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Molecular cloning of a cysteine proteinase cDNA from the cotton boll weevil Anthonomus grandis (Coleoptera: Curculionidae).

The cotton boll weevil (Anthonomus grandis) causes severe cotton crop losses in North and South America. This report describes the presence of cysteine proteinase activity in the cotton boll weevil. Cysteine proteinase inhibitors from different sources were assayed against total A. grandis proteinases but, unexpectedly, no inhibitor tested was particularly effective. In order to screen for active inhibitors against the boll weevil, a cysteine proteinase cDNA (Agcys1) was isolated from A. grandis larvae using degenerate primers and rapid amplification of cDNA ends (RACE) techniques. Sequence analysis showed significant homologies with other insect cysteine proteinases. Northern blot analysis indicated that the mRNA encoding the proteinase was transcribed mainly in the gut of larvae. No mRNA was detected in neonatal larvae, pupae, or in the gut of the adult insect, suggesting that Agcys1 is an important cysteine proteinase for larvae digestion. The isolated gene will facilitate the search for highly active inhibitors towards boll weevil larvae that may provide a new opportunity to control this important insect pest.

Animals↗

Evaluation of mechanical prototype machines for continuous separation of respirable cotton dust fractions.

The original cotton particulates analyzer used stationary screens to separate aerodynamically the coarse and fine dust fractions from small samples of cotton. By replacing stationary with rotary screens that were continuously cleaned, larger supplies of respirable dust were obtained without interruption. A ginned and a waste cotton were used to test two methods of separation: a rotating 38-micron screen and a tandem rotating 710- or 38- micron screen and cyclone. The fine dust fractions from both systems were captured on filters and examined gravimetrically, by Coulter counter, and scanning electron microscope. The dust passing 38 micron stationary or rotary screens contained particles of 15 micron maximum diameter whereas dust from the 710-gmm rotary screen and tandem cyclone exhibited particles of 10 micron maximum diameter and lint fragments. Dust fractions with particles less than 10 micron diameter and free of lint were obtained with a 38-micron rotary screen and tandem cyclone.

Air Pollutants, Occupational↗

Exposure of cotton workers in an experimental cardroom with reference to airborne endotoxins.

Workers from cotton mills were exposed to cotton dust during carding in an experimental cardroom. Cotton from different geographical locations with varying amounts of endotoxin were used. Exposure levels ranged from 0.6 to 3.6 mg dust/m3 (from a vertical elutriator) and from 0.1 to 8.0 micrograms/m3 of endotoxin. No relationship was found between the decrease in FEV1 over the workshift and the amount of airborne dust. Airborne endotoxin correlated with the decrease in FEV1 and the increase in blood neutrophils. The FEV1 decrease was more pronounced among smokers. The data suggest that the amount of airborne endotoxin determines the risk for development of the acute symptoms in the byssinosis syndrome.

Air Pollutants, Occupational↗

Pulmonary function among cotton textile workers. A study of variability in symptom reporting, across-shift drop in FEV1, and longitudinal change.

Longitudinal variability in respiratory responses, including symptom reporting and across-shift change in ventilatory function, were examined in relation to long-term loss of ventilatory function in a group of 447 cotton textile workers in Shanghai, China. The study used a standardized respiratory questionnaire and standardized spirometric testing before and after a work shift on the first day of the workweek. Prediction equations for FEV1 were generated from a group of silk textile workers from the same city. Environmental samples included both vertical elutriated cotton dust and endotoxin levels. There was considerable variability in symptom reporting between the baseline and 5-year follow-up survey for all symptoms. However, subjects who consistently reported symptoms had a significantly accelerated 5-year loss in FEV1 compared with those who never reported symptoms. Subjects with symptoms of chest tightness or dyspnea at one survey lost FEV1 at a rate intermediate between the never or both groups. Moreover, subjects with an across-shift change in FEV1 of more than 5 percent at both surveys had the greatest loss in FEV1 over 5 years (-267 ml) when compared with one-time responders (-224 ml), and nonresponders (-180 ml), though the differences were not significant. Workers with chest tightness and chronic bronchitis in both surveys were overrepresented in the high dust and endotoxin areas. Our results indicate that even with substantial survey-to-survey variability in responses, there is important information contained in both questionnaires and across-shift spirometry. Among cotton workers, consistent responders to either symptom questionnaire or across-shift FEV1 decrements of > or = 5 percent appear to be at increased risk for lung function impairment.

Adult↗

A longitudinal study of pulmonary function in cotton gin workers in the San Joaquin Valley.

In a previous study we found no significant effects of acute exposure to the cotton gin environment on FEV1 in San Joaquin Valley gin workers compared with concurrent control subjects, nor was there an excess of obstructive lung disease among the same gin workers. The present study looked at the mean annual decline in FEV1, FEV1/FVC, and FEF25-75% in San Joaquin Valley cotton gin workers vs concurrent control subjects over four years. The FEV1 of gin workers completing all four years of the study declined 27.3 ml vs 35.5 ml in control subjects (p = 0.37); FEV1/FVC declined 0.21 percent in gin workers and 0.23 percent in control subjects (p = 0.86); FEF25-75% declined 0.058 L/s in gin workers and 0.066 L/s in control subjects (p = 0.68). Also, the mean annual rate of change of FEV1 was less among the gin workers who did not complete all four years than the gin workers completing all four years. The presence of "byssinosis" symptoms did not significantly influence the results. This prospective, longitudinal study failed to find any detrimental effect of the cotton gin environment on the rate of decline of FEV1, FEV1/FVC, and FEF25-75%. This result was independent of "byssinosis" symptoms and the effect of "dropouts."

Adult↗

Cotton (Gossypium hirsutum L.).

Considering the economic importance of cotton in many developing and developed countries, there is an urgent need to accelerate the application of biotechnological tools to address the problems associated with the production of this crop and to improve the quality of fiber and seed. This requires a simple yet robust gene delivery/transformant recovery system. A protocol for the production of transgenic cotton plants was refined in our laboratory. It involves Agrobacterium-mediated transformation of cotton cells, selection of stable transgenic callus lines, and recovery of plants via somatic embryogenesis. A detailed description of the protocol is provided in this chapter.

Agrobacterium tumefaciens↗

Trichogramma exiguum (Hymenoptera: Trichogrammatidae) releases in North Carolina cotton: evaluation of heliothine pest suppression.

Field studies were conducted in 1996 and 1997 to reevaluate the use of augmentative releases of Trichogramma wasps for heliothine management in cotton. In 1996, nine releases of Trichogramma exiguum Pinto & Platner, spaced 3-4 d apart, were made into three 0.4-ha cotton plots. Six weekly releases were made in 1997, each containing two T. exiguum cohorts developmentally staggered by 45 degrees C degree-days. Field release rates, estimated from laboratory and field quality control data, averaged 108,357 T. exiguum female female per hectare per cohort per release in 1996 and 193,366 female female per hectare per cohort per release in 1997. In 1996, mean +/- SD adult emergence under laboratory conditions for released cohorts was 92 +/- 7%; 62 +/- 5% of emerged adults were females, 3 +/- 2% of females displayed brachyptery (nonfunctional wings), mean female longevity under laboratory conditions was 15 +/- 4 d, and mean +/- SD field emergence was 97 +/- 2%. Quality control measurements were similar in 1997. In 1996, mean +/- SD percent parasitism of heliothine eggs in field plots on the sampled dates ranged from 67 +/- 4 to 83 +/- 5% in T. exiguum release plots and 25 +/- 9 to 55 +/- 8% in control plots. In 1997, parasitism levels ranged from 74 +/- 4 to 89 +/- 5% in T. exiguum release plots and 18 +/- 18 to 69 +/- 11% in control plots. Despite increased parasitism levels in T. exiguum release plots, there were no significant differences in density of fifth instars, boll damage, or yield between T. exiguum release and control plots. Therefore, it is concluded that Trichogramma augmentation is not an effective heliothine management tool in North Carolina cotton.

Animals↗

Plant-toxin interactions in transgenic Bt cotton and their effect on mortality of Helicoverpa armigera (Lepidoptera: Noctuidae).

In Australia, transgenic cotton plants expressing the cry1Ac gene from Bacillus thuringiensis Berliner variety kurstaki are less toxic to first-instar Helicoverpa armigera (Hübner) after the plant is producing fruit. We developed two bioassay methods (leaf mush, leaf disk) to test if the physiological state of the plants explained changes in toxicity and a third method (diet incorporation) was developed to quantify the toxicity of Bt leaves when mixed in chickpea diet. Cry1Ac protein was less toxic to H. armigera larvae when the protein was mixed with leaves from fruiting versus presquare conventional cotton. Differences in LC50 varied from 2.4- to 726-fold, depending on the source of toxin and conventional plant material. These results suggest that plant-toxin interactions in fruiting cotton are reducing the toxicity of the Cry1Ac protein. The possible role of tannins in these changes is discussed.

Animals↗

Large-scale management of insect resistance to transgenic cotton in Arizona: can transgenic insecticidal crops be sustained?

A major challenge for agriculture is management of insect resistance to toxins from Bacillus thuringiensis (Bt) produced by transgenic crops. Here we describe how a large-scale program is being developed in Arizona for management of resistance to Bt cotton in the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), and other insect pests of cotton. Financial support from growers makes this program possible. Collaboration between the Arizona Cotton Research and Protection Council, the University of Arizona, and government agencies has led to development of resistance management guidelines, a remedial action plan, and tools for monitoring compliance with the proposed guidelines. Direct participation in development of resistance management policies is a strong incentive for growers to invest in resistance management research. However, more research, regularly updated regulations, and increased collaboration between stakeholders are urgently needed to maintain efficacy of Bt toxins in transgenic crops.

Animals↗

Impact of Bt cottons expressing one or two insecticidal proteins of Bacillus thuringiensis Berliner on growth and survival of noctuid (Lepidoptera) larvae.

A series of laboratory assays were performed to compare the relative impact of commercial and experimental cultivars of cotton, Gossypium hirsutum (L.), expressing zero, one, or two insecticidal proteins of Bacillus thuringiensis Berliner, on several lepidopteran pests. Assays in which larvae were fed fresh plant tissue indicated that dual-toxin B. thuringiensis (Bt) cultivars, expressing both Cry1Ac and Cry2Ab endotoxins of B. thuringiensis, were more toxic to bollworms, Helicoverpa zea (Boddie), fall armyworms, Spodoptera frugiperda (J. E. Smith), and beet armyworms, Spodoptera exigua (Hubner), than single-toxin cultivars expressing Cry1Ac. Assays in which lyophilized plant tissue was incorporated into artificial diet also indicated improved activity of the dual-toxin Bt cultivar compared with single-toxin plants. Both bollworm and tobacco budworm, Heliothis virescens (F.), growth was reduced by Bt cotton, particularly the dual-toxin cultivar. Although assays with lyophilized tissues were done using largely sublethal doses, bollworm survival was reduced by the dual-toxin cultivar. It appears that this newly developed Bt cotton expressing two toxins will be more effective and have a wider range of activity on these lepidopteran pests.

Animals↗

Field efficacy and seasonal expression profiles for terminal leaves of single and double Bacillus thuringiensis toxin cotton genotypes.

Examination of commercial Cry1Ac transgenic Bacillus thuringiensis Berliner (Bt) cotton varieties (Bollgard, Monsanto, St. Louis, MO) and an experimental Cry1Ac + Cry2Ab transgenic Bt cotton variety (Bollgard II, Monsanto) for lepidopteran field efficacy was conducted during the 2000 growing season. In addition, a commercially available (Envirologix, Portland, ME) quantification assay (ELISA) was used to measure and profile the expression levels of Cry proteins in two of these varieties ['DP50B, Bollgard'; 'DP50BII, Bollgard II' (Delta & Pine Land, Scott, MS)]. Populations of beet army worms, Spodoptera exigua (Hübner), and soybean loopers, Pseudoplusia includens (Walker), were significantly lower (P < 0.05) in Bollgard II plots compared with Bollgard. Population numbers for fall army worms, Spodoptera frugiperda (J. E. Smith), and salt marsh caterpillars, Estigmene acrea (Drury), were lower in Bollgard II plots compared with Bollgard but means did not differ significantly (P > 0.05). Single and dual-toxin genotypes remained superior (P < 0.05) compared with conventional cotton against the tobacco budworm, Heliothis virescens (F.). The addition of Cry2Ab had no significant (P > 0.05) impact on Cry1Ac expression in Bollgard II compared with Cry1Ac expression in Bollgard. Furthermore, throughout the season Cry2Ab was present at much higher levels in the plant compared with Cry1Ac for Bollgard II plants. Possible species-specific reasons for increased efficacy of Bollgard II over Bollgard are discussed.

Animals↗

Effects of planting dates on boll weevils (Coleoptera: Curculionidae) and cotton fruit in the subtropics.

The effects of planting dates 2-3-wk apart on boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), field-level populations, and feeding and oviposition damage to cotton, Gossypium hirsutum L., squares and bolls, were studied during 2002 and 2003 in the Lower Rio Grande Valley of Texas. Squares were 44-56% more abundant in some later planted treatments than in the earlier planted treatments, but mean cumulative numbers of oviposition- and feeding-damaged squares were 2.7 - 4.8-fold greater in some later planted treatments than in earlier treatments. Increased square production in later planted cotton was offset by boll weevil infestations that occurred when squares are most vulnerable and contribute most toward the pest's reproduction. Early planting avoided boll weevil population buildups in the field when large squares were abundant. Lint yields in 2002 did not differ significantly between the planting date treatments, but in 2003, mean yield in the middle treatment was 23% greater than in the early and late-planted treatments. Insecticide sprays in the earliest planted treatment of each year, based on the 10% damaged squares threshold, were >33% and >43% fewer than in the corresponding middle and latest planting treatments, respectively. Delayed planting, relative to the onset of favorable cotton-growing weather, at the field level, even when not applied uniformly on an areawide scale, is more cost-effective than planting too early or too late.

Agriculture↗

Selecting for efficacy of Bollgard cotton cultivars against various Lepidoptera using forward breeding techniques.

Studies during the past 5 yr have shown that the overall level of protein (Cry1Ac) produced from the cry1Ac transgene (Monsanto Co., St. Louis, MO) differ among commercial Bollgard cotton, Gossypium hirsutum L., cultivars. These differences between cultivars are under genetic control and have been correlated with efficacy of certain lepidopteran pests. Previous studies have shown that the parental background (i.e., non-Cry1Ac conventional cultivar) has a significant influence on the amount of Cry1Ac protein in Bollgard cultivars. Unlike the backcross technique commonly used to acquire commercial Bollgard cultivars, we used forward breeding to obtain cultivars of Bollgard cotton that were selected for various levels of Cry1Ac. These differences in the amount of Cry1Ac were correlated with growth and survival of two lepidopteran pests of cotton. Implications for effective resistance management as well as relative ease of this procedure are discussed.

Animals↗

Thermal comfort properties of cotton and nonwoven surgical gowns with dual functional finish.

The purpose of this study was to evaluate thermal comfort properties of surgical gowns made of dual functional finish cotton and nonwoven fabrics which have barrier properties of blood and micro-organism. Four types of surgical gowns, which were made of nonwoven fabrics with finish or without and were made of cotton with finish or without, were tested. The thermal insulations of four surgical gowns were measured with thermal manikin. Subjective experiments on thermal comfort, skin temperature and clothing microclimate were conducted. Six male subjects, between 26 and 28 years age old, participated in the wear trials tests. Typical activities for surgeons in the operation theater were simulated during the experimental sessions. Air temperature in a climate chamber was kept at 22 degrees C and its humidity was 60% RH. Air velocity was controlled at less than 0.15 m/s. Inner radiant temperature was almost equal to the air temperature. Basic thermal insulation of the dual functional finished nonwoven surgical gown was 0.87 clo, which was slightly higher than that of untreated (0.84 clo). However, the skin temperature of the subject wearing a dual functional finished surgical gown was significantly lower at P < .05. When the subject wears the dual functional finished gown, the amount of sweating was less than that when wearing untreated. Microclimate temperature and humidity of dual functional finished surgical gown were lower than untreated and it was statistically significant. There was no significant difference in subjective humid and overall comfort sensation between finished and untreated ones. Thermal sensation of dual functional finished one was significantly different from untreated one only during the first exercise. The results of this study indicate that the dual functional finish surgical gown allowed heat to be transferred from the skin of subject to the atmosphere better than untreated. The nonwoven surgical gown showed no difference in comfort properties from cotton one.

Adult↗

Simulating endosulfan transport in runoff from cotton fields in Australia with the GLEAMS model.

Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9methano-2,4,3-benzodioxathiepin 3-oxide), a pesticide that is highly toxic to aquatic organisms, is widely used in the cotton (Gossypium hirsutum L.) industry in Australia and is a risk to the downstream riverine environment. We used the GLEAMS model to evaluate the effectiveness of a range of management scenarios aimed at minimizing endosulfan transport in runoff at the field scale. The field management scenarios simulated were (i) Conventional, bare soil at the beginning of the cotton season and seven irrigations per season; (ii) Improved Irrigation, irrigation amounts reduced and frequency increased to reduce runoff from excess irrigation; (iii) Dryland, no irrigation; (iv) Stubble Retained, increased soil cover created by retaining residue from the previous crop or a specially planted winter cover crop; and (v) Reduced Sprays, a fewer number of sprays. Stubble Retained was the most effective scenario for minimizing endosulfan transport because infiltration was increased and erosion reduced, and the stubble intercepted and neutralized a proportion of the applied endosulfan. Reducing excess irrigation reduced annual export rates by 80 to 90%, but transport in larger storm events was still high. Reducing the number of pesticide applications only reduced transport when three or fewer sprays were applied. We conclude that endosulfan transport from cotton farms can be minimized with a combination of field management practices that reduce excess irrigation and concentration of pesticide on the soil at any point in time; however, discharges, probably with endosulfan concentrations exceeding guideline values, will still occur in storm events.

Agriculture↗

Narrow-waveband reflectance ratios for remote estimation of nitrogen status in cotton.

Tailoring nitrogen (N) fertilizer applications to cotton (Gossypium hirsutum L.) in response to leaf N status may optimize N use efficiency and reduce off-site effects of excessive fertilizer use. This study compared leaf and canopy reflectance within the 350 to 950 nm range in order to identify reflectance ratios sensitive to leaf chlorophyll (Chl), and hence N status, in cotton. Plants were grown outdoors in large pots using half-strength Hoagland's (control) solution until some three-row plots received a restricted supply of N. Treatments comprised control, 20% of control N at first flower bud (square) onward; 0 and 20% of control N at first flower onward; and 0% of control N at fruit-filling onward. Despite leaf N values ranging from 51 to 19 g kg-1 across treatments and sampling dates, a weak correlation was obtained between Chl and N (r2 = 0.32, df = 70). In general, N stress led to increased reflectance at 695 +/- 2.5 nm (R695) and decreased reflectance at R410, and changes in leaf N were best correlated with either R695 or R755 in leaves and either R410 or R700 in canopies. The strongest associations between leaf constituent and canopy reflectance ratio were Chl vs. R415/R695 (r2 = 0.72), carotenoids vs. R415/R685 (r2 = 0.79), and N vs. R415/R710 (r2 = 0.70). The R415 measure appears to be a more stable spectral feature under N stress, as compared with more pronounced changes along the reflectance red edge (690-730 nm). Multiple regression identified a three-waveband canopy reflectance model that explained 80% of the variability in leaf N. Results indicate that remote sensing of N status in cotton is feasible using narrow-waveband reflectance ratios that involve the violet or blue region of the spectrum (400 to 450 nm) and the more commonly featured red-edge region.

Chlorophyll↗

The cotton rat as an experimental model of human parainfluenza virus type 3 disease.

Intranasal or aerosol inoculation of cotton rats (Sigmodon hispidus) with human parainfluenza virus type 3 (P3) produces pulmonary changes which closely resemble human disease due to this agent. Active viral replication resulted in peak titers by day 2 of infection. Viral antigen was localized by immunofluorescence in bronchial epithelial cells alone (aerosol inoculation) or epithelial and alveolar cells (intranasal instillation). Cytopathic effects in epithelial cells was noted as early as day 2, and marked cellular proliferation occurred by day 7. Intranasal instillation of virus resulted in development of patchy interstitial pneumonitis. Mononuclear cell infiltration in the peribronchial and perivascular areas peaked on days 6 and 7. A brisk systemic antibody response was measured both by complement fixation and virus neutralization techniques. These data establish the value of the cotton rat as an experimental model for the study of bronchiolitis and pneumonia due to P3. Since previous work has demonstrated that respiratory syncytial virus and Mycoplasma pneumoniae also will replicate and cause pulmonary disease in cotton rats, comparative studies in a single animal species are facilitated.

Animals↗

Use of anti-TNF-alpha antiserum to investigate toxic alveolitis arising from cotton dust exposure.

Cotton dust has been associated with byssinosis and toxic alveolitis. A murine animal model has been developed with which to investigate the pathogenesis of these disorders. Studies with the model have reproduced the neutrophilic inflammation characteristic of the alveolitis, and have shown the presence of tumor necrosis factor-alpha (TNF-alpha) in the bronchoalveolar lavage (BAL) fluid. The current study investigated the role of TNF-alpha in the inflammatory response by use of a polyclonal antiserum to recombinant murine TNF-alpha. Following a 4-h exposure to cotton dust, experimental animals showed a 40-fold increase in BAL cells with 92% neutrophils. There was a 24-fold increase in TNF-alpha in the BAL fluid. Up regulation of TNF-alpha mRNA expression was detected in BAL cells. Mice pretreated with anti-TNA-alpha antiserum displayed a marked attenuation of the neutrophilic inflammation; however, the level of TNF-alpha mRNA expression was not reduced in these mice. These studies support a major role of TNF-alpha in the toxic alveolitis induced by cotton dust inhalation.

Alveolitis, Extrinsic Allergic↗