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Land spreading of olive mill wastewater: effects on soil microbial activity and potential phytotoxicity.

Extremely high organic load and the toxic nature of olive mill wastewater (OMW) prevent their direct discharge into domestic wastewater treatment systems. In addition to the various treatment schemes designed for such wastewater, controlled land spreading of untreated OMW has been suggested as an alternative mean of disposal. A field study was conducted between October 2004 and September 2005 to assess possible effects of OMW on soil microbial activity and potential phytotoxicity. The experiment was carried out in an organic orchard located on a Vertisol-type soil (Jezre'el Valley, Israel) and included two application levels of OMW (36 and 72m(3)ha(-1)). Total microbial counts, and to less extent the hydrolytic activity and soil respiration were increased following the high OMW application level. A bench-scale lab experiment showed that the rate of OMW mineralization was mainly dependent on the general status of soil activity and was not related to previous acclimatization of the soil microflora to OMW. Soil phytotoxicity (% germination and root elongation) was assessed in soil extracts of samples collected before and after each OMW application, using germinating cress (Lepidium sativum L.) seeds. We found direct short-term effect of OMW application on soil phytotoxicity. However, the soil was partly or completely recovered between successive applications. No further phytotoxicity was observed in treated soils as compared with control soil, 3 months after OMW application. Such short-term phytotoxicity was not in correlation with measured EC and total polyphenols in the soil extracts. Overall, the results of this study further support a safe controlled OMW spreading on lands that are not associated with sensitive aquifers.

Biodegradation, Environmental↗

Enumeration of purified suspensions of Mycobacterium leprae.

Previously described methods of counting noncultivable bacteria have a number of drawbacks including unpredictable variation due to differential staining, low reproducibility between replicate sample smears, and inexact estimations of the bacillary population due to the non-normal distribution of bacilli across the counting field. A simple method is described which minimizes the disadvantages of previous methods and allows the application of a stratified sample technique which improves the accuracy of the count by partially compensating for the uneven distribution of bacilli across the counting area. This method of analysis also allows an estimation of the optimum sampling rate in each strata of the counting area and the determination of a sample variance. Different fixation and staining techniques have been compared, and the methenamine silver method which is more likely to visualize either the entire population or a constant proportion of the bacilli is recommended.

Humans↗

Must initiators come first? Tumorigenic and carcinogenic effects on skin of 3-methylcholanthrene and TPA in various sequences.

Groups of hairless mice were treated with 4 skin applications of 470 nmol 3-methylcholanthrene (MCA) in benzene and 4 of 20 nmol 12-O-tetradecanoylphorbol-13-acetate (TPA) in various sequences, twice a week, together and separately. Three days after the last application, cell kinetic investigations were made comprising the counting of basal and suprabasal cells, the assessment of hyperplasia, the mitotic rate by the stathmokinetic method, the labelling index and the specific activity of DNA after injection of a dose of [3H]dT, and the determination of percentage of cells in each cell-cycle phase by flow cytometry. These studies showed that various treatment schedules with 4 applications stimulated proliferation and caused epidermal hyperplasia, but there was no significant difference between the groups in degree of growth stimulation. There was a significantly higher tumour production by all the combinations than by MCA alone. It was of no significant importance for the tumour production whether the 4 applications of MCA came before or after the 4 of TPA. Alternating treatment (MCA-TPA, etc.) seemed to give a higher tumour risk than the other treatment sequences. The consequences of these results for the two-stage theory of carcinogenesis (stating that initiation must come first) are discussed, and it is concluded that (at least under the experimental conditions used here) initiation does not need to come first for a good tumour yield.

Animals↗

Platelet volume: laboratory measurement and clinical application.

Modern cell analyzers routinely produce platelet indices, but many clinicians do not utilize them in clinical decision-making. In part this can be attributed to difficulties with their laboratory measurement. The mean platelet volume is dependent on a number of variables, including time of analysis after venepuncture, method of analysis, anticoagulant used and specimen storage temperature. The influence of these laboratory variables is significant and reproducible mean platelet volumes are dependent on standardized laboratory methodology. When pre-analysis factors are controlled, alterations in platelet volume can be demonstrated in a number of disease states and assessment of platelet volume can be useful in the monitoring and diagnosis of patients. An understanding of the pathophysiology of alterations in platelet volume and of the inverse relationship between platelet volume and count is a prerequisite for the successful clinical application of platelet volume measurements.

Anticoagulants↗

The effect of sustained-release varnish of chlorhexidine in dental plastic shells on salivary Streptococcus mutans.

The suppression of S. mutans is a predominate factor in preventing tooth decay. Sustained-release-delivery varnish of chlorhexidine was applied to dental plastic shells and administered to eight volunteers to be worn while sleeping. Salivary bacterial samples were taken on days 0, 7, 14, 21, 28, 35. A statistically significant reduction in S. mutans counts was recorded during the course of the study while a non-significant reduction in the total bacterial counts was found. Our results indicate that the application of chlorhexidine in the form of slow-release varnish in plastic shells is an effective intraoral drug delivery system resulting in reduction of S. mutans.

Chlorhexidine↗

Liming poultry manures to decrease soluble phosphorus and suppress the bacteria population.

Stabilizing phosphorus (P) in poultry waste to reduce P losses from manured soils is important to protect surface waters, while pathogens in manures are an emerging issue. This study was conducted to evaluate CaO and Ca(OH)2 for killing manure bacterial populations (pathogens) and stabilizing P in poultry wastes and to investigate the influence on soils following amendment with the treated wastes. Layer manure and broiler litter varying in moisture content were treated with CaO and Ca(OH)2 at rates of 2.5, 5, 10, and 15% by weight. All treated wastes were analyzed for microbial plate counts, pH, and water-soluble phosphorus (WSP), while a few selected layer manures were analyzed by phosphorus X-ray absorption near edge structure (XANES). A loamy sand and a silt loam were amended with broiler litter and layer manure treated with CaO at rates of 0, 2.5, 5, 10, and 15% and soil WSP and pH were measured at times 1, 8, and 29 d. Liming reduced bacterial populations, with greater rates of lime leading to greater reductions; for example 10% CaO applied to 20% solids broiler litter reduced the plate counts from 793,000 to 6500 mL-1. Liming also reduced the WSP in the manures by over 90% in all cases where at least 10% CaO was added. Liming the manures also reduced WSP in soils immediately following application and raised soil pH. The liming process used successfully reduced plate counts and concerns about P losses in runoff following land application of these limed products due to decreased WSP.

Animals↗

The sympathetic efferent innervation of the cutaneous and muscle veins in cats. A comparative study using retrograde localization with horseradish peroxidase.

Previous studies using electrical stimulation, tension recording and fluorescence histochemical methods indicate that variations in the sympathoadrenergic innervation exist in regional veins of the limb. In the present experiment, we used horseradish peroxidase (HRP) as a retrograde tracer to localize and quantitate the sympathetic innervations to the saphenous, femoral and muscle veins in the cat. The animal was anesthetized with pentobarbital. In three groups of cats, a segment of saphenous (n = 8), femoral (n = 8) and muscle (n = 10) vein was isolated. HRP was applied on the outer vein wall for 3-4 h to allow uptake into the nerve endings. The paravertebral sympathetic chain on the same side of HRP application was dissected after the animal was killed and fixed 60 h following the application of tracer. HRP-labeled neurons were counted in each sympathetic ganglion from L1 to S1. The average number of neurons (mean +/- SE) were 913 +/- 99, 732 +/- 70 and 234 +/- 32, respectively, for saphenous, femoral and muscle vein. There was no statistical difference between the saphenous and femoral vein (P > 0.1). The muscle vein was far less innervated (P < 0.001). When the surface area (mm2) of the vein segment for HRP application was taken into account, the neurons per mm2 were 44.1 +/- 4.8 for saphenous vein, 24.6 +/- 1.8 for femoral vein and 10.2 +/- 1.3 for muscle vein. The neuron density was significantly different among the three groups (P < 0.01). In a single ganglion, the distribution of HRP-neurons appeared to be scattering in pattern.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Blood rheology as a marker of venous hypertension in patients with venous disease.

During chronic venous insufficiency (CVI), several microvascular functional abnormalities, due to venous hypertension, develop. To look for blood rheological consequences of venous hypertension "VH", whole blood viscosity and its main determinants were measured in 11 normal controls and 36 patients with CVI exposed to a short-term experimental VH. Patients were subdivided into 2 groups according to the severity of their disease. Blood was taken from a foot vein before and after VH, which was induced by appling a pneumatic tourniquet to 100 mmHg for 15 minutes. Whole blood viscosity at low and high shear rates, red blood cell (RBC) aggregation, RBC rigidity, plasma viscosity and proteins as well as red and white blood cell (WBC) counts were recorded. Patients at baseline, i.e., before application of the tourniquet, showed several hemorheological abnormalities such as an increased RBC aggregation, increased low shear rate viscosity, and a significant elevation in plasma fibrinogen level. Patients with more severe CVI had more marked hemorheological changes. The short term VH in patients led to further aggravation of these changes. There were also at baseline lower values, however not significantly, of hematocrit and RBC count, suggesting that hemoconcentration is not a feature of CVI. These same parameters were slightly, however not significantly, increased after VH, indicating a fluid escape into the extravascular space. A significant fall in WBC count was also observed after VH, in keeping with the white cell trapping hypothesis. In conclusion, even a short-term VH is able to induce several hemorheological impairments, which are probably involved in the failure of the microcirculation and hence the initiation of tissue damage in patients with CVI.

Biomarkers↗

Evaluation of focal mollusciciding in the Rahad Irrigation Scheme, Sudan.

Since the establishment of the Rahad Irrigation Scheme in 1979, the prevalence of schistosomiasis has increased to high levels in some villages despite the efforts of the Blue Nile Health Project to prevent this. Although a programme of focal snail control has been implemented since 1979, when the first intermediate host snails were detected, infected snails are abundant in human water contact near villages where transmission of both S. mansoni and S. haematobium takes place. The objective of the present study was to assess the feasibility and effectiveness of the focal snail control operations in irrigation canals, in terms of reductions in snail density and repopulation rates of treated sites. The efficacy of focal mollusciciding was evaluated by (1) evaluation of the routine molluscicide operation done by the Blue Nile Health Project, (2) experimental application of various dosages of the molluscicide and (3) study of snail repopulation rates. Efficiency was measured by mortality among caged snails placed in the target stretch and from pre- and post-treatment counts of relative snail densities. The short-time application of niclosamide can be very effective in reducing the density of the intermediate host snails in the target stretch of canal. An efficient exposure seems to be approximately 3 ppm (a.i.) for 30-40 min when water flow is slow, while 1 ppm is efficient when water velocity is very slow, i.e. less than 0.04 m S-1, and 0.6 ppm is efficient when the water is virtually stagnant. Turbidity seems not to impair the effect of the treatment and vegetation does not reduce the efficiency of niclosamide, provided the current speed is low and the chemical is applied to the entire width of the canal. However, the results also clearly demonstrate some of the shortcomings of the standard procedure of niclosamide application. A high water velocity and discharge will rapidly wash the chemical through the target canal stretch and the snails will be exposed only for a short period if the routine procedure is followed. In addition, the chemical will not mix sufficiently with water within standards of aquatic macrophytes. Recolonization of treated canal stretches is fast, especially during the irrigation period when water velocities are high and juvenile snails are abundant. Snail density may attain pretreatment levels three weeks after an effective application. Since the routine procedure is most effective at low current speeds, it is suggested that water velocity could be controlled during the application in coordination with the Irrigation Department.

Animals↗

Methods of quantitative autoradiography using incident light microphotometry.

Several different approaches of automated grain counting of microautoradiographic grain densities have been reported in the literature. Application of grain counters to cell biology is limited, however, primarily due to shortage of methods allowing the interpretation of grain counts on a molecular basis. Two suitable methods of quantitative autoradiography at the cellular level are reviewed, developed for the isotopes 14C and 125 I. They permit evaluation of absolute radioactivity in autoradiographs and, thus, determination of synthesis processes such as deoxyribonucleic acid synthesis, and of antigen densities on cell surfaces. In this approach towards quantitative autoradiography, grain densities are compared photometrically over labeled cells and over a standard source on the same autoradiograph. Allowance has to be made for the specific geometric factors of the isotope used. This can be advantageously done with an integrating type of measurement using incident light bright-field. With this type of recording, there is an exponential dependence of the photometric values on the radioactive dose. As an example of application, results are presented of the deoxyribonucleic acid synthesis rate of human myelocytes in aplastic anemia and of the immunoglobulin G density on lymphocyte membranes in the normal state and in chronic lymphocytic leukemia.

Animals↗

Evaluation of fipronil spot-on in the treatment of flea allergic dermatitis in dogs.

The purpose of this study was to evaluate the effectiveness of treatment with 10 per cent fipronil solution for controlling signs of flea allergic dermatitis in dogs under field conditions. Thirty-one client-owned dogs with flea allergic dermatitis were treated with three monthly applications of 10 per cent fipronil solution. Flea counts and pruritus were significantly reduced at all post-treatment visits. At the final visit, on day 90, flea counts were reduced by 98 per cent, and pruritus was reduced or eliminated in 84 per cent of the study dogs. Dermatological lesion scores for erythema, crusts, scales and papules were also significantly improved by the final visit. The overall assessment of efficacy on day 90 was 'excellent' to 'good' for 87 per cent of the study dogs. The results demonstrate that treatment with monthly topical applications of 10 per cent fipronil solution is effective in reducing the prevalence and severity of signs of flea allergic dermatitis in dogs.

Administration, Cutaneous↗

Henna (Lawsonia inermis Linn.) inducing haemolysis among G6PD-deficient newborns. A new clinical observation.

Over a 10-year period, 15 glucose-6-phosphate dehydrogenase (G6PD)-deficient male newborns were admitted to Al-Jahra Hospital with acute haemolysis a few days after applying henna dye over the body, which is a unique Bedouin tribal practice to celebrate the arrival of the first-born boy. Laboratory investigations revealed significant anaemia, reticulocytosis and indirect hyperbilirubinaemia among the index newborns as compared with controls (p < 0.001). The mean (SD) haemoglobin concentration in index patients was 113.4 (13.4) g/l vs 171.2 (11.2) g/l in controls, reticulocytic count 13.8% (4.2%) vs 1.4% (0.74%), and indirect bilirubin 382.8 (58.7) mumol/l vs 63.7 (61.4) mumol/l. G6PD-deficient newborns with haemolysis linked to henna application had delayed age at presentation and a higher reticulocytic count and hyperbilirubinaemia compared with non-henna-induced haemolysis (p < 0.05). Percutaneous henna absorption is well recognized and clinical findings support the harmful effect of henna on G6PD-deficient red blood cells. A local health education programme has been established to prevent the use of henna dye in infancy.

Administration, Topical↗

Five years experiences with microinjection of testicular spermatozoa into oocytes in Hungary.

The aim of the study was to summarize our five years experience (1996-2000) of testicular spermatozoa for intracytoplasmic sperm injection in Hungary. The influence of sperm count, maternal age, number of transferred embryos, and application of assisted hatching on outcome was investigated. Testicular spermatozoa were retrieved by microsurgical testicular sperm extraction. Samples were classified depending on the number of spermatozoa. Indication for testicular sperm extraction in conjunction with intracytoplasmic sperm injection was severe azoospermia or azoospermia combined with tubal origin infertility. Ovarian stimulation was carried out using an ultrashort protocol with GnRH agonist and gonadotrophin. Intracytoplasmic sperm injection was performed without PVP. Embryos were cultured for 48 or 72 h before embryo transfer. Indications for assisted hatching included elevated maternal age, increased zona thickness or at least two previous unsuccessful IVF cycles. Testicular spermatozoa were successfully retrieved in 218 out of 273 cases. Extreme low sperm count was found more frequently in cases of nonobstructive azoospermia. No significant differences were observed in fertilization rate (61.1% vs. 51.7%) or clinical pregnancy rate (29.0% vs. 26.7%) between patients with obstructive or nonobstructive azoospermia. Maternal age, number of transferred embryos and application of assisted hatching had a significant effect on outcome. A total of 55 clinical pregnancies were achieved, including 14 sets of twins, three sets of triplets and two sets of quadruplets. It is concluded that testicular sperm extraction is an efficient way of obtaining testicular spermatozoa, allowing not only successful fertilization by ICSI, but also freezing of testicular spermatozoa for use in subsequent cycles.

Female↗

Application of antimicrobial-producing lactic acid bacteria to control pathogens in ready-to-use vegetables.

Five psychrotrophic strains of lactic acid bacteria (Lactobacillus casei, Lact. plantarum and Pediococcus spp.) were isolated from 22 samples of commercial salads. These strains were shown to inhibit Aeromonas hydrophila, Listeria monocytogenes, Salmonella typhimurium and Staphylococcus aureus on MRS agar, in salads and in juice prepared from vegetable salads. Lactobacillus casei IMPCLC34 was most effective in reducing total mesophilic bacteria and the coliform group; Aer. hydrophila, Salm. typhimurium and Staph. aureus disappeared after 6 d of storage, while the counts for L. monocytogenes remained constant. The potential application of antimicrobial-producing lactic acid bacteria as biopreservatives of ready-to-use vegetables is suggested.

Antibiosis↗

Effect of a quaternary ammonium sanitizer for hatching eggs on their contamination, permeability, water loss, and hatchability.

The present study tested the biocidal effectiveness of the quaternary ammonium Hatching Egg Sanitizer Spray (HES Spray) in disinfecting broiler hatching eggs and in terms of its effects on eggshell permeability, water loss, and hatchability. The application of HES at a 1.5% or a 3.0% concentration resulted in significant reductions in the total aerobic counts on the egg surface of 98.1% and 99.9%, respectively, within 30 min of application. Molds and yeasts were significantly reduced by 3.0% HES at 14 days of incubation. Significant reductions in total aerobic counts on the egg surface due to 1.5% and 3.0% HES were also observed on eggs that were allowed to "sweat." The 3.0% HES concentration also reduced coliforms on egg surfaces. Hatchability of fertile eggs from a 32-wk-old flock was significantly increased, over 6.0%, by spraying 1.5% or 3.0% HES in comparison to controls that were not sprayed, with no significant difference in hatchability due to treatment observed in eggs from flocks that were 36, 42, 46, or 62 wk of age. This change in hatchability associated with spraying HES may be due to a change in eggshell permeability (respiration) caused by an interaction of HES with the eggshell cuticle.

Animals↗

Reduced bactericidal activity in neutrophils from scorbutic animals and the effect of ascorbic acid on these target bacteria in vivo and in vitro.

Actinomycetes, involved in oral and periodontal diseases, cause serious infections in immunocompromised hosts. Severely scorbutic guinea pig leukocytes killed only 12% of phagocytosed actinomycetes, had distorted nuclear morphology, had 16 times less ascorbate, and had no chemotactic responses in vitro. Ascorbate reversed these indices and also prevented nitrosamine formation by oral organisms. Degranulating leukocytes release lactoferrin and ascorbate that chelate iron, essential for microorganisms. Ascorbic acid, 2,2'-bipyridine and 1,10-phenanthroline were bactericidal to several bacterial pathogens at millimolar concentrations. Iron alone reversed this effect. In in vivo experiments an Actinomyces viscosus monoflora was implanted in rhesus monkeys. Plaque and serum samples showed decreased (by six orders of magnitude) bacterial counts and decreased actinomycete antibody titers in animals given 1 g ascorbate/d. Removing ascorbate returned counts and titers to preascorbate concentrations. Fifteen marmosets, receiving twice daily topical applications of ascorbate or water, had comparatively lower gingival, calculus, and plaque indices and only slightly lowered actinomycete counts.

Actinomyces viscosus↗

Cytomat-R: a computer-controlled multiple laser source multiparameter flow cytophotometer system.

A multiple illumination wavelength multiparameter flow cytophotometer system, using laser sources and controlled by a small, general-purpose digital computer, has been produced for use in the development of new flow cytometric techniques. Three different laser wave-lengths can be used simultaneously to illuminate different regions of the flow chamber; as many as five measurements of light scattering at various angles, extinction, and fluorescence at one or more wavelengths can be made at each illuminated station. Cells in suspension may be examined at rates of 1000 cells/sec, with seven correlated optical measurements being recorded for each cell. A library of programs for data manipulation and statistical analysis make it possible to use the system to develop and implement cell characterization, counting and classification procedures for basic and clinical research applications.

Blood Cells↗