Dietary stress does not strengthen selection against single deleterious mutations in Drosophila melanogaster.

Stress is generally thought to increase the strength of selection, although empirical results are mixed and general conclusions are difficult because data are limited. Here we compare the fitness effects of nine independent recessive mutations in Drosophila melanogaster in a high- and low-dietary-stress environment, estimating the strength of selection on these mutations arising from both a competitive measure of male reproductive success and productivity (female fecundity and the subsequent survival to adulthood of her offspring). The effect of stress on male reproductive success has not been addressed previously for individual loci and is of particular interest with respect to the alignment of natural and sexual selection. Our results do not support the hypothesis that stress increases the efficacy of selection arising from either fitness component. Results concerning the alignment of natural and sexual selection were mixed, although data are limited. In the low-stress environment, selection on mating success and productivity were concordant for five of nine mutations (four out of four when restricted to those with significant or near-significant productivity effects), whereas in the high-stress environment, selection aligned for seven of nine mutations (two out of two when restricted to those having significant productivity effects). General conclusions as to the effects of stress on the strength of selection and the alignment of natural and sexual selection await data from additional mutations, fitness components and stressors.

Soil radon measurements in the Canadian cities.

Radon has been identified as the second leading cause of lung cancer after tobacco smoking. Information on indoor radon concentrations is required to assess the lung cancer burden due to radon exposure. Since radon in soil is believed to be the main source of radon in homes, measurements of soil gas radon concentrations can be used to estimate variations in radon potential of indoor environments. This study reports surveys of natural background variation in soil radon levels in four cities, Montreal, Gatineau, Kingston and the largest Canadian city of Toronto. A total of 212 sites were surveyed. The average soil gas radon concentrations varied significantly from site to site, and ranged from below detection limit to 157 kBq m(-3). For each site, the soil radon potential (SRP) index was determined with the average soil radon concentration and average soil permeability measured. The average SRP indexes are 20±16, 12±11, 8±9 and 12±10 for Montreal, Gatineau, Kingston and Toronto, respectively. The results provide additional data for the validation of an association between indoor and soil radon potentials and for the development of radon potential map of Canada.

Preservation of protein globules and peptidoglycan in the mineralized cell wall of nitrate-reducing, iron(II)-oxidizing bacteria: a cryo-electron microscopy study.

Iron-oxidizing bacteria are important actors of the geochemical cycle of iron in modern environments and may have played a key role all over Earth's history. However, in order to better assess that role on the modern and the past Earth, there is a need for better understanding the mechanisms of bacterial iron oxidation and for defining potential biosignatures to be looked for in the geologic record. In this study, we investigated experimentally and at the nanometre scale the mineralization of iron-oxidizing bacteria with a combination of synchrotron-based scanning transmission X-ray microscopy (STXM), scanning transmission electron microscopy (STEM) and cryo-transmission electron microscopy (cryo-TEM). We show that the use of cryo-TEM instead of conventional microscopy provides detailed information of the successive iron biomineralization stages in anaerobic nitrate-reducing iron-oxidizing bacteria. These results suggest the existence of preferential Fe-binding and Fe-oxidizing sites on the outer face of the plasma membrane leading to the nucleation and growth of Fe minerals within the periplasm of these cells that eventually become completely encrusted. In contrast, the septa of dividing cells remain nonmineralized. In addition, the use of cryo-TEM offers a detailed view of the exceptional preservation of protein globules and the peptidoglycan within the Fe-mineralized cell walls of these bacteria. These organic molecules and ultrastructural details might be protected from further degradation by entrapment in the mineral matrix down to the nanometre scale. This is discussed in the light of previous studies on the properties of Fe-organic interactions and more generally on the fossilization of mineral-organic assemblies.

Proximal interphalangeal joint arthrodesis in 34 horses using two parallel 5.5-mm cortical bone screws.

OBJECTIVE: To report clinical experience with arthrodesis of the proximal interphalangeal joint in horses using two parallel 5.5-mm cortical bone screws placed in lag fashion. STUDY DESIGN: Retrospective, clinical study. ANIMALS: Thirty-four horses, aged 1 to 19 years. METHODS: Medical records for all horses admitted (1991-1997) for pastern arthrodesis using two 5.5-mm ASIF cortical bone screws, in parallel orientation, and placed in lag fashion by use of a combined aiming device to facilitate accuracy were reviewed. Signalment, lameness diagnosis, duration of lameness, limb(s) involved, and outcome were recorded. Criteria for successful outcome were determined as return to previous level of function or future intended athletic use. RESULTS: Thirty-nine proximal interphalangeal joint arthrodeses were performed on 34 horses. One horse was euthanatized in the recovery room and was excluded from data analysis. Successful outcome occurred in 85% of frontlimbs and 89% of hindlimbs. Failure occurred in 5 joints; 1 horse had lameness directly associated with surgery, whereas 4 horses had unrelated lameness. CONCLUSION AND CLINICAL RELEVANCE: Age, breed, and initial disease did not affect outcome. Arthrodesis of the proximal interphalangeal joint by use of two 5.5-mm ASIF cortical bone screws, in parallel orientation, placed in lag fashion by use of a combined aiming device, resulted in sound use of the limb in >85% of the joints with shortened postoperative coaptation.

Processing postmortem specimens with C18-carboxypropylbetaine and analysis by PCR to develop an antemortem test for Mycobacterium avium infections in ducks.

Mycobacterium avium is the causative agent of the avian mycobacteriosis commonly known as avian tuberculosis (ATB). This infection causes disseminated disease, is difficult to diagnose, and is of serious concern because it causes significant mortality in birds. A new method was developed for processing specimens for an antemortem screening test for ATB. This novel method uses the zwitterionic detergent C18-carboxypropylbetaine (CB-18). Blood, bone marrow, bursa, and fecal specimens from 28 ducks and swabs of 20 lesions were processed with CB-18 for analysis by smear, culture, and polymerase chain reaction (PCR). Postmortem examination confirmed nine of these birds as either positive or highly suspect for disseminated disease. The sensitivities of smear, culture, and PCR, relative to postmortem analysis and independent of specimen type, were 44.4%, 88.9%, and 100%, respectively, and the specificities were 84.2%, 57.9%, and 15.8%, respectively. Reductions in specificity were due primarily to results among fecal specimens. However, these results were clustered among a subset of birds, suggesting that these tests actually identified birds in early stages of the disease. Restriction fragment length polymorphism mapping identified one strain of M. avium (serotype 1) that was isolated from lesions, bursa, bone marrow, blood, and feces of all but three of the culture-positive birds. In birds with confirmed disease, blood had the lowest sensitivity and the highest specificity by all diagnostic methods. Swabs of lesions provided the highest sensitivity by smear and culture (33.3% and 77.8%, respectively), whereas fecal specimens had the highest sensitivity by PCR (77.8%). The results of this study indicate that processing fecal specimens with CB-18, followed by PCR analysis, may provide a valuable first step for monitoring the presence of ATB in birds.

In vitro comparison of NALC-NaOH, tween 80, and C18-carboxypropylbetaine for processing of specimens for recovery of mycobacteria.

A recent article (C. G. Thornton et al., J. Clin. Microbiol. 36:1996-2003, 1998) reported a new specimen-processing method for improved recovery of mycobacteria. This method used C18-carboxypropylbetaine (CB-18) and increased both smear and culture sensitivity. The companion article (C. G. Thornton et al., J. Clin. Microbiol. 36:2004-2013, 1998) described initial improvements to this method. Additional significant parameters of the CB-18 processing method are identified herein. First, eliminating the incubation step was shown to further improve culture sensitivity. Subsequently, recovery of several mycobacterial isolates by the CB-18 method was compared to a contemporary processing method that combines NALC and NaOH (NALC-NaOH) and a Tween 80-based method. Recovery of the tuberculous isolates following NALC-NaOH processing averaged 20% and ranged from 1.6 to 45%, whereas recovery of the nontuberculous isolates averaged 11% and ranged from 0.1 to 55%. Recovery of the tuberculous and nontuberculous isolates by the Tween 80-based method ranged from 22 to 92% and 27 to 93%, respectively, with averages of 58 and 65%, respectively. Recovery of the tuberculous and nontuberculous mycobacteria following CB-18 processing averaged 86 and 73%, respectively, with ranges from 61 to over 100% and from 43 to over 100%, respectively. Other parameters of the CB-18 method were also examined, including recovery versus CB-18 concentration and the relationship between CB-18 concentration and the tuberculocidal effect. The tuberculocidal effect was time dependent but independent of concentration, whereas recovery was directly proportional to concentration. Increasing the CB-18 concentration to 4 mM provided quantitative recovery on solid medium; however, higher concentrations of CB-18 were not compatible with liquid culture. Examination of the relationship between increasing CB-18 and lecithin concentrations suggested that lecithin could not overcome the deleterious effects of CB-18 in liquid culture at these higher concentrations.

Novel method for processing respiratory specimens for detection of mycobacteria by using C18-carboxypropylbetaine: blinded study.

A novel method for processing respiratory specimens to improve culture and acid-fast staining of mycobacteria is introduced. This new method utilized N,N-dimethyl-N-(n-octadecyl)-N-(3-carboxypropyl)ammonium inner salt (Chemical Abstract Service no. 78195-27-4), also known as C18-carboxypropylbetaine (CB-18). In a blinded, five-center study, CB-18-based processing was compared to the standard method combining NALC and NaOH (NALC/NaOH). A total of 573 respiratory specimens were tested. Individual specimens were split approximately equally; the host institutions processed half of each specimen by the NALC/NaOH method, while the other half was processed with CB-18 at Quest Diagnostics--Baltimore. A total of 106 specimens were culture positive for acid-fast bacilli (AFB). Replacement of the primary decontamination agent with CB-18 caused changes in all diagnostic parameters. Aggregate culture sensitivity improved by approximately 43% (P < 0.01), and smear sensitivity improved by approximately 58% (P < 0.01). The sensitivity of smear relative to that of M. tuberculosis isolates exceeded 93% (P < 0.01) when specimens were processed with CB-18. The average times to a positive result were reduced by 7.3 days in liquid culture (P < 0.01) and 5.3 days on solid media (P < 0.05); however, the CB-18 method had a 20.8% contamination rate in liquid culture versus a rate of approximately 7.5% with NALC/NaOH processing. There were also unusual reductions in liquid culture sensitivity and smear specificity among CB-18-processed specimens. The characteristics of the latter parameters suggested that refinement of the CB-18 processing method should allow further improvements in culture sensitivity. This study showed that the CB-18 method has the potential to improve both smear and culture detection for these important human pathogens.

Processing respiratory specimens with C18-carboxypropylbetaine: development of a sediment resuspension buffer that contains lytic enzymes to reduce the contamination rate and lecithin to alleviate toxicity.

The C18-carboxypropylbetaine (CB-18) procedure for processing respiratory specimens for the detection of mycobacteria was shown to provide significant increases in sensitivity by smear and culture. However, the procedure also produced increased contamination, a loss in liquid culture sensitivity, and a reduction in smear specificity. Because of these observations, the toxicity of CB-18 and the nature of the contamination were characterized. Preincubation in 1 mM CB-18 impacted viability in a time-dependent fashion, but the magnitude of the loss was species and isolate dependent. Mycobacterium tuberculosis isolates were the most susceptible, losing 20 to 30% of the CFU within 30 min and 30 to 60% after 3 h, whereas Mycobacterium avium and Mycobacterium fortuitum isolates were unaffected by CB-18. In liquid culture, when the concentration of CB-18 exceeded 5 microg/ml, there was an impact on growth characteristics for the most susceptible M. tuberculosis isolate. In contrast, M. fortuitum isolates were able to grow in 100 microg of CB-18 per ml. In liquid culture, the deleterious effects of CB-18 were enhanced in the presence of antibiotics, whereas growth on solid media was not similarly affected. Supplementation of the resuspension buffer with 0.15% lecithin alleviated toxicity. Initial attempts to modify the CB-18 procedure to control contamination incorporated acids or alkalis; however, losses in culture sensitivity occurred. Studies to identify these contaminants led to the development of a sediment resuspension buffer that contained lytic enzymes to combat contamination and lecithin to alleviate toxicity. This formulation included lysozyme, zymolyase, and Cytophaga and Trichoderma extracts and was seen to reduce contamination to acceptable levels (<5%).

Behavior of captive american kestrels hatched from o,p'-dicofol exposed females.

A two-generation laboratory study was conducted on captive American kestrels (Falco sparverius) to investigate the possible behavioral alterations from the compound o,p'-dicofol. Paired females were exposed to three levels of dicofol by oral gavage. Adults of the second generation, exposed only via the maternal dose of o,p'-dicofol at 5- and 20-mg/kg, displayed negatively correlated changes in reproductive behaviors. Second generation females showed a significant difference (p < 0.05) from control birds in the miscellaneous behavior category, where more time was spent picking at the rope perch, watching flies, laying down on the perch and other such displacement activities. Second generation 5-mg/kg males completed significantly fewer copulations (p < 0.05) than the corresponding control birds. The number of attempted or failed food transfers (a pair bonding activity) was also significantly higher (p < 0.05) in the second generation 5-mg/kg male group. Trials were conducted on second generation males to determine aggressiveness of these individuals when placed in a competitive arena. Primary perch sites and food items were obtained by control birds significantly (p < 0.05) more often than dicofol-exposed males. A bird's maternal dose had a significant negative effect (p < 0.05) on its ability to achieve a high rank order position.

Lag screw fixation of an extensor process fracture in a foal with flexural deformity.

A 4-month-old colt with stage I flexural deformity of the distal interphalangeal joint was diagnosed as having a type IV nondisplaced fracture of the extensor process of the 3rd phalanx. An inferior check ligament desmotomy and internal fixation of the fracture were performed. Favorable long-term results for internal fixation of extensor process fractures are presented.

Reproductive and morphological effects of o,p'-dicofol on two generations of captive American kestrels.

A two-generation laboratory study was conducted on a captive population of American kestrels (Falco sparverius) to investigate the possible reproductive and teratogenic effects of the pesticide dicofol. Paired females were exposed to three levels of dicofol: 0, 5, and 20 mg/kg. Integrity of the reproductive tract of the resulting embryos was examined. Viable eggs were hatched, and these birds were permitted to breed the following year. Breeding performance for these birds was measured based on their ability to form pair bonds and exhibit normal behavior in the presence of a mate. Clutch completion, fertility, hatchability, and number of hatchlings reared to the fledging were used as reproductive parameters. Females dosed with 20 mg/kg of dicofol laid eggs with shells that were significantly (p < 0.05) thinner than those of the control birds. Residue levels of dicofol in the form of dichlorobenzophenone were detected in the first and second clutch eggs of the 20-mg/kg dose group only. Male embryos from females dosed with 5 and 20 mg/kg of dicofol had gonads that were significantly different (p < 0.05) from the control chicks. Feminization of male embryos was confirmed by the presence of primordial germ cells in the male gonad. Second-generation adult 5-mg/kg females showed a significantly (p < 0.05) greater number of eggs and hatched chicks lost when compared to second-generation control females. Similar results were found in second-generation 5-mg/kg males paired with normal females and had a significant (p < 0.05) number of chicks die posthatching. Results of second-generation breeding parameters indicate a negative effect on reproductive behavior.

Subchronic toxicity of 3,3',4,4'-tetrachlorobiphenyl in the rat liver: an electron microscope study.

Ultrastructural effects of 3,3'4,4'-tetrachlorobiphenyl (PCB) congener #77 on the liver were evaluated following its feeding to Sprague-Dawley weanling rats. Treatment diets were prepared by dissolving the congener in 4% corn oil. Ten animals, either male or female, in each group were placed on the respective diets containing 10, 100, 1,000 and 10,000 ppb congener for 13 weeks. Ten animals of each sex served as the control that had only the oil added to the diets. In the congener-exposed animals the alterations consisted of a marked increase in the profiles of smooth endoplasmic reticulum, and in the heightened number of lipid droplets in many parenchymal cells. Several mitochondria showed abnormalities such as dumbbell shapes, and in others, the cristae were oriented parallel to the long axis of the organelle. Peroxisomes were numerous in the 10 ppb group and apparently had increased numerically in the liver of animals from the higher dose groups. Females were notably more affected by the congener when compared to their male counterparts. The results indicated that the compound is mildly toxic, and alteration in structure and function can be noted at the lowest dose used (10 ppb congener exposure). It is concluded that congener #77 may be moderately toxic and it may affect the overall health of the exposed animal.

Toxicity of 2,2',4,4',5,5'-hexachlorobiphenyl in the rat liver: an electron microscope study.

Morphological effects on the liver of Sprague-Dawley rats administered orally 2,2',4,4',5,5'-hexachlorobiphenyl (PCB), congener #153, were analyzed. Treatment diets were prepared by dissolving the congener in 4% corn oil. Ten animals of either sex in each group were placed on the respective diets that contained 50, 500, 5,000, or 50,000 ppb congener. Ten animals of each sex served as the control that had only the oil added to the diets. Thirteen weeks after the onset of dosing, the animals were euthanized and liver samples were obtained from the animals and prepared for electron microscopy. Animals exposed to the congener showed (in a dose-related manner) a marked increase in smooth endoplasmic reticulum profiles, and in the number of lipid droplets in many parenchymal cells. Mitochondrial abnormalities such as dumbbell shapes, and in others, the cristae that were oriented parallel to the long axis of the organelle were present. The magnitude of morphologic alterations did not reveal gender differences. The results indicate mild hepatotoxicity of the congener in the rat.

Subchronic toxicity of pentachlorobiphenyl congeners n. 126 or 118 in the rat liver. An electron microscope study.

3,3',4,4',5-Pentachlorobiphenyl (PCB) or congener n. 126 and 2,3',4,4',5-pentachlorobiphenyl or congener n. 118 were given independently to male and female Sprague-Dawley weanling rats. Experimental diets were prepared by dissolving the congeners in 4% corn oil. The congeners were administered as follows: congener n. 126--groups of three animals, either male or female, in each group were placed on the respective diets containing 0.1, 1.0, 10.0 ppb congener, 5.0 micrograms/kg bw loading dose + 10.0, or 100 ppb; congener n. 118--the females were dosed with 2, 20, 200, and 2,000 ppb congener, and the males received 10, 100, 1,000, 10,000 ppb. Thirteen weeks after the start of dosing with the two congeners, liver samples were obtained from all the animals and prepared for electron microscopy. In the congener n. 126-exposed animals, the alterations noted in a dose-related fashion consisted of a marked increase in the profiles of smooth endoplasmic reticulum (SER), and in the heightened number of lipid droplets in many parenchymal cells. Mitochondria showed abnormalities such as dumb-bell shapes, and the cristae parallel to the long axis of the organelle. Lipofuscin granules were numerous in the liver of animals that received 100 ppb of the congener; notably the females of the treatment group expressed this trait more abundantly than the males of the group. We conclude that the compound is mildly toxic. In the animals administered congener n. 118, the alterations were revealed in the liver of both male and female animals in a dose-related manner, also the most evident hepatocyte architectural modifications included an augmentation of SER profiles, mitochondrial aberrations, and an elevated number of lysosomal elements and lipid droplets. Abnormal shapes, and cristae in atypical orientation comprised mitochondrial aberrations. Alterations in the liver morphology of the females were qualitatively similar to those in the males; however, the dose levels used in the latter were five-folds of that which were given to the females. We conclude that the females are more sensitive than the males of the species to congener n. 118. We further conclude that congener n. 118 is less toxic than n. 126 since the lesions were induced by several-folds high dose levels used for the former.