A comparative study on the different staining methods and number of specimens for the detection of acid fast bacilli

The presence of acid fast bacilli in multiple specimens was investigated comparatively with Ziehl-Neelsen (ZN) and fluorescence microscopy (FM) staining in order to determine sensitivity in detecting tuberculosis (TB). A total of 465 specimens obtained from 295 patients were analysed at Harran University Medical School Hospital between March 1998 and March 2000. The culture was employed as the reference method. Sixty-eight patients (23.1%) were diagnosed as having TB by culture. The ZN and FM staining sensitivities were 67.6% (46/68) and 85.2% (58/68) respectively. Two hundred and one patients (68.1%) submitted one specimen to the laboratory. TB positivity was detected in 42 (20.9%) of these patients by culture. The sensitivities of ZN and FM stains were found to be 61% and 83% in these patients. However, in 18 patients (6.1%) who submitted two specimens to the laboratory, the TB was positive in six of them (33.3%) and ZN and FM sensitivities were 66% and 83% respectively. When three specimens or more were collected from the patients (76 patients, 25.8%), TB positivity was determined in 20 of them (26.3%) and the sensitivities were 80% and 92% in the ZN- and FM-stained smears, respectively. Our data indicate that in the diagnosis of TB, FM has greater sensitivity than ZN. In particular, in the case of a single specimen, the diagnostic value of FM is quite significant. It is, therefore, possible to conclude that both ZN and FM staining can be used for the diagnosis of TB when there are more than two specimens. However, if only one or two specimens are available, FM staining is preferable.

Laboratory evaluation of the effects of triflumuron on the development of Rhodnius prolixus nymph

Studies were carried out to evaluate the efficacy of the growth regulator, triflumuron (TFM) (Starycide® sc 480 Bayer), for disrupting the development of Rhodnius prolixus fifth-instar nymph by oral, topical or continuous treatment. All treatments were able to induce high levels of mortality, delay development and molt inhibition. Oral treatment induced molt inhibition in all insects that survived at doses of 0.25, 0.50 and 5.0 mg/mL of a blood meal. The highest levels of both mortality in 24 h and molt inhibition were always observed after topical treatment. The lowest doses needed to obtain considerable biological effects were always observed after continuous treatment. In this way, the highest levels of mortality within 30 days were detected after continuous treatment, which also induced an extended inter-molting period, a lower number of over-aged nymphs and the highest level of molting in nymphs that survived. Moreover, the effects of TFM on insects were often displayed in a dose response manner. These results indicate that TFM acts as a potent growth inhibitor of R. prolixus nymphs and has the potential to be used in integrated vector control programs against hematophagous triatomine species.

Effect of serum sample inactivation on the performance of latex agglutination test for paracoccidioidomycosis serodiagnosis

Paracoccidioidomycosis is diagnosed from the direct observation of the causative agent, but serology can facilitate and decrease the time required for diagnosis. The objective of this study was to determine the influence of serum sample inactivation on the performance of the latex agglutination test (LAT) for detecting antibodies against Paracoccidioides brasiliensis. The sensitivity of LAT from inactivated or non-inactivated samples was 73% and 83%, respectively and the LAT selectivity was 79% and 90%, respectively. The LAT evaluated here was no more specific than the double-immunodiffusion assay. We suggest the investigation of other methods for improving the LAT, such as the use of deglycosylated antigen.

Prohexadione calcium on the growth and quality of eggplant seedlings

Abstract:The objective of this work was to evaluate the effects of prohexadione calcium concentrations on the growth and quality of eggplant (Solanum melongena) seedlings. The effects of prohexadione calcium concentrations of 0, 50, 100, or 150 mg L-1 on seedling growth parameters were evaluated in a greenhouse experiment. After the greenhouse experiment, the seedlings were transplanted to the field. During the field experiment, the number of days to flowering, plant height, number of fruits per plant, average fruit weight and yield were evaluated. Both experiments were carried out in a completely randomized design with four replicates. All prohexadione calcium concentrations significantly reduced shoot height and internode length, when compared to the control. The concentrations of 50, 100, and 150 mg L-1 prohexadione calcium reduced shoot height by 27, 32, and 38%, respectively. Prohexadione calcium treatments (except the one with 50 mg L-1) enhanced relative chlorophyll content of leaves in comparison to the control. There were no delays in flowering and no significant differences in number of fruits per plant among treatments with prohexadione calcium. The concentrations of 100 and 150 mg L-1 prohexadione calcium significantly reduced yield per plant and total fruit yield, whereas the concentration of 50 mg L-1 did not cause any change in yield compared to the control. The lowest prohexadione calcium concentration can be used to control excessive elongation of eggplant seedlings without yield loss.

Occurence of thrips on Niagara table grape and its control with the insecticides thiacloprid and methiocarb associated with Metarhizium anisopliae

Thrips are reported as important pests on table grapes in United States and several countries of Europe. Damage caused by thrips, particulary Frankliniella occidentalis, was observed on niagara table grape crop in Limeira-SP, Brazil. During the blooming period, high thrips densities were observed feeding on pollen and small berries. The symptoms left were more visible after the development of the berries and were characterized by dark scars and suberized surface on berries, sometimes causing the berry to crack, and the seed to prolapse. The effect of insecticides thiacloprid or methiocarb, associated or not with the entomopathogenic fungus Metarhizium anisopliae were evaluated during the blooming period. For evaluation of thrips damage on fruits, the treatments were applied three additional times, 7, 14 and 21 days after the first application. The treatments were: a) M. anisopliae (strain 1037) 1x10(7) conidia/mL; b) thiacloprid 20mL/100L; c-d) methiocarb 100 and 150mL/100L; e) methiocarb 100mL/100L + M. anisopliae 1x10(7) conidia/mL. Only methiocarb, associated or not with the fungus, was effective in reducing thrips infestation, and no phytotoxic damage was observed. The efficiency of methiocarb 150mL/100L and the insecticide associated with the fungus for the control of the thrips population was 84.2 and 95.5%, respectively. In both cases, there was a reduction of approximately 70% in the number of berries with scars symptoms. For control of thrips on table grapes, chemical insecticides associated or not with M. anisopliae should be applied during the blooming period of the crop.

Use of botanical extracts, cassava wastewater and nematicide for the control of root-knot nematode on carrot

Aqueous extracts of several plant species have shown promising in controlling root-knot nematode, Meloidogyne incognita (Kofoid & White), one of the most limiting agents for carrot cultivation. The current study evaluated the effect of aqueous extracts from seven botanical species applied to 40, 50, 60, 70 and 80 days after sowing 'Nantes' carrots in soil infested with root-knot nematode. Three other treatments included cassava wastewater, distilled water (control), which were applied in the same periods of the extracts application, in addition to carbofuran 50G (80Kg/ha), which was applied once at 60 days after carrot sowing. Evaluations were performed at 90 days after inoculation to determine shoot and root fresh weight, as well as the diameter and the length of principal roots and the number of galls on primary and secondary roots. Plants treated with cassava wastewater, extracts of Ricinus communis L. seeds, Crotalaria juncea L. seeds, R. communis leaves + branches + fruits, Chenopodium ambrosioides L. leaves + branches + inflorescences and Azadirachta indica A. Juss. seeds showed the highest rates of total weight (root + shoot) and shoot weight. The extract of R. communis leaves + branches + fruits provides the highest total root weight and principal root diameter. Cassava wastewater and extracts of R. communis seeds provided the highest principal root weight. The extract of R. communis seeds and cassava wastewater can be considered promising for the alternative control of M. incognita.

Influence of distance, geometry and number of control stations on quality of local geodetic networks for the purpose of georeferencing of rural properties

The implementation of local geodetic networks for georeferencing of rural properties has become a requirement after publication of the Georeferencing Technical Standard by INCRA. According to this standard, the maximum distance of baselines to GNSS L1 receivers is of 20 km. Besides the length of the baseline, the geometry and the number of geodetic control stations are other factors to be considered in the implementation of geodetic networks. Thus, this research aimed to examine the influence of baseline lengths higher than the regulated limit of 20 km, the geometry and the number of control stations on quality of local geodetic networks for georeferencing, and also to demonstrate the importance of using specific tests to evaluate the solution of ambiguities and on the quality of the adjustment. The results indicated that the increasing number of control stations has improved the quality of the network, the geometry has not influenced on the quality and the baseline length has influenced on the quality; however, lengths higher than 20 km has not interrupted the implementation, with GPS L1 receiver, of the local geodetic network for the purpose of georeferencing. Also, the use of different statistical tests, both for the evaluation of the resolution of ambiguities and for the adjustment, have enabled greater clearness in analyzing the results, which allow that unsuitable observations may be eliminated.

Effects of laparoscopy on the cardiorespiratory system of brown brocket deer (Mazama gouazoubira) anesthetized with ketamine/ xylazine/ midazolam combination and isoflurane

Laparoscopy is not widely used as a tool to perform assisted reproduction techniques in South American cervids; thus, scarce information in literature is available regarding its effects and appropriate anesthetic protocols to perform it. This study evaluated the effect of laparoscopy on heart rate (HR), respiration rate (RR), saturation of oxyhemoglobin (SpO2) and rectal temperature (RT) of six female brown brocket deer (Mazama gouazoubira) anesthetized with ketamine (5mg/kg), xylazine (0.3mg/kg), midazolam (0.5mg/kg) combination i.v. and isoflurane. Twelve laparoscopies were performed and each animal was used twice with a 40-day interval. After anesthetized, the animals were placed in dorsal recumbency to perform laparoscopy procedure using abdominal CO2 insufflations (14.2 ± 2.39mmHg; M ± SE). The main events of the laparoscopy procedure were divided into three periods: animal without (P1) and with abdominal insufflation (P2) and abdominal insufflation with the hips raised at 45º (P3). As a control, the animals were anesthetized again 40 days after the last laparoscopy, and were maintained in a dorsal recumbency for the same average duration of the previous anesthesia and no laparoscopy procedure was conducted. The period of anesthesia for the controls was also divided into P1, P2, and P3 considering the average duration of these periods in previous laparoscopies performed. Data were analyzed through the (ANOVA) variance analysis followed by Tukey test and values at P<0.05 were considered significant. No significant differences were observed in the parameters evaluated at P1, P2 and P3 between the animals submitted to laparoscopy and control. However, the RR mean between P1 (38.8 ± 4.42) and P3 (32.7 ± 4.81); and the RT mean between the P1 (38.2ºC ± 0.17), P2 (37.6ºC ± 0.19) and P3 (37.0ºC ± 0.21) varied significantly, independent of the laparoscopy. These data indicated that laparoscopy didn't cause any significant alterations in the cardiorespiratory parameters evaluated, even though the anesthetic protocol used can cause a reduction in the RT contributing to development of hypothermia during anesthesia.

Grass Weeds Interfering with Eucalypt: Effects of the Distance of Coexistence on the Initial Plant Growth

Two experiments were carried out to evaluate the initial plant growth of Eucalyptus urograndis growing in coexistence with Urochloa decumbens and U. ruziziensis. In 100-L box, one plant of U. decumbens or U. ruziziensis grew in coexistence with one plant of E. urograndis clones C219H or H15, respectively, in the distances of 0, 5, 10, 15, 20, 25, 30, 35, and 40 cm from the crop. After 30, 60, 90 (both clones), and 150 days (just for H15), growth characteristics were evaluated. Plants of both clones, growing in weed-free situations, showed a better growth and development than plants that grew in weedy situations, independently of the distance, having the highest plant height, stem diameter, dry mass of stem, and dry mass of leaves. As the same way, the number of branches, number of leaves, and leaf area of the clone C219H were similarly affected. Urochloa ruziziensis reduced the dry mass accumulation of stem and leaves by the rate of 0.06 and 0.32 g per plant, respectively, per each centimeter growing nearest to the crop, while U. decumbens reduced by 0.03 and 0.14 g per plant. The interference of U. decumbens and U. ruziziensis with E. urograndis is more intense when weedy plants grow in short distances from the crop.

Interaction between the mineral content and the occurrence number of aquatic fungi in leaves submerged in a stream in the Atlantic rainforest, São Paulo, Brazil

Leaves of Ficus microcarpa L. f., Quercus robur L., and Alchornea triplinervia (Spreng.) Muell. Arg., submerged in a stream of the Atlantic rainforest in the "Reserva Biológica do Alto da Serra de Paranapiacaba", State of São Paulo, Brazil, were collected monthly, from April to November 1990, in order to determine the number of fungal occurrences (zoosporic fungi and aquatic Hyphomycetes), and the content of total N (%), total P (%), K+ (%), Ca+2 (%), Mg+2 (%), S+3 (%), Fe+3 (ppm), Cu+3 (ppm), Mn+2 (ppm), Zn+2 (ppm), Bo (ppm), Na+2 (ppm) and Al+3 (ppm). According to the tests of Kruskal-Wallis, Mann-Whitney and Wilcoxon, the means of the mineral content of the three types of leaves were significantly different, except for Mg+2 (%), Mn+2 (ppm), Zn+2 (ppm) and Na+2 (ppm). On comparing the mineral content with the number of fungal occurrence, an independence test showed a positive correlation between the presence of zoosporic fungi on the leaves of A. triplinervia and the total nitrogen, phosphorus and S+3 content, whereas the aquatic Hyphomycetes depended on the amount of Ca+2 available. Regarding leaves of F. microcarpa, the occurrence of zoosporic fungi was linked to the S+3 level, and the presence of aquatic Hyphomycetes, to the content of K+, Ca+2, S+3 and Bo. On Q. robur leaves, zoosporic fungi showed a positive correlation with the Ca+2 content, but a negative one with Fe+3 and Al+3 levels, while the occurrence of aquatic Hyphomycetes was influenced by the content of Ca+2, Mg+2, Fe+3, Al+3, Mn+2, Zn+2 and Na+2. The correlation between the occurrence number of aquatic Hyphomycetes and a high mineral content indicates that their nutritional requirements may be more complex than those of zoosporic fungi. Further studies are still required to understand the implications of this tendency on the diversity of aquatic native mycota.

Effects of bromocriptine on serum prolactin levels, pituitary weight and immunoreactive prolactin cells in estradiol-treated ovariectomized rats: an experimental model of estrogen-dependent hyperprolactinemia

The present study was designed to assess the effects of bromocriptine, a dopamine agonist, on pituitary wet weight, number of immunoreactive prolactin cells and serum prolactin concentrations in estradiol-treated rats. Ovariectomized Wistar rats were injected subcutaneously with sunflower oil vehicle or estradiol valerate (50 or 300 µg rat-1 week-1) for 2, 4 or 10 weeks. Bromocriptine (0.2 or 0.6 mg rat-1 day-1) was injected daily during the last 5 or 12 days of estrogen treatment. Data were compared with those obtained for intact control rats. Administration of both doses of estrogen increased serum prolactin levels. No difference in the number of prolactin cells in rats treated with 50 µg estradiol valerate was observed compared to intact adult animals. In contrast, rats treated with 300 µg estradiol valerate showed a significant increase in the number of prolactin cells (P<0.05). Therefore, the increase in serum prolactin levels observed in rats treated with 50 µg estradiol valerate, in the absence of morphological changes in the pituitary cells, suggests a "functional" estrogen-induced hyperprolactinemia. Bromocriptine decreased prolactin levels in all estrogen-treated rats. The administration of this drug to rats previously treated with 300 µg estradiol valerate also resulted in a significant decrease in pituitary weight and number of prolactin cells when compared to the group treated with estradiol alone. The general antiprolactinemic and antiproliferative pituitary effects of bromocriptine treatment reported here validate the experimental model of estrogen-induced hyperprolactinemic rats

Influence of lactation on motor activity and elevated plus maze behavior

Lactating rats show less noise-induced freezing and fewer inhibitory responses on the 6th day post-delivery when submitted to water and food deprivation in a classical conflict paradigm. Lactating mice go more often to the illuminated chamber in a light-dark cage and stay longer in it than virgin females. The present study was designed to assess the influence of this physiological state, i.e. lactation, on the elevated plus maze (EPM) and open-field behavior in adult female rats. Total (TL) and central (CL) locomotion and rearing (RF) frequencies were measured in an open-field. Number of entries into the open and closed arms as well as the time spent in each of these arms were measured in the EPM. Percent time spent and number of entries into the open arms were calculated and compared. In the open-field, TL was significantly decreased (115 ± 10.6 vs 150 ± 11.6) while CL and RF did not differ from those presented by virgin rats. In the EPM, lactating rats displayed a significant reduction in percent time spent (10.9 ± 1.5 vs 17.4 ± 2.3) in the open arms as well as a tendency to a reduction in percent entries into the open arms (35.7 ± 4.7 vs 45.7 ± 4.3). These results show that the physiological state of lactation modulates the open-field and EPM behaviors in rats

Aversive stimulation during the stress-hyporesponsive period does not affect the number of corticotroph cells in neonatal male rats

Immunohistochemistry was used to evaluate the effects of neonatal handling and aversive stimulation during the first 10 days of life on the number of corticotrophs in the anterior lobe of the pituitary of 11-day-old male Wistar rats. Since adult rats handled during infancy respond with reduced corticosterone secretion in response to stressors and with less behavior inhibition in novel environments, we assumed that neonatal stimulation could affect pituitary morphology during this critical period of cell differentiation. Three groups of animals were studied: intact (no manipulation, N = 5), handled (N = 5) and stimulated (submitted to 3 different aversive stimuli, N = 5). The percentage of ACTH-immunoreactive cells in the anterior lobe of the pituitary (number of ACTH-stained cells divided by total number of cells) was determined by examining three slices per pituitary in which a minimum of 200 cells were counted by two independent researchers. Although animals during the neonatal period are less reactive to stress-like stimulation in terms of ACTH and corticosterone secretion, results showed that the relative number of ACTH-stained cells of neonatal handled (0.25 ± 0.01) and aversive stimulated (0.29 ± 0.03) rats was not significantly different from intact (0.30 ± 0.03) animals. Neonatal stimulation may have a differential effect on the various subpopulations of corticotroph cells in the anterior pituitary

A priori estimation of accuracy and of the number of wells to be employed in limiting dilution assays

The use of limiting dilution assay (LDA) for assessing the frequency of responders in a cell population is a method extensively used by immunologists. A series of studies addressing the statistical method of choice in an LDA have been published. However, none of these studies has addressed the point of how many wells should be employed in a given assay. The objective of this study was to demonstrate how a researcher can predict the number of wells that should be employed in order to obtain results with a given accuracy, and, therefore, to help in choosing a better experimental design to fulfill one's expectations. We present the rationale underlying the expected relative error computation based on simple binomial distributions. A series of simulated in machina experiments were performed to test the validity of the a priori computation of expected errors, thus confirming the predictions. The step-by-step procedure of the relative error estimation is given. We also discuss the constraints under which an LDA must be performed.