Differential responses of nematode communities to soybean genotypes resistant and susceptible to Heterodera glycines race 3

Heterodera glycines and Helicotylenchus dihystera were the two most abundant plant-parasitic nematodes found in two H. glycines race 3-infested fields, Chapadão do Céu, MS and Campo Alegre, MG. These fields had been planted with resistant (R) and susceptible (S) plants to cyst nematodes. In the first field, soybean (Glycine max) FT-Cristalina (S) was susceptible to H. glycines but resistant to H. dihystera, while GOBR93 122243 (R) was just the opposite. In the second field, M-Soy 8400 (R) was more resistant to the spiral nematode than M-Soy8411 (S), but the resistance to the cyst nematode was not different between the two genotypes. The total abundance of nematodes was not different between the susceptible and resistant plants in the two fields, suggesting that H. dihystera and/or bacterial feeders and other trophic groups replaced the reduced abundance of the cyst nematodes in resistant plants. Bacterial feeders acted as a compensatory factor to plant-parasitic nematodes in ecological function. The populations of fungal feeders were higher in GOBR93 122243 (R) than in susceptible FT-Cristalina (S) in Chapadão do Céu, but lower in M-Soy 8400 (R) than in M-Soy 8411 (S) in Campo Alegre. This is being attributed to the different periods of soil samplings that were made at the florescent period in the first field, and at the final growing cycle in the second field. Only four nematodes, H. glycines, H. dihystera, Acrobeles sp. and Panagrolaimus sp. dominated the nematode resistant community GOBR93 122243 (R) in Chapadão do Céu, but dominance was shared by ten genera in Campo Alegre, which explains why the five diversity indexes (S, d, Ds, H' and T) were higher in resistant plants than in susceptible plants in field two.

Management of crotalaria and pigeon pea for control of yam nematode diseases

Management of plant-parasitic nematodes with the use of nematicides has not been recommended for small farmers that grow yam in the Northeastern region of Brazil, due to its high cost and residue toxicity. The use of plants with antagonistic effect to nematodes and green manure which improves soil chemical, physical and biological characteristics can be a viable and low cost alternative to control parasitic nematodes. This work aimed to evaluate the effect of crotalaria (Crotalaria juncea) and pigeon pea (Cajanus cajan) plants on the control of yam nematodes. Three experiments were carried out. The first was conducted under in vitro conditions to evaluate the nematostatic and nematicide effect of extracts from fresh and dry matter of the above ground parts of crotalaria, pigeon pea, and the combination of both. The second experiment was carried out under greenhouse conditions to evaluate the effect of soil amendment with crotalaria, pigeon pea, and the combination of both in the infectivity of Scutellonema bradys, using tomato plants as the host plant. The third experiment was conducted under field conditions to evaluate the effect of crotalaria, pigeon pea, and the combination of both, cultivated between yam planting rows and incorporated to soil surface, on yam nematodes. The aqueous extract obtained form fresh matter of crotalaria had a nematicide effect of 100% for S. bradys. Extracts from dry matter of both crotalaria and pigeon pea did not have any nematicide effect, but had a nematostatic effect. Incorporation of crotalaria to soil inhibited infectivity of S. bradys in tomato seedlings. These results showed that planting crotalaria alone or in combination with pigeon pea, between the yam planting rows, is an efficient method for controlling S. bradys and Rotylenchulus reniformis associated with yams. Crotalaria can be used for controlling these plant-parasitic nematodes in soil.

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.

Nematicidal effect of volatile organic compounds (VOCs) on the plant-parasitic nematode Meloidogyne javanica

Previous studies have demonstrated that volatile organic compounds (VOCs), produced by the yeast Saccharomyces cerevisiae, were able to inhibit the development of phytopathogenic fungi. In this context, the nematicidal potential of the synthetic mixture of VOCs, constituted of alcohols and esters, was evaluated for the control of the root-knot nematode Meloidogyne javanica, which causes losses to crops of high economic value. The fumigation of substrate containing second-stage juveniles with VOCs exhibited nematicidal effect higher than 30% for the lowest concentration tested (33.3 µL g-1 substrate), whereas at 66.6 and 133.3 µL g-1 substrate, the nematode mortality was 100%. The present results stimulate other studies on VOCs for nematode management.

Comparison of Two- and Three-dimensional Ultrasonography in the Evaluation of Lesion Level in Fetuses with Spina Bifida

Purpose To evaluate the precision of both two- and three-dimensional ultrasonography in determining vertebral lesion level (the first open vertebra) in patients with spina bifida. Methods This was a prospective longitudinal study comprising of fetuses with open spina bifida who were treated in the fetal medicine division of the department of obstetrics of Hospital das Clínicas of the Universidade de São Paulo between 2004 and 2013. Vertebral lesion level was established by using both two- and three-dimensional ultrasonography in 50 fetuses (two examiners in each method). The lesion level in the neonatal period was established by radiological assessment of the spine. All pregnancies were followed in our hospital prenatally, and delivery was scheduled to allow immediate postnatal surgical correction. Results Two-dimensional sonography precisely estimated the spina bifida level in 53% of the cases. The estimate error was within one vertebra in 80% of the cases, in up to two vertebrae in 89%, and in up to three vertebrae in 100%, showing a good interobserver agreement. Three-dimensional ultrasonography precisely estimated the lesion level in 50% of the cases. The estimate error was within one vertebra in 82% of the cases, in up to two vertebrae in 90%, and in up to three vertebrae in 100%, also showing good interobserver agreement. Whenever an estimate error was observed, both two- and three-dimensional ultrasonography scans tended to underestimate the true lesion level (55.3% and 62% of the cases, respectively). Conclusions No relevant difference in diagnostic performance was observed between the two- and three-dimensional ultrasonography. The use of three-dimensional ultrasonography showed no additional benefit in diagnosing the lesion level in the fetuses with spina bifida. Errors in both methods showed a tendency to underestimate lesion level.

High Risk HPV E6/E7 Oncoprotein Expression in Women with High Grade Squamous Intraepithelial Lesion

Purpose To correlate the expression of high-risk HPV E6 mRNA with pap smear, colposcopy, and biopsy results in women with high grade squamous intraepithelial lesion (HSIL). Methods A cross-sectional study was performed on women referred for primary care services after cytological diagnosis of HSIL. We evaluated the expression of E6/E7 mRNA of HPV types 16,18,31,33, and 45 and correlated the results with those of Pap smear, colposcopy, and biopsy. For amplification/detection of mRNA E6 / E7 we used NucliSENSEasyQ kit to detect HPV mRNA by polymerase chain reaction with primers/ probes for HPV types 16, 18, 31, 33, and 45. Results Out of 128 valid tests, the results of 30 (23.4%) tests were negative and 98 (70%) tests were positive. Only one type of HPV was detected in 87.7% of the E6/E7 mRNA positive cases. HPV16 was detected in 61.2% of the cases, followed by HPV33 (26.5%), HPV31 (17.3%), HPV18 (10%), and HPV45 (4.08%). Pap smear tests revealed that the E6/E7 test was positive in 107 (83.8%) women with atypical squamous cells - high grade (ASC-H), HSIL, or higher. The E6/E7 test was positive in 69 (57.5%) specimens presenting negative cytology results. When analyzing the association with colposcopy results, the frequency of positive E6/E7 results increased with the severity of the injury, ranging from 57.1% in women without colposcopy-detected injury to 86.5% in those with higher levels of colposcopy findings. Of the 111 women who underwent biopsy and E6/E7 testing, the E6/E7 test was positive in 84.7% of the women who presented with lesions of cervical intraepithelial neoplasia (CIN) grade 2 or higher. Finally, 41.2% of women with a negative biopsy presented a positive E6/E7 test. Conclusions E6/E7mRNA expression was higher in women with HSIL and CIN grade 2 or higher.

Fibrinonecrotic enteritis of piglets in a commercial farm: a postmortem study of the prevalence and the role of lesion associated agents Isospora suis and Clostridium perfringens

The objectives were to determine the prevalence of fibrinonecrotic enteritis (FNE) on a farrow-to-finish farm of 1,000 sows, to categorize the pathological changes, and to to investigate the lesion associated agents Isospora suis and Clostridium perfringens. Causes of preweaning mortality (PWM) were classified into 8 categories including FNE. Obtained data were evaluated for statistical significance by adjusted Chi-square analysis. Samples of FNE were taken for complementary studies including a PCR technique for genotyping toxin genes of Clostridium perfringens from gut samples fixed in 10% neutral formalin. From 3,153 piglets examined, less than 1% was classified as FNE. FNE prevalence increased progressively from the first to the third week, the last differing statistically from the others. Eighty percent of gut samples with FNE lesions were positive to Isospora suis, when examined by PCR from 9 severe FNE lesions detected 7 positive samples only for a toxin gene, characteristic of C. perfringens type-A.

Experimental osteoarthritis in rabbits: lesion progression

The aim of this study was to evaluate the progression of lesions in different stages of osteoarthritis (OA) experimental by radiography (RX), computed tomography (CT), macroscopic and histopathology, linking these different diagnostic methods, helped to provide information that helps the best time for the therapeutic approach. Four experimental periods were delineated at 3, 6, 9 and 12 weeks after induction of OA, known as PI, PII, PIII and PIV, respectively, each with six animals. We evaluated the five compartments of the femorotibial joint: medial femoral condyle (MFC), lateral femoral condyle (LFC), medial tibial plateau (MTP), lateral tibial plateau (LTP) and femoral trochlea (FT). Therefore we established an index by compartment (IC) and by adding such an index was estimated joint femorotibial (IFT). It was observed that the CFM was the compartment with the highest IC also differed significantly (p<0.05) from other compartments. Compartments showed no significant difference (p>0.05) between the PI and PII, however contrary fact occurred between the PII and PIII (p<0.05), PIII and PIV (p<0.01) and between PI and PIV (p<0.001). Similarly the IFT, showed a significant difference in the animals of PIV compared to PI (p<0.001), PII (p<0.001) and PIII (p<0.01), and there was no statistical difference (p> 0.05) between the PI and PII. In the variation of the average interval between periods, there was a higher value between the PIII PIV and for the other intervals of time periods (PI, PII, and PIII-PII). However, these intervals showed no statistically significant difference (p>0.05). Through the RX, CT, macroscopic and histopathological findings, we found similar patterns among individuals within the same period demonstrating a gradual progression of the disease. These results show that between 3 and 6 weeks progression of the lesion is slower and probably also can be reversed in comparison to other ranges where proved further progression between 9 and 12 weeks after induction of trauma OA. These results may provide a better therapeutic approach aimed at reversing the lesions in early stages of OA. We conclude that the interconnection of the four diagnostic methods individually classified into scores, which were unified in both indices in the evaluation by the femorotibial joint compartment and may represent a diagnostic condition closer to the true condition of the injury and its progression.

Variability in the fecal egg count and the parasitic burden of hair sheep after grazing in nematode infected paddocks

This study aimed to evaluate the variability in the fecal egg count (FEC) and the parasitic burden of naive hair sheep after grazing in nematode infected paddocks. The research was carried out in Tabasco, Mexico, during two periods (August and December). In each period 32 lambs were grazed for one month on African star grass (Cynodon plectostachyus) contaminated with gastrointestinal parasitic nematodes. FEC, packed cell volume (PCV) and body weight (BW) were recorded. Gastrointestinal worms were recovered at necropsy. Data were analyzed with the MIXED procedure of SAS using a model of repeated measurements over time. A higher number of Haemonchus contortus worms was found in December (2814±838) than in August (1166±305). The opposite occurred with Cooperia curticei (2167±393 and 3638±441, respectively). The FEC and correlation coefficient in respect to the worm burden were higher in December (6516 ± 1599, r=0.83, respectively) than in August (4364±771, r=0.44, respectively). A high variability in resistance-susceptibility to gastrointestinal nematodes (GIN) occurred in Katahdin × Pelibuey lambs after grazing.

Host suitability of weeds and forage species to root-knot nematode meloidogyne graminicola as a funcion of irrigation management

The irrigated rice production can be limited by various phytopathogenic agents, including root-knot nematodes (Meloidogyne spp.). Thus, the aim of this research was to check the host suitability of plant species most often found off-season and during rice cultivation, to root-knot nematode Meloidogyne graminicola, under two irrigation managements. Two experiments were conducted in a completely randomized design. In the first experiment seven plant species that occur in an area of rice cultivation, in fallow, off-season were evaluated. For the second experiment nine weed species infesting the irrigated rice culture were tested in rainfed and flooding conditions. The sixteen species, kept individually in pots with sterilized substrate, were inoculated with 5,000 eggs and second stage juveniles (J2) of nematode. BRS 410 IRGA rice plants inoculated with M.graminicola were used as control. Two months after inoculation, the root system of each plant was evaluated for number of galls and nematode reproduction factor. It was verified that the species of off-season of rice cultivation Sida rhombifolia, Raphanus raphanistrum, Spergula arvensis, Lotus corniculatus and Trifolium repens, and, during the cycle of rice cultivation, Aeschynomene denticulata, Leersia hexandra, are immune to nematode. The plant species off-season, Avena strigosa and Lolium multiflorum and of cultivation, Alternanthera philoxeroides, red rice, Echinochloa crusgalli, Cyperus difformis, Cyperus esculentus, Cyperus iria and Fimbristylis miliacea would behave as hosts of M.graminicola, mostly under rainfed conditions.

Faster gastric emptying of a liquid meal in rats after hypothalamic dorsomedial nucleus lesion

The effects of dorsomedial hypothalamic (DMH) nucleus lesion on body weight, plasma glucose levels, and the gastric emptying of a liquid meal were investigated in male Wistar rats (170-250 g). DMH lesions were produced stereotaxically by delivering a 2.0-mA current for 20 s through nichrome electrodes (0.3-mm tip exposure). In a second set of experiments, the DMH and the ventromedial hypothalamic (VMH) nucleus were lesioned with a 1.0-mA current for 10 s (0.1-mm tip exposure). The medial hypothalamus (MH) was also lesioned separately using a nichrome electrode (0.3-mm tip exposure) with a 2.0-mA current for 20 s. Gastric emptying was measured following the orogastric infusion of a liquid test meal consisting of physiological saline (0.9% NaCl, w/v) plus phenol red dye (6 mg/dl) as a marker. Plasma glucose levels were determined after an 18-h fast before the lesion and on the 7th and 15th postoperative day. Body weight was determined before lesioning and before sacrificing the rats. The DMH-lesioned rats showed a significantly faster (P<0.05) gastric emptying (24.7% gastric retention, N = 11) than control (33.0% gastric retention, N = 8) and sham-lesioned (33.5% gastric retention, N = 12) rats, with a transient hypoglycemia on the 7th postoperative day which returned to normal by the 15th postoperative day. In all cases, weight gain was slower among lesioned rats. Additional experiments using a smaller current to induce lesions confirmed that DMH-lesioned rats had a faster gastric emptying (25.1% gastric retention, N = 7) than control (33.4% gastric retention, N = 17) and VMH-lesioned (34.6% gastric retention, N = 7) rats. MH lesions resulted in an even slower gastric emptying (43.7% gastric retention, N = 7) than in the latter two groups. We conclude that although DMH lesions reduce weight gain, they do not produce consistent changes in plasma glucose levels. These lesions also promote faster gastric emptying of an inert liquid meal, thus suggesting a role for the DMH in the regulation of gastric motility

Isoenzyme analysis of Arthrobotrys, a nematode-trapping fungus

Extraction and isoenzyme analysis of four isolates of Arthrobotrys including A. musiformis, A. robusta and A. conoides were conducted. Among the 14 enzymes studied by starch gel electrophoresis, using morpholine-citrate as gel/electrode buffer, the following nine enzymes showed interpretable banding patterns: a-esterase, fumarase, hexokinase, isocitrate dehydrogenase, leucine aminopeptidase, malate dehydrogenase, 6-phosphogluconate dehydrogenase, phosphoglucomutase and phosphoglucoisomerase. All isolates studied displayed typical isoenzyme phenotypes for each species. Two isolates of A. conoides differed in their a-isoesterase banding patterns, but no differences were observed for the other enzymes. The assay was satisfactory for enzyme extraction and resolution of Arthrobotrys and could be used in future taxonomic and genetic studies of this organism

Effects of septal cholinergic lesion on rat exploratory behavior in an open-field

The medial septum participates in the modulation of exploratory behavior triggered by novelty. Also, selective lesions of the cholinergic component of the septohippocampal system alter the habituation of rats to an elevated plus-maze without modifying anxiety indices. We investigated the effects of the intraseptal injection of the cholinergic immunotoxin 192 IgG-saporin (SAP) on the behavior of rats in an open-field. Thirty-nine male Wistar rats (weight: 194-230 g) were divided into three groups, non-injected controls and rats injected with either saline (0.5 µl) or SAP (237.5 ng/0.5 µl). Twelve days after surgery, the animals were placed in a square open-field (120 cm) and allowed to freely explore for 5 min. After the test, the rats were killed by decapitation and the septum, hippocampus and frontal cortex were removed and assayed for acetylcholinesterase activity. SAP increased acetylcholinesterase activity in the septum, hippocampus and frontal cortex and decreased the total distance run (9.15 ± 1.51 m) in comparison to controls (13.49 ± 0.91 m). The time spent in the center and at the periphery was not altered by SAP but the distance run was reduced during the first and second minutes (2.43 ± 0.36 and 1.75 ± 0.34 m) compared to controls (4.18 ± 0.26 and 3.14 ± 0.25 m). SAP-treated rats showed decreased but persistent exploration throughout the session. These results suggest that septohippocampal cholinergic mechanisms contribute to at least two critical processes, one related to the motivation to explore new environments and the other to the acquisition and storage of spatial information (i.e., spatial memory).

Decreased gastric tone and delayed gastric emptying precede neutrophil infiltration and mucosal lesion formation in indomethacin-induced gastric damage in rats

Gastric antral dysmotility has been implicated in the pathogenesis of indomethacin-induced gastric damage, but the relationship between gastric motor abnormalities and mucosal lesions has not been extensively studied. We investigated whether changes in gastric tone and gastric retention correlate with mucosal lesions and neutrophil migration in indomethacin-induced gastric damage in rats. Indomethacin, either 5 or 20 mg/kg (INDO-5 and INDO-20), was instilled into the stomach, and then gastric damage, neutrophil migration, gastric tone and gastric retention were assessed 1 or 3 h later. Gastric damage was calculated as the sum of the lengths of all mucosal lesions, and neutrophil migration was measured by assaying myeloperoxidase activity. Gastric tone was determined by a plethysmometric method, and gastric retention of either saline or Sustacal® was evaluated by a scintigraphic method. Gastric damage was detectable 3 h after either INDO-5 or INDO-20, but not after 1 h. Neutrophil migration was significantly higher 3 h after INDO-20 as compared with INDO-5 or control group, but not after 1 h. Values of gastric tone 1 and 3 h after either INDO-5 (1 h = 1.73 ± 0.07 ml; 3 h = 1.87 ± 0.03 ml) or INDO-20 (1 h = 1.70 ± 0.02 ml; 3 h = 1.79 ± 0.03 ml) were significantly lower than in controls (1 h = 1.48 ± 0.05 ml; 3 h = 1.60 ± 0.06 ml). Gastric retention of saline was higher 1 h after INDO-5 (58.9 ± 3.3%) or INDO-20 (56.1 ± 3.1%) compared to control (45.5 ± 1.7%), but not after 3 h. There were no differences concerning gastric retention of Sustacal® between the various groups. Indomethacin induced decreased gastric tone and delayed gastric emptying, which precede mucosal lesion and neutrophil infiltration. These results indicate that there is no relationship between these gastric motor abnormalities and mucosal lesion in indomethacin-induced gastropathy.

Effect of electrolytic lesion of the dorsal raphe nucleus on water intake and sodium appetite

The present study determined the effect of an electrolytic lesion of the dorsal raphe nucleus (DRN) on water intake and sodium appetite. Male Wistar rats weighing 290-320 g with a lesion of the DRN (L-DRN), performed two days before experiments and confirmed by histology at the end of the experiments, presented increased sensitivity to the dehydration induced by fluid deprivation. The cumulative water intake of L-DRN rats reached 23.3 ± 1.9 ml (a 79% increase, N = 9) while sham-lesioned rats (SL-DRN) did not exceed 13.0 ± 1.0 ml (N = 11, P < 0.0001) after 5 h. The L-DRN rats treated with isoproterenol (300 µg kg-1 ml-1, sc) exhibited an increase in water intake that persisted throughout the experimental period (L-DRN, 15.7 ± 1.47 ml, N = 9 vs SL-DRN, 9.3 ± 1.8 ml, N = 11, P < 0.05). The L-DRN rats also showed an increased spontaneous sodium appetite during the entire period of assessment. The intake of 0.3 M NaCl after 12, 24, 36 and 72 h by the L-DRN rats was always higher than 20.2 ± 4.45 ml (N = 10), while the intake by SL-DRN was always lower than 2.45 ± 0.86 ml (N = 10, P < 0.00001). Sodium- and water-depleted L-DRN rats also exhibited an increased sodium appetite (13.9 ± 2.0 ml, N = 11) compared to SL-DRN (4.6 ± 0.64 ml, N = 11) after 120 min of observation (P < 0.02). The sodium preference of L-DRN rats in both conditions was always higher than that of SL-DRN rats. These results suggest that electrolytic lesion of the DRN overcomes a tonic inhibitory component of sodium appetite.