Chlorothalonil Treatment Research Articles

Adverse outcome pathway-based approach to reveal the mechanisms of skin sensitization and long-term aging effects of chlorothalonil

Chlorothalonil (CHT) is a widely used antifungal agent and is reported to be a sensitizer that can cause allergic contact dermatitis (ACD). ACD initiation is associated with various innate immune cell contributions and is usually accompanied by persistent inflammation, which is a potential contributing factor to skin damage. However, detailed information on the mechanisms by which CHT induces skin sensitization and damage is still insufficient. This study focused on investigating the possible sensitization process and mechanism of CHT and the adverse effects of repeated CHT exposure. CHT activates dendritic cells and promotes the proliferation of lymph cells in the skin sensitization phase, causing severe inflammation. Keratinocytes activate the NLRP3 inflammasome pathway to cause inflammation during CHT treatment, and macrophages also secrete inflammatory cytokines. In addition, CHT-induced inflammation triggered skin wrinkles, decreased epidermal thickness and decreased collagen. Cell experiments also showed that repeated exposure to CHT led to cell proliferation inhibition and senescence, and CHT-induced autophagy dysfunction was not only the reason for inflammation but also for senescence. This study defined the possible process through which CHT is involved in the skin sensitization phase and elucidated the mechanism of CHT-induced inflammation in innate immune responses. We also determined that repeated CHT exposure caused persistent inflammation, ultimately leading to skin aging.

Chlorothalonil drives the antibiotic resistome in earthworm guts

Earthworms are recognized as carriers of pollutants; however, how fungicide residues affect microbiota and antibiotic resistance genes (ARGs) in earthworm guts has remained unclear. In this work, changes in the earthworm gut microbiome and resistome were investigated after chlorothalonil (CTL) application. Earthworm activity accelerated the dissipation of CTL in soil, while metagenomic analysis revealed that CTL altered the ARG profile, leading to an increased abundance of ARGs in earthworm guts, particularly with respect to ARG subtypes CRP and OXA-427. CTL also reduced bacterial diversity and elevated the relative abundance of the phylum Proteobacteria, including a potential ARG host, Aeromonas, which is a known pathogen. Various bacterial genera from the Actinobacteria and Proteobacteria phyla were identified as broad-spectrum hosts for ARGs in earthworm guts. CTL could increase the abundance of multidrug efflux pump genes and enhance the abundance of mobile genetic elements, especially plasmids. Various co-occurrence patterns between plasmids and ARGs were also found after CTL treatments. It is concluded that CTL may act as a selective stress for ARGs and lead to an increase in their abundance by facilitating the proliferation of potential ARG hosts and enhancing plasmid-mediated horizontal transfer frequency of ARGs in earthworm guts.

Influence of Agaricus bisporus establishment and fungicidal treatments on casing soil metataxonomy during mushroom cultivation

The cultivation of edible mushroom is an emerging sector with a potential yet to be discovered. Unlike plants, it is a less developed agriculture where many studies are lacking to optimize the cultivation. In this work we have employed high-throughput techniques by next generation sequencing to screen the microbial structure of casing soil employed in mushroom cultivation (Agaricus bisporus) while sequencing V3-V4 of the 16S rRNA gene for bacteria and the ITS2 region of rRNA for. In addition, the microbiota dynamics and evolution (bacterial and fungal communities) in peat-based casing along the process of incubation of A. bisporus have been studied, while comparing the effect of fungicide treatment (chlorothalonil and metrafenone). Statistically significant changes in populations of bacteria and fungi were observed. Microbial composition differed significantly based on incubation day, changing radically from the original communities in the raw material to a specific microbial composition driven by the A. bisporus mycelium growth. Chlorothalonil treatment seems to delay casing colonization by A. bisporus. Proteobacteria and Bacteroidota appeared as the most dominant bacterial phyla. We observed a great change in the structure of the bacteria populations between day 0 and the following days. Fungi populations changed more gradually, with A. bisporus displacing the rest of the species as the cultivation cycle progresses. A better understanding of the microbial communities in the casing will hopefully allow us to increase the biological efficiency of the crop.

Effects of Chlorothalonil Application on the Physio-Biochemical Properties and Microbial Community of a Yellow–Brown Loam Soil

To gain better knowledge of the effects of residual chlorothalonil on soil characteristics and soil microbial communities, we evaluated the dissipation of chlorothalonil and the effects of different chlorothalonil concentrations on soil respiration, enzyme activities, and microbial community structure in yellow–brown loam soils. Bacterial and fungal soil communities were examined using traditional plate counting and polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE) methods. Soil properties and the results of DGGE band analysis were both used to estimate the status of the soil microbial ecosystem. The results show that residual chlorothalonil has considerable effects on soil respiration, enzymatic activities, and microbial community structure. In particular, soil respiration and phosphatase activities were increased, while saccharase activity, microbial biomass, and microbial community diversity were decreased by increasing levels of chlorothalonil treatment. Correlation analyses revealed that the application of chlorothalonil was significantly correlated with the change of the soil respiration, urease activity, sucrase activity, soil culturable bacteria, and culturable fungi biomass. We conclude that residual chlorothalonil is directly related to soil respiration, enzyme activities, and microbial community structure.

Chlorothalonil alters the gut microbiota and reduces the survival of immature honey bees reared in vitro.

Chlorothalonil is a nonsystemic fungicide, and it is one of the most widely detected pesticides in bee hives. The effect of chlorothalonil on the survival, weight, and gut microbiota of immature Apis mellifera L. reared in vitro was studied. Larvae were fed 1, 2, 4, 8, and 16 μg/mL chlorothalonil and compared with larvae fed the negative control (diet without any additives), positive control (45 mg/L dimethoate), and solvent control (2% acetone). Compared with the control groups, the survival of the 2, 4, 8, and 16 μg/mL chlorothalonil treatments was significantly reduced. The no-observed-adverse-effect concentration of chlorothalonil was 1μg/mL. Chlorothalonil had no significant effect on larval weight. The gut bacterial community composition of newly emerged bees was determined by PacBio 16S rDNA gene sequencing. linear discriminant analysis effect size (LEFSe) analysis showed that Pseudomonadales and Burkholderiales were affected by exposure to chlorothalonil. Chlorothalonil reduced the survival of honey bee larvae and altered the gut microbiota of newly emerged bees. The risk of pesticides to honey bees is related to their toxicity and exposure dose.

Dissipation of chlorothalonil in the presence of chlortetracycline and ciprofloxacin and their combined effects on soil enzyme activity.

The long-term application of substantial amounts of fungicides and antibiotic-polluted organic manure (OM) in greenhouse has caused the co-existence of fungicides and antibiotics in soils. However, little is known about the effects of antibiotics on the persistence of fungicides in soils or their combined effects on soil enzyme activity. In this study, fungicide chlorothalonil (CTL) alone and in combination with antibiotic chlortetracycline (CTC) or ciprofloxacin (CIP) were repeatedly added to OM-amended soil to investigate the changes in the residual characteristics of CTL and in soil dehydrogenase and urease activity. The results showed that CTL rapidly dissipated in soils with the corresponding half-lives of 0.9-3.2, which initially increased, then decreased and finally stabilized with an increased treatment frequency. The dissipation of CTL was inhibited by CTC and CIP during the first several treatments. The soil dehydrogenase and urease activity in CTL-treated soils was inhibited during the first six treatments and then recovered afterwards. Compared with the OM-amended soil+CTL treatment, the OM-amended soil+CTL+CTC and OM-amended soil+CTL+CIP treatments had stronger inhibitory effects on soil enzyme activity during the first six repeated treatments but exhibited slight stimulating effects afterwards. Therefore, the results obtained in this study suggested that the long-term co-existence of CTL, CTC, and CIP altered the dissipation characteristics of CTL in soil and affected the soil enzyme activity levels. The prudent application of large and frequent of fungicides and OM-containing antibiotic residues in greenhouses should therefore be carefully considered in order to reduce the long-term combined pollution in soils.

Azoxystrobin dissipation and its effect on soil microbial community structure and function in the presence of chlorothalonil, chlortetracycline and ciprofloxacin

The residual characteristics and the adsorption-desorption behaviors of azoxystrobin (AZO) as well as the soil ecological effects in the individual repeated treatments of AZO and its combination with chlorothalonil (CTL), chlortetracycline (CTC) and ciprofloxacin (CIP) were systematically studied in organic manure (OM)-amended soil under laboratory conditions. The presence of CTL, CTC, and CIP, both individually and combined, decreased the sorption affinity of AZO with the Freundlich adsorption and desorption coefficient decreasing by 0.3–24.2%, and CTC and CIP exhibited greater adverse effects than CTL. AZO dissipated slowly and the residues significantly accumulated during ten repeated treatments. The dissipation of AZO was inhibited to different degrees in the combined treatments. Biolog analysis revealed that the soil microbial functional diversity in the OM-soil + AZO and OM-soil + AZO + CTL treatments was higher than that in the OM-soil treatment during the former three repeated treatments, but which was inhibited during the latter seven repeated treatments. The soil microbial functional diversity in the OM-soil + AZO + CTC, OM-soil + AZO + CIP and OM-soil + AZO + CTL + CTC + CIP treatments was inhibited during the ten repeated treatments compared with OM-soil treatment. Metagenomic results showed that all repeated treatments significantly increased the relative abundance of Actinobacteria, but significantly decreased that of Proteobacteria and Firmicutes during the ten repeated treatments. Furthermore, the relative abundance of soil dominant bacterial genera Rhodococcus, Mycobacterium and Arthrobacter in all the repeated treatments significantly increased by 1.5–1283.9% compared with the OM-soil treatment. It is concluded that coexistence of CTL, CTC and CIP, both individually and combined, with AZO can inhibit the dissipation of AZO, reduce the adsorption affinity of AZO on soil, and alter the soil microbial community structure and functional diversity.

Chlorothalonil induces oxidative stress and reduces enzymatic activities of Na+/K+-ATPase and acetylcholinesterase in gill tissues of marine bivalves.

Chlorothalonil is a thiol-reactive antifoulant that disperses widely and has been found in the marine environment. However, there is limited information on the deleterious effects of chlorothalonil in marine mollusks. In this study, we evaluated the effects of chlorothalonil on the gill tissues of the Pacific oyster, Crassostrea gigas and the blue mussel, Mytilus edulis after exposure to different concentrations of chlorothalonil (0.1, 1, and 10 μg L−1) for 96 h. Following exposure to 1 and/or 10 μg L−1 of chlorothalonil, malondialdehyde (MDA) levels significantly increased in the gill tissues of C. gigas and M. edulis compared to that in the control group at 96 h. Similarly, glutathione (GSH) levels were significantly affected in both bivalves after chlorothalonil exposure. The chlorothalonil treatment caused a significant time- and concentration-dependent increase in the activity of enzymes, such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR), in the antioxidant defense system. Furthermore, 10 μg L−1 of chlorothalonil resulted in significant inhibitions in the enzymatic activity of Na+/K+-ATPase and acetylcholinesterase (AChE). These results suggest that chlorothalonil induces potential oxidative stress and changes in osmoregulation and the cholinergic system in bivalve gill tissues. This information will be a useful reference for the potential toxicity of chlorothalonil in marine bivalves.

Fungicides for Control of Downy Mildew on Pickling Cucumber in Michigan

Michigan growers rely on fungicides to limit cucurbit downy mildew (CDM), incited by Pseudoperonospora cubensis; resistance of the pathogen to fungicides is a concern. We evaluated fungicides against CDM in Michigan field studies from 2015 to 2017. According to the relative area under the disease progress curve (rAUDPC), in 2015, mandipropamid, propamocarb, fluxapyroxad/pyraclostrobin, copper octanoate, and dimethomorph resulted in disease levels similar to the control. These treatments, along with cymoxanil, were similar to the control in 2016. Fungicides that were ineffective during 2015 and 2016 did not limit CDM in 2017. Famoxadone/cymoxanil and fluopicolide did not limit CDM in 2017. Each year, the following treatments were similar for disease based on rAUDPC data: oxathiapiprolin applied alone or premixed with chlorothalonil or mandipropamid, ametoctradin/dimethomorph, fluazinam, mancozeb/zoxamide, cyazofamid, and ethaboxam. An exception occurred in 2017, when ethaboxam was less effective than fluazinam, oxathiapiprolin/chlorothalonil, and oxathiapiprolin/mandipropamid. Mancozeb and chlorothalonil treatments were similar in 2015 and 2017, according to rAUDPC data. In 2017, yields were increased for oxathiapiprolin/chlorothalonil, oxathiapiprolin/mandipropamid, mancozeb, ametoctradin/dimethomorph, mancozeb/zoxamide, ethaboxam, cyazofamid, chlorothalonil, and fluazinam compared with the untreated control.

Exogenous 24-epibrassinolide regulates antioxidant and pesticide detoxification systems in grapevine after chlorothalonil treatment

The influence of 24-epibrassinolide (EBR) on the regulation of antioxidant and pesticide detoxification systems in grape seedlings (Vitis vinifera L.) was investigated. The grapevine leaves were pretreated with different concentrations of EBR (0.10, 0.21 and 0.42 μM) before spraying with chlorothalonil (CHT) (600 times diluent). Compared with the control, CHT treatment showed an adverse effect on osmoregulation substances (soluble protein and free proline) and increased the content of malondialdehyde (MDA), activities of antioxidant enzymes and detoxification enzymes as well as the content of some non-enzymatic antioxidants. Compared to the CHT treatment alone, application of three concentrations of EBR resulted in an overall enhancement of these enzyme activities, soluble protein and free proline contents and reduction in MDA content. In addition, the expression of some key genes involved in CHT degradation including glutathione synthetase (GSH), glutathione S-transferase (GST), glutathione reductase (GR), P450 monooxygenase (P450) and MRP-type transporter (MRP) was analyzed. The transcriptional level of these genes increased in response to CHT treatment with the exception of GSH. EBR treatment of 0.10 μM could effectively promote expression of most genes and accelerate CHT degradation by 29 % as compared to only CHT treatment in grapevine leaves.

Biocontrol of <I>Rhizoctonia solani</I> Root Rot of Chilli by <I>Bacillus subtilis</I> formulations under Pot Conditions

The chilli crop suffers massive yield loss due to root rot caused by Rhizoctonia solani . An increase of 10 × 10 5 root colonizing units/cm was obtained as a result of Bacillus subtilis in vitro root colonisation assay post germination. Shelf life studies of the formulations revealed stable population level of the biocontrol agent upto 180 th day (30°C - 1.6 × 10 8 ; 4°C - 1.9 × 10 8 ) in talc and upto 150 th day in lignite (30°C - 1.5 × 10 8 ; 4°C - 1.3 × 10 8 ). Soil, seed, foliar spray and dip treatment methods of B. subtilis and chlorothalonil brought about a considerable enhancement of all biometric parameters and reduced disease incidence compared to the untreated control. In comparison to the untreated control (50 g and 21 g fresh and dry weight, respectively), highest plant fresh weight (76.84 g) and dry weight (34.17 g) was achieved by the Seed application method. Comparison of plant height revealed maximum values 70 cm (soil application) and 77 cm (dip treatment) with B. subtilis application which was analogous to chlorothalonil treatment (56.5 cm with soil application and 70.33 cm with dip treatment) which was considerably superior to the untreated control (58.2 cm with dip treatment and 61 cm with soil application, respectively). Root dip treatment showed considerable increase in root length with B. subtilis (33 cm) and chlorothalonil (28.5 cm) when compared to untreated control (15 cm). Growth promotion was better with Root dip application while disease control was achieved better with seed application. A 66% and 84% reduction in incitation of disease was noticed with soil and seed application methods, respectively.

Biocontrol of <I>Rhizoctonia solani</I> Root Rot of Chilli by <I>Bacillus subtilis</I> formulations under Pot Conditions

The chilli crop suffers massive yield loss due to root rot caused by <em>Rhizoctonia solani</em>. An increase of 10 × 10<sup>5</sup> root colonizing units/cm was obtained as a result of <em>Bacillus subtilis</em> in vitro root colonisation assay post germination. Shelf life studies of the formulations revealed stable population level of the biocontrol agent upto 180<sup>th</sup> day (30°C - 1.6 × 10<sup>8</sup>; 4°C - 1.9 × 10<sup>8</sup>) in talc and upto 150<sup>th</sup> day in lignite (30°C - 1.5 × 10<sup>8</sup>; 4°C - 1.3 × 10<sup>8</sup>). Soil, seed, foliar spray and dip treatment methods of <em>B. subtilis</em> and chlorothalonil brought about a considerable enhancement of all biometric parameters and reduced disease incidence compared to the untreated control. In comparison to the untreated control (50 g and 21 g fresh and dry weight, respectively), highest plant fresh weight (76.84 g) and dry weight (34.17 g) was achieved by the Seed application method. Comparison of plant height revealed maximum values 70 cm (soil application) and 77 cm (dip treatment) with <em>B. subtilis</em> application which was analogous to chlorothalonil treatment (56.5 cm with soil application and 70.33 cm with dip treatment) which was considerably superior to the untreated control (58.2 cm with dip treatment and 61 cm with soil application, respectively). Root dip treatment showed considerable increase in root length with <em>B. subtilis</em> (33 cm) and chlorothalonil (28.5 cm) when compared to untreated control (15 cm). Growth promotion was better with Root dip application while disease control was achieved better with seed application. A 66% and 84% reduction in incitation of disease was noticed with soil and seed application methods, respectively.

Root colonization by mycorrhizal INIFAP<sup>MR</sup> in barley seed treated with fungicide

The effect of barley seed treated with chlorothalonil fungicide on mycorrhizal root colonization was evaluated. The treatments were: 1) Seed with Mycorrhizal INIFAP?, one, two and three doses, and uninoculated seed; 2) Seed with and without fungicide. Monthly seed plantings were performed with different storage time, looking forward to assessing the colonization degree of the roots. As revealed by the results, the seed treated with chlorothalonil did not lower the root colonization by Mycorrhizal INIFAP?; the chlorothalonil treatment had a 32.63% average root colonization, whereas without chlorothalonil, it was 36.46%. When the seed was treated with root colonization by Mycorrhizal INIFAP?, this was lower in the treatment with one dose; no significant difference was revealed by the treatments with either double or triple doses. The colonization percentage was progressively decreased by seed storage. The root colonization by Mycorrhizal INIFAP?, throughout the inoculated seed storage time, remained constant and unchanged for the first six months, but then decreased in 50% within a 10-month period for both treated and not treated fungicide seed.

Glutathione biosynthesis and regeneration play an important role in the metabolism of chlorothalonil in tomato

Glutathione is one of the major endogenous antioxidants produced by cells. In plants, glutathione is crucial for both abiotic and biotic stress resistance, and also involved in the detoxification of xenobiotics in many organisms. However, as in vivo evidences of glutathione function are still lacking so far, its roles in plants are still poorly understood. In this study, we investigated the changes of thiols, glutathione homeostasis and transcripts of genes potentially involved in chlorothalonil (CHT) metabolism in tomato (Solanum lycopersicum L.). Two genes (GSH1, GSH2) encoding γ-glutamylcysteine synthetase and glutathione synthetase, respectively, and a gene for glutathione reductase (GR1) involved in glutathione regeneration were silenced by virus induced gene silencing (VIGS) approach. Silencing of GSH1, GSH2 and GR1 decreased glutathione contents and the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG), but increased CHT residues in plant tissues. The GSH1 and GR1 silenced plants showed the lowest GSH level and ratio of GSH/GSSG, respectively. Transcripts of P450, GST and ABC transporter genes as well as glutathione S-transferase (GST) activity were induced after CHT treatment. However, the increases of these transcripts were compromised in GSH1, GSH2 and GR1 silenced plants. This study indicates that glutathione not only serves as a substrate for CHT conjugation, but is also involved in regulation of transcripts of gene in pesticide metabolism via controlling redox homeostasis.

Effects of Azoxystrobin Application Rate and Treatment Interval on the Control of Rhabdocline pseudotsugae on Douglas-fir Christmas Trees

Rhabdocline needlecast caused by Rhabdocline pseudotsugae is the primary disease limiting Douglas-fir Christmas tree production in the northeastern United States. Azoxystrobin (Quadris) was recently registered for control of needlecasts on conifers, but little is known about its efficacy. In 2002, azoxystrobin was applied to field-grown Lincoln N.F. Douglas-fir Christmas trees at 0.14 or 0.28 g a.i./liter as the first or second spray of a chlorothalonil-based control program. In 2003, four sequential sprays of azoxystrobin at 0.28, 0.55, and 1.10 g a.i./liter were compared to chlorothalonil treatments. Untreated trees at both Pennsylvania test sites in both years were heavily infected, confirming high inoculum levels and environmental conditions favorable for infection. The standard program that consisted of 1.29 g a.i./liter chlorothalonil applications was very effective in controlling Rhabdocline in all experiments. In 2002, application interval had a significant effect on efficacy. Trees sprayed with 0.14 and 0.28 g a.i./liter azoxystrobin 9 days prior to an infection period had a higher disease index rating than those sprayed 2 days prior to the same infection period. Application rate did not appear to have an effect on efficacy. Trees receiving 1.10 g a.i./liter azoxystrobin had the same disease index rating as trees receiving the 0.28 g a.i./liter rate. In all cases, azoxystrobin treatments had significantly higher infection rates than the standard chlorothalonil treatment. The level of natural inoculum present at each site also appeared to play a role in azoxystrobin efficacy. Azoxystrobin is more than twice the cost of chlorothalonil and the data presented does not support any cost incentive for its inclusion in a Rhabdocline control program. Accepted for publication 19 May 2005. Published 17 June 2005.

How the use of chlorothalonil on golf courses impacts Paenibacillus sp., a pathogen of Ataenius spretulus (Coleoptera: Scarabaeidae).

One of the most frequently used fungicides on golf courses, chlorothalonil, was applied to field plots at four different golf courses to determine how Ataenius spretulus Haldeman, Aphodius granarius L., carabids, staphylinids, formicids and Paenibacillus sp. were affected. Chlorothalonil had no influence on the incidence of Paenibacillus sp. infection of A. spretulus larvae in golf course fairways. The incidence of Paenibacillus sp. infection of A. spretulus larvae was much greater in the rough (47.4%) compared with the fairway (26.4%) at Cattails Golf Club despite a more dense population (fourfold) of larvae in the fairway. Chlorothalonil treatment of turf plots had no influence on the number of staphylinids, carabids, formicids, or A. spretulus adults caught in pitfall traps. It also did not affect the density of A. spretulus larvae. Data from this study suggest that the widespread use of chlorothalonil on fairways is not likely to cause outbreaks of A. spretulus on golf courses.

Response of field pea cultivars to chlorothalonil in the control of mycosphaerella blight

The efficacy of chlorothalonil in controlling mycosphaerella blight in field pea was evaluated from 1998 to 2000 using 10 cultivars representing different leaf types, market classes, seed sizes, plant heights, and maturities. Significant differences ( P < 0.05) were observed among cultivars in disease severity, yield, 1000-seed weight (TSW), and seed infection each year. Cultivar × fungicide treatment interactions for these parameters were not significant in any of the 3 yr. Under both chlorothalonil treated and untreated conditions, Radley was the most resistant, while Carrera and Keoma were equally the most susceptible cultivars. A higher efficacy of chlorothalonil treatments was observed in 1999 when disease severity was greater than in 1998 or 2000. On average, chlorothalonil treatments reduced disease severity significantly in 1999 and 2000, but not in 1998. Overall, the fungicide increased yield by 6.4%, and TSW by 0.9–5.1%, which were significant in 1998 and 1999, but not in 2000. Based on 2-yr assessments of seed infection by Mycosphaerella pinodes, chlorothalonil reduced seed infection by 19.2%, which was significant in 1999, but not in 2000. Chlorothalonil treatments provide a greater benefit in years when disease pressure and yield potential are high. Key words: Field pea, Pisum sativum, mycosphaerella blight, Mycosphaerella pinodes, chlorothalonil

The impact of chlorothalonil application on soil bacterial and fungal populations as assessed by denaturing gradient gel electrophoresis

The impact of the fungicide chlorothalonil on dominant bacterial and fungal populations following application to turfgrass, forest, and agricultural soils was investigated. Chlorothalonil was applied to each soil at three rates, representing 0.2, 1 and 5 times the recommended label rate for turfgrass, and incubated for a 2-week period. Changes to the microbial community caused by the chlorothalonil application were assessed following DNA extraction, PCR-amplification using both bacteria domain- and fungal-specific primers, then separation with denaturing gradient gel electrophoresis (DGGE). Digitized DGGE images were used to determine two parameters: the number of bands per lane and the Shannon–Wiener index of diversity ( H′), both of which were used only for comparison of the different treatments, and not as true diversity measurements. Bands appearing to be either enhanced or inhibited as a result of the chlorothalonil treatment were excised and sequenced. Increased rates of chlorothalonil impacted eight bacterial populations (two inhibitions, four enhancements, and two non-specific responses) and four fungal populations (all inhibitions). Band number and H′ indicated an altered but not significantly different ( P<0.05) bacterial and fungal community structure following chlorothalonil application. Sequencing of excised DGGE bands indicated an impact on several groups of bacteria ( Cytophaga–Flavobacterium–Bacteroides, α-, β-, γ-, and δ-proteobacteria) and two fungal taxa (zygomycota and ascomycota). Although changes to the overall community structure of dominant species were non-significant, we conclude that following a single chlorothalonil application and a short incubation period, community changes including both enhancement and inhibition of a variety of dominant organisms can occur.

Coccinellids in Cotton: Population Response to Pesticide Application and Feeding Response to Cotton Aphids (Homoptera: Aphididae)

Four treatments were evaluated during 1997 and 1998 to determine the effects of pesticides on coccinellid densities: (1) untreated control, (2) foliar applications of a systemic insecticide, imidacloprid (Provado 1.6 F), when spray thresholds of aphid numbers were exceeded in all plots, (3) weekly foliar applications of a fungicide, chlorothalonil (Bravo 720), and (4) weekly foliar applications of imidacloprid (Provado 1.6 F). The coccinellids observed in Georgia cotton during the study included Hippodamia convergens Guerin-Meneville, Scymnus spp., Harmonia axyri- dis (Pallas), Coccinella septempunctata L., and Coleomegilla maculata (Mulsant). During 1997, H. convergens and Scymnus spp. comprised 75 and 15%, respectively, of all coccinellids observed during the study. During 1998, Scymnus spp. comprised 44% of all coccinellids, and H. convergens comprised 33%. Coccinellid population densities closely tracked those of cotton aphids during both years. Cotton aphid and coccinellid densities were greatest in the chlorothalonil treatment during both years of the study. In functional feeding response experiments, fourth-instar and adult Scynmus creperus exhibited a type II functional response to A. gossypii density under laboratory conditions. Fourth instars exhibited a higher search rate and shorter handling time than adult S. creperus. Collectively, coccinellids are a valuable component of the cotton aphid's natural enemy complex.

Management of Blackspot on Three Rose Cultivars Using Antitranspirants in Combination with Chlorothalonil

Abstract The efficacy of reducing fungicide use, by combining chlorothalonil with film-forming antitranspirant products and lengthening intervals between applications, was evaluated for control of blackspot of rose. Antitranspirants included three highly refined pine oil (pinolene) products (Nu Film 17, Vapor Gard, and Wilt Pruf) and two latex products (Stressguard and Transfilm), each of which were applied in combination with chlorothalonil fungicide. Antitranspirant solutions were applied at 2- or 3-week intervals, depending on recommended use of the antitranspirant. Blackspot severity, based on proportion of symptomatic foliage, did not differ among treatments in either year of the study. In both study years, defoliation was lower on plants treated weekly with the fungicide alone, with Stressguard (applied at 0.25% in combination with chlorothalonil every 2 weeks), or with Vapor Gard (applied at 2% in solution with chlorothalonil every 3 weeks) compared to nontreated plants. Overall, the above fungicide and antitranspirant solutions reduced season-long fungicide use by 40-69%, without a decrease in disease control, compared to the weekly chlorothalonil treatment. Plant vigor and/or growth, however, was lower with the antitranspirant treatments compared to chlorothalonil alone.