Antibiotic Uptake by Antibiotic Uptake by Plants from Soil Plants from Soil

Fertilized with Animal Fertilized with Animal ManureManure

K.Kumar, S.c. Gupta, SK.Baidoo,Y.Chander,and C.J.RosenK.Kumar, S.c. Gupta, SK.Baidoo,Y.Chander,and C.J.Rosen

Presented by Ju Hwan OhPresented by Ju Hwan Oh

Soil Science Lab.Soil Science Lab.

Published in J. Environ. Qual 34:2082-2085 (2005)Published in J. Environ. Qual 34:2082-2085 (2005)

Antibiotics are compounds produced by various living organisms, such as yeast or fungi.

Treating infectious diseases (human, animal)

Antibiotics?

Tylosin

Chlortetracycline

feces and urine

Common antibiotics tetracyclines (oxatetracycline and chlortetracycline)

tylosin

sulfamethazine

amprolium

menensin

virginiamycin

penicillin

nicarbazine

(Webb and Fontenot, 1975; De Liguoro et al., 2003;Kumar et al., 2004, 2005)

Manure applied as raw manure (fresh or

dried) or composted, no guidelines on

the presence of contaminants such as

hormone and antibiotics.

Absorption of antibiotics in the animal gut is not complete and as a result substantial amounts of antibiotics are excreted in urine and feces that end up in manure.

Manure is used worldwide not only as a source of plant nutrients but also as a source of organic matter to improve soil quality especially in organic and sustainable agriculture.

Tetracyclines are broad - spectrum bacteriostatic agents and act by inhibiting p

rotein synthesis by blocking the binding of aminoacyl tRNA (transfer

RNA) to the mRNA(messenger RNA) ribosome complex.Antiacne agent (topical) - probably due to their antibacterial activity.

Topical tetracyclines are thought to suppress the growth of propionibacterium acne

s, an anaerobe found in sebaceous glands and follicles. P. acnes produces protea

ses hyaluronidases, lipases, and chemotactic factors, alI of which can produce infl

ammatory components or inflammation directly.

Action of tetracycline ?

The objective of the study

To evaluate whether or not plants take up antibiotics from manure-soil mixtures, and if so to what extent?

Materials and methods

Experiment 1: Plant Uptake of Antibiotics from Soil spiked with Antibiotic

1. Soil : Hubbard loamy sand soil

2. Crop : Green onion(4/pot) and Cabbage(2/pot)

3. Added 50ml each of chlortetracycline and tylosin (20ug L-1 conc.).

4. Watered with 100ml of water twice a week and harvested at 3 and 6 wk after

transplanting.

5. Fresh plant material grounding and extraction.

6. Antibiotics analyzed on the same day

Materials and methods

Experiment 2: Plant Uptake of Antibiotics from Manure-Applied Soils

1. Treatment – i) control ii) with antibiotics (100g chlortetracycline, 100g sulfamethazine,

and 50g penicillin per ton feed)

2. At last week, manure collected

3. Treatments in the uptake study

i) C-M (control manure) and Ab-M (antibiotic manure)

ii) C-M + antibiotic and Ab-M + antibiotics (additional 100mg of chlortetracycline and tylosin per kg of dry manure)

iii) Crop : corn, green onion and cabbage

4. Manure : 200 kg N ha-1

5. Antibiotic analysis in plant : ELISA method ( Kumar et al., 2004 )

Results and discussion

Green onionGreen onion CabbageCabbage

AntibioticAntibiotic 3 wk3 wk 6wk6wk 3 wk3 wk 6wk6wk

--------------- ng g--------------- ng g-1-1 fresh weight --------------- fresh weight ---------------

TylosinTylosin ND*ND* NDND NDND NDND

ChlortetracyclineChlortetracycline 14.4±2.314.4±2.3 12.8±1.712.8±1.7 11.4±2.111.4±2.1 10.0±1.810.0±1.8

*Not detected

Table 1. Concentration of antibiotics in plant tissues after 3 and 6 wk.

Treatment*Treatment* ChlortetracyclineChlortetracycline TyrosinTyrosin

---------- ug pot---------- ug pot-1 -1 --------------------

C-MC-M 00 00

C-M + antibioticsC-M + antibiotics 10001000 10001000

Ab-MAb-M 587587 00

Ab-M + antibioticsAb-M + antibiotics 15871587 10001000

Results and discussion

Table 2. Amount of antibiotics present per pot in various treatment

* See “Experiment 2” section of Materials and Methods for treatment descriptions.

RecoveryRecovery

C-MC-M Ab-MAb-M C-M + C-M + antibioticsantibiotics

Ab-M + aAb-M + antibioticsntibiotics

------------------ % ------------------------------------ % ------------------

Green onionGreen onion 00 0.340.34 0.300.30 0.210.21

CabbageCabbage 00 0.640.64 0.800.80 0.620.62

CornCorn 00 1.041.04 1.021.02 0.800.80

Results and discussion

Table 3. Mean amount of chlortetracycline recovered in plant tops in various treatment.*

* See “Experiment 2” section of Materials and Methods for treatment descriptions.

Results and discussion

Ch

lort

etra

cyc

lin

e co

nce

ntr

atio

n (

ng

g-1 f

resh

so

il)

Chlortetracycline ( ug pot-1)

Figure 1. Concentration of chlortetracycline in plant tops in relation to the amount of chlortetracycline present in the soil-manure mixture in a pot.

Antibiotic (class)Antibiotic (class) MassMass Water solubility*Water solubility* Log KLog Kowow****

------------- mg L------------- mg L-1 -1 --------------------------

Chlortetracycline (tChlortetracycline (tetracyclines)etracyclines) 479479 230 to 52000230 to 52000 -1.3 to 0.05-1.3 to 0.05

Tylosin (macrolideTylosin (macrolides)s) 916916 0.45 to 150.45 to 15 1.6 to 3.11.6 to 3.1

*Range provided for the whole class of antibiotics.

** Octanol-water partition coefficient

Results and discussion

Table 4. Characteristics of chlortetracycline and tylosin antibiotics.

Conclusion

The adverse impacts of ingesting antibiotics present in plants by humans are not known at this stage.

A few adverse impacts of consuming antibiotics in fresh vegetables (sweet corn) and friuts are speculated :

- Alleric or toxic ( Patterson et al., 1995 and Basaraba et al., 1999)

- Antibiotic resistance ( Selim and Cullor, 1997; Langford et al., 2003)

There is an urgent need study

i) The fate of different antibiotic present in manure

ii) which antibiotics and their degradation products may be taken up by plants

iii) whether or not antibiotics present in food degrade when cooked

iv) whether or not antibiotics or their degradation products are bioactive to impart antibiotic resistance to gut bacteria or cause adverse immunological reaction in humans.

Further study