Release of Release of CryptosporidiumCryptosporidium and and GiardiaGiardia ((OoOo)cysts )cysts From Dairy Calf Manure: Impact of Solution Salinity From Dairy Calf Manure: Impact of Solution Salinity

Scott A. BradfordScott A. Bradford11 and Jack F. Schijvenand Jack F. Schijven22

11George E. Brown,George E. Brown, JrJr., Salinity Laboratory., Salinity LaboratoryUSDA, ARSUSDA, ARS

22National Institute of Public Health and the Environment (RIVM)National Institute of Public Health and the Environment (RIVM)BilthovenBilthoven, The Netherlands, The Netherlands

Prevalence in herds of veal calvesPrevalence in herds of veal calvesversusversus age of veal calvesage of veal calves

0%

20%

40%

60%

80%

100%

0 - 6 7 - 12 13 - 18 19 - 24 25 - 35Age [weeks]

Perc

enta

ge o

f pos

itive

her

ds

Cryptosporidium Giardia

Average concentrations in positive samples of Average concentrations in positive samples of fresh manure from veal calvesfresh manure from veal calves versusversus age age

Cryptosporidium

0.001

0.1

10

1000

0-6 7-12 13-18 19-24 25-35Age [weeks]

Nx106/kg

Giardia

0-6 7-12 13-18 19-24 25-35Age [weeks]

Major pathways of Major pathways of CryptosporidumCryptosporidum and and GiardiaGiardiato surface waters in The Netherlandsto surface waters in The Netherlands

ManMan Farm animalsFarm animalsWildWild

mammalsmammalsand birdsand birds

DomesticDomesticwastewaterwastewater ManureManure

Treated and untreated dischargesTreated and untreated discharges

Import from abroadImport from abroad(rivers Rhine and Meuse)(rivers Rhine and Meuse)

Surface watersSurface watersin The Netherlandsin The Netherlands

LandLand

WastewaterWastewater

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++

++

++ ++

++

++

??

????

??

IntroductionIntroduction

••Cattle manure, especially from calves, form an Cattle manure, especially from calves, form an enormous source of enormous source of CryptosporidiumCryptosporidium and and GiardiaGiardia..

••Due to animal waste application to agricultural Due to animal waste application to agricultural land, large numbers of land, large numbers of ((oooo)cysts)cysts may reach may reach groundwater and surface water. groundwater and surface water.

••Release rates of (Release rates of (oooo)cysts from manure are )cysts from manure are important boundary conditions for loading rates important boundary conditions for loading rates into the environment.into the environment.

••Manure is exposed to a wide range of solution Manure is exposed to a wide range of solution salinitiessalinities (rain or urine).(rain or urine).

Holstein dairy cattleHolstein dairy cattle ((ChinoChino, CA), CA)

Manure phase concentrations of (oo)cysts, mip, N g-1.Cryptosporidium Giardia

Average 7.1 × 104 9.5 × 104

S.D. 1.7 × 104 3.0 × 104

N 6 12

00--3 3 monthsmonths 33--5 5 monthsmonthsManureManure samplesample

Manure Drip ExperimentsManure Drip Experiments Aerosol or droplets

Manure disk

105 µm sieve

Funnel

Funnel

Sample collection

T = 0 min T = 250 min

Experimental conditionsExperimental conditionsManure Water

applicationQ

[ml min-1]EC

[dS m-1]Temp.[°C]

Calf Mist 10 0.3 5Calf Mist 2.9 0.3 5Calf Drip 2.4 0.3 5Calf + Cow 1:10 Drip 2.7 0.3 5Calf + Cow 1:1 Drip 3.0 0.3 5Calf Drip 2.6 0.3 23Calf Drip 2.0 0.3 23Calf Drip 2.5 5.0 23Calf Drip 2.0 9.5 23Calf Drip 2.1 15 23ρi =1.1 g cm-3 from Vm (34.4 ml) and initial weight of manure disk (g).

Detection of Cryptosporidium and Giardia• FITC - Monoclonal Antibody• Direct Enumeration via Microscopy

Manure ReleaseManure Release

Cm & OD660

y = 7.8289xR2 = 0.9832

0369

1215

0 0.5 1 1.5 2OD660

Cm

(g/l)

0

2

4

6

8

10

0 100 200 300Time (min)

Cm

(g/l)

0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

0

1

2

3

4

5

0 100 200 300Time (min)

Cum

ulat

ive

Man

ure

(g)

0.3 dS/m5 dS/m9.5 dS/m14.75 dS/m

GiardiaGiardia ReleaseRelease

0

100000

200000

300000

400000

500000

600000

0 100 200 300Time (min)

Cum

ulat

ive

Gia

rdia

(N)

0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

0

1000

2000

3000

4000

5000

0 100 200 300Time (min)

Gia

rdia

Rel

ease

Rat

e (N

/min

)

0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

0

200

400

600

800

1000

1200

0 200 400 600 800

Cm*Cio (N/ml)

Gia

rdia

(N/m

l) 14.75 dS/m9.5 dS/m5.0 dS/m0.3 dS/mPrediction

CryptosporidiumCryptosporidium ReleaseRelease

0

50000

100000

150000

200000

250000

300000

0 100 200 300Time (min)

Cum

ulat

ive

Cry

pto

(N)

0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

0

500

1000

1500

2000

2500

0 100 200 300Time (min)

Cry

pto

Rel

ease

Rat

e (N

/min

)

0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

0100200300400500600700800900

0 200 400 600

Cm*Cio (N/ml)

Cry

pto

(N/m

l) 14.75 dS/m9.5 dS/m5.0 dS/m0.3 dS/mPrediction

Aqueous PhaseManure Phase

Boundary Layermi

meq

m

Distance

0

2

4

6

8

10

12

14

0 100 200 300

Time (min)

Cm

(g/l) DATA

FIT

0

1

10

100

1 10 100 1000 10000

Time (min)

Cm

(g/l) DATA

FIT

y = 3.84e0.1158x

R2 = 0.8955

010

2030

0 5 10 15 20

EC (dS/m)

β

FITModel

y = 0.0005x-0.1318

R2 = 0.9895

0.00000.00020.00040.00060.0008

0 5 10 15 20

EC (dS/m)

αFITModel

0

2

4

6

8

10

0 100 200 300Time (min)

Cm

(g/l)

y = -0.0443x + 1.0923R2 = 0.700

0.0

0.5

1.0

1.5

0 5 10 15 20EC (dS/m)

Erog FIT

Model

y = 0.164Ln(x) + 0.9165R2 = 0.977

0.0

0.5

1.0

1.5

0 0.25 0.5 0.75 11-(EC/15)

Eroc FIT

Model

0

100000

200000

300000

400000

500000

600000

0 100 200 300Time (min)

Cum

ulat

ive

Gia

rdia

(N) 0.3 dS/m

5.0 dS/m9.5 dS/m14.75 dS/m0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

0

50000

100000

150000

200000

250000

300000

0 100 200 300Time (min)

Cum

ulat

ive

Cry

pto

(N) 0.3 dS/m

5.0 dS/m9.5 dS/m14.75 dS/m0.3 dS/m5.0 dS/m9.5 dS/m14.75 dS/m

ConclusionsConclusionsEffluent concentrations of manure and (Effluent concentrations of manure and (oooo)cysts were )cysts were initially several orders of magnitude below their initial initially several orders of magnitude below their initial concentration in the manure.concentration in the manure.Increasing the solution salinity tended to decrease the Increasing the solution salinity tended to decrease the manure and (manure and (oooo)cyst concentrations and, hence, the )cyst concentrations and, hence, the cumulative amount released into the aqueous phase.cumulative amount released into the aqueous phase.Increasing salinity was hypothesized to stabilize the manure Increasing salinity was hypothesized to stabilize the manure by compression of the diffuse double layer thickness by compression of the diffuse double layer thickness between negatively charged colloidal material in the between negatively charged colloidal material in the manure. manure. ((OoOo)cyst release efficiencies tend to decrease with )cyst release efficiencies tend to decrease with increasing salinity.increasing salinity.A conceptual model was developed to predict manure and A conceptual model was developed to predict manure and ((oooo)cyst release and loading rates from manure. )cyst release and loading rates from manure.