Humic acid-vitamin agar,a new medium for the selective isolation of soil actinomycetes

A new medium, designated HV agar, containing soil humic acid as the sole source of carbon and nitrogen was developed.The HV agar was superior to other currently used media, including colloidal chitin agar, glycerol-arginine agar and starch-casein-nitrate agar, for the isolation and enumeration of soil actinomycetes: It allowed the growth of the largest numbers of actinomycete colonies belonging to each genus of Streptomyces, Micromonospora, Microbispora, Streptosporangium, Nocardia, Dactylosporangium, Microtetraspora and Thermomonospora on the plate, while restricting the development of true bacteria. The HV agar supported adequate growth and good sporulation for these actinomycetes.Even when spore suspensions were used as the inoculum, the HV agar produced remarkably larger numbers of actinomycetes, especially strains of the genera Micromonospora, Microbispora, Streptosporangium, Dactylosporangium and Saccharomonospora, than did glycerol-arginine agar. It was found that the spores of these actinomycetes were activated upon germination by treatment at 20°C for 30 min with a O.2% solution of humic acid prior to incubation.     

Colonization of plant rhizosphere by actinomycetes of different genera

The survival of environmental isolates of actinomycetes introduced with the seeds of agricultural plants in root-free soil and in the rhizosphere and rhizoplane was studied. Different strategies of colonization of the rhizosphere were revealed for the representatives of the genera Streptomyces, Micromonospora, and Streptosporangium, organisms typical for the moderate climate rhizosphere. The plants of winter rye (Secale cereale L.) inoculated with actinomycetes were shown to have growth advantages, while the cow clover plants (Trifolium pratense L.) had no growth advantages compared to uninoculated plants. The role of the plant component in the interaction with mycelial prokaryotes is discussed.     

Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv. glycine

In this study, actinomycetes from roots and rhizospheric soils of leguminous plants were isolated using starch casein agar supplemented with antifungal and antibacterial antibiotics. Three hundred and seventeen actinomycetes were isolated with 77 isolates obtained from plant roots and 240 isolates from rhizospheric soils. Analysis of whole-organism hydrolysates showed that 289 strains were rich in the LL-isomer of diaminopimelic acid, a result consistent with their assignment to the streptomycetes. The remaining 28 strains were assigned to non-streptomycetes based on the presence of meso-isomer of diaminopimelic acid in cell wall. Sixty-four isolates (20.2 %) showed antagonistic activity against soybean pathogen Xanthomonas campestris pv. glycine by agar overlay method. Isolate RM 365 showed the highest activity with an inhibition ratio of 3.79, with no inhibitory activity on the growth of Rhizobium japonicum TISTR 079, Rhizobium sp. TISTR 061 and Rhizobium sp. TISTR 063. The 16S rRNA gene sequence analysis revealed that isolate RM 365 shared 99.28 % similarity to Streptomyces caeruleatus GIMN4^T (GQ329712). In addition, isolates which contained meso-DAP were also identified by 16S rRNA gene sequence analysis. The results showed that they were members of the genus Amycolatopsis, Isoptericola, Micromonospora, Microbispora, Nocardia, Nonomuraea, Promicromonospora and Pseudonocardia.     

Haloalkaliphilic actinomycetes in soils of Mongolian desert steppes

The abundance of actinomycetes isolated from the soils of Mongolian desert steppes varies from several thousand to hundreds of thousands of CFU/g soil, depending on soil type and isolation medium. Eight actinomycete genera have been found in these soils: Streptomyces, Micromonospora, Saccharopolyspora, Actinomadura, Microtetraspora, Thermomonospora, Nocardia, and Dactylosporangium. The streptomycete complexes of brown desert-steppe and gray-brown desert alkaline soils include halophilic, alkaliphilic, and haloalkaliphilic species that grow most successfully on the media with a salt concentration of 5% and pH 8–9.     

Oligotrophy is Helpful for the Isolation of Bioactive Actinomycetes

It is necessary to develop new methods for the isolation of unknown actinomycetes from soils. To evaluate the effects of oligotrophic medium on the isolation of soil actinomycetes and develop a new isolation method, the Gause’s synthetic medium was diluted to one tenth the recommended concentration in the present study. Soil dilution plate technique was used to isolate actinomycetes from the soil samples. Oligotrophy decreased actinomycete and streptomycete counts, as well as the number of antagonistic actinomycete species. Oligotrophy also decreased the number of actinomycete species in five samples. Some actinomycete species were cultured only on the oligotrophic medium, whereas other species could not be cultured. Oligotrophy decreased actinomycete counts more significantly for soils with organic matter content >40 g/kg. We used 16S rRNA sequence analysis to identify 22 actinomycete species that were only cultured on the oligotrophic medium. Oligotrophic medium was helpful for the isolation of Streptomyces spp., Micromonospora spp. and Streptosporangium spp. Slightly more than 80 % of the identified actinomycete species were biologically active. Therefore, we could draw a conclusion that oligotrophic medium could be helpful for the discovery of new antibiotic producers and the exploitation and utilization of new, biologically active compounds.     

Microbiological differences between limed and unlimed soils and their relationship with cavity spot disease of carrots (Daucus carota L.) caused by Pythium coloratum in Western Australia

Application of lime (4000 kg ha^-1) to a soil used for commercial carrot production (pH 6.9) significantly (p<0.05) reduced the incidence of cavity spot disease of carrots compared to unlimed soil (pH 5.1). It significantly (p<0.01) increased soil microbial activity as measured by the hydrolysis of fluorescein diacetate and arginine ammonification. The application of lime resulted in a significant (p<0.01) increase in the total numbers of colony forming units (efu) of aerobic bacteria, fluorescent pseudomonads, Gram negative bacteria, actinomycetes and a significant (p<0.01) decrease in the cfu of filamentous fungi and yeasts compared to unlimed soil. Liming also increased the cfu of non-streptomycete actinomycetes rarely reported in similar studies. These non-streptomycete actinomycetes were estimated and isolated using polyvalent Streptomyces phages and the dry heat technique to reduce the dominance of streptomycetes on isolation plates. The non-streptomycete actinomycetes isolated included species of Actinoplanes, Micromonospora, Streptoverticillium, Nocardia, Rhodococcus, Microbispora, Actinomadura, Dactylosporangium and Streptosporangium. The numbers of actinomycetes antagonistic to Pythium coloratum, a causal agent of cavity spot disease of carrots increased in soil amended with lime. Application of lime also reduced the isolation frequency of P. coloratum from asymptomatic carrot roots grown in soil artificially infested with the pathogen, 3, 4 and 5 weeks after sowing.     

Culturable actinomycetes from desert ecosystem in northeast of Qinghai-Tibet Plateau

In this study, 53 actinomycetes strains were isolated from desert ecosystems located in northeast of Qinghai-Tibet Plateau and grouped into four RFLP patterns. Twenty-six actinomycetes with obvious morphology differences were chosen for phylogenetic diversity study and antimicrobial testing. As a result, the 16S rRNA gene sequencing showed that these strains belonged to Streptomyces, Micromonospora, Saccharothrix, Streptosporangium and Cellulomonas, and that most of the strains had antimicrobial bioactivity. The PKS and NRPS genes detection indicated diversified potential bioactive products of actinomycetes in this ecosystem. Among these strains, Sd-31 was chosen to study the bioactive products using HPLC-MS because of its optimum antimicrobial bioactivity. The result showed that it might produce Granatomycin A, Granatomycin C, and an unknown compound.     

Actinomycetes isolated from medicinal plant rhizosphere soils: diversity and screening of antifungal compounds,indole-3-acetic acid and siderophore production

A total of 445 actinomycete isolates were obtained from 16 medicinal plant rhizosphere soils. Morphological and chemotaxonomic studies indicated that 89% of the isolates belonged to the genus Streptomyces, 11% were non-Streptomycetes: Actinomadura sp., Microbispora sp., Micromonospora sp., Nocardia sp, Nonomurea sp. and three isolates were unclassified. The highest number and diversity of actinomycetes were isolated from Curcuma mangga rhizosphere soil. Twenty-three Streptomyces isolates showed activity against at least one of the five phytopathogenic fungi: Alternaria brassicicola, Collectotrichum gloeosporioides, Fusarium oxysporum, Penicillium digitatum and Sclerotium rolfsii. Thirty-six actinomycete isolates showed abilities to produce indole-3-acetic acid (IAA) and 75 isolates produced siderophores on chrome azurol S (CAS) agar. Streptomyces CMU-PA101 and Streptomyces CMU-SK126 had high ability to produced antifungal compounds, IAA and siderophores.     

Identification ofStreptomyces sp. Tc022, an endophyte inAlpinia galanga, and the isolation of actinomycin D

Some endophytic actinomycetes (120) were isolated from the roots ofAlpinia galanga. Identification of these endophytes was based on their morphology and amino acid composition of the whole-cell extract. Most isolates were classified aStreptomyces sp. (82), with the remainder belonging toNocardia sp. (11),Microbispora sp. (3) andMicromonospora sp. (2). Eight isolates were unclassified and 14 were lost during subculture. The strain identified as endophyticStreptomyces sp. Tc022 strongly inhibitedColletotrichum musae andCandida albicans. This endophyte was cultured, the agar was extracted with organic solvent and the extract was purified on a column of silica gel to give a major component, which was identified to be actinomycin D on the basis of spectroscopic dat Actinomycin D showed antifungal activity againstColletotrichum musae andCandida albicans with the MIC of 10 and 20 mg ml^?1, respectively.     

Efficacy of artificial humic acid as a selective nutrient in HV agar used for the isolation of soil actinomycetes

The efficacies of various kinds of humic acid, as the source of carbon and nitrogen in HV agar reported in the previous paper, were compared to the selective isolation of soil actinomycetes. The 4 types of natural humic acid, A, B, P and Rp were prepared from different soils, and 3 kinds of artificial humic acid were made from carbohydrates and urea in our laboratory.Among the natural humic acids, type Rp, which has been reported to be associated with an initial state of humification in natural conditions, showed the greatest efficacy.However, one of the artificial humic acids, which was prepared from glucose and urea, was considered to be superior to the Rp natural humic acid: 1) The HV agar containing this artificial humic acid (HV-glucose HA agar) produced the same large number of actinomycete colonies on the plate as that of the HV agar with type Rp soil humic acid (HV-Rp agar). 2) The HV-glucose HA agar restricted the number of bacterial colonies on the plates to one-half of that on HV-Rp agar plates. 3) The quality of natural himic acids varies, whereas artificial ones are more constant and can be made in any laboratory.     

Thermotolerant and thermophilic actinomycetes from soils of Mongolia desert steppe zone

In the actinomycete complexes of Mongolian desert soils, thermotolerant and thermophilic actinomycetes were found in high abundance, exceeding that of the mesophilic forms. Among the thermotolerant members of the order Actinomycetales, Streptomyces, Micromonospora, Actinomadura, and Streptosporangium species were most widespread in desert soils. Experiments with soil microcosms demonstrated that thermophilic actinomycetes in desert soils grew, developed, and formed mycelia of the length comparable to that of the mesophilic forms of actinomycetes. Molecular biological investigation of the samples of desert steppe soils by denaturing gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization (FISH) revealed members of the phylum Actinobacteria. FISH analysis revealed that the biomass of the metabolically active mycelial actinobacteria in the prokaryotic community of Mongolian desert soils exceeded that of the unicellular Actinobacteria.     

Isolation of endophytic actinomycetes from selected plants and their antifungal activity

The isolation of endophytic actinomycetes from surface-sterilized tissues of 36 plant species was made using humic acid–vitamin (HV) agar as a selection medium. Of the 330 isolates recovered, 212 were from roots, 97 from leaves and 21 isolates from stems with a prevalence of 3.9, 1.7 and 0.3%, respectively. Identification of endophytic actinomycetes was based on their morphology and the amino acid composition of the whole-cell extract. Most isolates were classified as Streptomyces sp. (n = 277); with the remainder belonging to Microbispora sp. (n = 14), Nocardia sp. (n = 8) and Micromonospora sp. (n = 4). Four isolates were unclassified and 23 were lost during subculture. The most prevalent group of isolates were the Streptomyces sp. occurring in 6.4% of the tissue samples of Zingiber officinale. Scanning electron microscopy investigation of this plant revealed that 7.5% of the root and 5% of the leaf samples contained endophytes. Three of the Streptomyces sp. isolates strongly inhibited Colletotrichum musae, five were very active against Fusarium oxysporum and two strongly inhibited growth of both test fungi.     

Application of a method incorporating differential centrifugation for selective isolation of motile actinomycetes in soil and plant litter

The present paper describes a simple enrichment technique which enables rapid and selective isolation of diverse zoosporic actinomycete genera directly from soil and plant litter. This technique, designated the rehydration and centrifugation (RC) method, consists of immersing the air-dried source material in 10 mM phosphate buffer containing 10% soil extract, letting the preparation stand at 30 °C for 90 min, followed by centrifugation of the fluid at 1,500×g for 20 min. Portions of the supernatant containing actinomycete zoospores are plated on the humic acid-vitamin agar which is supplemented with nalidixic acid and trimethoprim as the selective inhibitors for Gram-negative bacteria and bacilli. The phosphate buffer-soil extract solution significantly promoted liberation of motile zoospores from the source material. The centrifugation stage greatly eliminated streptomycetes and other non-motile actinomycetes from the liquid phase, thereby facilitating selective growth of rare, motile actinomycetes on the isolation plates subsequent to inoculation. Ten different soil and leaf-litter samples, taken from fields, forests, and stream banks, were examined. The RC method consistently achieved preferential isolation of motile actinomycetes in all samples, which accounted for 37–86% of the total microbial population recovered. The most frequently isolated motile actinomycetes were Actinoplanes and Dactylosporangium. Strains of Actinokineospora, Catenuloplanes and Kineosporia were also recovered, depending on the nature of the samples examined. Other motile actinomycetes that were occasionally isolated in small numbers included Actinosynnema, Geodermatophilus and Sporichthya.     

半干旱黄土丘陵区纯林土壤腐殖质异化特征及与其他性质的关系

纯林长期生长或多代连栽必然会导致土壤腐殖质含量和构成发生异化,探究这种异化特征及其与土壤其他性质的关系可以为纯林管理或混交改造提供科学依据。通过对半干旱黄土丘陵区南泥湾林场8种典型纯林土壤腐殖质及其他性质进行系统检测,结果表明:(1)侧柏林土壤腐殖质含量最高(34.61 g/kg),腐殖化程度和稳定性一般;白榆和白桦林土壤的腐殖质含量中等(19.69—23.58 g/kg)、腐殖化程度和稳定性最佳;茶条槭和小叶杨林土壤的腐殖质含量(20.59—22.53 g/kg)和构成均为中等水平;油松、沙棘和刺槐林土壤的腐殖质质量较低(11.77—13.81 g/kg),且腐殖化程度较低,稳定性相对最差;(2)与胡敏酸含量存在显著相互促进作用(P0.05)的土壤性质为CEC、N、微生物量和蛋白酶活性(相关系数0.769—0.926,下同),存在显著相互抑制作用的为有效Cu(-0.793);与富啡酸存在显著相互促进作用的为N、CEC、微生物量、蔗糖酶和磷酸酶活性(0.836—0.955),存在显著相互抑制作用的为有效Cu(-0.822);与胡敏素存在显著相互促进作用的为N、CEC、微生物量、磷酸酶活性和有效Zn(0.766—0.951),存在显著相互抑制作用的为脱氢酶活性(-0.784)。(3)腐殖质构成与其他性质的相关性均不显著(P0.05),其中相对有利于提高胡敏酸/腐殖酸含量之比的土壤性质为蛋白酶、蔗糖酶和过氧化氢酶活性,而不利的是脱氢酶活性;相对有利于提高胡敏酸/富啡酸含量之比的为速效K、CEC和脲酶活性,而不利的是脱氢酶活性。(4)总体而言土壤腐殖质含量较之腐殖质构成与其他性质之间具有更大的相关性;向土壤增施N肥可以促进腐殖质的形成,增加K肥则有利于腐殖质构成的改善,而通过混交改造或增加林下植被是促进纯林土壤腐殖质化过程和解决土壤退化的根本措施。     

Growth Promotion of Fluorescent Pseudomonads and Control of Potato Common Scab in Field Soil with Non-antibiotic Actinomycete-biofertilizer

Field studies on the control of potato common scab were done with non-antibiotic Actinomycete biofertilizer, which was manufactured from swine feces with coprophilous actinomycetes and acidified to pH 5.0 with sulfuric acid. The field soil at pH 5.0 was mixed with 0.56 kg of the acidified biofertilizer per subplot (1.0 x 1.2 m) to a depth of the 15 cm. The scab severity, in terms of the percent coverage of the total surface area of potato tubers, was 10.7%, in the control, while the lesions were slight in the biofertilizer subplot, scab severity being only 2.30/0, and the potato production was increased 3 times with the biofertilizer. The viable counts of Streptomyces scabies in the biofertilizer subplot decreased to 1 × 10³ per gram soil from 5 × 10⁵ by 12 weeks, while the inhabitant fluorescent pseudomonads increased to 2.4 × 10⁷ per gram soil from 3 × 10⁴, and the increased fluorescent pseudomonads were antagonistic to the pathogenic strains.     

Catalytic properties of three lactate dehydrogenases from potato tubers (Solanum tuberosum)

Two l-lactate dehydrogenase isoenzymes and one dl-lactate dehydrogenase could be separated from potato tubers by polyacrylamide-gel electrophoresis. The enzymes are specific for lactate, while β-hydroxybutyric acid, glycolic acid, and glyoxylic acid are not oxidized. Their pH optima are pH 6.9 for the oxidation and 8.0 for the reduction reaction.The K_m values for l-lactate for the two isoenzymes are 2.00 × 10^?2 and 1.82 × 10^?2, m. In the reverse reaction the affinities for pyruvate are 3.24 × 10^?4 and 3.34 × 10^?4, m. Both enzymes have similar affinities for NAD and NADH (3.00 × 10^?4; 4.00 × 10^?4, and 8.35 × 10^?4; 5.25 × 10^?4, m).The dl-lactate oxidoreductase may transfer electrons either to NAD or N-methyl-phenazinemethosulfate. The K_m values of this enzyme for l-lactate are 4.5 × 10^?2, m and for d-lactate 3.34 × 10^?2, m. Its affinity for pyruvate is 4.75 × 10^?4, m. The enzyme is inhibited by excess NAD (K_m = 1.54 × 10^?4, M) and has an affinity toward NADH (K_m = 5.00 × 10^?3, M) which is about one tenth of that of the two isoenzymes of l-lactate dehydrogenase.     

Spore formation in the Actinoplanaceae (Actinomycetales)

Spore development in four genera, Actinoplanes, Dactylosporangium, Planomonospora, and Streptosporangium, was studied by transmission and scanning electron microscopy. Actinoplanes and Streptosporangium formed spores by fragmentation of a hypha within its expanded outer sheath, as do many other actinomycetes. Dactylosporangium and Planomonospora formed spores endogenously by development of wall material within the parent hypha. In this respect, they resembled the genera Actinobifida and Thermoactinomyces. The term sporangium has therefore been used to describe structures which are not homologous. It was suggested that the term should be confined to structures in which endogenous spore formation occurs.     

Changes in methane oxidation activity and methanotrophic community composition in saline alkaline soils

The soil of the former Lake Texcoco is a saline alkaline environment where anthropogenic drainage in some areas has reduced salt content and pH. Potential methane (CH₄) consumption rates were measured in three soils of the former Lake Texcoco with different electrolytic conductivity (EC) and pH, i.e. Tex-S1 a >18 years drained soil (EC 0.7 dS m^?1, pH 8.5), Tex-S2 drained for ~10 years (EC 9.0 dS m^?1, pH 10.3) and the undrained Tex-S3 (EC 84.8 dS m^?1, pH 10.3). An arable soil from Alcholoya (EC 0.7 dS m^?1, pH 6.7), located nearby Lake Texcoco was used as control. Methane oxidation in the soil Tex-S1 (lowest EC and pH) was similar to that in the arable soil from Alcholoya (32.5 and 34.7 mg CH₄ kg^?1 dry soil day^?1, respectively). Meanwhile, in soils Tex-S2 and Tex-S3, the potential CH₄ oxidation rates were only 15.0 and 12.8 mg CH₄ kg^?1 dry soil day^?1, respectively. Differences in CH₄ oxidation were also related to changes in the methane-oxidizing communities in these soils. Sequence analysis of pmoA gene showed that soils differed in the identity and number of methanotrophic phylotypes. The Alcholoya soil and Tex-S1 contained phylotypes grouped within the upland soil cluster gamma and the Jasper Ridge, California JR-2 clade. In soil Tex-S3, a phylotype related to Methylomicrobium alcaliphilum was detected.     

Remediation of Explosive-Contaminated Soils: Alkaline Hydrolysis and Subcritical Water Degradation

Alkaline hydrolysis and subcritical water degradation were investigated as ex-situ remediation processes to treat explosive-contaminated soils from military training sites in South Korea. The addition of NaOH solution to the contaminated soils resulted in rapid degradation of the explosives. The degradation of explosives via alkaline hydrolysis was greatly enhanced at pH ≥12. Estimated pseudo-first-order rate constants for the alkaline hydrolysis of 2,4-dinitrotoluene (DNT), 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in contaminated soil at pH 13 were (9.6?±?0.1)×10^?2, (2.2?±?0.1)×10^?1, and (1.7?±?0.2)×10^?2 min^?1, respectively. In the case of subcritical water degradation, the three explosives were completely removed at 200–300°C due to oxidation at high temperatures and pressures. The degradation rate increased as temperature increased. The pseudo-first-order rate constants for DNT, TNT, and RDX at 300°C were (9.4?±?0.8)×10^?2, (22.8?±?0.3)×10^?2, and (16.4?±?1.0)×10^?2, respectively. When the soil-to-water ratio was more than 1:5, the extent of alkaline hydrolysis and subcritical water degradation was significantly inhibited.     

Impact of commonly used agrochemicals on bacterial diversity in cultivated soils

The effects of three selected agrochemicals on bacterial diversity in cultivated soil have been studied. The selected agrochemicals are Cerox (an insecticide), Ceresate and Paraquat (both herbicides). The effect on bacterial population was studied by looking at the total heterotrophic bacteria presence and the effect of the agrochemicals on some selected soil microbes. The soil type used was loamy with pH of 6.0–7.0. The soil was placed in opaque pots and bambara bean (Vigna subterranean) seeds cultivated in them. The agrochemicals were applied two weeks after germination of seeds at concentrations based on manufacturer’s recommendation. Plant growth was assessed by weekly measurement of plant height, foliage appearance and number of nodules formed after one month. The results indicated that the diversity index (Di) among the bacteria populations in untreated soil and that of Cerox-treated soils were high with mean diversity index above 0.95. Mean Di for Ceresate-treated soil was 0.88, and that for Paraquattreated soil was 0.85 indicating low bacterial populations in these treatment-type soils. The study also showed that application of the agrochemicals caused reduction in the number of total heterotrophic bacteria population sizes in the soil. Ceresate caused 82.50% reduction in bacteria number from a mean of 40 × 10⁵ cfu g⁻¹ of soil sample to 70 × 10⁴ cfu g⁻¹. Paraquat-treated soil showed 92.86% reduction, from a mean of 56 × 10⁵ cfu g⁻¹ to 40 × 10⁴ cfu g⁻¹. Application of Cerox to the soil did not have any remarkable reduction in bacterial population number. Total viable cell count studies using Congo red yeast-extract mannitol agar indicated reduction in the number of Rhizobium spp. after application of the agrochemicals. Mean number of Rhizobium population numbers per gram of soil was 180 × 10⁴ for the untreated soil. Cerox-treated soil recorded mean number of 138 × 10⁴ rhizobial cfu g⁻¹ of soil, a 23.33% reduction. Ceresate- and Paraquat-treated soils recorded 20 × 10⁴ and 12 × 10⁴ cfu g⁻¹ of soil, respectively, representing 88.89% and 93.33% reduction in Rhizobium population numbers. Correspondingly, the mean number of nodules per plant was 44 for the growth in untreated soil, 30 for the plant in the Cerox-treated soil, 8 for the plant in Paraquat-treated soil and 3 for the plant in Ceresate-treated soil. The study has confirmed detrimental effect of insecticide on bacterial populations in the soil. Total heterotrophic counts, rhizobial counts as well as the number of nodules of all samples taken from the chemically treated soils were all low as compared to values obtained for the untreated soil. However, the effect of the insecticide was minimal in all cases as compared to the effects of the herbicides on the soil fauna. Indiscriminate use of agrochemicals on farms can therefore affect soil flora and subsequently food production.