Presentation at the 25 SETAC Europe Barcellona 3-7 May

Monitoring of fluoroquinolone antibiotics in piggery wastewater and their mobility in solid materials after the

application of the slurry as fertilizer

Vassilis Litskas, Andreas Parpounas, Evroula Hapeshi, Costas Michael and Despo Fatta-Kassinos

Civil and Environmental Engineering Department and Nireas, International Water Research Center, University of Cyprus, P.O. Box 20537, 1678, Nicosia,

Cyprus E-mail contact: [email protected]

GAIA-Laboratory of Environmental

Engineering

University of Cyprus

SETAC Europe 25 Annual Meeting, Barcelona 3-7 MAY 2015

mailto:[email protected]

Scope of the study • Develop an analytical method for the simultaneous

determination of Enrofloxacin (ENR) and Ciprofloxacin (CIP) in piggery wastewater.

• Monitor the antibiotics in piggery wastewater.

• Determine their adsorption in solid materials (e.g. soil, piggery sludge).

• Assess their presence and mobility in the agricultural environment.

GAIA-Laboratory of Environmental Engineering



Matrix FQs Residue (mg/kg) Sludge CIP, NOR 1.40-2.42, 1.54-2.37

Soil CIP, NOR 3-5.8, 6.2-9.8 Sewage ENR 3.4-15 Manure CIP, ENR 0.053-8.3

Soil ENR 0.02-0.05 Compost CIP, NOR 0.0005, 0.022 Manure OFL 0.59

Soil ENR, MAR 0.0113-0.020 Sediment CIP, OFL 0.00595-0.01203

Table 1 Residual concentrations of fluoroquinolone (FQs) . Adopted from Speltini et al. (2011).




ENR/CIP environmental presence

DIGESTER

LAGOONS

FARM


Nursery Collection tank VT = 200 x 103 L

(max)

Swine CAFO facilities

V1 = 15,000 L/day V2 = 75,000 L/day

Digester VT = 3000 x 103 L

(max)

Vinfluent = 100,000 L/day

Veffluent =100,000 L/day

10,000 L/day Co-substrates

ENR 5mg per piglet




Parameter Influent Effluent

Q (L x 103/day) 100 100

BOD5 (mg/L) 9520 5806

COD (mg/L) 26550 10071

TN (mg/L) 3500 1600

pH 7 – 7.5 6.5 – 8.5

Methods

• Selection of the appropriate solvent for ENR and CIP extraction from piggery wastewater.

• Optimization of the solid phase extraction (SPE) procedure (pH and EDTA, elution solvent). Antibiotics were quantified with UPLC-MS/MS (LODs << μg/L).

• Collection of composite wastewater samples and urine and faeces samples from new born piglets (March and October 2014).

• Batch equilibrium experiments (OECD 106) to determine adsorption to solid materials.




Results • ENR and CIP extraction from wastewater




0

25

50

75

100

Re

cove

ry %

Enrofloxacin Ciprofloxacin

MeOH/H2O ACN (2% AA) Control Mg(NO3)2,NH3 MeOH/ACN/H2O

Results • Optimum sample pH and EDTA concentration prior SPE loading

0

25

50

75

100

0.0 17.8 35.6 53.5 106.9

EDTA concentration (mg/L)

ENR

CIP

0

25

50

75

100

1.5 2.2 3.5 5 7

Re

cove

ry %

Sample pH

ENR

CIP




Results • Optimum SPE elution solvent pH




0

10

20

30

40

50

60

70

80

90

100

2.5 3.5 4.5 5.5 6.5 9.5

Re

cove

ry %

MeOH pH (for SPE elution)

ENR CIP

Results • FQs in piglets’ urine and faeces

0

100

200

300

400

500

600

8 h 24 h

FQs

con

cen

trat

ion

(μ

g/L

)

Sampling time after ENR application

enr cip




Results • FQs in piggery wastewater

LOQ

0

5

10

15

20

25

30

35

40

Wastewatercollection system

Primary collectiontank

After digester

FQ c

on

cen

trat

ion

(μ

g/L

)

ENR CIP




Nursery

Collection tank VT = 200 000 L (max)

Swine CAFO facilities

V1= 15,000 L/day

V2 = 75,000 L/day

Digester VT = 3,000,000 L (max)

Vinfluent = 100,000 L/day

Veffluent =100,000 L/day

10,000 L Co-substrates

[ENR], [CIP] = 0

[ENR], [CIP] = 0

[ENR] = 31.40 μg/L [CIP] = 16.05 μg/L

[ENR] = 3.12 μg/L [CIP] = 1.84 μg/L

[ENR] = LOQ [CIP] = LOQ




Fertilizing fields If piggery wastewater is not used for CH4 production and is directly applied as fertilizer: • TN in piggery wastewater : 3.5 g/L • Fertilization rate : 170 kg N/ha • Piggery wastewater applied as fertilizer : 4,857 L/ha • ENR and CIP in piggery wastewater : 3.14 and 1.84 μg/L • Total ENR and applied per ha agricultural land : 15.15 and 8.94 mg,

respectively. • 1 ha = 10,000 m2, 1.25 g/cm3 bulk density and 0.25 m soil depth

3,125,000 kg soil/ha • 4.84 ng/kg soil ENR and 2.86 ng/kg soil CIP (estimated concentrations)




FQs adsorption in solid materials

Solid material ENR Kdads CIP Kd

ads

Quartz sand 412 (74) 155 (29)

Clay soil 1786 (92) 1054 (44)

Sandy loam soil 731 (102) 606 (57)

Wastewater solid material 5075 (254) 3365 (201)

Table 4: Adsorption distribution coefficient average value (n=3) for ENR and CIP in different solid materials. The standard error value is given in the parenthesis.




Conclusions



• The development of a fast and accurate method for the analysis of FQs in urine, faeces and piggery wastewater, using off-line solid phase extraction (SPE) followed by LC-MS/MS, resulted in the detection and quantification of ENR and CIP in piggery wastewater.

• The antibiotics were present in the wastewater, even when ENR is administered only in new-born piglets. The major ENR metabolite, CIP, a frequently used antibiotic in human medicine was also quantified in the wastewater.


Conclusions



• It is probable that these antibiotics could be released in trace amounts in the agricultural environment.

• The batch equilibrium experiments indicated that ENR and CIP could be tightly adsorbed in soils and other solid materials. Adsorption determines ENR and CIP mobility and fate in the terrestrial environment.


Thank you for your attention !!




Documents

Presentation at the 25 SETAC Europe Barcellona 3-7 May