Effect of some herbicides on activities of microorganisms and enzymes in soil

Abstract

Laboratory tests were conducted with eight herbicides, atrazine, butylate, ethalfluralin, imazethapyr, linuron, metolachlor, metribuzin and trifluralin, applied to a loamy sand at rate of 10 μg/g to determine if these materials caused any serious effects on microbial and enzymatic activities related to soil fertility. Some herbicides showed an effect on bacteria and fungi for the first week of incubation, but, subsequently, the populations returned to levels similar to those obtained in the controls. After several herbicide treatments there appeared to cause a slight depression of nitrification. Sulfur oxidation was better than that obtained with untreated soil in all treatments. Oxygen consumption was increased significantly after 96 hr incubation with atrazine. The soil dehydrogenase and amylase activities were inhibited by ethalfluralin treatment respectively for 1 wk and 1 day, and p‐nitrophenol liberation was inhibited for 2 hrs by all herbicide treatments. Results indicated that the herbicidal treatments at the level tested were not drastic enough to be considered deleterious to soil microbial and enzymatic activities which are important to soil fertility.     

Effect of tracer dyes on initial deposits and persistence of bacillus thuringiensis subsp. kurstaki toxin after application of two commercial formulations onto spruce trees

Abstract

The effect of two tracer dyes [Erio Acid Red (EAR) and Acid Black 48 (AB‐48)] on initial deposits and persistence of Bacillus thuringiensis subsp. kurstaki (Btk) toxin (delta‐endotoxin) was studied after spraying two commercial formulations, Foray® 48B and Foray® 76B, over potted white spruce [Picea glauca (Moench) Voss] seedlings, at a dosage rate of 30 billion international units (BIU) per ha. Spray was applied using a spinning disc atomizer calibrated to deliver droplet sizes similar to those utilized in ultra‐low‐volume (ULV) treatments in operational insect control programs. The sprayed seedlings were left outdoors at the Sault Ste. Marie laboratory for 18 days under natural conditions of sunlight, wind and rainfall. Initial deposits and persistence of delta‐endotoxin protein in spruce foliage were determined by immunoassay [enzyme linked immunosorbent assay (ELISA)] quantification of the delta‐endotoxin. The total protein (inactive plus active) and delta‐endotoxin (active protein) concentrations in the two formulations were determined by a gravimetric procedure and by ELISA respectively.

The initial deposit levels of the toxin on foliage were not markedly affected by the addition of either of the two tracer dyes, and showed only a narrow range of 1521 to 1625 ng/g foliage (fresh weight) for Foray 48B, and 1789 to 2056 ng/g for Foray 76B. However, the persistence of the toxin was significantly influenced by the presence of the dyes. The toxin persisted in foliage only for 7 d post‐spray When the EAR dye was added to Foray 48B, compared to 10 d when no dye was added. The average half‐life (DT₅₀) of disappearance was 17.4 h for Foray 48B with EAR, and 20.9 h when no dye was present. In contrast, the situation was reversed in Foray 76B, since the duration of persistence was 10 d when EAR was added to Foray 76B, compared to 7 d when no dye was added. The average DT₅₀ was 27.9 h for Foray 76B with EAR, and 22.2 h without the dye. Persistence was the longest (14 d) when the AB‐48 dye was added to Foray 76B, and the DT₅₀ was 44.9 h.     

Defending the scientific integrity of conservation‐policy processes

Government agencies faced with politically controversial decisions often discount or ignore scientific information, whether from agency staff or nongovernmental scientists. Recent developments in scientific integrity (the ability to perform, use, communicate, and publish science free from censorship or political interference) in Canada, Australia, and the United States demonstrate a similar trajectory. A perceived increase in scientific‐integrity abuses provokes concerted pressure by the scientific community, leading to efforts to improve scientific‐integrity protections under a new administration. However, protections are often inconsistently applied and are at risk of reversal under administrations publicly hostile to evidence‐based policy. We compared recent challenges to scientific integrity to determine what aspects of scientific input into conservation policy are most at risk of political distortion and what can be done to strengthen safeguards against such abuses. To ensure the integrity of outbound communications from government scientists to the public, we suggest governments strengthen scientific integrity policies, include scientists’ right to speak freely in collective‐bargaining agreements, guarantee public access to scientific information, and strengthen agency culture supporting scientific integrity. To ensure the transparency and integrity with which information from nongovernmental scientists (e.g., submitted comments or formal policy reviews) informs the policy process, we suggest governments broaden the scope of independent reviews, ensure greater diversity of expert input and transparency regarding conflicts of interest, require a substantive response to input from agencies, and engage proactively with scientific societies. For their part, scientists and scientific societies have a responsibility to engage with the public to affirm that science is a crucial resource for developing evidence‐based policy and regulations in the public interest.     

Progress in the genetic manipulation of crops aimed at changing starch structure and increasing starch accumulation

The starch content and its composition have important consequences for the yield of the harvested crop and the materials extracted from it. The functional properties of the foods or other processed materials derived from these crops are also affected by the structure and composition of the starch. Recently, genetic engineering has been used to produce plants with an elevated starch content, achieved by transforming the plant with a mutated bacterial gene coding for an ADPglucose pyrophosphorylase that is active in the presence of metabolites which inhibit the plant enzyme. Besides the practical implications of these results, this experiment provided direct evidence for the regulatory role of the ADPglucose pyrophosphorylase in starch synthesis. Other bacterial enzymes, such as glycogen synthase and branching enzyme, could be introduced in order to modify starch structure. However, a more elegant (but longer-term) approach would be to learn enough about the structure-function relationships of the plant enzymes so that the product of their action could be changed. To achieve this objective, much more will have to be learned about the enzymes involved in the biosynthesis of starch than is presently known. Here, the basic properties of starch and the current research approaches to understanding its biosynthesis are described, together with a perspective of how genetic manipulation of starch structure may be achieved.Paper presented at the Bio/Environmentally Degradable Polymer Society—Third National Meeting, June 6–8, 1994, Boston, Massachusetts.     

Purification and characterization of extracellular poly(3-hydroxybutyrate) depolymerases

Five extracellular PHB depolymerases of bacteria isolated from various sources were purified to electrophoretic homogeneity and compared with known extracellular PHB depolymerase fromAlcaligenes faecalis T1. The molecular mass of these enzymes were all around 40–50 kDa. Nonionic detergent, diisopropylfluorophosphate and dithiothreitol inhibited the PHB depolymerase activity of all these enzymes. Trypsin abolished PHB depolymerase activity, but not theD-3-hydroxybutyric acid dimer hydrolase activity of all the enzymes. These results showed that the basic properties of these PHB depolymerases resemble those of theA. faecalis T1 enzyme. Analysis ofN-terminal amino acid sequence of the purified enzymes revealed that these enzymes includingA. faecalis T1 enzyme fall into three groups.     

The Role Played by Acetyl Group Distribution Patterns in Substituted Starch Toward Enzymatic De-acetylation and Chain Fragmentation

The nature and distribution of the acetylated groups were evaluated by ¹³C-NMR and ¹H-NMR. The starch substrate with a DS of 1.5 comprises only two patterns: -(14)-d-glucopyranose and 2,3,6-tri-O-acetyl--(14)-d-glucopyranose. The starch with a DS of 3.0 also comprises two patterns: 2,3,4,6-tetra-O-acetyl--(14)-d-glucopyranose and 2,3,6-tri-O-acetyl--(14)-d-glucopyranose; whereas starch (DS = 1.9) contains 4 patterns: 2,3,6-tri-O-acetyl--(14)-d-glucopyranose, 2,3,4,6-tetra-O-acetyl--(14)-d-glucopyranose terminal, 2,6-di-O-acetyl--(14)-d-glucopyranose, and 3,6-di-O-acetyl--(14)-d-glucopyranose. Using esterase from Viscozyme, it has been possible to hydrolyze up to 7% of the DS 3.0 starch. An -amylase (Fungamyl 800) was then added to these acetylesterases. With a 2.4 FAU/mL fraction of -amylase and 2.4 U/mL from the Viscozyme's acetylesterase, 28% of the acetylated end groups were hydrolyzed for the starch substrates with DS 3.0. Moreover, a synergic action between -amylase and acetylesterase was noticed, allowing fragmentation of 32% for DS 1.5, 30% for DS 1.9, and 11% for DS 3.0.     

HUMAN BEHAVIOR AND RESPONSE TOWARDS STORM HAZARD IN WEST VANCOUVER,BRITISH COLUMBIA1

ABSTRACT: Human behavior and response towards storm hazard are examined in this study. One-hundred and twenty flood victims in West Vancouver, British Columbia, were administered a sentence completion test to ascertain how these victims behaved and responded to the flash flood of July 12, 1972. The sentence completion test consisted of 11 sentence stems. Seven of these dealt with the respondent's behavior toward the storm experience, and four with the issue of internal vs. external locus of control. Completions of the seven sentence stems at three time situations showed that West Vancouverites expressed the normal reactions of fear and anxiety before the storm; showed negative emotions during the storm; and displayed a feeling of mutual cooperation after the storm. Completions of the remaining four sentence stems revealed that West Vancouverites emphasized the importance of internal factors, viz., autonomy, drive and hard work, in controlling their lives. They recognized God as a benevolent - protective and almighty - powerful figure. Forty percent of the respondents did not believe in luck, while 8 percent regarded it as important in their lives. One might conclude that West Vancouverites could cope with natural hazardous threats better than those who rely on external factors in directing their lives.     

Ammonium,Nitrate and Nitrite Nitrogen and Nitrate Reductase Enzyme Activity in Groundnut (Arachis hypogaea L.) Fields After Diazinon,Imidacloprid and Lindane Treatments

Impacts of diazinon (O,O-diethyl O-2-isopropyl-6-methylpyrimidin-4-yl phosphorothioate), imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine] and lindane (1,2,3,4,5.6-hexachlorocyclohexane) treatments on ammonium, nitrate, and nitrite nitrogen and nitrate reductase enzyme activities were determined in groundnut (Arachis hypogaea L.) field for three consecutive years (1997 to 1999). Diazinon was applied for both seed- and soil-treatments but imidacloprid and lindane were used for seed treatments only at recommended rates. Diazinon residues persisted for 60 days in both the cases. Average half-lives (t_1/2) of diazinon were found 29.3 and 34.8 days respectively in seed and soil treatments. In diazinon seed treatment, NH₄ ⁺, NO₃ ^?, and NO₂ ^? nitrogen and nitrate reductase activity were not affected. Whereas, diazinon soil treatment indicated significant increase in NH₄ ⁺-N in a 1-day sample, which continued until 90 days. Some declines in NO₃ ^?N were found from 15 to 60 days. Along with this decline, significant increases in NO₂ ^?N and nitrate reductase activity were found between 1 and 30 days. Imidacloprid and lindane persisted for 90 and 120 days with average half-lives (t_1/2) of 40.9 and 53.3 days, respectively. Within 90 days, imidacloprid residues lost by 73.17% to 82.49% while such losses for lindane residues were found 78.19% to 79.86 % within 120 days. In imidacloprid seed-treated field, stimulation of NO₃ ^?N and the decline in NH₄ ⁺NO₂ ^?-N and nitrate reductase enzyme activity were observed between 15 to 90 days. However, lindane seed treatment indicated significant increases in NH₄ ⁺-N, NO₂ ^?-N and nitrate reductase activity and some adverse effects on NO₃ ^?N between 15 and 90 days.     

Spore inhibition-based enzyme substrate assay for monitoring of aflatoxin M1 in milk

A spore germination-based concept and its transformation into a field level prototype for monitoring aflatoxin M₁ (AFM₁) in milk was developed. Initially, 15 strains of Bacillus spp. procured from different culture collection were screened for AFM₁ sensitivity using spot assay and marker strain showing inhibition at 0.5 ppb was selected based upon maximum zone of inhibition. The selected strain B. megaterium 2949 was further screened for different enzymes activities and subsequently its spores were produced to an extent of 73.13% ± 3.197% in newly developed sporulation medium containing beef extract (0.0075% ± 0.0004%), yeast extract (0.015% ± 0.001%), peptone (0.0375% ± 0.0016%), and sodium chloride (0.0375% ± 0.0018%). A spore germination-based concept/ assay was optimized by immobilizing spores in eppendorf with pretreated milk (80°C/15 min) containing germinant and chromogenic substrate followed by incubation at 37°C. The appearance of sky blue color within real time of 45 min indicated spores germination and release of specific marker enzyme such as acetyl esterase and its specific action on chromogenic substrate which demonstrates absence of AFM₁ in milk. However, if there was no color change, presence of AFM₁ at 0.5 ppb MRL was denoted by Codex. The developed concept on AFM₁ detection was validated and a correlation of 0.97 was established with AOAC approved Charm 6602 and ELISA at Codex MRL with minimal false positive and negative results. The cost effective test has potential application in dairy farms, manufacturing, and R&D units for routine monitoring of AFM₁ in milk.