CHITINASE AS THE MOST IMPORTANT SECONDARY METABOLITES OF STREPTOMYCES BACTERISIJSIT Editor
Fungal phytopathogens pose serious problems worldwide in the cultivation of economi cally
important plants.
Chemical fungicides are extensively used in current agriculture.However, excessive use of chemical
fungicides in agriculture has led to deteriorating human health , environmental pollution, damaged to
ecosystem and development of pathogen resistance to fungicide.
Because of the worsening problems in fungal disease control , a serious search is needed to identify
alternative methods for plant protection, which are less dependent on chemicals and are more
environmentally friendly. Microbial antagonists are widely used for the biocontrol of fungal plant diseases.
Many species of actinomycates, particulary those belonging to the genus sterptomyces, are well known as
antifungal biocontrol agents that inhibit several plant pathogenic fungi.
Another way biological control has been developed as an alternative of chemicals to tock with plant
pathogenic fungi. Considering high presence of chitin in fungal cell wall, chitinase enzyme is camped as an
effective biocontrol agent against phytopathogenic fungi. Streptomyces bacteria are able to produce various chitinase enzymes, chitinases produced by streptomyces belong to the families 18 and 19 glycosyl hydrolases.
The antifungal activity is mostly shown by fomily 19 Chitinases. In comparison with bacterial family 18
chitinases, the specific hydrolyzing activity of chitinase 19 against soluble and in soluble chitinous substrates
has been markedly higher. Considering the importance of family to investigate antifungal potential of
streptomyces bacteria isolated from east Azarbijan region soils based on molecular identification of family 19
chitinase. encoding gene in these bacteria.
To aim the purpose 110 soil samples were collected from East Azarbaijan and 310 strepomyces
isolates were selected using macroscopic and microscopic observations. DNA genomic of all of the isolates
were extracted and PCR reactions was done using chitinase 19 designed primers as marker.
Totally isolates were selected with molecular selection and antagonistic test were done. One of the isolates
exhibit the most strong antifungal activity.
The strain was identified using 16srDNA gene, and the chitinase encoding gene were amplified partially to
prove the PCR selection. Finally the bacterium were introduced as potentially biological fertilizer.
MOLECULAR ANALYSIS OF BACTERIAL GENE CODING CHITINASE ENZYMES, FAMILY 19 STR...IJSIT Editor
Fungal phytopathogens pose serious problems worldwide in the cultivation of economically
important plants.
Chemical fungicides are extensively used in current agriculture.However, excessive use of chemical
fungicides in agriculture has led to deteriorating human health , environmental pollution, damaged to
ecosystem and development of pathogen resistance to fungicide.
Because of the worsening problems in fungal disease control , a serious search is needed to identify
alternative methods for plant protection, which are less dependent on chemicals and are more
environmentally friendly. Microbial antagonists are widely used for the biocontrol of fungal plant diseases.
Many species of actinomycates, particulary those belonging to the genus sterptomyces, are well known as
antifungal biocontrol agents that inhibit several plant pathogenic fungi.
Another way biological control has been developed as an alternative of chemicals to tock with plant
pathogenic fungi. Considering high presence of chitin in fungal cell wall, chitinase enzyme is camped as an
effective biocontrol agent against phytopathogenic fungi. Streptomyces bacteria are able to produce various chitinase enzymes, chitinases produced by streptomyces belong to the families 18 and 19 glycosyl hydrolases.
The antifungal activity is mostly shown by fomily 19 Chitinases. In comparison with bacterial family 18
chitinases, the specific hydrolyzing activity of chitinase 19 against soluble and in soluble chitinous substrates
has been markedly higher. Considering the importance of family to investigate antifungal potential of
streptomyces bacteria isolated from east Azarbijan region soils based on molecular identification of family 19
chitinase. encoding gene in these bacteria.
To aim the purpose 110 soil samples were collected from East Azarbaijan and 310 strepomyces
isolates were selected using macroscopic and microscopic observations. DNA genomic of all of the isolates
were extracted and PCR reactions was done using chitinase 19 designed primers as marker.
Totally isolates were selected with molecular selection and antagonistic test were done. One of the isolates
exhibit the most strong antifungal activity.
The strain was identified using 16srDNA gene, and the chitinase encoding gene were amplified partially to
prove the PCR selection. Finally the bacterium were introduced as potentially biological fertilizer.
Biofumigation: A Potential Aspect for Suppression of Plant-Parasitic NematodesIJEABJ
Plant-parasitic nematode cause economic loss to crops throughout the world. Biofumigation is the environmental friendly control option for the suppression of plant-parasitic as well as other pathogenic soil microbes. Glucosinolates are the main active compound present in some plants which are responsible for biofumigation process. To increase the efficiency of biofumigation selection of varieties containing more glucosinolates is highly desirable. Plant growth stage, soil temperature, soil texture, moisture, soil depth and soil microbes play important role in efficient biofumigation.
Effect of plant growth promoting rhizobacterial (PGPR) inoculation on growth ...IJEAB
Plant Growth promoting rhizobacteria are a heterogeneous group of bacteria that can be found in the rhizosphere, at root surfaces and in association with roots. They benefit plants through Production of plant hormones, such as auxins, asymbiotic N2 fixation, solubilization of mineral phosphates, antagonism against phytopathogenic microorganisms by production of antibiotics, siderophroes, Chitinase and other nutrients ability to effectively colonize roots are responsible for plant growth promotion. An experiment was conducted in the field of National Institute of Agronomic Research of Meknes. Morocco. The experiment was a completely randomized design with six replicates. There were four treatments viz. T1: (control; N0 -PGPR), T2: (N0 +2027-2), T3: (N0 +2066-7) and T4: (N0+2025-1). The results indicated that a remarkable increase in root growth, namely length, the diameter of the rod and the total chlorophyll. A total of three different bacteria colonies were isolated and proceed with in vitro screening for plant growth promoting activities; phosphate solubilization, nitrogen fixation, indole acetic acid (IAA), ammonia production and antimicrobial enzymes (cellulose, chitinase and protease) activity. Among the three bacterial strains, all bacterial strains are able to produce ammonia, IAA production and nitrogen fixation activity, one strain phosphate solubilizing activity, two strain are able to produce cellulase syntheses, Protease activity and Chitinase activity.
The objectives of this study are: (i): To investigate and recognize the internal and abnormalities impacts induced by phytoplasma infection in the tomato host according to recent studies have shown that the association between plants and phytoplasmas can result in anatomical alteration in phloem tissues of infected plants, and great differences between healthy and diseased samples using microscopic examination of longitudinal, cross or ultra-thin sections of leaf blade, leaf petiole and stem. (ii): To determine the efficiency of different techniques toward production of phytoplasma-free tomato plantlets and mitigation of phytoplasma disease.
Prevalence, occurrence and biochemical characterization of Xanthomonas campes...INNS PUBNET
Xanthomonas campestris pv. vesicatoria the causal organism of bacterial spot in tomato results in heavy losses both in the form of quality and. In this study a survey was carried out to report the incidence of bacterial spot disease of tomato in district Swat. We reported maximum disease incidence in tehsil Kabal (71.66%), followed by Charbagh (61.66%) and Barikot (58.33%). For resistant screening a total of 13 tomato germplasms were screened against the disease. The foliar severity ranged from 3.33% to 73.33%, while severity for fruits was ranged from 18.33% to 30.66%. In case of phenotypic data the highest numbers of fruits obtained were 34, plant height 79.5cm and fruit weight was 470 grams/ten tomatoes. While the lowest average numbers of fruits were 6.67, plant height 45.7cm and fruit weight recorded was 215.67 grams/ten tomatoes. Line 1288 showed highest level of resistance followed by Red-stone. However, line 9708 showed highest susceptibility when exposed to artificial inoculation. Our study showed that bacterial spot is a major issue in some part of Pakistan and germplasm screening are linked to increased host resistance and could offer an important contribution to future integrated bacterial spot management programs.
CHITINASE AS THE MOST IMPORTANT SECONDARY METABOLITES OF STREPTOMYCES BACTERISIJSIT Editor
Fungal phytopathogens pose serious problems worldwide in the cultivation of economi cally
important plants.
Chemical fungicides are extensively used in current agriculture.However, excessive use of chemical
fungicides in agriculture has led to deteriorating human health , environmental pollution, damaged to
ecosystem and development of pathogen resistance to fungicide.
Because of the worsening problems in fungal disease control , a serious search is needed to identify
alternative methods for plant protection, which are less dependent on chemicals and are more
environmentally friendly. Microbial antagonists are widely used for the biocontrol of fungal plant diseases.
Many species of actinomycates, particulary those belonging to the genus sterptomyces, are well known as
antifungal biocontrol agents that inhibit several plant pathogenic fungi.
Another way biological control has been developed as an alternative of chemicals to tock with plant
pathogenic fungi. Considering high presence of chitin in fungal cell wall, chitinase enzyme is camped as an
effective biocontrol agent against phytopathogenic fungi. Streptomyces bacteria are able to produce various chitinase enzymes, chitinases produced by streptomyces belong to the families 18 and 19 glycosyl hydrolases.
The antifungal activity is mostly shown by fomily 19 Chitinases. In comparison with bacterial family 18
chitinases, the specific hydrolyzing activity of chitinase 19 against soluble and in soluble chitinous substrates
has been markedly higher. Considering the importance of family to investigate antifungal potential of
streptomyces bacteria isolated from east Azarbijan region soils based on molecular identification of family 19
chitinase. encoding gene in these bacteria.
To aim the purpose 110 soil samples were collected from East Azarbaijan and 310 strepomyces
isolates were selected using macroscopic and microscopic observations. DNA genomic of all of the isolates
were extracted and PCR reactions was done using chitinase 19 designed primers as marker.
Totally isolates were selected with molecular selection and antagonistic test were done. One of the isolates
exhibit the most strong antifungal activity.
The strain was identified using 16srDNA gene, and the chitinase encoding gene were amplified partially to
prove the PCR selection. Finally the bacterium were introduced as potentially biological fertilizer.
MOLECULAR ANALYSIS OF BACTERIAL GENE CODING CHITINASE ENZYMES, FAMILY 19 STR...IJSIT Editor
Fungal phytopathogens pose serious problems worldwide in the cultivation of economically
important plants.
Chemical fungicides are extensively used in current agriculture.However, excessive use of chemical
fungicides in agriculture has led to deteriorating human health , environmental pollution, damaged to
ecosystem and development of pathogen resistance to fungicide.
Because of the worsening problems in fungal disease control , a serious search is needed to identify
alternative methods for plant protection, which are less dependent on chemicals and are more
environmentally friendly. Microbial antagonists are widely used for the biocontrol of fungal plant diseases.
Many species of actinomycates, particulary those belonging to the genus sterptomyces, are well known as
antifungal biocontrol agents that inhibit several plant pathogenic fungi.
Another way biological control has been developed as an alternative of chemicals to tock with plant
pathogenic fungi. Considering high presence of chitin in fungal cell wall, chitinase enzyme is camped as an
effective biocontrol agent against phytopathogenic fungi. Streptomyces bacteria are able to produce various chitinase enzymes, chitinases produced by streptomyces belong to the families 18 and 19 glycosyl hydrolases.
The antifungal activity is mostly shown by fomily 19 Chitinases. In comparison with bacterial family 18
chitinases, the specific hydrolyzing activity of chitinase 19 against soluble and in soluble chitinous substrates
has been markedly higher. Considering the importance of family to investigate antifungal potential of
streptomyces bacteria isolated from east Azarbijan region soils based on molecular identification of family 19
chitinase. encoding gene in these bacteria.
To aim the purpose 110 soil samples were collected from East Azarbaijan and 310 strepomyces
isolates were selected using macroscopic and microscopic observations. DNA genomic of all of the isolates
were extracted and PCR reactions was done using chitinase 19 designed primers as marker.
Totally isolates were selected with molecular selection and antagonistic test were done. One of the isolates
exhibit the most strong antifungal activity.
The strain was identified using 16srDNA gene, and the chitinase encoding gene were amplified partially to
prove the PCR selection. Finally the bacterium were introduced as potentially biological fertilizer.
Biofumigation: A Potential Aspect for Suppression of Plant-Parasitic NematodesIJEABJ
Plant-parasitic nematode cause economic loss to crops throughout the world. Biofumigation is the environmental friendly control option for the suppression of plant-parasitic as well as other pathogenic soil microbes. Glucosinolates are the main active compound present in some plants which are responsible for biofumigation process. To increase the efficiency of biofumigation selection of varieties containing more glucosinolates is highly desirable. Plant growth stage, soil temperature, soil texture, moisture, soil depth and soil microbes play important role in efficient biofumigation.
Effect of plant growth promoting rhizobacterial (PGPR) inoculation on growth ...IJEAB
Plant Growth promoting rhizobacteria are a heterogeneous group of bacteria that can be found in the rhizosphere, at root surfaces and in association with roots. They benefit plants through Production of plant hormones, such as auxins, asymbiotic N2 fixation, solubilization of mineral phosphates, antagonism against phytopathogenic microorganisms by production of antibiotics, siderophroes, Chitinase and other nutrients ability to effectively colonize roots are responsible for plant growth promotion. An experiment was conducted in the field of National Institute of Agronomic Research of Meknes. Morocco. The experiment was a completely randomized design with six replicates. There were four treatments viz. T1: (control; N0 -PGPR), T2: (N0 +2027-2), T3: (N0 +2066-7) and T4: (N0+2025-1). The results indicated that a remarkable increase in root growth, namely length, the diameter of the rod and the total chlorophyll. A total of three different bacteria colonies were isolated and proceed with in vitro screening for plant growth promoting activities; phosphate solubilization, nitrogen fixation, indole acetic acid (IAA), ammonia production and antimicrobial enzymes (cellulose, chitinase and protease) activity. Among the three bacterial strains, all bacterial strains are able to produce ammonia, IAA production and nitrogen fixation activity, one strain phosphate solubilizing activity, two strain are able to produce cellulase syntheses, Protease activity and Chitinase activity.
The objectives of this study are: (i): To investigate and recognize the internal and abnormalities impacts induced by phytoplasma infection in the tomato host according to recent studies have shown that the association between plants and phytoplasmas can result in anatomical alteration in phloem tissues of infected plants, and great differences between healthy and diseased samples using microscopic examination of longitudinal, cross or ultra-thin sections of leaf blade, leaf petiole and stem. (ii): To determine the efficiency of different techniques toward production of phytoplasma-free tomato plantlets and mitigation of phytoplasma disease.
Prevalence, occurrence and biochemical characterization of Xanthomonas campes...INNS PUBNET
Xanthomonas campestris pv. vesicatoria the causal organism of bacterial spot in tomato results in heavy losses both in the form of quality and. In this study a survey was carried out to report the incidence of bacterial spot disease of tomato in district Swat. We reported maximum disease incidence in tehsil Kabal (71.66%), followed by Charbagh (61.66%) and Barikot (58.33%). For resistant screening a total of 13 tomato germplasms were screened against the disease. The foliar severity ranged from 3.33% to 73.33%, while severity for fruits was ranged from 18.33% to 30.66%. In case of phenotypic data the highest numbers of fruits obtained were 34, plant height 79.5cm and fruit weight was 470 grams/ten tomatoes. While the lowest average numbers of fruits were 6.67, plant height 45.7cm and fruit weight recorded was 215.67 grams/ten tomatoes. Line 1288 showed highest level of resistance followed by Red-stone. However, line 9708 showed highest susceptibility when exposed to artificial inoculation. Our study showed that bacterial spot is a major issue in some part of Pakistan and germplasm screening are linked to increased host resistance and could offer an important contribution to future integrated bacterial spot management programs.
Two field experiment were run at the farms of Omar AL-Mokhtar university, in order to knew the effect of some methods used to control tomato pests on soil arthropod systemic groups in organic cultivated tomato field such as (BT, algifol, methyl salicylate, neem oil), Results indicated that application of B. thuringensis and Algifol achieved the highest performance showing 100% reduction in mites and other soil arthropods systemic groups, and gave reduction 55.5 and 54.96% in both insect and total population of soil arthropods collected with pitfall traps. As far as Neemazone and Methyl salicylate increased the insect population with percentages 132.5 and 310% and total soil arthropods with 133.2% and 308.6% respectively and reduced the mites and the other arthropods population with 100%. Also results showed that the pesticides (avermectin, indoxacarb, & neemazone) increased the average number of the total soil arthropod population groups, and these pesticides showed selective effect, so that, it will be recommended that in order to preserve the beneficial predators, carnivorous and parasitoids these biological and selective pesticides in conventional tomato field should be applied.
According to the results obtained from this work, the changes in diversity and equitability and decrease or increase of percent population of soil arthropods groups were differed according to the soil arthropod groups, sampling period, applied plant protection products and system of agricultural.
Effect of glyphosate herbicide on the behavior of soil arthropods in non-orga...Salah Hussein
The objective of this study was to assess the effect of the glyphosate
application on the population of soil arthropods (collected with pitfall and
Berlese funnels) and their diversity after application of herbicide. Results
of soil arthropods collected with pitfall traps showed that glyphosate
herbicide has played a great role in detecting the activity of different
groups of soil arthropods with different reduction percentages. In insects
caused -23.7%, mites +48.8%, other arthropods -66.7% and total
population -23.3% reduction. Also glyphosate herbicide reduced the
diversity from 2.2 (ShanoonWiner values) to 1.6, as well as the equitability
%, from 46.6 % in pretreatment to 28.5% in post treatment. While it
caused a highly significant increase in the insect's population with
+55.26%, the other arthropods with +38.4%, and the total population of
soil arthropods with +54.04% in systemic groups of soil arthropods
extracted with Berlese funnel. However the population of mites extracted
with Berlese funnel was reduced with 80%. From results it could
concluded that glyphosate herbicide reduced all soil arthropods systemic
groups except mites collected with pitfall traps. However, it considerable
reduction effect was observed in mite populations collected with Berlese
funnel after application of glyphosate in tomato non organic system. This
reduction may be as a result of less food availability (e.g. plant roots) and
decreased green plant cover.
— The microbiological content of Lettuce (a vegetable), commonly vended in the Benin metropolis of Edo state were evaluated. Five vending locations were chosen for the study. Whole and soft rot samples were purchased and analysed for microbiological composition. Results showed high counts in soft rot samples in lettuce. Nutrient agar plated lettuce samples had bacterial counts in the range of 2.0x 103 to 4.7x10 7. Pseudomonas species was the dominant species found in lettuce samples. Bacillus species was isolated from one location in the lettuce samples. Mac Conkey agar plated lettuce plated had bacterial counts in the range of 2.3 x 10 3 to 5.7x 10 7. Enterobacter species, E. coli, and Klebsiella species were the dominant species isolated. Though, Proteus species was isolated from lettuce samples obtained from location five only. The study observes that consuming soft rot samples could pose a risk of introducing pathogens to the consumer due to their high microbial counts and could be detrimental to the health of the consumer.
Potential of Entomopathogenic Bacillus thuringiensis as Plant Growth Promotin...Agriculture Journal IJOEAR
Abstract— Bacillus thuringiensis has been used as an effective bioinsecticide because it produces the proteins Cry and Cyt, which are highly toxic to insects in certain situations. However, recently, B. thuringiensis was used as a biological control agent that can suppress plant disease. In this study, the antagonistic activities of B. thuringiensisAS17 japonensisand AS18 kurstaki against the fungal pathogen Fusarium oxysporum f.sp. lycopersici (FOL) were examined using a dual culture technique. Furthermore, B. thuringiensis strains suppressed the development of wilt symptoms caused by FOL in tomato plants. After inoculating six strains of B. thuringiensis suspension following inoculation of FOL, the development of wilt symptoms became less than control, especially with B. thuringiensisAS17 japonensis and AS20 CR371-H. Furthermore, we proved that B. thuringiensis strains are plant growth promoting rhizobacteria (PGPR) that can promote plant growth. Seed germination and shoot elongation were promoted by treating the tomato seeds with a bacterial culture filtrate and a bacterial suspension.
Effect of Trichoderma Fortified Compost on Disease Suppression, Growth and Yi...IJEAB
Trichoderma species are commonly used as effective biological control agents against phytopathogens especially the soil-borne fungi while some isolates are able to ameliorate plant growth. In the present study, Trichoderma fortified compost with different substrates were evaluated to reduce the pre-emergence and post-emergence seedling mortality, diseases of stem and root of chickpea caused by several soil-borne fungal pathogens, including Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii at different growth stages in the field under natural epiphytotic conditions. Among the twenty isolates of T. harzianum, Co-7 showed the most effective antagonist against the test pathogens in dual culture method. In field experiment, subsequently it was used for inoculum preparation with colonized wheat grain and mixed with well-matured decomposed composting materials like, saw dust, cow dung, tea waste, water hyacinth and poultry manure. Trichoderma fortified compost with poultry manure was found significantly effective in reducing pre-emergence and post emergence seedling mortality, disease incidence and disease severity of chickpea in the field. Interestingly, all the treatments significantly increased but Trichoderma fortified compost with poultry manure was the best to boost seed yield and quality.
in vitro screening of larvicidal and insecticidal activity of methanolic extr...IJEAB
The aim of this work was to evaluate the larvicidal and insecticidal activity of the selected plants namelyArtocarpus altilis, Piper betle and Artocarpus heterophyllus. The leaves of Artocarpus altilis and Artocarpus heterophyllus and roots of Piper betle were subjected to methanolic solvent extraction for the isolation of various bioactive constituents. The evaluation of larvicidal activity was carried out using late third instar larvae of Drosophila melanogaster. The insecticidal activity of extracts was studied against adult Bruchus pisorum, Tribolium castaneum, Sitophilus oryzae and was evaluated by direct contact application method .Nucleic acids and protein contents are regarded as important biomarkers of the metabolic potential of cells, as these play the main role in regulating the different activities of cells. Piper betle and Artocarpus heterophyllusextractshad a reducing effect on the nucleic acid and protein content in the larvae in a dose dependent manner whereas Artocarpus altilis extract did not exhibit any significant larvicidal activity. Piper betle and Artocarpus heterophyllusextractsshowed good insecticidal activity whereas A.altilis extract showed poor insecticidal activity. The results of the present study clearly indicate that Piper betle and Artocarpus heterophyllus extracts can be developed as ecofriendly larvicides and were also quite effective as insecticides for providing a better and excellent alternate for the control of insects.
Identification and Evaluation of Antifungal Compounds from Botanicals for th...researchagriculture
Red rot is a devastating disease in sugarcane caused by fungus,
Colletotrichum
falcatum
. In this study, eighteen different botanicals were screened for
identifying effective antifungal compound against
C.
falcatum.
Among the plants
screened, 15 per cent aqueous leaf extract of
Psoralea corylifolia
alone inhibited 100
per cent growth of both mycelium as well as spore germination under
in vitro
conditions. The extract did not exhibit any inhibitory effect to the beneficial microbes
viz.
,
Pseudomonas fluorescens
,
Bacillus megaterium
and
Gluconacetobacter
diazotrophicus
which are normally used in sugarcane. The effective plant extracts
exhibiting 100 per cent antifungal activity was subjected to TLC, HPLC and GC
-
MS
analysis to identify the bioactive antifungal compound. It revealed the
presence of
7H
-
furo [3,2
-
G] (1) benzopyran
-
7
-
one as main bioactive compound which is thought to be
the intermediate of antifungal compound, 8
–
methoxypsoralen formed during
biosynthesis.
Relative toxicity of selected insecticides against adult whitefly, t. vaporar...Sachin U.S
The present experiment was conducted to assess relative toxicity of selected insecticides against whitefly in the Entomology laboratory at College of Horticulture, Mudigere during the year 2014-2015. Among the eleven treatments, imidachloprid, thiamethoxam and cyantraniliprole were highly toxic to adults which recorded 100 per cent mortality, four days after treatment. Cyantraniliprole recorded the highest adult mortality comparatively early than imidachloprid and thiamethoxam. Fipronil 80% WG, recorded 100 percent mortality five days after treatment which was followed by emamectin benzoate 5% SG at sixth day after treatment and recorded as next best treatments under laboratory condition against whitefly. Acephate 75% SP and azadirhactin 10000 ppm recorded highest level of mortality (100%) at seventh and eighth day after treatment, respectively which was followed by triazophos 40% EC, spinosad 480% SC and buprofezin 25% SC at eleventh day after treatment. Considering the result, cyantraniliprole, thiamethoxam and imidacloprid were recommended for effective control of sucking pests in cotton ecosystem.
In vitro evaluation of Trichoderma viride and Trichoderma harzianum for its e...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Two field experiment were run at the farms of Omar AL-Mokhtar university, in order to knew the effect of some methods used to control tomato pests on soil arthropod systemic groups in organic cultivated tomato field such as (BT, algifol, methyl salicylate, neem oil), Results indicated that application of B. thuringensis and Algifol achieved the highest performance showing 100% reduction in mites and other soil arthropods systemic groups, and gave reduction 55.5 and 54.96% in both insect and total population of soil arthropods collected with pitfall traps. As far as Neemazone and Methyl salicylate increased the insect population with percentages 132.5 and 310% and total soil arthropods with 133.2% and 308.6% respectively and reduced the mites and the other arthropods population with 100%. Also results showed that the pesticides (avermectin, indoxacarb, & neemazone) increased the average number of the total soil arthropod population groups, and these pesticides showed selective effect, so that, it will be recommended that in order to preserve the beneficial predators, carnivorous and parasitoids these biological and selective pesticides in conventional tomato field should be applied.
According to the results obtained from this work, the changes in diversity and equitability and decrease or increase of percent population of soil arthropods groups were differed according to the soil arthropod groups, sampling period, applied plant protection products and system of agricultural.
Effect of glyphosate herbicide on the behavior of soil arthropods in non-orga...Salah Hussein
The objective of this study was to assess the effect of the glyphosate
application on the population of soil arthropods (collected with pitfall and
Berlese funnels) and their diversity after application of herbicide. Results
of soil arthropods collected with pitfall traps showed that glyphosate
herbicide has played a great role in detecting the activity of different
groups of soil arthropods with different reduction percentages. In insects
caused -23.7%, mites +48.8%, other arthropods -66.7% and total
population -23.3% reduction. Also glyphosate herbicide reduced the
diversity from 2.2 (ShanoonWiner values) to 1.6, as well as the equitability
%, from 46.6 % in pretreatment to 28.5% in post treatment. While it
caused a highly significant increase in the insect's population with
+55.26%, the other arthropods with +38.4%, and the total population of
soil arthropods with +54.04% in systemic groups of soil arthropods
extracted with Berlese funnel. However the population of mites extracted
with Berlese funnel was reduced with 80%. From results it could
concluded that glyphosate herbicide reduced all soil arthropods systemic
groups except mites collected with pitfall traps. However, it considerable
reduction effect was observed in mite populations collected with Berlese
funnel after application of glyphosate in tomato non organic system. This
reduction may be as a result of less food availability (e.g. plant roots) and
decreased green plant cover.
— The microbiological content of Lettuce (a vegetable), commonly vended in the Benin metropolis of Edo state were evaluated. Five vending locations were chosen for the study. Whole and soft rot samples were purchased and analysed for microbiological composition. Results showed high counts in soft rot samples in lettuce. Nutrient agar plated lettuce samples had bacterial counts in the range of 2.0x 103 to 4.7x10 7. Pseudomonas species was the dominant species found in lettuce samples. Bacillus species was isolated from one location in the lettuce samples. Mac Conkey agar plated lettuce plated had bacterial counts in the range of 2.3 x 10 3 to 5.7x 10 7. Enterobacter species, E. coli, and Klebsiella species were the dominant species isolated. Though, Proteus species was isolated from lettuce samples obtained from location five only. The study observes that consuming soft rot samples could pose a risk of introducing pathogens to the consumer due to their high microbial counts and could be detrimental to the health of the consumer.
Potential of Entomopathogenic Bacillus thuringiensis as Plant Growth Promotin...Agriculture Journal IJOEAR
Abstract— Bacillus thuringiensis has been used as an effective bioinsecticide because it produces the proteins Cry and Cyt, which are highly toxic to insects in certain situations. However, recently, B. thuringiensis was used as a biological control agent that can suppress plant disease. In this study, the antagonistic activities of B. thuringiensisAS17 japonensisand AS18 kurstaki against the fungal pathogen Fusarium oxysporum f.sp. lycopersici (FOL) were examined using a dual culture technique. Furthermore, B. thuringiensis strains suppressed the development of wilt symptoms caused by FOL in tomato plants. After inoculating six strains of B. thuringiensis suspension following inoculation of FOL, the development of wilt symptoms became less than control, especially with B. thuringiensisAS17 japonensis and AS20 CR371-H. Furthermore, we proved that B. thuringiensis strains are plant growth promoting rhizobacteria (PGPR) that can promote plant growth. Seed germination and shoot elongation were promoted by treating the tomato seeds with a bacterial culture filtrate and a bacterial suspension.
Effect of Trichoderma Fortified Compost on Disease Suppression, Growth and Yi...IJEAB
Trichoderma species are commonly used as effective biological control agents against phytopathogens especially the soil-borne fungi while some isolates are able to ameliorate plant growth. In the present study, Trichoderma fortified compost with different substrates were evaluated to reduce the pre-emergence and post-emergence seedling mortality, diseases of stem and root of chickpea caused by several soil-borne fungal pathogens, including Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii at different growth stages in the field under natural epiphytotic conditions. Among the twenty isolates of T. harzianum, Co-7 showed the most effective antagonist against the test pathogens in dual culture method. In field experiment, subsequently it was used for inoculum preparation with colonized wheat grain and mixed with well-matured decomposed composting materials like, saw dust, cow dung, tea waste, water hyacinth and poultry manure. Trichoderma fortified compost with poultry manure was found significantly effective in reducing pre-emergence and post emergence seedling mortality, disease incidence and disease severity of chickpea in the field. Interestingly, all the treatments significantly increased but Trichoderma fortified compost with poultry manure was the best to boost seed yield and quality.
in vitro screening of larvicidal and insecticidal activity of methanolic extr...IJEAB
The aim of this work was to evaluate the larvicidal and insecticidal activity of the selected plants namelyArtocarpus altilis, Piper betle and Artocarpus heterophyllus. The leaves of Artocarpus altilis and Artocarpus heterophyllus and roots of Piper betle were subjected to methanolic solvent extraction for the isolation of various bioactive constituents. The evaluation of larvicidal activity was carried out using late third instar larvae of Drosophila melanogaster. The insecticidal activity of extracts was studied against adult Bruchus pisorum, Tribolium castaneum, Sitophilus oryzae and was evaluated by direct contact application method .Nucleic acids and protein contents are regarded as important biomarkers of the metabolic potential of cells, as these play the main role in regulating the different activities of cells. Piper betle and Artocarpus heterophyllusextractshad a reducing effect on the nucleic acid and protein content in the larvae in a dose dependent manner whereas Artocarpus altilis extract did not exhibit any significant larvicidal activity. Piper betle and Artocarpus heterophyllusextractsshowed good insecticidal activity whereas A.altilis extract showed poor insecticidal activity. The results of the present study clearly indicate that Piper betle and Artocarpus heterophyllus extracts can be developed as ecofriendly larvicides and were also quite effective as insecticides for providing a better and excellent alternate for the control of insects.
Identification and Evaluation of Antifungal Compounds from Botanicals for th...researchagriculture
Red rot is a devastating disease in sugarcane caused by fungus,
Colletotrichum
falcatum
. In this study, eighteen different botanicals were screened for
identifying effective antifungal compound against
C.
falcatum.
Among the plants
screened, 15 per cent aqueous leaf extract of
Psoralea corylifolia
alone inhibited 100
per cent growth of both mycelium as well as spore germination under
in vitro
conditions. The extract did not exhibit any inhibitory effect to the beneficial microbes
viz.
,
Pseudomonas fluorescens
,
Bacillus megaterium
and
Gluconacetobacter
diazotrophicus
which are normally used in sugarcane. The effective plant extracts
exhibiting 100 per cent antifungal activity was subjected to TLC, HPLC and GC
-
MS
analysis to identify the bioactive antifungal compound. It revealed the
presence of
7H
-
furo [3,2
-
G] (1) benzopyran
-
7
-
one as main bioactive compound which is thought to be
the intermediate of antifungal compound, 8
–
methoxypsoralen formed during
biosynthesis.
Relative toxicity of selected insecticides against adult whitefly, t. vaporar...Sachin U.S
The present experiment was conducted to assess relative toxicity of selected insecticides against whitefly in the Entomology laboratory at College of Horticulture, Mudigere during the year 2014-2015. Among the eleven treatments, imidachloprid, thiamethoxam and cyantraniliprole were highly toxic to adults which recorded 100 per cent mortality, four days after treatment. Cyantraniliprole recorded the highest adult mortality comparatively early than imidachloprid and thiamethoxam. Fipronil 80% WG, recorded 100 percent mortality five days after treatment which was followed by emamectin benzoate 5% SG at sixth day after treatment and recorded as next best treatments under laboratory condition against whitefly. Acephate 75% SP and azadirhactin 10000 ppm recorded highest level of mortality (100%) at seventh and eighth day after treatment, respectively which was followed by triazophos 40% EC, spinosad 480% SC and buprofezin 25% SC at eleventh day after treatment. Considering the result, cyantraniliprole, thiamethoxam and imidacloprid were recommended for effective control of sucking pests in cotton ecosystem.
In vitro evaluation of Trichoderma viride and Trichoderma harzianum for its e...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Bio-fertilizers are being essential component of organic farming are the preparations containing live or latent cells of efficient strains of nitrogen fixing, phosphate solubilizing or cellulolytic micro-organisms used for application to seed, soil or composting areas with the objective of increasing number of such micro-organisms and accelerate those microbial processes which augment the availability of nutrients that can be easily assimilated by plants. Biofertilizers play a very significant role in improving soil fertility by fixing atmospheric nitrogen, both, in association with plant roots and without it, solubilise insoluble soil phosphates and produces plant growth substances in the soil.
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The study was carried out with the aim of sourcing for bacteria from the natural environment having antifungal capabilities to control and inhibit postharvest fungal spoilage of fruits and vegetables caused by Botrytis cinerea. Soil and water samples were collected from Heriot Watt University environment and Dr Ruth Fowler’s garden and inoculated using the spread plate technique; identification was carried out using Microbact Identification kits; and isolates assayed for antifungal activities against Botrytis cinerea. Forty eight bacteria species were isolated out of which sixteen (16) belonging to genera Pseudomonas, Bacillus, Escherichia, Burkholderia, Staphylococcus, Streptococcus, and Proteus showed antifungal activities. Bacteria species Pseudomonas stutzeri and Burkholderia cepacia had the highest zones of inhibition with average radii of 3.06 and 3.20 cm respectively. The bacteria had the potential to inhibit mycelial and spore growth at varying levels thus making them possible candidates for further tests and studies. Considering the aim of the study, further research into identifying these antifungal isolates inhibitory compounds and metabolites is highly recommended.
Synergetic effects of various plant extracts as bio-pesticide against wheat a...Muhammad Qasim
Human health and environment are greatly affected by extensive used of synthetic insecticide, which is why the alternative way of pesticides, such as botanical pesticide consumption increase with the passage of time to control of insect pests. The insecticidal impact of Moringa (Moringa oleifera), Neem (Azadirachta indica) and Euclyptus (Eucalyptus globules) leaves extracts were investigated on Diurophous noxia (Hemiptera: Aphididae). This study was aimed to evaluate the efficiency of bio-pesticide instead of synthetic chemicals against D. noxia. Experimental results showed that the percentage of mortality fluctuate with the fluctuation of concentration with time intervals and have great impact on the population of D. noxia. Single, doubled and tripled combinations of leaves extract were tested at two different concentrations (25 and 50%). The results suggested and concluded that from all the application of single botanicals A. indica give better result as compared to other single application while application of double combination M. olerifera with A. indica give significance result instead of other double mixture, meanwhile the triple combination (M. olerifera, A. indica and E. globules) performed excellent result as the others triple mixtures. The study revealed that combine used of M. olerifera, A. indica and E. globules leaves extract were very effective against D. noxia and surge as an alternative way of pesticides instead of synthetic chemicals.
Identification and evaluation of antifungal compounds from botanicals for the...researchagriculture
Red rot is a devastating disease in sugarcane caused by fungus, Colletotrichum falcatum. In this study, eighteen different botanicals were screened for identifying effective antifungal compound against C. falcatum. Among the plants screened, 15 per cent aqueous leaf extract of Psoralea corylifolia alone inhibited 100 per cent growth of both mycelium as well as spore germination under in vitro conditions. The extract did not exhibit any inhibitory effect to the beneficial microbes viz., Pseudomonas fluorescens, Bacillus megaterium and Gluconacetobacter diazotrophicus which are normally used in sugarcane. The effective plant extracts exhibiting 100 per cent antifungal activity was subjected to TLC, HPLC and GC-MS analysis to identify the bioactive antifungal compound. It revealed the presence of 7H-furo [3,2-G] (1) benzopyran-7-one as main bioactive compound which is thought to be the intermediate of antifungal compound, 8 – methoxypsoralen formed during biosynthesis.
Article Citation:
Rajkumar D and Murugesan R.
Identification and Evaluation of Antifungal Compounds from Botanicals for the Control of Sugarcane Red Rot Pathogen, Colletotrichum falcatum.
Journal of Research in Agriculture (2013) 2(1): 164-172.
Full Text:
http://www.jagri.info/documents/AG0044.pdf
Efficiency of bio fertilizers and compost on soil arthropod diversity and spe...Salah Hussein
The effect of three types of bio fertilizers ( N, P and K), plant and animal compost on soil arthropod diversity and species abundance were studied in organic tomato field. Nitrogen fixation bacteria increased the insects population with 16.26%, and the total population of soil arthropods with 7.3% and caused reduction % in mites and other arthropods population with 66.53 and 100 % reduction. Fertilization with potassium soluble bacteria (K) increased the population of insects, mites, and the total population of soil arthropods systemic groups with 175, 1353.8 and 741.23 % respectively. Similar results were achieved in the treatments with phosphorous soluble bacteria (P)
The treatments with animal compost increased all soil arthropods systemic groups, insects, mites and the total population of soil arthropods systemic groups gave 486.48, 356.36 and 347% increasing percentages compared with pretreatment. While this treatment caused reduction in the other soil arthropods with 66.6%.
The two types of compost increased both the terrestrial soil arthropods systemic groups, which collected by pitfall traps , and the superficial soil arthropods systemic groups that found in the upper surface layer, until (30cm depth), which collected by Berlese funnel methods, with percentage arranged from 47% to 61% in animal compost.
Generally steep decline occurred in other arthropods in tomato organic cultivated after application of different bio-fertilizers and increased in insect population, mites and total population of soil arthropods collected with pitfall traps and Berlese funnel.
Keywords : (organic tomato system, soil arthropods, agro ecosystem, Diversity and equitability, biofertilizers, plant and animal compost).
Abstract— The indiscriminate use of agricultural inputs, such as fertilizers e and synthetic pesticides, can cause high levels of toxic residues in food, biological imbalance, environmental contamination, intoxication of humans and animals, and other direct and indirect effects. The use of plant extracts as alternative insecticides is a way to minimize the problems caused by synthetic insecticides. Insecticides are in the second position in the trading market of agrotoxics in Brazil. The species Tagetes erecta L. and Tagetes patula L. have antioxidant properties, larvicidal, fungicidal, antimicrobial, nematicide and insecticide. In order to verify the possibility of Tagetes sp. extracts be suitable alternative to the use of synthetic insecticides, the bioassays laboratorial were designed through of insect mortality test of Sitophilus zeamais. We evaluated the antioxidant activity by the test of DPPH, in addition to screen the chromatographic profile of the extracts. It is concluded that the extracts evaluated are efficient in insect mortality, checking still considerable amount of antioxidant compounds, as identified also flavonoids, terpenes and alkaloids in extracts. According to these results we conclude that T. erecta and T. patula has phytotoxic compounds that can promote and expand its use as a natural insecticide.
Bioremediating Effect of Glomus Hoi and Pseudomonas Aeruginosa on the Organic...IJEAB
This study analyzed the degrading effect of Glomus hoi and Pseudomonas aeruginosa on the organic content and heavy metals of oil refinery effluent polluted soil using Amaranthus cruentus as the test plant. This study was carried out to determine if agricultural activities can be improved using any or both of the micoorganisms. Eight different treatment layouts were used with three replicates for each level of pollution in the treatment layout. Ninety six (96) pots, each containing three kilograms of soil from both sterilized and unsterilized soil were used for the study. Fifty (50) grams of soil inoculum from propagated Arbuscular mycorrhiza was inoculated to a set of twenty four (24) experimental pots containing both sterilized and unsterilized soil before A. cruentus seedlings were transplanted to them. Another set of twenty four (24) pots containing both sterilized and unsterilized soil were injected with thirty (30) mL of P. aeruginosa inoculum solution before transplanting A. cruentus seedlings to them. The third set of twenty four (24) pots received dual inoculation of both fifty (50) grams of soil inoculum containing G. hoi and thirty (30) mL of P. aeruginosa inoculum solution before A. cruentus were transplanted to them. The residual twenty four (24) pots served as the control. Thereafter, pot preparation was arranged in the screenhouse in a randomized block design. The A. cruentus seedlings were raised in nursery for a period of two weeks before they were transplanted to the pots, seedlings were left for 3 days to overcome transplanting shock before contaminating the soil with refinery effluent at various concentrations of 0%, 2%, 4% and 6% v/w. The seedlings were allowed to grow for eight weeks before the termination of the experiment. The pre planting analysis of soil showed that heavy metals analyses (zinc and iron) of sterilized soil had a lower concentration to the unsterilized. The soil pH ranged from 6.3 to 6.8. It also revealed that organic matter and organic carbon content ranged from 0.8% to 1.3% and 0.4% to 1.7%. However, after the experiment, it was discovered in this study that treatments without any microorganism inoculation in sterilized and unsterilized soil had a higher level of % organic carbon and % organic matter content compared to the other treatments that were inoculated with one or two micro-organisms across all the levels of effluent concentration. Heavy metals of soil in all the soil samples were found to increase as the petrochemical effluent increased in concentration. The results obtained were analyzed using Duncan Multiple Range Test (DMRT) and other descriptive statistics. This study opined that the combined use of G. hoi and P. aeruginosa was more effective in improving the organic contentand the reduce heavy metals of oil refinery effluent polluted soil than when either is used singly.
International Journal of Advances in Biology (IJAB)ijcsity
International Journal of Advances in Biology (IJAB) is a peer-reviewed, open access journal, addresses the impacts and challenges of Biology. The journal documents practical and theoretical results which make a fundamental contribution for the development of Biological sciences and applications.
RESIDUAL ESTIMATION OF ISOPROTURON, ATRAZINE AND GRAIN PROTECTANTS IN STORED ...ijabjournal
This study was carried out to see the occurrence and level of pesticide residues in stored wheat grain samples. Wheat grains were collected from different godowns located in Faisalabad district. Weedicide residues in wheat grains were analyzed by using thin plate liquid chromatography and validated high
performance liquid chromatography. On the basis of extent of inhibition of the Hill reaction, it was found that the level of atrazine was (0-0.028 ngkg-1) and the level of isopproturon was (0-0.024 ngkg-1) in test samples. The calculated values were below the Maximum Residue Limit (MEL) i.e. 100 ngkg-1 for atrazine and 2200 ngkg-1 for isoproturon. It was found that presence of herbicide residues in stored wheat was negligible. Further, on wheat grain samples placed in jars and treated with deltamethrin and cypermethrin @0.4 and 0.8% concentration. The persistency data was calculated after 1 hour, 7, 15 and 30 days of treatment. The grain protectants were continuously degraded with the passage of time and after the 30 days
the amount of residues of the grain protectants was found to be very small. It was concluded that the use of the grain protectants in the powder form in combination with other botanicals on stored wheat samples is feasible to reduce the insect pest in stored grains.
Isolation, identification of antagonistic rhizobacterial strains obtained fro...Shazia Shahzaman
Plant growth promoting rhizobacteria (PGPR), are associated with roots, found in the rhizosphere and can directly or indirectly enhance the plant growth. In this study soil was collected from rhizosphere of chickpea fields of different areas of Rawalpindi division of Pakistan. PGPR were isolated, screened and characterized. Eight isolates of rhizobacteria (RHA, RPG, RFJ, RC, RTR, RT and RK) were isolated from Rawalpindi division and were characterized. The antagonistic activity of these PGPR isolates against root infecting fungi (Fusarium oxysporum and Verticillium spp.,) was done and production of indole acetic acid (IAA), siderophore and P-solubilization was evaluated. The isolates RHA, RPG, RFJ, RC, RRD and RT were found to be positive in producing siderophore, IAA and P-solubilization. Furthermore, most of the isolates showed antifungal activity against Fusarium oxysporum, and Verticillium spp. The rhizobacterial isolates RHA, RPG, RFJ, RC, RRD, RTR, RT and RK were used as bio-inoculants that might be beneficial for chickpea cultivation as the rhizobacterial isolates possessed the plant growth promoting characters i.e. siderophore, IAA production, phosphate solubilization. In in vitro tests, Pseudomonas sp. and Bacillus spp. inhibited the mycelial growth of the fungal root pathogens. The isolates (RHA and RPG) also significantly increased (60-70%) seed germination, shoot length, root length of the chickpea. The incidence of fungi was reduced by the colonization of RHA and RPG which enhanced the seedling vigor index and seed germination. The observations revealed that isolates RHA and RPG is quite effective to reduce the fungal root infection in greenhouse, and also increases seed yields significantly. These rhizobacterial isolates appear to be efficient yield increasing as well as effective biocontrol agent against fungal root pathogen.
Pesticidal efficacy of crude aqueous extracts of Tephrosia vogelii L., Allium...researchagriculture
Cabbage aphid (Brevicoryne brassicae L.) is one of the most problematic pests in smallholder vegetable production, causing significant yield losses in heavy infestations. Current control strategy focuses on use of synthetic pesticides that consequently lead to decimation of natural enemies, development of insect resistance and resurgence and upset biodiversity. Botanical pesticides have been used widely in smallholder farmers but not much documented literature exists on efficacy of these products. A field trial was done to assess the efficacy of crude aqueous extracts of Tephrosia vogelii, Allium sativum and Solanum incanum in controlling Brevicoryne brassicae in Brassica napus production. The trial was laid in a randomized complete block design (RCBD) with five treatments replicated four times. The five treatments used in the experiment were T. vogelii, A. sativum, S. incanum, dimethoate and control. Wingless adult female aphids were inoculated three weeks after transplanting of seedlings. Spraying and data collection were done weekly for four weeks. Data was collected on aphid nymph and adult counts on the third leaf from the aerial plant part of randomly selected plants from each treatment for 24 hours after the application of treatments and total plant fresh weight per each treatment. There were significant differences (p<0.05)><0.05) on the yield of rape. It was concluded that T. vogelii, S. incanum and A. sativum aqueous crude extracts have some pesticidal effects on aphid in rape production.
Article Citation:
Shepherd Mudzingwa, Simbarashe Muzemu and James Chitamba.
Pesticidal efficacy of crude aqueous extracts of Tephrosia vogelii L., Allium sativum L. and Solanum incanum L. in controlling aphids (Brevicoryne brassicae L.) in rape (Brassica napus L.)
Journal of Research in Agriculture (2013) 2(1): 157-163.
Full Text:
http://www.jagri.info/documents/AG0040.pdf
Similar to Effects of six selected antibiotics on plant growth and soil microbial and enzymatic activities (20)
Effects of six selected antibiotics on plant growth and soil microbial and enzymatic activities
1. Effects of six selected antibiotics on plant growth and soil microbial and
enzymatic activities
Feng Liu a
, Guang-Guo Ying a,*, Ran Tao a
, Jian-Liang Zhao a
, Ji-Feng Yang a
, Lan-Feng Zhao b
a
State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640, China
b
College of Resource and Environmental Science, South China Agricultural University, Guangzhou 510642, China
Terrestrial ecotoxicological effects of antibiotics are related to their sorption and degradation behavior in soil.
a r t i c l e i n f o
Article history:
Received 20 July 2008
Received in revised form
15 December 2008
Accepted 17 December 2008
Keywords:
Antibiotics
Phytotoxicity
Soil microbial activity
Respiration
Phosphatase
Behavior
a b s t r a c t
The potential impact of six antibiotics (chlortetracycline, tetracycline and tylosin; sulfamethoxazole,
sulfamethazine and trimethoprim) on plant growth and soil quality was studied by using seed germi-
nation test on filter paper and plant growth test in soil, soil respiration and phosphatase activity tests.
The phytotoxic effects varied between the antibiotics and between plant species (sweet oat, rice and
cucumber). Rice was most sensitive to sulfamethoxazole with the EC10 value of 0.1 mg/L. The antibiotics
tested inhibited soil phosphatase activity during the 22 days’ incubation. Significant effects on soil
respiration were found for the two sulfonamides (sulfamethoxazole and sulfamethazine) and trimeth-
oprim, whereas little effects were observed for the two tetracyclines and tylosin. The effective concen-
trations (EC10 values) for soil respiration in the first 2 days were 7 mg/kg for sulfamethoxazole, 13 mg/kg
for sulfamethazine and 20 mg/kg for trimethoprim. Antibiotic residues in manure and soils may affect
soil microbial and enzyme activities.
Ó 2008 Elsevier Ltd. All rights reserved.
1. Introduction
Tons of pharmacologically active substances are used annually
in human and animal medicines for treatment and prevention of
illness (Dı´az-Cruz et al., 2003; Sarmah et al., 2006). Antibiotics are
specifically designed to control bacteria in human or animals and
help to protect their health. After treatment, most antibiotics are
excreted from the treated body, either unaltered or as metabolites,
some of which are still bioactive (Sarmah et al., 2006). Obviously
this makes them potentially hazardous to bacteria and other
organisms in the environment (Baguer et al., 2000). Different types
of drugs have different anticipated exposure routes to the envi-
ronment (Jørgensen and Halling-Sørensen, 2000). The dominant
pathway for antibiotic release in the terrestrial environment is via
the application of animal manure and biosolids containing excreted
antibiotics to agricultural land as fertilizer (Jørgensen and Halling-
Sørensen, 2000; Dı´az-Cruz et al., 2003, 2006; Golet et al., 2003;
Go¨bel et al., 2005; Kemper, 2008). Antibiotics can also be intro-
duced to agricultural land through irrigation with reclaimed
wastewater, since they have been frequently detected in the raw
and treated sewage wastewaters (Renew and Huang, 2004; Yang
et al., 2005; Gulkowska et al., 2008). Therefore, it is necessary to
understand the environmental impact of antibiotics associated
with application of animal manure, biosolids and wastewater on
agricultural land.
Unlike pesticides used on agricultural land, antibiotics have not
aroused attention as potential pollutants until fairly recently
(Halling-Sørensen et al., 1998; Ku¨mmerer, 2001). Bacterial resis-
tance has been a big issue in terms of human and animal health;
however, antibiotic ecotoxicological relevance is scarcely known
because the potential effects of antibiotics in the environment are
very limited (Pang et al., 1994; Rooklidge, 2004).
When antibiotics get into the arable land, they could possibly
impact vegetation growth and development as well as soil micro-
bial activity (Jjemba, 2002a,b). Phytotoxicity of a chemical can be
assayed using seed germination and plant growth tests. Limited
studies have been conducted to investigate the phytotoxicity of
some antibiotics (e.g. sulphadimethoxine, enrofloxacin and
oxytetracycline) to crop plants (Migliore et al., 1998, 2003; Kong
et al., 2007). The effects of antibiotics on plants in soils were found
different between compounds and between plant species (Batch-
elder, 1982; Jjemba, 2002a; Farkas et al., 2007). Tetracyclines
increased radish yields, but decreased pinto bean yields (Batch-
elder, 1982). When grown in chlortetracycline-treated soil,
a significant increase in the activities of the plant stress proteins
glutathione S-transferases and peroxidases was observed in maize
plants, but not in pinto beans (Farkas et al., 2007).
* Corresponding author. Tel./fax: þ86 20 85290200.
E-mail address: guangguo.ying@gmail.com (G.-G. Ying).
Contents lists available at ScienceDirect
Environmental Pollution
journal homepage: www.elsevier.com/locate/envpol
0269-7491/$ – see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envpol.2008.12.021
Environmental Pollution 157 (2009) 1636–1642
2. As antibiotics are designed to be biologically active toward
microorganisms, it would be interesting to understand the poten-
tial effects on soil microbial activity. However, previous reports on
the effects of pharmaceutical antibiotics on soil microorganisms are
scarce and inconsistent (Patten et al., 1980; Thiele-Bruhn and Beck,
2005; Kong et al., 2006; Zielezny et al., 2006; Kotzerke et al., 2008).
A number of soil microbiological parameters, including microbial
biomass carbon and basal respiration, have been suggested as
possible indicators of soil environmental monitoring programs (Yao
et al., 2000; Winding et al., 2005).
In the present study, two classes of antibiotics were chosen to
study the effects on plant growth and soil microbial activity. The
antibiotics used in the study were: tetracyclines (chlortetracycline
and tetracycline, as well as tylosin commonly used in combination
with tetracyclines) and sulfonamides (sulfamethoxazole and sul-
famethazine, as well as trimethoprim commonly used in
combination with sulfonamides). The phytotoxicity was assayed
using seed germination tests on filter paper and plant growth tests
in soil. Soil microbial activity was assessed by measuring soil
microbial respiration and phosphatase activity.
2. Materials and methods
2.1. Chemicals
Chlortetracycline (98% purity), tetracycline (98% purity), tylosin (90% purity), sulfa-
methazine (98% purity), sulfamethoxazole (98% purity), and trimethoprim (96% purity)
were purchased from DeBioChem Reagents & Instruments Co. Ltd. (Nanjing, China). All
the reagents used in the following tests were purchased from Qianhui Reagents &
Instruments Co. Ltd. (Guangzhou, China) and they were of analytical grade.
2.2. Seeds and soil
Seeds of rice (Oryza sativa L.) were obtained as a gift from South China Agri-
cultural University, while seeds of cucumber (Cucumis sativus L.) and sweet oat
(Cichaorium endivia) were purchased from Seeds Collection, Guangdong Academy of
Agricultural Sciences, China. Preliminary incubation showed that all the seeds used
in this study had more than 90% germination rates.
An agricultural soil (0–20 cm deep) was collected from a rice paddy in an
experimental station, Guangdong Academy of Agricultural Sciences, China. The soil
type is classified as Anthrosol based on its properties. The soil was air-dried until
water content reached about 20% of the maximum water-holding capacity (MWHC).
After removal of large pieces of plant materials and soil animals by screening
through a 2 mm sieve, the soil was mixed well and stored at 4 C until use. The soil
had a silt loam texture with a pH value of 5.7 and total carbon content of 18.2 g/kg,
total nitrogen content of 0.959 g/kg, and total phosphate content of 0.215 g/kg.
2.3. Seed germination test
Laboratory tests to evaluate the effects of antibiotics on seed germination of
three plants (rice, cucumber and sweet oat) were carried out using the filter paper
method according to the International Seed Testing Association (ISTA) test protocols
(ISTA,1985). After having been sterilized using 0.1% NaClO and pretreated by soaking
in distilled water for six hours, seeds of cucumber (15), rice (20) and sweet oat (20),
which depended on the size of the seeds, were placed on a filter paper (9 cm
diameter) kept in each Petri dish (10 cm diameter).
For each antibiotic compound, the filter papers in Petri dishes were treated with
5 mL of the antibiotic solution at different concentrations and covered before placing
in an incubator. Seeds were germinated in the incubator under the conditions of 25 C
temperature, 80% humidity and darkness. The seed germination was evaluated using
root length of seedlings as endpoint (primary root ! 5 mm) after 4–5 days (Tiquia
et al., 1996). Except trimethoprim, each antibiotic test had 8 treatments with chlor-
tetracycline concentrations of 0 (CK), 0.1, 1, 10, 50, 100, 200, 500 mg/L, tetracycline
concentrations of 0 (CK), 0.1, 1, 10, 30, 50, 100, 300 mg/L, sulfamethazine or sulfa-
methoxazole concentrations of 0 (CK), 1, 10, 30, 50, 70, 100, 300 mg/L and tylosin
concentrations of 0 (CK),1,10, 30, 50,100, 300, 500 mg/L. The trimethoprim test had 9
treatments with concentrations of 0 (CK), 5% acetone/water solution,1,10, 30, 50,100,
300, 500 mg/L. Each treatment including controls was carried out in three replicates.
2.4. Plant growth test
The effects of the antibiotics on plant growth were assayed in a silt loam soil
using the method modified from the literature (OECD, 1984; Batchelder, 1982;
Baguer et al., 2000). Chlortetracycline and tylosin were directly added in an aqueous
solution to the soil, while the other antibiotics were added by spiking into fine
quartz sand due to their low water solubility. The tests had 8 treatments with
concentrations of chlortetracycline, tetracycline, sulfamethazine, sulfamethoxazole
and trimethoprim at 0 (CK), 1, 10, 30, 50, 70,100 and 300 mg/kg in soil, and tylosin at
0 (CK), 1, 10, 30, 50, 100, 300, 500 mg/kg in soil. Each treatment including controls
was carried out in triplicates. The detailed procedure was described briefly as
follows. The test was conducted in plastic cups (150 mL with a diameter of 7.5 cm).
Each antibiotic was spiked into 100 g of the test soil in each cup in dark, and the soil
was mixed in a shaker for 12 h. Two plant seeds cucumber and rice were chosen in
the phytotoxicity tests. All these seeds were treated in the same way as the seeds
used in the filter paper method. Into each plastic cup 10 seeds of rice or 8 seeds of
cucumber were sown at a depth of 0.5 cm. Then the soil moisture in each plastic cup
was adjusted to 50% of maximum water-holding capacity (MWHC). The treated
plastic cups were placed in a climate chamber at a temperature of 25 C and
humidity of 80% under darkness. After the seeds in the cups were all germinated, the
test conditions of the chamber were changed to a controlled photoperiod (12 h
light:12 h dark). Seven days later following seed germination, plant seedlings were
thinned to five rice plants or four cucumber plants per cup. During the test period,
the soil water moisture was maintained everyday by adding appropriate amount of
water. The plants in the cups were harvested at the 20th day and their shoot and root
lengths were measured.
2.5. Soil respiration and phosphatase activity tests
Soil respiration and phosphatase activity were assayed and used as the indica-
tors of soil microbial activity. The effect of antibiotics on soil microbial respiration
was assayed by the direct absorption method using sodium hydroxide (Wang et al.,
2005; Diao et al., 2006; Yao et al., 2006). The test had 8 treatments with sulfame-
thazine or sulfamethoxazole concentrations of 0 (CK), 1, 10, 40, 70 and 100 mg/kg in
soil and other antibiotics concentrations of 0 (CK), 1, 10, 40, 70, 100 and 300 mg/kg in
soil. Each treatment was conducted in triplicates. The experimental procedure is
described briefly as follows. Each antibiotic was spiked to the test soil (50 g) in each
cup, and 1 mL of 0.1 M glucose solution was also added. Then 10 mL of pure water
was added to obtain soil moisture level at 25% MWHC. The spiked soils were mixed
and left overnight to be acclimatized in the fume hood. After the soil moisture was
adjusted to 60% MWHC, the plastic cups were put into 1 L air-tight plastic jars with
a little cup holding 20 mL of 0.15 N sodium hydroxide in the bottom of each jar and
incubated at 25 C in the darkness. Two blanks without soil but with 20 mL of 0.15 N
sodium hydroxide were also included in the test. The CO2 was determined by
titration of the NaOH solution. At different time intervals (2, 4, 6, 9, 12, 16 and 22
days), the sodium hydroxide in each jar was titrated with 0.1 N hydrochloric acid and
a new 20 mL of 0.15 N sodium hydroxide was placed in the jar. The intensity of soil
respiration was calculated by the following formula: Respiration value (mgCO2 gÀ1
dry soil) ¼ (blank-titer) Â 0.1 Â44/50, where (blank-titer) in the formula is the blank
titration volume of hydrochloric acid in the treatment without soil subtract the
titration volume of hydrochloric acid in the treatments with soil, 0.1 means
concentration of hydrochloric acid, and 50 means weight of dry soil.
Phosphatase activity was assayed using 0.1 M acetate buffer (pH 5) and 0.5%
disodium phenyl phosphate substrate according to the method described by Guan
(1983). In this acidic phosphatase activity test, the soils were treated in the same
way as in the soil respiration test and incubated at 25 C in the darkness. At different
time intervals (2nd, 5th, 9th, 14th, 19th and 23rd days following treatment), 3 g of
soil was randomly sampled from each container to measure the soil phosphatase
activity. Phosphatase activity was expressed as mg phenol per kg of soil within 1 h
incubation time.
2.6. Extraction and analysis
Antibiotic residues in the soil during the soil respiration and enzymatic tests
were monitored by using high performance liquid chromatography with a diode-
array detector (HPLC-DAD). Sulfamethazine, sulfamethoxazole, trimethoprim and
tylosin in soil samples were extracted with acetonitrile for three times using soni-
cation, while chlortetracycline and tetracycline were extracted with 90% methanol
with 0.8 M oxalic acid and 0.85 M citric acid. After extraction, the extracts were
reconstituted in the initial mobile phase solution. The injection volume was 20 mL
and the column temperature was set at 30 C. Different mobile phases and gradient
programs were applied for the six antibiotics. For the two sulfonamides, acetonitrile
and 0.1% formic acid aqueous solution were used as mobile phase at a flow rate of
0.75 mL/min: 15% at 0 min to 60% of acetonitrile at 10 min, back to 15% at 12 min
which was kept for 3 min. The ultraviolet wavelength (UV) was set at 270 nm. For
trimethoprim, acetonitrile and Milli-Q water were used as mobile phase at a flow
rate of 0.75 mL/min: 30% at 0 min to 60% of acetonitrile at 10 min, back to 30% at
12 min which was kept for 5 min. The UV wavelength for trimethoprim was 230 nm.
For tylosin, acetonitrile and 0.08% acetic acid solution (9 mM ammonium acetate)
were used as mobile phase at a flow rate of 1 mL/min: 35% at 0 min to 60% of
acetonitrile at 15 min, further to 90% at 16 min, back to 35% at 17 min which was
kept for 5 min. The UV wavelength for tylosin was 285 nm. For the two tetracyclines,
acetonitrile and 10 mM oxalic acid solution were used as mobile phase at 0.6 mL/
min: 25% of acetonitrile at 0–8 min, increase to 90% at 10 min, back to 25% at 12 min
F. Liu et al. / Environmental Pollution 157 (2009) 1636–1642 1637
3. which was kept for 5 min. The UV wavelength was set at 370 nm for chlortetracy-
cline and tetracycline.
The instrumental detection limits were 0.30, 0.23, 4.71, 12.6, 14.1 and 66.5 mg/L
for sulfamethazine, sulfamethoxazole, trimethoprim, chlortetracycline, tetracycline
and tylosin, respectively; while their recoveries at the spiking concentration of
10 mg/kg were 99%, 96%, 72%, 70%, 73% and 77%, respectively.
2.7. Data analysis
Unless specified, all reported data were compared by using Duncan’s new
multiple range test at the 5% level. Differences between values at p 0.05 were
considered statistically significant. EC50 values (the concentration causing 50%
effect) as well as EC10 values (the concentration causing 10% effect) of the tested
antibiotics were calculated by plotting logged concentrations versus seed germi-
nation rate or plant growth endpoints (shoot height and root length) by using EC50
calculator program developed by CSIRO, Australia.
After the data had been tested for normality and homogeneity of variance, the
no-observed-effect concentration (NOEC, highest concentration to cause no signif-
icant effect) and the lowest-observed-effect concentration (LOEC, lowest concen-
tration to cause a significant effect) were estimated by SAS 8.2 using Dunnett’s
multiple comparison test to determine which treatments differed significantly from
the controls (1-tailed, p 0.05).
3. Results and discussion
3.1. Phytotoxicity of antibiotics
In seed germination tests, root length instead of number of
germinated seeds was used as the endpoint in statistical analysis,
which is consistent with the approaches used by previous studies
for metals and organic contaminants (Mishra and Choudhuri, 1999;
Martı´ et al., 2007). Table 1 lists EC10, EC50, NOEC and LOEC values
for the antibiotics tested on three plant seeds (sweet oat, rice and
cucumber). The results (EC50 values) showed that sweet oat and
rice seeds presented more susceptibility to the antibiotics, while
cucumber seeds were less sensitive to all antibiotics. Rice was most
sensitive to sulfamethoxazole with the EC10 value of 0.1 mg/L.
Chlortetracycline and tetracycline inhibited germination of the
three plant seeds with EC50 values less than 300 mg/L. Tylosin was
the least toxic compound, especially toward rice and cucumber
seeds with EC50 values more than 300 mg/L. Sulfamethoxazole and
sulfamethazine also inhibited seed germination of the three plants
with the EC50 values for the two sulfonamides being less than
100 mg/L.
The seed germination tests demonstrated that antibiotics could
negatively affect plant seed germination, but the effects varied
between the plant species and between the antibiotics used in the
tests. Among the three plants, sweet oat was the most sensitive
plant to the six antibiotics although with varying toxicity values.
Tetracyclines and sulfonamides were more toxic to plant seed
germination while tylosin and trimethoprim were less toxic to seed
germination.
In plant growth tests, only sulfonamides (sulfamethoxazole and
sulfamethazine) strongly affected rice growth in soil (Table 2). No
obvious rice growth inhibition was observed when treated with the
other antibiotics. This is in contrast with the results from the seed
germination tests, which showed inhibitory effects by tetracyclines
and sulfonamides. The results from the present study are consistent
with those of previous studies (Batchelder, 1982; Norman, 1955).
Norman (1955) found that root growth of several crops was
inhibited by 5–10 mg/L of oxytetracycline in solution, but the
effects were not observed in soil. The lesser inhibitory effects of
tetracyclines in the soil than in the solution might be attributed to
their strong adsorption onto soil components (clay and organic
matter) (Tolls, 2001; Figueroa et al., 2004; Kulshrestha et al., 2004;
Figueroa and Mackay, 2005; Mackay and Canterbury, 2005; Pils and
Laird, 2007). Sorption coefficients of sulfonamides are very low in
soil (Boxall et al., 2002), which indicates that sulfonamides are
more bioavailable.
As found in the seed germination tests, cucumber was less
sensitive to the antibiotics than rice in terms of plant growth in soil
(Table 2). For sulfonamides, the EC50 values for rice were less than
Table 1
Toxicity data from seed germination tests for sweet oat, rice and cucumber (endpoint:root length).
Compound EC10 EC50 NOEC LOEC
oat rice cucumber oat rice cucumber oat rice cucumber oat rice cucumber
Chlortetracycline 0.2a
8 0.7 16 39 48 0.1 1 0.1 0.1 10 1
Tetracycline 14 16 8 57 69 203 1 1 1 10 10 10
Tylosin 19 500 217 141 500 500 1 500 100 10 500 300
Sulfamethoxazole 16 0.1 300 69 8 300 1 0.1 300 10 1 300
Sulfamethazine 2 6 6 37 45 300 0.1 1 1 1 10 10
Trimethoprim 24 23 2 86 118 300 1 0.1 1 1 1 10
a
All concentrations are in mg/L.
Table 2
Toxicity data from plant growth tests in soil for rice and cucumber.
Compound Endpoint EC10 EC50 NOEC LOEC
rice cucumber rice cucumber rice cucumber rice cucumber
Chlortetracycline Seedling height 300a
19 300 300 300 100 300 300
Root length 300 300 300 300 300 70 300 100
Tetracycline Seedling height 300 300 300 300 300 300 300 300
Root length 300 300 300 300 300 300 300 300
Tylosin Seedling height 500 90 500 343 500 50 500 100
Root length 500 35 500 500 500 50 500 100
Sulfamethoxazole Seedling height 25 85 38 300 30 100 50 300
Root length 2 66 13 300 1 100 10 300
Sulfamethazine Seedling height 92 249 220 300 70 100 100 300
Root length 1 120 43 300 1 100 10 300
Trimethoprim Seedling height 300 0.7 300 300 300 1 300 10
Root length 300 85 300 300 300 100 300 300
a
All concentrations are in mg/kg dry soil weight.
F. Liu et al. / Environmental Pollution 157 (2009) 1636–16421638
4. 300 mg/kg whereas the EC50 values for cucumber were all near or
more than 300 mg/kg. Similar results of sulfonamides’ effects on
cucumber were observed by Migliore et al. (1998). Sulphame-
thoxine at a concentration of 300 mg/kg significantly depressed the
growth of Amaranthus restroflexus, Plantago major, Rumex acetosella,
and Zea mays in vitro, as well as Hordeum disthicum both in vitro
and in soil (Migliore et al., 1998). Species variability was also found
in previous studies (Batchelder, 1982; Farkas et al., 2007). The
growth of radish and wheat was enhanced in the presence of
chlortetracycline and oxytetracycline whereas the growth of corn
was unaffected by these antibiotics (Batchelder, 1982). Chlortetra-
cycline was found to significantly increase the activities of the plant
stress proteins glutathione S-transferases and peroxidases in maize
plants, but not in pinto beans (Farkas et al., 2007).
The concentration of sulfonamides in manure ranged between
10 mg/kg and 91 mg/kg (Pfeifer et al., 2002; Christian et al., 2003;
Jacobsen and Halling-Sørensen, 2006; Martı´nez-Carballo et al., 2007).
In sludge, sulfonamides were also detected with concentrations even
up to 197 mg/kg for sulfapyridine and 73 mg/kg for sulfamethoxazole in
Swiss wastewater treatment plants (Go¨bel et al., 2005; Dı´az-Cruz et al.,
2006). Sukul and Spiteller (2006) proposed that with manure slurry
being applied in the field as fertilizer with a maximum dose rate of
50 m3
/ha,sulfonamideresiduesinsoilcouldreach1 kg/ha,whichisthe
same order of magnitude as the application rate of modern pesticides.
Trimethoprim is used as a synergist to sulfonamides and was detected
with concentrations upto 17 mg/kg in chicken and turkey dung but not
in pig manure (Martı´nez-Carballo et al., 2007). In pig manure, up to
46 mg/kg chlortetracycline, 29 mg/kg oxytetracycline and 23 mg/kg
Chlortetracycline
*
**
*
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0 ˜ 2 2 ˜ 4 4 ˜ 6 6 ˜ 9 9 ˜ 12 12 ˜16 16 ˜ 21
Incubation time (day)
Soilrespiration
(CO2mg/gdrysoil)
0 1 10 40 70 100 300
Tetracycline
*
*
*
*
0.0
0.5
1.0
1.5
2.0
2.5
3.0
0 ˜ 2 2 ˜ 4 4 ˜ 6 6 ˜ 9 9 ˜ 13 13 ˜ 17 17 ˜ 22
Incubation time (day)
Soilrespiration
(CO2mg/gdrysoil)
Tylosin
*
*
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0 ˜ 2 2 ˜ 4 4 ˜ 6 6 ˜ 9 9 ˜ 12 12 ˜ 16 16 ˜ 21
Incubation time (day)
Soilrespiration
(CO2mg/gdrysoil)
0 1 10 40 70 100 300
0 1 10 40 70 100 300
Fig. 1. Effects of chlortetracycline, tetracycline and tylosin on soil respiration measured as the cumulative CO2 generated within different incubation periods. The error bars are the
standard deviation (n ¼ 3). The asterisk (*) indicates a significant difference compared to the controls without addition of antibiotics (p 0.05).
F. Liu et al. / Environmental Pollution 157 (2009) 1636–1642 1639
5. tetracycline were reported in Austria (Martı´nez-Carballo et al., 2007).
Tylosin was not detected in swine manure and may have been
degraded during mixing of the manure (Jacobsen and Halling-Sør-
ensen, 2006). Based on the NOEC values in Table 2, only sulfonamides
may affect growth of the plants, especially rice.
3.2. Antibiotic effects on soil microbial and enzyme activity
Fig. 1 shows little effects of tetracyclines and tylosin on soil
microbial respiration, with statistically significant variations only
observed at the higher concentration levels. In contrast,
sulfonamides and trimethoprim were found to cause significant
decreases in soil respiration within the first 4 days (Fig. 2). Kotzerke
et al. (2008) also observed reduced microbial activity by antibiotic
sulfadiazine in manure for up to 4 days after manure application.
Soil respiration measured as CO2 decreased significantly with
increasing concentrations of sulfamethoxazole and sulfamethazine
as well as trimethoprim in the soil. The effective concentrations
(EC10 values) in the first 2 days were calculated to be 7 mg/kg for
sulfamethoxazole, 13 mg/kg for sulfamethazine and 20 mg/kg for
trimethoprim. Increased soil respiration activity with antibiotic
concentrations was observed for the two sulfonamides and
Sulfamethazine
*
* *
*
*
* **
*
*
*
*
*
* * *
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Incubation time (day)
Soilrespiration
(CO2mg/gdrysoil)
Sulfamethoxazole
*
*
*
* * *
*
*
*
*
*
**
*
*
*
*
*
*
*
*
*
*
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Incubation time (day)
Soilrespiration
(CO2mg/gdrysoil)
Trimethoprim
*
*
**
*
*
*
**
*
*
**
*
*
*
*
*
*
*
*
*
* *
*
* *
* *
*
*
0.0
0.5
1.0
1.5
2.0
2.5
3.0
9 ˜ 13 13 ˜ 17 17 ˜ 22
Incubation time (day)
Soilrespiration
(CO2mg/gdrysoil)
300
0 ˜ 2 2 ˜ 4 4 ˜ 6 6 ˜ 8 8 ˜ 12 12 ˜16 16 ˜ 21
0 ˜ 2 2 ˜ 4 4 ˜ 6 6 ˜ 8
0 ˜ 2 2 ˜ 4 4 ˜ 6 6 ˜ 9
8 ˜ 12 12 ˜16 16 ˜ 21
0 1 10 40 70 100
0 1 10 40 70 100
0 1 10 40 70 100
Fig. 2. Effects of sulfamethazine, sulfamethoxazole and trimethoprim on soil respiration measured as the cumulative CO2 generated within different incubation periods. The error
bars are the standard deviation (n ¼ 3). The asterisk (*) indicates a significant difference compared to the controls without addition of antibiotics (p 0.05).
F. Liu et al. / Environmental Pollution 157 (2009) 1636–16421640
6. trimethoprim at certain stages after the first 4 days. In the later
incubation periods, a decreasing respiration activity was followed
in comparison with the activity in the first few days. This indicates
that the effect of these antibiotics (sulfamethoxazole, sulfametha-
zine and trimethoprim) on soil microbial respiration was time
dependent. The increased soil respiratory activity was also reported
in the previous studies (Fru¨ nd et al., 2000; Ingerslev and Halling-
Sørensen, 2000; Halling-Sørensen et al., 2003; Schmitt et al., 2004).
The recovery and increase of soil respiration could be attributed to
a decrease in the bioavailable antibiotic fraction, and an increasing
adaptation and resistance of the microorganisms (Thiele-Bruhn
and Beck, 2005). Chemical monitoring of the soil samples showed
DT50 values (dissipation half-lives) for the three compounds (sul-
famethoxazole, sulfamethazine and trimethoprim) ranged
between 2 and 5 days; therefore, the recovery of soil respiration
after the first 4 days was partially due to the significant loss of these
antibiotics in the soil. Based on the concentrations (up to 91 mg/kg)
detected in manure and soils (Pfeifer et al., 2002; Christian et al.,
2003; Jacobsen and Halling-Sørensen, 2006; Martı´nez-Carballo
et al., 2007) and the EC10 values in the present study, sulfonamides
and trimethoprim have the potential to affect soil respiration in
those lands applied with animal manure and biosolids.
In the present study, no obvious effects of tetracycline, chlortet-
racycline and tylosin on soil respiration could be observed. Sorption
and degradation processes played certain roles in reducing the
effects of these antibiotics. These three compounds exhibited strong
adsorption onto soil, suggesting they are less bioavailable (Sarmah
et al., 2006). Previous studies found that tylosinwas not persistent in
soil and its DT50 was no more than 1 week (Teeter and Meyerhoff,
2003; Hu and Coats,2007). In the presentstudy, tylosinhad a DT50of
8 days in the soil. So tylosin will not accumulate in soil and pose very
little risk to soil microbial respirationprocess (Blackwell et al., 2007).
Tetracyclines had DT50 values of more than20 days in the soilused in
the present study. Moreover, tetracyclines have strong adsorption
and complexation with cations such as calcium in soil (Kemper,
2008; Pils and Laird, 2007; Wessels et al.,1998; Zielezny et al., 2006).
This could significantly reduce the bioavailability and effects of
tetracyclines on soil microbial respiration.
Fig. 3 shows inhibition rates of soil phosphatase activity with
addition of antibiotics. The inhibition rates were very variable
during the various incubation periods (22 days). This could be
caused by the heterogeneous nature of soil. The present study
suggests that addition of antibiotics to soil at the concentration
used (1–300 mg/kg) can significantly affect soil phosphatase
activity (p 0.05). The EC10 values calculated for the six antibiotics
ranged from 1 mg/kg for sulfamethazine to 406 mg/kg for tetra-
cycline. Comparing with antibiotic concentrations (up to 91 mg/kg)
in the manure and soils (Pfeifer et al., 2002; Christian et al., 2003;
Jacobsen and Halling-Sørensen, 2006; Martı´nez-Carballo et al.,
2007), inhibition effects may be expected from some antibiotics
such as sulfonamides in real environment. Boleas et al. (2005) also
observed significant effects of oxytetracycline on soil microbial
enzymatic activities (phosphatase and dehydrogenase). However,
Thiele-Bruhn and Beck (2005) found no effects on dehydrogenase
activity even at a concentration of 1000 mg/kg of sulfapyridine and
oxytetracycline. Phosphatase activity was not measured in their
study. The reason behind the inconsistent results on dehydroge-
nase activity remains unclear. Microbial parameters such as enzy-
matic activities could be influenced by various factors and they may
not be specific for antibiotic effects in soil.
4. Conclusion
The different terrestrial toxicological effects of six antibiotics
were observed through using a series of bioassays including plant
seed germination and growth tests in soil, soil respiration and
phosphatase activity tests. Root elongation was found to be
a sensitive endpoint for plant germination and growth tests. The
two sulfonamides (sulfamethoxazole and sulfamethazine) and
trimethoprim were found to be most toxic to plant growth in soil.
Sweet oat and rice were more sensitive to the antibiotic compounds
than cucumber. In comparison with the controls, all antibiotics
tested inhibited soil phosphatase activity at the concentration
range used. Sulfamethoxazole, sulfamethazine and trimethoprim
had temporal effects on soil respiration whereas tetracycline,
chlortetracycline and tylosin had little effects on soil respiration.
The different toxic effects between the two groups of antibiotic
compounds were due to their different behavior in the soil; sorp-
tion, degradation and chelating with metals played important roles
for tetracyclines and tylosin. Considering the environmental levels
and fate of these antibiotics in soil, we would expect low toxic
effects on plant growth and soil microbial activities following
application of wastes with antibiotics such as sulfonamides and
trimethoprim and also a quick recovery from the stress due to the
loss and/or binding of the antibiotics onto soil components.
Acknowledgements
The authors would like to acknowledge the financial support
from the National Natural Science Foundation of China (NSFC
40688001, 40771180 and 40821003) and partial support from
Guangdong Natural Science Foundation. This is contribution No. IS
1026 from GIGCAS.
0
5
10
15
20
25
30
35
0
5
10
15
20
25
30
35
inhibitionrateof
phosphataseactivity(%)
inhibitionrateof
phosphataseactivity(%)
0 10 40 70 100 300
Antibiotic Concentration (mg/kg)
1
0 10 40 70 100 300
Antibiotic Concentration (mg/kg)
1
22 days
CTC
TC
TYL
TMP
SMZ
SMX
2 days
CTC
TC
TYL
TMP
SMZ
SMX
Fig. 3. Inhibition of phosphatase activity by six antibiotics compared to the controls
without addition of antibiotics at different times (2 days and 22 days) during the
incubation. CTC: chlortetracycline, TC: tetracycline, TYL: tylosin, SMZ: sulfamethazine,
SMX: sulfamethoxazole and TMP: trimethoprim.
F. Liu et al. / Environmental Pollution 157 (2009) 1636–1642 1641
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