Ilias G. Eleftherohorinos

Pesticide Exposure, Safety Issues, and Risk Assessment Indicators

Pesticides are widely used in agricultural production to prevent or control pests, diseases, weeds, and other plant pathogens in an effort to reduce or eliminate yield losses and maintain high product quality. Although pesticides are developed through very strict regulation processes to function with reasonable certainty and minimal impact on human health and the environment, serious concerns have been raised about health risks resulting from occupational exposure and from residues in food and drinking water. Occupational exposure to pesticides often occurs in the case of agricultural workers in open fields and greenhouses, workers in the pesticide industry, and exterminators of house pests. Exposure of the general population to pesticides occurs primarily through eating food and drinking water contaminated with pesticide residues, whereas substantial exposure can also occur in or around the home. Regarding the adverse effects on the environment (water, soil and air contamination from leaching, runoff, and spray drift, as well as the detrimental effects on wildlife, fish, plants, and other non-target organisms), many of these effects depend on the toxicity of the pesticide, the measures taken during its application, the dosage applied, the adsorption on soil colloids, the weather conditions prevailing after application, and how long the pesticide persists in the environment. Therefore, the risk assessment of the impact of pesticides either on human health or on the environment is not an easy and particularly accurate process because of differences in the periods and levels of exposure, the types of pesticides used (regarding toxicity and persistence), and the environmental characteristics of the areas where pesticides are usually applied. Also, the number of the criteria used and the method of their implementation to assess the adverse effects of pesticides on human health could affect risk assessment and would possibly affect the characterization of the already approved pesticides and the approval of the new compounds in the near future. Thus, new tools or techniques with greater reliability than those already existing are needed to predict the potential hazards of pesticides and thus contribute to reduction of the adverse effects on human health and the environment. On the other hand, the implementation of alternative cropping systems that are less dependent on pesticides, the development of new pesticides with novel modes of action and improved safety profiles, and the improvement of the already used pesticide formulations towards safer formulations (e.g., microcapsule suspensions) could reduce the adverse effects of farming and particularly the toxic effects of pesticides. In addition, the use of appropriate and well-maintained spraying equipment along with taking all precautions that are required in all stages of pesticide handling could minimize human exposure to pesticides and their potential adverse effects on the environment.

Influence of nitrogen on competition between winter cereals and sterile oat

Abstract Field experiments were carried out in northern Greece during 1994, 1995, and 1996 to study the effect of nitrogen fertilization on competition between sterile oat and wheat, barley, and triticale. Dry weight of all crops was not affected until early March by sterile oats (110 plants m−2), but wheat and triticale dry weight were significantly reduced by sterile oats competition after that time. Grain yield of both wheat and triticale was equally reduced by 61% due to the presence of sterile oats, whereas the reduction for barley grain yield was 9%. Nitrogen fertilization (150 kg N ha−1) slightly increased yield of all crops grown without weed competition compared to the control (0 kg N), whereas the same treatment increased sterile oats dry weight as well as its competitive ability against wheat and triticale. Split application of nitrogen (50 kg N ha−1 before planting and 100 kg N ha−1 in early March) caused a slightly higher increase in sterile oats dry weight compared to the control or one application (150 kg N ha−1) before planting, when grown with wheat and triticale. However, dry weight of sterile oats grown with barley was severely reduced by the interference of the crop. Total nitrogen content of all crop plants grown without sterile oats increased with nitrogen fertilization compared to the control. However, total nitrogen in crop plants grown with sterile oats was reduced compared to the weed-free control; percent reduction was greater in plants grown in plots treated with nitrogen than in the control. These results indicate that barley could be used for limiting sterile oats interference in areas where winter cereals are grown; time of nitrogen application could also be used for a slight reduction of sterile oats competitive ability against wheat or triticale. Nomenclature: Barley, Hordeum vulgare = distichum L. ‘Carina’; sterile oat, Avena sterilis L. AVEST; triticale, × Triticosecale ‘Dada’; wheat, Triticum aestivum L. ‘Yecora’.

Weeds in greenhouses and tobacco fields are differentially infected by Tomato spotted wilt virus and infested by its vector species.

A survey was conducted in the Macedonia region of Greece to determine the reservoir hosts of Tomato spotted wilt virus (TSWV) in three tobacco fields and in a greenhouse complex in which lettuce and the ornamentals chrysanthemum, gerbera, aster, and anemone were grown. Assays for TSWV infection were made by enzyme-linked immunosorbent assay on 6,172 plant samples, 3,909 from tobacco fields and 2,263 from the greenhouse complex, comprising plants of 208 species in 137 genera of 42 families. Plants of 86 species out of 63 genera of 27 families were infected of which 39 species are newly reported hosts of TSWV. An infection index was developed to evaluate the relative potential of each weed species as a virus source in both systems. Seventeen species in the tobacco fields and nine in the greenhouses had an infection index higher than one. Most species with infected plants were found in the Compositae family. Plants of some species occurring both in tobacco fields and in greenhouses were infected at only one of these sites. Frankliniella occidentalis was the common thrips species on weeds and crops in the greenhouses, while Thrips tabaci was the only vector on tobacco plants and weeds in the tobacco fields. This observation strongly suggests that the occurrence of species with infected plants and their number have to be attributed to the vector species prevailing in the greenhouse complex or tobacco fields, supporting the conclusion that TSWV is spread in two different epidemiological processes in Greece.

Dicamba and Atrazine Antagonism on Sulfonylurea Herbicides Used for Johnsongrass (Sorghum halepense) Control in Corn (Zea mays)1

Abstract: Field experiments were carried out during 1997 and 1998 in northern Greece to investigate the effects of tank mixing rimsulfuron and primisulfuron with atrazine or dicamba against johnsongrass in corn. Sequential applications, where the johnsongrass herbicides were applied 5 d after the broadleaf herbicides, were also evaluated. Rimsulfuron applied alone at 10 g ai/ha gave very good control (91%) of johnsongrass, which was significantly higher than that provided by 30 g ai/ha of primisulfuron (43%). Rimsulfuron applied in tank mixture with atrazine (1.0 kg ai/ha) or dicamba (0.28 kg ai/ha) gave 12 and 17% lower johnsongrass control, respectively, than of rimsulfuron applied alone, whereas the corresponding reduction for primisulfuron was 18 and 43%. Efficacy of rimsulfuron applied 5 d after the application of atrazine or dicamba was similar to that applied alone; however, this was not the case for primisulfuron, where reduced antagonism was observed compared to that produced by its tank mixture treatments. Again, primisulfuron was affected more by dicamba than by atrazine. Corn yield with rimsulfuron and primisulfuron applied alone was more than double that of the untreated control and similar to that of the weed-free control. Also, rimsulfuron applied with atrazine or dicamba (in tank mixture or sequentially) produced similar corn yield to that applied alone. However, primisulfuron applied in tank mixture or sequentially with dicamba gave 22 and 14% lower yield, respectively, than when applied alone, and slightly lower when applied with atrazine. Nomenclature: Atrazine, dicamba, primisulfuron, rimsulfuron, johnsongrass, Sorghum halepense (L.) Pers. #3 SORHA; corn, Zea mays L. ‘Pioneer Costanza’. Additional index words: Sequential applications, tank mixtures. Abbreviations: DAT, days after treatment.

Interference of red rice in rice grown in Greece

Abstract Field studies were conducted in northern Greece, in 1997 and 1998, to investigate the effect of nitrogen fertilization and red rice density on interference between red rice and two rice cultivars (Thaibonnet, Ariette). Interference between rice and red rice began 3 wk after rice emergence, but was not affected by the increasing nitrogen rate from 100 to 150 kg N ha–1. Dry weight of both rice cultivars was proportionally reduced with increasing red rice interference duration and density, but dry weight of Thaibonnet was reduced more than that of Ariette. At harvest, grain yield of Thaibonnet was reduced by 58% because of the occurrence of 40 red rice plants m–2, whereas that of Ariette was reduced by 46%. Red rice interference affected panicle number more than 1,000 grain weight in both rice cultivars. The reduction of all yield components was greater in Thaibonnet than in Ariette. Dry weight and stem or panicle number of red rice plants grown with either of the two rice cultivars increased with increasing red rice density and were greater most of the time when grown with Thaibonnet than with Ariette. Ten weeks after rice emergence, red rice plants were 14 and 35 cm taller than the Ariette and Thaibonnet plants, respectively. Shattering of red rice plants ranged from 63 to 79% and was greater grown with Thaibonnet than with Ariette, but it was not affected by nitrogen fertilization and red rice density. Nomenclature: Red rice, Oryza sativa L. ORYSA; rice, Oryza sativa L. ‘Ariette’, ‘Thaibonnet’.

Critical periods of weed competition in cotton in Greece

Four experiments were conducted in central Greece during 1997 and 1998 to determine the late-season presence of weeds in cotton (Gossypium hirsutum L.) and the critical times for removing weeds. Experiments were conducted in natural, heavily infested cropland. The presence of weeds for more than 3 weeks after crop emergence caused significant reductions in crop growth and lint yields. However, weeds that emerged 11 weeks or more after crop emergence did not adversely impact yields. Total weed biomass increased with increasing time prior to weed removal. A weed-free period of 11 weeks after crop emergence was needed to prevent significant reductions in cotton height, biomass, number of squares, and yield. These results indicated that postemergence herbicides or other control measures should be initiated within 2 weeks after crop emergence to avoid significant yield reduction. For greater efficiency, soil-applied herbicides in cotton should provide effective weed control for at least 11 weeks. Curvilinear regression equations were derived to describe the relationship between critical periods of weed presence and cotton growth and fruit development.

Interference between corn and johnsongrass (Sorghum halepense) from seed or rhizomes

Abstract Field experiments were conducted in 1999 and 2000 in northern Greece to study interference between johnsongrass from seed or rhizomes and three corn hybrids (‘Dunia’, ‘Papea’, and ‘Costanza’). Fresh weight of each corn hybrid was lower with greater duration of johnsongrass interference, and it was lower for corn grown with johnsongrass from rhizomes than from seed. Corn-silage yield and corn-grain yield were 10 to 14% less than weed-free corn yield when grown with johnsongrass plants from rhizomes 4 wk after sowing (WAS) or from seed 6 WAS. Regression analysis indicated that a corn-grain yield loss of 10% was associated with an interference period of 2.6 to 2.7 WAS for johnsongrass from rhizomes and 4.3 to 5.1 WAS for johnsongrass from seed compared with weed-free corn. Corn-silage yield with season-long interference by johnsongrass from rhizomes or from seed was 83 and 62% lower, respectively, than was weed-free corn yield. Similarly, corn-grain yield with season-long interference by johnsongrass from rhizomes or from seed was 88 and 57% lower, respectively, than was the yield from weed-free corn. Corn-ear length of each hybrid was more affected by johnsongrass interference than was 1,000-seed weight. Johnsongrass plants from rhizomes emerged earlier, grew faster, and produced greater fresh weight than did plants from seed. However, fresh weight and stem number of johnsongrass plants from either seed or rhizomes were not affected by corn hybrids. Nomenclature: Johnsongrass, Sorghum halepense (L.) Pers. SORHA; corn, Zea mays L. ‘Pioneer Costanza’, ‘Pioneer Dunia’, ‘Pioneer Papea’.

Corn poppy (Papaver rhoeas) cross-resistance to ALS-inhibiting herbicides.

BACKGROUND Papaver rhoeas (L.) has evolved resistance to tribenuron in winter wheat fields in northern Greece owing to multiple Pro(197) substitutions. Therefore, the cross-resistance pattern to other sulfonylurea and non-sulfonylurea ALS-inhibiting herbicides of the tribenuron resistant (R) and susceptible (S) corn poppy populations was studied by using whole-plant trials and in vitro ALS catalytic activity assays. RESULTS The whole-plant trials revealed that tribenuron R populations were also cross-resistant to sulfonylureas mesosulfuron + iodosulfuron, chlorsulfuron and triasulfuron. The whole-plant resistance factors (RFs) calculated for pyrithiobac, imazamox and florasulam ranged from 12.4 to > 88, from 1.5 to 28.3 and from 5.6 to 25.4, respectively, and were lower than the respective tribenuron RF values (137 to > 2400). The ALS activity assay showed higher resistance of the ALS enzyme to sulfonylurea herbicides (tribenuron > chlorsulfuron) and lower resistance to non-sulfonylurea ALS-inhibiting herbicides (pyrithiobac > florasulam ≈ imazamox). CONCLUSION These findings indicate that Pro(197) substitution by Ala, Ser, Arg or Thr in corn poppy results in a less sensitive ALS enzyme to sulfonylurea herbicides than to other ALS-inhibiting herbicides. The continued use of sulfonylurea herbicides led to cross-resistance to all ALS-inhibiting herbicides, making their use impossible in corn poppy resistance management programmes.

Multiple Pro197 Substitutions in the Acetolactate Synthase of Corn Poppy (Papaver rhoeas) Confer Resistance to Tribenuron

Abstract Variations in the acetolactate synthase (ALS) gene sequence were determined from 28 populations of corn poppy resistant (R) to tribenuron and from 6 populations susceptible (S) to this herbicide. The ALS gene fragment (634 bp) sequence revealed in R populations five point mutations at the codon Pro197, and among them the substitution of Pro197 by Ala was the most common. The sequencing chromatograms revealed that nine R individuals had only the mutant ALS gene and were homozygous (RR), 18 R individuals had both the wild type and the mutant ALS gene and were heterozygous (RS), whereas one R individual was heterozygous but with two different mutant ALS alleles (R1R2). The use of restriction digestion profile analysis to verify the DNA sequence results by detecting the existence of point mutations at the codon 197 managed to distinguish the R and S alleles and confirmed the results obtained from the sequencing chromatograms analysis. The secondary protein structure prediction suggested the formation of novel β-strands for each of the five mentioned amino acid substitutions that was not present in wild type ALS around the mutant site. These findings support the hypothesis that the substitution of Pro197 by Ser, Thr, Ala, Arg, or Leu resulted in altered secondary structure, which stabilizes an ALS tertiary conformation that prevents tribenuron binding and thus confers resistance to this herbicide. Nomenclature: Tribenuron; corn poppy, Papaver rhoeas L. PAPRH.

COMPETITIVE ABILITY OF WINTER CEREAL–COMMON VETCH INTERCROPS AGAINST STERILE OAT

Intercropping cereals with legumes for forage or food production is extensively used as a cropping practice in many parts of the world. A 2-year field study was conducted using common vetch ( Vicia sativa ), winter wheat ( Triticum aestivum ), triticale (× Triticosecale ), barley ( Hordeum vulgare ) and oat ( Avena sativa ) sole crops as well as common vetch intercrops with each of these cereals in a 35:65 cereal:common vetch ratio based on seed numbers, to determine their ability to compete with sterile oat ( Avena sterilis spp. sterilis ). At nine weeks after planting, fewer sterile oat plants emerged in common vetch sole crop than in cereal sole crops. Intercropping of cereals with common vetch generally did not affect sterile oat stem number and biomass compared with cereal sole crops. At harvest, cereal sole crops provided greater total dry biomass (DB) than the common vetch sole crop. However, triticale and oat produced more DB than winter wheat and barley. In most cases intercropping reduced total DB compared with cereal sole crops. The results of this study indicated that intercropping of the four winter cereals with common vetch did not provide any significant competitive advantage against sterile oat. However, common vetch sole crop showed the greatest suppressive ability against sterile oat among the sole crops or intercrops studied.