KAMPAI ~TEAM 2 PAST RESEARCH~
HealthEconomic

Visualization
Quantification

Veterinary Medicine~Dean Dr. Kennedy Choongo~
Kaampwe Muzandu

Andrew Kataba

John Yabe

Shouta MMNakayama

VeterinaryMedicine

Haruya Toyomaki

Yared Beyene

Economics
MasatoHiwatari

Daiju
NaritaBona
ChitahChrispin
MphukaMedicine & Nursing
Takeshi Saito

Nosiku Sipilanyambe Munyinda

Wakako Fujita

Nancy Zyongwe

Harukazu Tohyama

Sandra Shanunga

Takahiko Yoshida

Education
Robert Serphell

Beatrice Matafwali

Joe Kalima

Gabliel Walubita

Background -Mining areas in Zambia The rapid economic progress in Africa has contributed to theproblem of environmental pollution.
 Major economic activity in Zambia is mining, and this is mainly
done in the Copperbelt region (Cu-Co) and Kabwe (Pb-Zn).

Mine in the Copperbelt region

Mining wastes are directly discarded into theKafue River in the Copperbelt region.
Copperbelt

Kabwe
Mine in Kabwe

Background -Kabwe City Kabwe city re-ranked among ‘the 10 most polluted
places in the world’ (Blacksmith Institute, 2014).
 However, no studies have been done to assess metal
pollution levels in edible tissues of chicken andcattle from Kabwe.
 No recent study is conducted in human.
Scrap metal
scavenging at mine

Kabwe

The purpose of the study isTo assess the metal pollution levels in
soil, domestic animals and children in
Kabwe.

Clarify if soil Pb contamination issource for animals and human, using
Stable Pb isotopes.

Research achievement

PAST

PRESENT

FUTUREOutline of PAST study
2009~2011

Pb pollution inchildren?

Pb & Cd

contamination inchicken goat

2012~2015

Study Outline
Soil contaminationand diffusion

Pb & Cd
contamination

2008~2009
Kabwe Pb & Zn Miningincattle

Soil sampling location in Kabwe (n=101, May 2009)• GPS coordination• Microwave digestion(HNO3)• Measured by AAS or
ICP-MS(Modified from Google Earth)Zn, Pb and Cu concentrations in Kabwe soils

Zn, Cu and Pb
concentrations
in soilexceeded theFAO Limits.
Zn

PbCu

Zn

Pb

Cu

Range

5 – 91,000

9 – 51,000

2 – 5,000

Median61028040

FAO Limits

500

150100Cd and As in Kabwe soil
Cd and As
concentrations
also exceed theFAO Limits.

CdAsCdAsRange

0.01 - 1400.04 - 140Median17FAO Limits

1.618

Itai-itai disease (public hazard in Japan)→ Soil Cd: 25-50 ppm

GIS (Geographic Information System) analysis
Low concentration metals

High concentration metals

CdPbLow

CrPb-Zn
mine
Zn

CoAsHighCu← Diffusion from mine

Ni

Southern part is morecontaminated than Northern part.

Occurred naturally →(ArcGIS ver 9.3, ESRI)

Outline of PAST study

Pb pollution inchildren?

Pb & Cd
contamination

inchicken
2009~2011

Soil contaminationand diffusion

Pb & Cd
contamination

inKabwe Pb & Zn Miningcattle

6 towns were selected for cattle samplingRegional difference of Pb in cattle liver in Zambia• Kabwe higher levels ofPb in the liver• Chibombo Pb levels weresimilar to Kabwe• Other towns
 Lower Pb levels• Asterisk: p < 0.001

Regional difference of Cd in cattle kidney in Zambia• Kabwe Higher levels of
Cd in kidneys• Chibombo
 Cd levels weresimilar toKabwe• Other towns
 Lower Cd levels• Asterisk: p < 0.001(Dunettee test)

High Pb and Cd levels in muscle and offalof Free-range chickens (n=17) in Kabwe
AA

mg/kg (wet weight)

Permissible limit(Offal: 0.5, muscle: 0.1)

AA

BCCC

mg/kg (wet weight)

CdPb

Permissible limit(Liver: 0.5, Kidney: 1.0)
BDCFE

F

(Tukey test)

Scavenging
Free-range
chickenExceeded the permissible limitsPb and Cd comparison in liver ofBroiler (n=32) and Free-range (n=17) in Kabwe
Broiler

Kept indoors and fed commercial feed

Freerange

Roam and scavenge for food in themine townships
PbBroiler*Cd*Free-range

Broiler

Free-range

Outline of PAST study

Pb pollution inchildren?

Pb & Cd
contamination

inchicken

Soil contaminationand diffusion

2012~2015

Pb & Cd
contamination

inKabwe Pb & Zn Mining

cattleHuman sample collection from Kabwe mine area
was done in May 2012
Map of Kabwe

 We collected samples
from local communities
around Pb-Zn mine.
 0-7 years old children
were main participants.

Makululu(n=129)Central 3 Health Centres(Makululu, Kasanda,Chowa area)

Chowa (n=70)

 299 blood
 250 urine 250 feces

Kasanda (n=100)
Pb-Znmine

Mutwe Wansofu(It was difficult to collect urine
and fecal samples from allparticipants)

One of the Health Centres where
we collected samples

Some children arebarefootMetal exposure
through road dust ?

Scrap metal
scavenging at mine

Blood Pb concentration in children in Kabwe

5 μg/dL in human blood, Pb
causes toxicological effects onhuman (especially risk is high forchildren).

5Concerned level by CDC

Kasanda (n=100)

Makululu (n=129)

Blood Pb concentrations (ug/dL)

Almost
100 % ofchildren
exceeded
5 ug/dL

5 ug/dL
Number of children and percent

BPb: Kasanda vs Makululu(Male and female analyzed separately )FemaleMale**

Kasanda was significantly higher in both maleand female.

Sex difference (boys vs. girls)
Kasanda（No difference）

Makululu（Male was high）

Blood Pb (µg/dL)

Blood Pb (µg/dL)*

Age difference (0-3 years vs 4-7 years)*

Makululu*
Blood Pb (µg/dL)

Blood Pb (µg/dL)

KasandaCorrelation between Age & blood Pb
Fit each mean value
Linear Fit(BLL = 107.8 - 9.5*Age, p < 0.0001)

Blood Pb (µg/dL)

Polynominal Fit, Degree=2[BLL = 103.5 – 9.4*Age + 0.92*(Age-4.2)^2,p < 0.0001]

1(N=24)2(N=39)3(N=20)4(N=38)5(N=33)Age (sample size)

6(N=35)7(N=34)

Summary of PAST study
AA

Permissible limit(Offal: 0.5, muscle: 0.1)

AA

B

CCC

Cd

mg/kg (wet weight)

mg/kg (wet weight)

Pb

Permissible limit(Liver: 0.5, Kidney: 1.0)

Pb pollution inchildren

BDCFEF2012~2015

Pb & Cd
contamination
inchicken

Study Outline

2009~2011

2008~2009
Soil contaminationand diffusionPb & Cdcontamination

incattle

Kabwe Pb & Zn MiningPublications on PAST research
1. Nakata H & Nakayama SMM et al. Reliability of stable Pb isotopes to identify Pb sources and verifying biologicalfractionation of Pb isotopes in goats and chickens. Environ Pollut. 208(Pt B):395-403 (2016)

2. Yabe J et al. Lead poisoning in children from townships in the vicinity of a lead-zinc mine in Kabwe, Zambia. Chemosphere119:941-7(2015)
3. M’kandawire E et al. Molecular characterization and mRNA expression of cytochrome P450 1A1 and cytochrome P450 3A in liver ofKafue Lechwe (Kobus leche kafuensis) as potential biomarkers of pollution of the Kafue River basin, Zambia. Engineering 6:51-58(2014)

4. Nakayama SMM, et al. Accumulation and biological effects of metals in wild rats in mining areas of Zambia. Environmental
Monitoring and Assessment 185:4907-4918 (2013)
5. Yabe J et al. Metal distribution in tissues of free-range chickens near a lead-zinc mine in Kabwe, Zambia. Environ Toxicol Chem. 2013
Jan;32(1):189-192.
6. Nakayama SMM, et al. GIS-based source estimation of Cu pollution in Lake Itezhi-tezhi and metal accumulation profiles in
Oreochromis spp. from both field and laboratory studies. Arch Environ Contam Toxicol. 2013 Jan;64(1):119-129.
7. Nakayama S.M.M, et al. Metal contaminated soil from mining area caused metal accumulation and biological responses in rats. JJVR,
61:S77-S79 (2013)
8. Yabe J et al. Accumulation of metals in the liver and kidneys of cattle from agricultural areas in Lusaka, Zambia. J Vet Med Sci. 2012
Oct;74(10):1345-7.
9. M’kandawire E, et al. The nucleotide sequence of metallothioneins (MT) in liver of the Kafue lechwe (Kobus leche kafuensis) andtheir potential as biomarkers of heavy metal pollution of the Kafue River. Gene. 2012 Sep 15;506(2):310-6.
10. Ikenaka Y et al. Effects of environmental lead contamination on cattle in a lead/zinc mining area: changes in cattle immune systems
on exposure to lead in vivo and in vitro. Environ Toxicol Chem. 2012 Oct;31(10):2300-5.
11. Nakayama SMM, et al. Metal and metalloid levels and bio-accumulation characteristics in soil, sediment, land plants andhippopotami (Hippopotamus amphibius L) from the South Luangwa National Park, Zambia. Ecotox Environ Saf. 2012 Jun;80:333-8.
12. Yabe J et al. Uptake of lead, cadmium, and other metals in the liver and kidneys of cattle near a lead-zinc mine in Kabwe, Zambia.
Environ Toxicol Chem. 2011 Aug;30(8):1892-7.
13. Nakayama SMM et al. Metal and metalloid contamination in roadside soil and wild rats around a Pb-Zn mine in Kabwe,Zambia.Environmental Pollution 159 (2011) 175-18.
14. Ikenaka Y, et al. Heavy metal contamination of soil and sediment in Zambia. African Journal of Environmental Science andTechnology 4 2010（11）：729-739.
15. Yabe J, et al. Current levels of heavy metal pollution in Africa. J Vet Med Sci. 2010 Oct;72(10):1257-63.
16. Nakayama , et al. Heavy metal accumulation in lake sediments, fish (Oreochromis niloticus and Serranochromis thumbergi) andcrayfish (Cherax quadricarinatus) in Lake Itezhi-tezhi and Lake Kariba, Zambia. Arch Environ Contam Toxicol. 2010 Aug;59(2):291-300.

Research achievement

PASTPRESENTFUTURE

Research activities
JUNE 17th ~ JULY 7th 2016Exposure Pathway ?

Blood Pb levelexceeding thelimit in 100% ofChildren

Soil contaminationand diffusion

Watercontamination

Plant/Vegetablecontamination

High Pb and Cd levels in muscle and offal
of Free-range chickens (n=17) in Kabwe
AA

mg/kg (wet weight)

Permissible limit(Offal: 0.5, muscle: 0.1)

AA

mg/kg (wet weight)

CdPb

Permissible limit(Liver: 0.5, Kidney: 1.0)
B

We did not pay attention to Eggs
BCCCDCFE

F(Tukey test)

Scavenging
Free-rangechicken

Exceeded the permissible limits

Eggs of Village Chicken
Commercial

Control site(Nakoli)

Mining area(Mutwe Wansofu)

SmallDog Blood Sampling•••Share homeenvironment
Pb exposure ?
Easy to collectHome visit with children to get
GPS coordinates of dog owners

Lecture of LEADCARE
by Dr. Jack (Pure Earth) in Kabwe

ONLY• 1 Drop of Blood• 3 minutes• Blood Pb level• Less invasive

Measured Blood Pb
on SITE (Kabwe)• 120 Dogs
• 85 Cattle• 25 Chickens

Research achievement

PAST

PRESENT

FUTURE

Co-Operation

Pure Earth ~Director of Research~
Jack Caravanos

Pure Earth ~Chowa area Remediation~
Gordon Binkhorst

KAMPAI PROJECT - OUTPUT 2

Objectives
Main objective• Assess lead (Pb) concentrations, stable Pb isotope
ratios and levels of selected biomarkers as well as
establish the health and socio-economical impact
Pb contamination in Kabwe
Specific objectives• Measure blood Pb concentrations in children and pregnant
mothers in communities around the mine in Kabwe• Measure Pb concentrations in breast milk
• Measure biomarkers to determine the effects of Pb exposure• Neurodevelopment assessment of Pb exposure in children• Assess the eStudy site
NEWS

Blood Pb concentrations in children, maternal
blood and breast milk
• Lead crosses the placenta Maternal and umbilical cord
blood Pb levels have a strong
correlation
 Blood Pb levels of the infant issimilar to that of the mother• Milk Pb levels
 Pb levels in breast milk increase
with level in maternal blood
 Concentrations in breast milk
indicates prenatal exposure• Fetuses and infants are at thehighest risk of Pb neurotoxicity

Brochin-et-al-2008-GUJHS-5-2

Blood Pb measurements

Lead Care IIICP-MS• Blood lead analysis will be conducted annually• Impact of remediation measures will be evaluatedStable Pb isotope ratios andbiomarkers• Stable Pb isotope ratios
ICP-MSTo clarify source of exposure• Blood biochemistry
COBAS Ready - blood chemical analyzer
To determine health effects• Plasma/Urine biomarker
HPLC-UVTo determine markers of exposure and effectNeurodevelopment Assessment• Children under 3 years of age
Infants and their mothers will be followed up for aperiod of 48 months at birth and thereafter everyyearThe Ages and Stages Questionnaire (ASQ) will be
used to measure the cognitive, motor, language
and behaviour of infants from one to 42 monthsof age

Neurodevelopment Assessment: 3-18 years old• Zambia Child Assessment Tool (ZamCAT)
 For preschool children to assess language, cognition and finemotor skills• Neuropsychological Assessment Tool (NEPSY) For 3-16 year old to assess subtle central nervous system
deficits (social, academic and behavioural difficulties)• Universal Nonverbal Intelligence Test (UNIT)
 For 5 to 17 year old to assess general intelligence using
nonverbal means• Kaufman's Assessment Battery for Children (KABC-II) For 7 through 18 year old to assess cognitive and mental
processes (Sequential Processing, Simultaneous processing,Learning and Planning Ability)

Economic impact assessment• Economic assessment is a new area in thisfield• The impact of lead in Kabwe will be“quantitatively” estimated to establish amodel for new assessment system

Ethical Considerations• The following specific standards will be upheld:
 Ethical clearance will be sort from UNZABREC and MOH
 Written informed consent will be obtained from parents
 Minimum risk will be upheld in the drawing of biologic
samples by the local medical personnel Samples (cord blood, breast milk, blood, hair and urine)
will be disposed off according to stipulated guidelines
 The presence of the parent/caregiver during theneurodevelopmental testing will also be a requirement
 Respondents will be assured of confidentiality of theresults Parents/guardians of affected children will be referred forfurther assessment and therapy for the child