Comparative study of cytotoxicity of Aeromonas spp. on four different cell lines

In vitro cytotoxicity is an important virulence property of motile mesophilic Aeromonas species. Cell-free supernatant prepared from 55 Aeromonas isolates including one A. hydrophila type strain (MTCC 646) were examined for their cytotoxic potential on four different cell lines (Vero, BHK-21, MDBK, B 95a). Results of the study revealed cytotoxic potential in 92.72% of the isolates. Analysis of data exposed significant variation among isolates in respect of their cytotoxicity. Vero cells proved to be most sensitive to aeromonal toxins and B 95a cells showed significantly (P<0.01) lower response compared to other cell lines. Sensitivities of BHK-21 and MDBK cell lines were in between Vero and B 95a.     

Predicting breeding values and accuracies from group in comparison to individual observations

Individual observations are routinely used in livestock evaluations. In some cases, pooled data representing the joint but not individual performance of a group of animals may be available. For example, pooled feed intake may be measured on a pen of livestock. The usual mixed model approach to genetic evaluation can still be applied as an exact method in this setting, provided incidence and residual variance-covariance matrices are suitably modified to account for the pooling. Approximate evaluations may be achieved by treating average performance as if it pertained to each individual in the pool. Theoretical accuracies can be obtained as a function of elements of the inverse coefficient matrix. A 3-generation data set representing 1,000 animals with feed intake observations from 49 sires and 200 maternal grand sires was simulated with heritability of 0.34. Individual records were pooled to represent circumstances in which animals with records were collectively measured. Animals were allocated into pens at random, by sire, or by maternal grand sire. Simulation was replicated with unique fixed effects for each pen. Following evaluation from each method, the empirical accuracy or product-moment correlation between true (simulated) and estimated merit could be quantified. The analysis of individual observations resulted in empirical accuracy of 0.63 for animals on test and 0.77 for their sires. Pooling the observations in pens of 2, 4, or 12 animals reduced empirical accuracies for animals on test to 0.50, 0.41, and 0.21 when pooling was at random and 0.53, 0.47, and 0.34 when pooling was by sire. Simulating a fixed pen effect representing 10% phenotypic variation, but ignoring that effect in the evaluation minimally reduced empirical accuracies to 0.52, 0.46, and 0.33 when pooling by sire. Theoretical accuracies were in close agreement with empirical accuracies when the exact method was used. The approximate method that treated averages of pooled data as if they were individually observed overstated accuracy and should not be used. Selection on the basis of pooled observations can be almost as effective as using individual observations when pool sizes are small. The exact method to account for pooled data is no more complex than conventional procedures.     

Quantitative analysis of animal model lens anatomy: accommodative range is related to fiber structure and organization

Purpose The results of a recent study on accommodation in humans and baboons has revealed that lens fiber structure and organization are key components of the mechanism of accommodation. Dynamic focusing involves the controlled displacement and replacement, or realignment, of cortical fiber-ends at sutures as the mechanism of accommodation at the fiber level. This emended explanation of the mechanism of accommodation raises the following question: as the structure of crystalline lenses are only similar, not identical between species, is accommodative amplitude related to differences in the structure and organization of fibers between species? To address this question, we have quantitatively examined the structure and organization of fibers in a number of the more commonly used animal models (mice, cattle, frogs, rabbits and chickens) for lens research. Materials and methods Lenses (a minimum of 12–18 lenses/species) from mice, cattle, frogs and rabbits were used for this study. Prior to fixation for structural analysis, measurements of the gross shape of the lenses (equatorial diameter, anterior and posterior minor radii [anterior + posterior minor radius = polar axis]) were taken directly through a stereo surgical dissecting microscope equipped with an ocular reticle. Lenses were then prepared for and examined by light (LM), transmission (TEM) and scanning electron microscopy (SEM). Scale computer-assisted drawings (CADs) of lenses and lens fibers were then constructed from quantitative data as described above and from quantitative data contained in micrographs. Results The differences in fiber structure and organization that effect accommodative range arise early in development and are continued throughout lifelong lens growth. In umbilical suture lenses (avian) secondary fibers develop with almost completely tapered anterior ends (85–90% reduction of their measures of width and thickness at the equator). By comparison, in lenses with line sutures (e.g. frogs and rabbits) secondary fibers develop with just a 50–60% reduction in anterior fiber taper. In lenses with Y sutures (mice and cattle), fiber width taper is only 25–40%. However, in all cases, while the taper of the posterior end width of fibers is just slightly less (approx. 15–20%) than that of anterior ends, posterior end thickness is only reduced by one half that of anterior thickness. Conclusions In humans, the mechanism of accommodation at the fiber level involves the controlled realignment of very flattened and flared, rather than tapered fiber-ends at sutures. In this manner, the simultaneous increase in lens thickness and surface curvature in the accommodated state is the result of fiber-ends being overlapped along multiple (9–12) suture branches covering the majority of the anterior and posterior surfaces. The results of this animal study strongly suggest that accommodative range is directly related to quantitative differences in fiber structure and organization in the different suture types. The very broad accommodative range in birds is made possible, at least in part, by the almost complete tapering of fiber-ends at umbilical sutures. In contrast, the essentially negligible accommodative range of animals that have line- and Y-suture lenses is at least partially the result of the fact that these lenses have fibers with very little end taper. Thus, the blunt ends of fibers in line- and Y-suture lenses precludes any significant overlap of end segments to effect accommodation.     

Identification and Quantification of Pathogenic Pythium spp. from Soils in Eastern Washington Using Real-Time Polymerase Chain Reaction

ABSTRACT Traditional methods of quantifying Pythium spp. rely on the use of selective media and dilution plating. However, high variability is inherent in this type of enumeration and counts may not be representative of the pathogenic population of Pythium spp. Variable regions of the internal transcribed spacer of the rDNA were used to design species-specific primers for detection and quantification of nine Pythium spp. from soils in eastern Washington. Primer pairs were designed for Pythium abappressorium, P. attrantheridium, P. heterothallicum, P. irregulare group I, P. irregulare group IV, P. paroecandrum, P. rostratifingens, P. sylvaticum, and P. ultimum and used with real-time polymerase chain reaction. Standard curves were generated for each of the species using SYBR Green I fluorescent dye for detection of amplification. Seventy-seven isolates of Pythium were screened to confirm specificity of each primer set. DNA was extracted from soil and standard curves were generated for P. irregulare group I, P. irregulare group IV, and P. ultimum to correlate populations of each species in the soil with quantities of DNA amplified from the same soil. Examination of raw field soils revealed results similar to those observed in previous studies. This new technique for the quantification of Pythium spp. is rapid and accurate, and will be a useful tool in the future study of these pathogenic Pythium spp.     

Skeletal changes in hemochromatosis of Salers cattle

Hemochromatosis is an inherited defect in Salers cattle, characterized by increased iron absorption and wasting in young animals. Loss of teeth and soft bones has been reported. This study details bone changes in a prospective study of an affected Salers heifer and an age-matched, half-sibling control. The heifer fell 2 weeks before euthanasia, developed non-weight bearing lameness, and then became recumbent. At necropsy, the heifer was smaller than the control and had fractures of the humerus, femur, and ribs. Distinctive circumferential laminations in the outer third of the cortex were seen radiographically as layers of variably decreased density. In midshaft cross sections, the cortex was wider in some long bones, and fluorescent labeling revealed irregular and intermittent mineralization in the outer cortical layers. Mineral apposition rates in osteons of the inner cortex were decreased as compared with the control, and longitudinal bone formation at the growth plate was markedly decreased. Histologically, there were buried layers of osteopenic, poorly mineralized plexiform bone in outer circumferential lamellae. There was stainable iron in woven bone cores of the matrix. Iron was found in occasional osteoclasts as well. The mandible was severely affected, with loss of cortical bone as well as trabecular bone around the teeth and resorption of dental roots. Bone analysis revealed iron levels in the affected calf that were 30-50 times greater than the control and decreased percent ash in the outer cortex. These results indicate that periosteal dysplasia and osteopenia are responsible for pathologic fractures and tooth loss.