, L. santarosai, L. noguchii and L. biflexa, to determine the detection threshold in semen samples and to evaluate the possibility of differentiation among serovars using 19 restriction endonucleases. The results showed that all serovars were amplified and the detection threshold in semen samples of a bull was 100 bacteria/ml. Using endonucleases we could classify the 26 serovars into eight groups. The present results show that PCR is a method of great potential for the detection of Leptospira spp, at bovine artificial insemination centers. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Durango apatite fission-track dating using length-based age corrections and neutron fluence measurements by natural thorium thin films and natural U-doped glasses calibrated through natural uranium thin films

In this work, we present an approach for neutron fluence measurements based on natural thorium thin films and natural uranium-doped glasses calibrated through natural uranium thin films to be used for dating with the Fission-Track Method (FTM). This neutron dosimetry approach allows the employment of FTM even when dating is carried out using low neutron themalization facilities. Besides, it makes possible the determination of the Th/U ratio of the mineral to be dated. Durango apatite which is often employed in FTM as an age standard was analyzed. This apatite presented a fairly high Th/U ratio, 29.9 +/- 1.7. Th fissions were 18%, 12% and 10% of the total for irradiations where thermal to fast neutron flux ratios were 2.4, 4.4 and 5,2, respectively. These results show that Th fission must be considered for this apatite, when not well-thermalized irradiation facilities are used. The ratio between spontaneous and induced track length, L(S)/L(1), close to 0.89, indicates a certain amount of rejuvenation of the age of Durango apatite. Therefore, its apparent age should be corrected, the application of a technique based on track-length measurements produced a corrected age of 29.7 +/- 1.1 Ma, consistent with the independent reference age of this apatite (31.4 +/- 0.5 Ma). This result represents a support for viability of the neutron dosimetry approach studied in this work for FTM.(C) 2002 Elsevier B.V. B.V. All rights reserved.

Influence of growth rate and length on fluoride detection in human nails

This study aimed to determine the lag time between increased fluoride (F) intake and F detection in human nails, as well as the influence of nails growth rate and length on this. Ten 20- to 35-year-old volunteers received 1.8 mg F daily, for 30 days. Nail growth rate and length were determined for all fingernails and toenails. Nail samples were collected at the beginning of the study and every 2 weeks (15 collections in all) and F concentrations were determined. The growth rate was statistically higher in fingernails than in toenails. No statistically significant differences were observed between right and left sides. Growth rate was significantly greater for big toenails than for the other toenails, but this pattern was not found for fingernails. The estimated mean lag times for F detection in fingernails and toenails were 101 and 123 days, respectively. An apparent increase in fingernail F concentrations was observed 84 days after the beginning of the study, although this was not statistically different from baseline. For toenails, statistically significant increases in F concentration in relation to baseline were observed 112 and 140 days after increased F ingestion. These increases occurred within the 95% confidence intervals for the calculated mean lag time for fluoride detection in nails. Considering the large amount of sample provided by the big toenails, together with their faster growth rate, as well as the fact that toenails are less prone to environmental contamination, our data suggest that big toenails are more suitable biomarkers of fluoride intake.

Evaluation of the influences of solution path length and additives concentrations on the solar photo-Fenton degradation of 4-chlorophenol using multivariate analysis

This study reports the photodegradation of 4-chlorophenol (4-CP) in aqueous solution by the photo-Fenton process using solar irradiation. The influence of solution path length, and Fe(NO3)(3) and H2O2 concentrations on the degradation of 4-CP is evaluated by response surface methodology. The degradation process was monitored by the removal of total organic carbon (TOC) and the release of chloride ion. The results showed a very important role of iron concentration either for TOC removal or dechlorination. on the other hand, a negative effect of increasing solution path length on mineralization was observed, which can be compensated by increasing the iron concentration. This permits an adjustment of the iron concentration according to the irradiation exposure area and path length (depth of a tank reactor). Under optimum conditions of 1.5 mM Fe(NO3)(3), 20.0 mM H2O2 and 4.5 cm solution path length, 17 min irradiation under solar light were sufficient to reduce a 72 mg C L-1 solution of 4-CP by 91 (c) 2006 Elsevier B.V. All rights reserved.

Effect of post length on the fatigue resistance of bovine teeth restored with bonded fiber posts: A pilot study

This study evaluated the influence of the cementation length of glass fiber-reinforced composite (FRC) on the fatigue resistance of bovine teeth restored with an adhesively cemented FRC. Thirty roots of single-rooted bovine teeth were allocated to 3 groups (n = 10), according to the ratio of crown length/root length (post cementation length): group 1 = 2/3, group 2 = 1/2, and group 3 = 1/1. The roots were prepared, the fiber posts (FRC Postec Plus) were cemented, and the specimens were submitted to 2 million mechanical cycles. After fatigue testing, a score was given based on the number of fatigue cycles until fracture, and data were submitted to statistical analysis. All specimens were resistant to fatigue. Taking into account the methodology and results of this study, the evaluated fiber posts can be cemented based on the ratio of crown/root at 1/1. Further clinical studies must be conducted to verify this ratio.

Use of PCR-RFLP (Polymerase Chain Reaction-Restricted Fragment Length Polymorphism) in the gene of the enzyme Stearoyl-CoA-Desaturase in Bubalus bubalis

The milk is an important food because it contents Conjugated Linoleic Acids (CIA). These fatty acids are synthesized in mammary gland under action of the enzyme Stearoyl CoA-Desaturase (SCD) and have showed some positive effects in human disease prevention and treatments. A variation of CLA in milk fat exists and can be partially explained by the different levels of expression of SCD. The aim was to study part of the encoding regions of SCD's gene using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Genomic DNA was extracted from lactating Murrah females. After this, PCR reactions were made by using primers Z (sic) (sic) D1 that encloses exon I, II and intron I. The fragments amplified are composed by 938 pb. Then, RFLP techniques were applied in the fragments using the restriction enzymes Pst I and Sma I. The enzyme Pst I has generated fragments of 788pb and 150bp and the Sma I has generated fragments of 693pb and 245pb. All the animals showed the same migration standard for both enzymes, characterizing a genetic monomorphism for this region of SCD gene. The analysis determined that there aren't genetic differences between these animals in the studied regions by using Pst I and Sma I enzymes.

Effect of grazing system on fetal development in Nellore cattle

Intensive grazing systems for beef females, based on abundant availability of high quality forages and supplementary concentrates, may affect fetal development. The objective of this study was to determine the effect of grazing system on length of gestation, fetal development, and characteristics of the calf at birth. Twenty-four pregnant (bred to Nellore bulls) Nellore females were allocated into two groups. The control group (G1) grazed Brachiaria decumbens (signal grass) in a traditional (extensive) grazing system and the second group (G2) were managed on Panicum maximumcv. Tanzania 1 (Tanzania grass) in an intensive grazing system. Fetal development was evaluated by ultrasonography on days 31, 45, 59, 94, 122, 220, and 255 of gestation. The diameter of the amniotic and allantoic cavities, crown-rump length, circumference, and diameter of the head and ocular orbit were determined. At birth, calves were weighed and height, length, thoracic circumference, and ocular orbit and bi-parietal diameters were measured. There were no differences (P > 0.05) in fetal development. The G1 cows had a longer gestation period (4.5 days; P < 0.05) and their calves had greater (P < 0.05) weight, height, length, and thoracic circumference at birth. In conclusion, Nellore females raised under intensive pasture management conditions, had significantly shorter gestation and smaller calves at birth than those raised under extensive pasture management conditions. Therefore, adoption of new management practices (e.g. intensive pasture management), should take into consideration animal behavior and productivity. (C) 2003 Elsevier B.V. All rights reserved.

A new polymerase chain reaction/restriction fragment length polymorphism protocol for Plasmodium vivax circumsporozoite protein genotype (VK210, VK247, and P-vivax-like) determination

For the molecular diagnosis of Plasmodium vivax variants (VK210, VK247, and P. vivax-like) using DNA amplification procedures in the laboratory, the choice of rapid and inexpensive identification products of the 3 different genotypes is an important prerequisite. We report here the standardization of a new polymerase chain reaction/restriction fragment length polymorphism technique to identify the 3 described P. vivax circumsporozoite protein (CSP) variants using amplification of the central immunodominant region of the CSP gene of this protozoan. The simplicity, specificity, and sensitivity of the system described here is important to determine the prevalence and the distribution of infection with these P. vivax genotypes in endemic and nonendemic malaria areas, enabling a better understanding of their phylogeny. (c) 2007 Published by Elsevier B.V.

Spermatogenic cycle length and spermatogenic efficiency in the gerbil (Meriones unguiculatus)

The gerbil (Meriones unguiculatus) is a rodent native of the and regions of Mongolia and China. Because the gerbil can be easily bred in laboratory conditions, this species has been largely used as an experimental model in biomedical research. However, there is still little information concerning the testis structure and function in the gerbil. In this regard, we performed a detailed morphofunctional analysis of the gerbil testis and estimated the spermatogenic cycle length utilizing H-3-thymidine as a marker for germ cell progression during their evolution through the spermatogenic process. The stage frequencies of the XII stages characterized according to the acrosome formation and development were (I-XII) 13.8, 10.1, 8.1, 7.8, 4.0, 11.2, 7.5, 7.1, 5.9, 7.6, 8.1, and 8.9. The mean duration of each seminiferous epithelium cycle was determined to be 10.6 +/- 1.0 days and the total duration of spermatogenesis, based on 4.5 cycles, was approximately 47.5 days. The volume density of tubular and interstitial compartments was approximately 92% and 8%, respectively. Based on the volume occupied by seminiferous tubules in the testis and the tubular diameter, about 9 and 18 m of seminiferous tubules were found per testis and per gram of testis, respectively. Twelve primary spermatocytes were formed from each type A1 spermatogonia. The meiotic index was 2.8, indicating that 30% of cell loss occurs during meiosis. The number of Leydig and Sertoli cells per gram of the testis was 28 million and each Sertoli cell was able to support approximately 13 spermatids. The daily sperm production per gram of testis (spermatogenic efficiency) was 33 million. Taken together, these data indicate that, mainly due to the high seminiferous tubule volume density and Sertoli cell support capacity for germ cells, the gerbil presents high spermatogenic efficiency compared with other mammalian species already investigated. The data obtained in the present study might provide the basis for future research involving the reproductive biology in this species.