Environmental temperature and growth

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 Articles

Environmental temperature and growth faltering in African

children: a cross-sectional study
Lucy S Tusting, John Bradley, Samir Bhatt, Harry S Gibson, Daniel J Weiss, Fiona C Shenton, Steve W Lindsay

Summary
Background Child growth faltering persists in sub-Saharan Africa despite the scale-up of nutrition, water, and Lancet Planet Health 2020;
sanitation interventions over the past 2 decades. High temperatures have been hypothesised to contribute to child 4: e116–23

growth faltering via an adaptive response to heat, reduced appetite, and the energetic cost of thermoregulation. We Department of Disease Control
(L S Tusting PhD) and MRC
did a cross-sectional study to assess whether child growth faltering is related to environmental temperature in Tropical Epidemiology Group
sub-Saharan Africa. (J Bradley PhD), London School
of Hygiene & Tropical Medicine,
Methods Data were extracted from 52 Demographic and Heath Surveys, dating from 2003 to 2016, that recorded London, UK; Big Data Institute,
Nuffield Department of
anthropometric data in children aged 0–5 years, and were linked with remotely sensed monthly mean daytime land Medicine, University of Oxford,
surface temperature for 2000–16. The odds of stunting (low height-for-age), wasting (low weight-for-height), and Oxford, UK (S Bhatt PhD);
underweight (low weight-for-age) relative to monthly mean daytime land surface temperature were determined using Department of Infectious
multivariable logistic regression. Disease Epidemiology, Imperial
College London, London, UK
(S Bhatt, H S Gibson PhD,
Findings The study population comprised 656 107 children resident in 373 012 households. Monthly mean daytime D J Weiss PhD); and Department
land surface temperature above 35°C was associated with increases in the odds of wasting (odds ratio 1·27, 95% CI of Biosciences, Durham
1·16–1·38; p<0·0001), underweight (1·09, 1·02–1·16; p=0·0073), and concurrent stunting with wasting (1·23, University, Durham, UK
(F C Shenton PhD,
1·07–1·41; p=0·0037), but a reduction in stunting (0·90, 0·85–0·96; p=0·00047) compared with a monthly mean Prof S W Lindsay PhD)
daytime land surface temperature of less than 30°C. Correspondence to:
Dr Lucy S Tusting, Department of
Interpretation Children living in hotter parts of sub-Saharan Africa are more likely to be wasted, underweight, and Disease Control, London School
concurrently stunted and wasted, but less likely to be stunted, than in cooler areas. Studies are needed to further of Hygiene & Tropical Medicine,
London WC1E 7HT, UK
investigate the relationship between temperature and child growth, including whether there is a direct effect not lucy.tusting@lshtm.ac.uk
mediated by food security, regional wealth, and other environmental variables. Rising temperature, linked to
anthropogenic climate change, might increase child growth faltering in sub-Saharan Africa.

Funding UK Medical Research Council and UK Global Challenges Research Fund.

Copyright © 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY
4.0 license.

Introduction water, and poor sanitation and hygiene, and nutritional

Childhood undernutrition, including growth faltering
and micronutrient deficiencies, is associated with
increased mortality and is estimated to contribute to
approximately 45% of deaths of children younger than
5 years globally.1 Despite reductions in stunting, wasting,
and underweight in nearly all African countries between
2000 and 2015, the prevalence of stunting remains as
high as 30% in some parts of sub-Saharan Africa.2
The WHO Global Nutrition Targets for 2012 to 2025 aim
to reduce the prevalence of stunting by 40% and the
prevalence of wasting to less than 5%. Many countries in
sub-Saharan Africa are on track to reach these targets by
2025, but few are expected to achieve Sustainable
Development Goal 2.2—to end malnutrition by 2030.3
Growth faltering is commonly attributed to prenatal,
genetic, and epigenetic factors, as well as environmental
exposures after birth, including inadequate diet, unclean
water and poor sanitation and hygiene, poor parental
education, repeat acute episodes of disease, and chronic
subclinical inflammation of the small intestine.4 Unclean
interventions have had a mixed effect on growth
outcomes,5 and reductions in child mortality and
diarrhoeal disease have not produced the expected
decrease in stunting.2 Evidence from rural parts of The
Gambia from the past 3 years suggests that major and
sustained improvements in living conditions, disease
reduction, diet, and health care might be needed to
eliminate undernutrition.2,6 The causal pathways remain
poorly understood, precluding effective inter­vention.
We explore the role of temperature in growth faltering
in sub-Saharan Africa. For thermeostasis, humans have
two responses to heat stress: decreasing food intake to
reduce metabolism and heat generation; and increasing
heat loss—eg, through increased sweating, a higher area-
to-mass ratio, and behavioural adaptations including
reduced activity. In an earlier study,7 Wells hypothesised
that these responses might contribute to growth faltering
among children living in hotter climates via three
mechanisms. First, decreasing food intake will reduce
growth and enhance heat loss through an increased

www.thelancet.com/planetary-health Vol 4 March 2020 e116

 Articles
Research in context
Evidence before this study
We searched PubMed between Jan 1, 1900, and Aug 1, 2019,
and Embase between Jan 1, 1980, and Aug 1, 2019, and
identified 45 unique studies with titles containing the terms
(“temperature*”, “climate” OR “heat”) AND (“*nutrition”,
“stunting”, “wasting”, “human growth”, “child growth”,
“growth failure”, OR “growth faltering”). One modelling study
proposed the hypothesis that child growth faltering is linked to
high environmental temperatures, via an adaptive response to
heat, reduced appetite, and the energetic cost of
thermoregulation. The study recommended empirical
investigation of this hypothesis, but we found no studies that
did so. Five studies explored the relationship between
environmental temperature and child growth, as mediated by
food prices, income, or agriculture, but none investigated a
direct biological effect.
Added value of this study
Our study is one of the first to consider a direct causal
relationship between high environmental temperature and
child growth faltering. We find evidence in support of this
area-to-mass ratio. Second, prolonged heat exposure
creates a selective pressure for reduced body size. Indeed,
there is evidence that Bergmann’s rule—that smaller
organisms are found in warmer regions—might apply to
modern humans where there is a difference between
populations greater than 50 degrees of latitude, or 30°C,
or both.8 Importantly, any effect on body size is likely to
manifest as wasting (low weight-for-height), which
increases area-to-mass ratio and cooling capacity, rather
than stunting (low height-for-age), which has little effect
on area-to-mass ratio.7 Third, heat stress incurs a high
energetic cost—eg, due to increased sweating. Together,
these effects are more pronounced in young children
who have a poorer heat tolerance than adults because of
their lower sweat rate and blood volume, among other
factors.
We assessed whether exposure to high temperatures in
sub-Saharan Africa is associated with child growth
faltering. Our study is, to our knowledge, one of the first
studies to explore a direct relationship between
environmental temperature and early childhood growth
outcomes. Our analysis is timely, given the predicted
increase in global mean surface temperature by
0·3–4·8°C by the end of this century, relative to the late
20th century,9 and the urgent need to understand the
effects of anthropogenic climate change on health.
Methods
Data sources
For growth outcomes in children aged 5 years or younger,
data were extracted from the Demographic and Health
Surveys (DHS) and Malaria Indicator Surveys (MIS).10
e117
relationship in sub-Saharan Africa, with major potential
implications for health. By analysing data for 656 107 children
aged 0–5 years in 29 countries across sub-Saharan Africa, a
region where growth faltering persists, we show that children
living in hotter parts of sub-Saharan Africa are more likely to be
wasted, underweight, and concurrently stunted and wasted,
but less likely to be stunted, than in cooler areas.
Implications of all the available evidence
Child growth faltering remains a major source of morbidity and
mortality in sub-Saharan Africa despite the scale-up of intensive
nutrition, water and sanitation interventions, and widespread
reductions in diarrhoea and all-cause mortality. To guide
effective intervention, the missing causes of growth faltering
urgently need to be understood. Our study suggests that
increasing temperature in sub-Saharan Africa might increase
the prevalence of wasting, underweight, and concurrent
stunting and wasting, but decrease the prevalence of stunting.
Future studies are needed to validate our findings, to
investigate causal pathways, and to find strategies to mitigate
against any effects of climate change on child growth.
The DHS and MIS collect health and sociodemographic
data using a two-stage random cluster sampling strategy,
where clusters are randomly selected from census files
and households are randomly selected in each cluster.
We included all georeferenced DHS and MIS done in
sub-Saharan Africa and available online by December,
2018, that collected anthropometric data and all
prespecified covariables. Stunting was defined as a
height-for-age Z score greater than two SDs below the
reference median, wasting as a weight-for-height Z score
less than –2, and underweight as a weight-for-age Z score
of less than –2.
To assess health care and household characteristics, the
following variables were extracted from DHS and MIS
datasets for each child: age, sex, insecticide-treated net use
the night before the survey, receipt of the third diphtheria-
pertussis-tetanus (DPT-3) vaccination, receipt of the first
measles vaccination (measles-1), any reported episode of
diarrhoea in the 2 weeks before the survey, improved or
unimproved drinking water source categorised using
WHO Joint Monitoring Programme (WHO-JMP) criteria,11
improved or unimproved sanitation facility categorised
using WHO-JMP crit­eria,11 education level of the household
head, urban or rural residence, and floor type (finished
[eg, parquet] vs unfinished or natural material [eg, earth]).
Household wealth index scores were calculated for
each survey using linear principal component analysis
(PCA). We applied inclusion criteria of fewer than 10%
missing values and population frequency between 5%
and 95% to the following assets: car, motorboat, scooter,
cart, bicycle, television, refrigerator, radio, watch, mobile
telephone, landline telephone, and electrification of the
www.thelancet.com/planetary-health Vol 4 March 2020
 Articles

50
Angola
Benin
Burkina Faso
40 Burundi
Cameroon
Chad
Comoros
Air temperature (°C)

30 DR Congo
Côte d’Ivoire
eSwatin
20 Gabon
Ghana
Guinea
Country Kenya
10 Liberia
Madagascar
Malawi
Mali
0
0 10 20 30 40 50 Mozambique
Land surface temperature (°C) Namibia
Nigeria
Figure 1: Monthly mean daytime air temperature versus monthly mean
Rwanda
daytime land surface temperature for 25 089 georeferenced clusters in
Senegal
52 surveys, 2000–16
Air temperature was determined from land surface temperature using Sierra Leone
a standard conversion.17 i
Tanzania
Togo
household. To condense these 12 assets into a single Uganda
dimensional index we tested isometric mapping, kernel Zambia
principle component analysis, t-distributed stochastic Zimbabwe 0 5 10 15 20 25 30 35 40 45
neighbour embedding, and linear PCA. We found Temperature (°C)
minimal differences between algorithms, so used linear
Figure 2: Monthly mean daytime land surface temperature for the
PCA because it is the most commonly used algorithm for 29 countries included in the analysis, 2000–16 (median, IQR, and range)
wealth indices.12 Outside values are not shown.
For each georeferenced survey cluster the following
data were extracted: synoptic mean monthly daytime land attendance of the household head at secondary education,
surface temperature (LST) from 2000 to 2016 (ie, the urban or rural survey cluster, type of household floor
average temperature for each month derived from material, household wealth, monthly mean EVI, mean
a multiyear time series);13 synoptic mean monthly total monthly precipitation, night-time lights, and
enhanced vegetation index (EVI) from 2000 to 2016, accessibility to large cities. Because growth outcomes
a measure of the proportion of photosynthetically active vary with age,18 we stratified the analysis by age group
radiation absorbed by vegetation, which is correlated with (age <2 vs 2–5 years). To allow for a non-linear relationship
vegetation density and active photosynthesis;13 synoptic between temperature and growth outcomes, LST was
total monthly precipitation from 2000 to 2016;14 night- modelled as a categorical variable (ie, <30°C, 30°C to
time lights, a proxy for regional wealth and urbanicity;15 <35°C, and ≥35°C). These cutoffs were chosen to give
and accessibility to large cities in 2015.16 a similar number of children in each temperature
category. Because of the hierarchical nature of the dataset
Data analysis in which children were sampled within households and
Logistic regression was used to estimate the odds of clusters, we used CIs that accounted for the highest level
stunting, wasting, underweight, and concurrent stunting of clustering (ie, survey cluster) and we included survey
with wasting at the level of the child, relative to synoptic as a fixed effect in the model.19 SEs were estimated using
mean monthly daytime LST from 2000 to 2016 for the Taylor linearisation.
survey cluster. An LST of 30°C approximately translates
into an air temperature of 27°C (figure 1),17 although this Role of the funding source
relationship is variable and depends on the month. The The funder of the study had no role in study design, data
analysis controlled for survey, age, and sex of the child, collection, data analysis, data interpretation, or writing of
insecticide-treated net use, DPT-3 vaccination, measles-1 the report. The corresponding author had full access to
vaccination, reported diarrhoea in the past 2 weeks, type all the data in the study and had final responsibility for
of drinking water source, type of sanitation facility, the decision to submit for publication.

www.thelancet.com/planetary-health Vol 4 March 2020 e118

 Articles

Stunting Wasting Underweight Stunting and wasting

Prevalence, n (%) OR* (95% CI) p value Prevalence, n (%) OR* (95% CI) p value Prevalence, n (%) OR* (95% CI) p value Prevalence, n (%) OR* (95% CI) p value
All children
<30°C 96 687 (30·7%) 1 (ref) 0·0009 94 587 (5·9%) 1 (ref) <0·0001 93 957 (19·4%) 1 (ref) 0·018 93 957 (1·4%) 1 (ref) 0·0058
30–34°C 106 552 (30·2%) 0·98 ·· 106 006 (7·9%) 1·05 ·· 104 995 (21·8%) 1·02 ·· 104 995 (2·0%) 1·17 ··
(0·94–1·01) (0·99–1·11) (0·98–1·06) (1·06–1·29)
≥35°C 66 668 (29·6%) 0·90 ·· 66 928 (12·5%) 1·27 ·· 66 343 (30·3%) 1·09 ·· 66 343 (3·6%) 1·23 ··
(0·85–0·96) (1·16–1·38) (1·02–1·16) (1·07-1·41)
Children aged <2 years
<30°C 42 442 (25·8%) 1 (ref) 0·080 41 589 (7·9%) 1 (ref) <0·0001 41 414 (18·5%) 1 (ref) 0·024 41 414 (1·7%) 1 (ref) 0·15
30–34°C 46 579 (25·9%) 0·98 ·· 46 197 (10·0%) 1·11 ·· 45 940 (20·5%) 1·07 ·· 45 940 (2·3%) 1·13 ··
(0·93–1·03) (1·04–1·19) (1·01–1·13) (0·99–1·28)
≥35°C 28 031 (24·0%) 0·92 ·· 27 986 (14·6%) 1·27 ·· 27 889 (26·7%) 1·11 ·· 27 889 (3·9%) 1·18 ··
(0·85–0·99) (1·15–1·40) (1·03–1·21) (0·98–1·41)
Children aged 2–5 years
<30°C 54 245 (34·5%) 1 (ref) 0·0013 52 998 (4·4%) 1 (ref) <0·0001 52 543 (20·2%) 1 (ref) 0·018 52 543 (1·2%) 1 (ref) 0·015
30–34°C 59 973 (33·5%) 0·98 ·· 59 809 (6·2%) 1·00 ·· 59 055 (22·8%) 0·99 ·· 59 055 (1·9%) 1·22 ··
(0·94–1·02) (0·92–1·09) (0·94–1·04) (1·06–1·41)
≥35°C 38 637 (33·7%) 0·89 ·· 38 942 (11·0%) 1·28 ·· 38 454 (33·0%) 1·07 ·· 38 454 (3·4%) 1·29 ··
(0·83–0·95) (1·13–1·45) (1·00–1·16) (1·06–1·57)
Temperatures are the monthly mean daytime land surface temperature, calculated from synoptic monthly means from 2000 to 2016. OR=odds ratio. *Adjusted for survey, age and sex of the child, type of
drinking water source, type of sanitation facility, third diphtheria-pertussis-tetanus immunisation, first measles immunisation, reported diarrhoea in the past 2 weeks, type of household floor material,
insecticide-treated net use, attendance of household heads at secondary education, urban or rural survey cluster, household wealth, mean monthly enhanced vegetation index, mean monthly precipitation,
urbanicity, night-time lights, and accessibility to large cities.

Table: Association between land surface temperature and growth outcomes in children aged 0–5 years in sub-Saharan Africa

Results [OR] 1·17, 95% CI 1·13–1·21; p<0·0001; appendix p 3).

Summary statistics are presented in the
See Online for appendix appendix (pp 1–2). Data were extracted for 121 DHS,
MIS, and AIDS Indicator surveys. Of these, 52 DHS
surveys that measured growth outcomes and all
prespecified covariables were included in the
multivariable analysis. The included surveys were done
from 2003 to 2016, in 29 countries. The total study
population comprised 656 107 children, aged 0–5 years,
resident in 373 012 households. The median cluster size
was 15 households (IQR 11–18). The mean age of
children was 2·5 years (95% CI 2·5–2·5) and
325 919 (49·7%) of 656 096 children were female.
During 2000–16, monthly mean LST in survey clusters
ranged from a median of 26·1°C (IQR 25·2–28·5) in
Gabon to 37·7°C (36·1–39·2) in Chad (figure 2).
Of the 336 299 children in 52 surveys for whom height
and age were recorded, 102 213 (30·4%) were stunted.
Stunting was more common among children aged
2–5 years (68 674 [33·5%] of 205 031) than children aged
younger than 2 years (33 539 [25·6%] of 131 268; p<0·0001),
and among boys (53 950 [31·9%] of 169 265) than girls
(48 263 [28·9%] of 167 033; p<0·0001). Stunting prevalence
ranged from 977 (15%) of 6637 in the Senegal 2016 DHS to
1758 (49%) of 3624 in the Burundi 2010 DHS
(appendix pp 1–2). In the unadjusted analysis, the odds of
stunting were 17% higher among children living in survey
clusters where monthly mean daytime LST exceeded
35°C, compared with those in clusters where monthly
mean daytime LST was less than 30°C (odds ratio
Controlling for the variables stated in the Methods section,
the odds of stunting were 10% lower among children
living in survey clusters where monthly mean daytime
LST exceeded 35°C, compared with those in clusters
where monthly mean daytime LST was less than 30°C
(0·90, 0·85–0·96; p=0·00047; table). The reduction in
stunting associated with an LST exceeding 35°C, compared
with an LST of less than 30°C, was greater in children
aged 2–5 years (0·89, 0·83–0·95; p=0·00081) than in
children younger than 2 years (0·92, 0·85–0·99; p=0·034).
Of the 335 835 children in 51 surveys for whom weight
and height were recorded, 26 264 (7·8%) were wasted.
Wasting was more common among children younger
than 2 years (13 351 [10·3%] of 130 054) than those aged
2–5 years (12 913 [6·3%] of 205 781; p<0·0001), and among
boys (14 038 [8·3%] of 169 013) than girls (12 226 [7·3%] of
166 820; p<0·0001). Wasting prevalence ranged from
67 (2%) of 3828 children in the Rwanda 2015 DHS to
3897 (15%) of 26 463 in the Nigeria 2013 DHS. In the
unadjusted analysis, the odds of wasting were 95% higher
in countries where monthly mean daytime LST exceeded
35°C, compared with countries in which monthly mean
daytime LST was less than 30°C (OR 1·95, 95% CI
1·85–2·05; p<0·0001; table). In the adjusted analysis, the
odds of wasting were 27% higher in countries where
monthly mean daytime LST exceeded 35°C, compared
with those where monthly mean daytime LST was less
than 30°C (1·27, 1·16–1·38; p<0·0001). The increase in
wasting associated with LST exceeding 35°C was similar

e119 www.thelancet.com/planetary-health Vol 4 March 2020


across age groups (age <2 years 1·27, 1·15–1·40, p<0·0001;
age 2–5 years 1·28, 1·13–1·45, p=0·0001).
For the 330 809 children in 51 surveys for whom
weight and age were recorded, 75 019 (22·7%) were
underweight. Underweight was more common among
children aged 2–5 years (47 587 [23·6%] of 201 562)
than those younger than 2 years (27 432 [21·2%] of
129 247 children; p<0·0001) and among boys (38 887
[23·4%] of 166 487) than girls (36 132 [22·0%] of 164 321;
p<0·0001). Underweight prevalence ranged from
203 (7%) of 2784 in the eSwatini 2006 DHS to 4017 (37%)
of 10 749 in the Chad 2014 DHS. In the unadjusted
analysis, the odds of underweight were 64% higher in
countries where monthly mean daytime LST exceeded
35°C, compared with countries where monthly mean
LST was less than 30°C (OR 1·64, 95% CI 1·57–1·70;
p<0·0001; table). In the adjusted analysis, the odds of
underweight were 9% higher among children living in
survey clusters where monthly mean daytime LST
exceeded 35°C, compared with children living in survey
clusters where monthly mean daytime LST was less
than 30°C (1·09, 1·02–1·16; p=0·0073). The increase in
underweight associated with LST exceeding 35°C was
slightly greater in children younger than 2 years (1·11,
1·03–1·21; p=0·011) compared with children aged
2–5 years (1·07, 1·00–1·16; p=0·063).
Height, age, and weight were recorded for
330 809 children in 51 surveys, of whom 6982 (2·1%)
were both stunted and wasted. Concurrent stunting
with wasting was more common in children younger
than 2 years (3185 [2·5%] of 129 247) than those aged
2–5 years (3797 [1·9%] of 201 562; p<0·0001), and among
boys (4039 [2·4%] of 166 487) than girls (2943 [1·8%] of
164 321; p<0·0001). The prevalence of concurrent
stunting with wasting ranged from 27 (0·4%) of 6142 in
the Zimbabwe 2015 DHS to 512 (5%) of 10 749 in the
Chad 2014 DHS. In the unadjusted analysis, the odds of
concurrent stunting with wasting were more than
two times higher in countries where monthly mean
daytime LST exceeded 35°C, compared with countries
where monthly mean daytime LST was less than 30°C
(OR 2·08, 95% CI 1·92–2·25; p<0·0001). In the adjusted
analysis, the odds of concurrent stunting with wasting
were 23% higher in countries where monthly mean
daytime LST exceeded 35°C, compared with countries
where monthly mean daytime LST was less than 30°C
(1·23, 1·07–1·41; p=0·0037). The increase in concurrent
stunting and wasting associated with LST exceeding
35°C was greater in children aged 2–5 years (1·29,
1·06–1·57; p=0·0099) compared with children aged
younger than 2 years (1·18, 0·98–1·41; p=0·073).
Discussion
We tested the hypothesis that young children in Africa
exposed to high temperatures will experience growth
faltering. By analysing data for children aged 0–5 years
from 52 national surveys in sub-Saharan Africa, we
www.thelancet.com/planetary-health Vol 4 March 2020
Articles
found that monthly mean daytime LSTs exceeding 35°C
were associated with a 27% increase in the odds of
wasting, a 9% increase in the odds of underweight, and a
23% increase in concurrent stunting with wasting, but a
10% reduction in stunting, compared with temperatures
averaging less than 30°C between 2000 and 2016.
Previous studies of a direct effect of temperature on child
growth are scarce. In a key study,7 Wells developed a
model of the relationship between body size and heat
production, showing that growth faltering can theor­
etically relieve heat stress in childhood and recom­
mending empirical investigation of this hypothesis.
Other studies20,21 support a role for environmental
temperature in early child growth, such as birthweight.
For example, Wells and Cole20 observed, across
108 populations, that a one unit increase in heat index
was associated with a 3% decrease in birthweight. To our
knowledge, ours is the first empirical investigation of a
direct relationship between temperature and early
childhood growth outcomes.
High temperatures are hypothesised to contribute to
growth faltering through three main mechanisms.7
First, wasting might be an adaptive response to high
temperatures. Arguably it is harder to avoid heat stress
in hot climates via behaviour change than in cooler
climates, creating a greater reliance on physiological
adaptations, particularly increased area-to-mass ratio.7
In the first 3 years of life, this pressure is exacerbated
because the greater overall body fat relative to the weight
of young children creates a low area-to-mass ratio and,
compared with adults, young children have a lower
sweat rate, greater blood volume per unit mass, higher
metabolic rate per unit mass, and higher metabolic cost
of locomotion, making it harder to keep cool.22 In
addition the heat stress experienced in hot climates
might be exacerbated in some African communities if a
baby is swaddled, the young child is carried on its
mother’s back (appendix p 4), or the child sleeps in a
metal-roofed house. In malnourished children, energy
expenditure might be higher still, because organ tissue
(which has a higher metabolic rate than normal tissue)
is proportionally larger than in healthy children.7
Second, linked to physiological adaptation, appetite
might be reduced in higher temperatures.23 If prolonged,
lower food intake would contribute to growth faltering.
Third, there is an energetic cost to keeping cool.
Approximately 40–50% of total energy expenditure in
mammals is used to maintain a constant core
temperature and a 1°C increase in body temperature is
associated with a 10–13% increase in metabolic rate.24
Children experiencing long-term heat stress might
therefore allocate less energy to growth.
Increased temperatures were associated with a greater
risk of wasting and underweight, but a decrease in
stunting. Although future studies are needed to replicate
this finding, several potential biological explanations
exist. First, ambient temperature is known to modulate
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bone elongation in mammals, with limbs of animals
raised in warmer ambient temperatures significantly and
permanently longer than those of siblings housed at
cooler temperatures.25 Thus, human limb length might
increase with heat exposure. Second, if wasting is
protective in high temperatures,7 stunted children might
have a higher mortality rate than wasted children under
heat stress. Wells’ model of temperature and body size
suggests that stunting and wasting both affect surface
area and energy expenditure, but stunting causes little
increase in area-to-mass ratio in children aged 0–2 years.7
In contrast, wasting increases area-to-mass ratio by at
least 15% in young children, promoting cooling capacity
in early infancy. We found stunted children to be older
than wasted children and the reduction in stunting
associated with higher temperatures to be most
pronounced among children aged 2–5 years. This finding
is consistent with a decline in prevalence of stunted
children in higher temperatures because of increased
mortality, although research is needed to understand this
further. There might be other, unknown, temperature-
related drivers making wasting a more likely outcome of
undernutrition than stunting as temperatures rise.
We hypothesise that a decline in stunting associated
with increased temperatures might also be linked to
genetic factors. Only 2% of our study population was
both stunted and wasted, similar to a pooled prevalence
of 3% in the same age group from a meta-analysis of
84 countries.26 This partly reflects the high mortality rate
in children affected by both,27 but also illustrates how
stunting and wasting largely affect different children.
Indeed, a high prevalence of stunting can be observed in
populations where wasting is uncommon.28 Human
height is approximately 80% genetically determined by at
least 600 genetic variants that are not yet fully
understood,29 with Africans having the highest levels of
within-population genetic diversity worldwide. It is
possible that height in African populations is influenced
by unknown epigenetic factors that interact with
environmental temperature. Studies of the prevalence of
stunting in relation to both ethnicity and environment in
sub-Saharan Africa might be insightful.
We found that the relationship between temperature
and growth outcomes varied with children’s age, with the
association between high temperature and growth
faltering being greater among children aged 2–5 years
than 0–2 years for stunting and concurrent stunting with
wasting, and the same association smaller among children
aged 2–5 years than 0–2 years for underweight. A previous
study showed systematically larger estimates of association
with risk factors for stunting in children aged 2–5 years
than younger than 2 years, but systematically smaller
effects in the older age group for wasting.18 This finding
might be because wasting rates are generally highest in
children aged younger than 2 years, but stunting can take
longer to manifest and is more prevalent after 2 years.18
Here we observed a similar pattern. Our analysis shows
e121
the importance of age disaggregation in observational
studies of child growth.
Previous studies investigating the causes of stunting,
wasting, and underweight have focused mainly on
nutrition, socioeconomic factors, exposure to infection
and maternal health.4 Yet the persistence of growth
faltering in sub-Saharan Africa despite the scale-up of
nutrition and hygiene-based interventions suggests
additional causes.2 The few studies30,31 to have explored the
role of climate have focused on the effect of environmental
temperature on food security, rather than a direct
biological effect, as we have done. Further research is
needed to validate and understand our findings, but a
direct, causal link between temperature and growth
faltering would have major implications for public health.
Mean annual temperatures are expected to rise by 2°C
across Africa by the end of this century, relative to the late
20th century, with some scenarios showing rises of
between 3°C and 6°C for the same period.9 Estimates of
future deaths attributable to climate change, as well as
public health strategies to mitigate against its effects
might therefore need to consider growth faltering.
Importantly, if wasting is an adaptive and arguably
protective response to high temperatures, it might be
helpful to counteract this effect by keeping children cool
while ensuring adequate nutrition. Although research
would be needed to identify appro­priate interventions,
one possible approach could be the promotion of cooler
housing in hotter parts of sub-Saharan Africa. Rather
than air conditioning, this can be achieved with buildings
with low thermal mass and good ventilation.32
We observed an association between environmental
temperature and child growth, but further research is
needed to understand the extent to which this is mediated
by food security and poverty, rather than a direct
biological effect. Hotter climates are generally associated
with greater levels of poverty, which is related to child
growth. For example, Chad was the hottest country
included in our analysis, but ranks near the lowest of the
global Human Development Index.33 Despite controlling
for local differences in wealth using a household-level
wealth index, education level of the household head,
urban–rural status and night-time lights, residual
confounding is possible. Higher temperatures can also
be associated with lower crop yields, affecting nutrition,
and we attempted to control for this using EVI and
precipitation. On the other hand, local climate is one of
many factors influencing food security, which is also
determined by food distribution, exchange and afford­
ability, the resources available to prepare food safely, and
the stability of such factors over time.34 Incorporating
survey as a fixed effect in our model helped to account for
national-level variation in such factors.
Our study has several limitations. First, our measure
of temperature was based on synoptic means from
2000 to 2016, and is therefore an imprecise measure of
climate conditions experienced by children of different
www.thelancet.com/planetary-health Vol 4 March 2020

ages and surveyed in different years. Nonetheless, child
growth is a long-term process that would be expected to
respond more slowly to environmental conditions than
other health outcomes, such as vector-borne disease.
Similarly, we used synoptic means for precipitation, and
EVI and static estimates for night-time lights and
accessibility. Second, LST does not directly translate into
ambient temperatures, although it is the most precise
metric of temperature available from remotely sensed
data.17 Third, the OR, although a valid measure in its
own right, might not be taken as approximation of the
risk ratio when the outcome of interest is common
(>10%). Fourth, as discussed, residual confounding by
social and environmental factors is possible. Last, the
causes of growth faltering are complex and our analysis
is only a preliminary exploration of the role of
temperature. Cross-sectional data, like all observational
data, cannot definitively determine causality. However,
because it is not feasible to randomise children to high
or low temperatures, it is the only method available to
investigate this question. Further studies are needed to
confirm our findings, such as the observed decrease in
stunting at higher temperatures. Research is also needed
to understand causal pathways, including whether the
persistence of growth faltering represents an adaption to
hot climates, and to understand the relationship between
heat exposure and growth over the human life course.
Although we did not adjust for birthweight in our model,
high environmental temperatures have previously been
shown to be associated with lower birthweight20,21 and
shorter gestational length.35 Overall, we aim to highlight
the potential role of environmental temperature in the
growth of African children and to provide a stimulus for
future research and discussion of this topic.
In conclusion, children aged 0–5 years living in areas of
higher temperatures in sub-Saharan Africa are more likely
to be wasted and underweight, but less likely to be stunted
as compared with those in lower temperatures. Future
studies should evaluate the relationship between heat
exposure and growth outcomes in children and elucidate
causal mechanisms. Any direct effect of temperature on
early childhood growth would need to be considered in
light of the likely increases in mean temperature that
Africans will experience this century, caused by climate
change.
Contributors
FCS and SWL conceived of the study. LST, SB, HSG, and DJW curated
data. LST, JB, DJW, and SWL designed the methods. LST and JB did the
formal analysis. LST, FCS, and SWL wrote the original draft of the
Article. LST, JB, SB, HSG, DJW, FCS, and SWL reviewed and edited
the Article.
Declaration of interests
We declare no competing interests.
Data sharing
All health data used in this analysis are available to download free of
charge by registered users from the Demographic and Health Surveys
Program. Registration, data, and full dataset access instructions are
available online. Remotely sensed data are available to download from
the cited sources.
www.thelancet.com/planetary-health Vol 4 March 2020
Articles
Acknowledgments
LST is a Skills Development Fellow (number N011570) jointly funded by
the UK Medical Research Council (MRC) and the UK Department for
International Development (DFID) under the MRC/DFID Concordat
agreement and is also part of the European and Developing Countries
Clinical Trials Partnership programme supported by the EU. SWL is
supported by the Global Challenges Research Fund funded by the
Biotechnology and Biological Sciences Research Council,
DFID, and MRC. JB received support from the UK MRC and DFID
(number MR/K012126/1) under the MRC/DFID Concordat agreement
and as part of the EDCTP2 programme supported by the EU. We thank
Maria J Carrasco-Tenezaca for the photograph in the appendix (p 4).
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www.thelancet.com/planetary-health Vol 4 March 2020
Another random document with
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titrated hydrochloric acid. For general purposes, however, this
determination is usually of little interest.
398. Müller’s Modified Kjeldahl Method.—Numerous
difficulties, as stated by Müller,[263] have attended the attempts to
apply the kjeldahl method for the estimation of nitrogen to samples
of soil, and he has modified the method to some extent and made
comparisons of the quantity of nitrogen by this modified method and
by the soda-lime method.
The principal difficulty encountered by him has been in the regular
heating of the mixture of fuming sulfuric acid and soil. The particles
of soil are deposited at the bottom of the flask and the result is that
the bottom layers become overheated, and, being poor conductors of
heat, fail to transmit a sufficient quantity of heat to penetrate to the
upper layers of the liquid to complete the reaction. In order to avoid
this difficulty Müller heats his flask in a small stove formed with a
straight vertical cylinder of iron or copper, the upper end of which is
covered with a sheet of iron pierced with a hole which allows the
neck of the flask to pass through, while the lower end is closed with a
piece of sheet iron furnished on its upper surface with a layer of
asbestos. This cylinder of metal is surrounded with a second one,
concentric with the first through which passes a current of heated
gases furnished by an ordinary bunsen. By heating the flask in this
stove or furnace an even distribution of the heat is secured to all
parts of the mixture, but the little drops of sulfuric acid, which are
condensed on the cold part of the neck, sometimes lead to the
fracture of the glass as they run down the sides of the flask to the hot
portions. To prevent the reflux of this condensed acid, which only
needs to be done near the end of the reaction, when it is necessary to
heat to a very high temperature, the neck of the flask is bent at the
point immediately above its emergence at the upper surface of the
furnace, and carried into a flask of about seventy-five cubic
centimeters capacity, which will receive the drops of sulfuric acid
condensed during the operation. The furnace has the following
dimensions; height, twelve centimeters; diameter of interior
cylinder, five and one-half centimeters; diameter of exterior cylinder,
seven and one-half centimeters.
It is supported on a triangle of large iron wire and is heated by an
ordinary bunsen, or by a concentric bunsen, according to the
temperature which it is necessary to obtain. The proportions which
should be observed between the amount of earth employed and the
sulfuric acid are about as follows:
Of the dry earth, fifteen grams; of the fuming sulfuric acid, thirty
cubic centimeters. There should also be added to the mixture about
three-tenths of a gram of pure stearic acid, or better, benzoic acid.
When the soil to be analyzed does not contain carbonate, the sulfuric
acid should be added in two portions. At first add about twenty cubic
centimeters of the acid, and after shaking it, the other ten cubic
centimeters, running it in from a burette or a pipette in such a
manner as to wash thoroughly the neck and sides of the flask. If the
earth contain carbonate, however, it is necessary to add the fuming
acid in small portions of about five cubic centimeters at a time,
waiting each time until the disengagement of the gas caused by the
previous addition has ceased. A soil which contains from thirty to
forty per cent of calcium carbonate should be carefully treated in a
porcelain capsule with a slight excess of sulfuric acid, pure and
dilute. The mixture is afterward to be evaporated to dryness upon a
sand-bath and the residue heated in a drying oven to 110°. The mass
is then pulverized, introduced into the flask, treated with three-
tenths of a gram of benzoic acid and thirty cubic centimeters of
fuming sulfuric acid, and treated as indicated above. In all cases it is
necessary to continue the heating until the contents of the flask are
colorless.
With soils containing considerable quantities of iron, however, a
slight red color will probably be observed which will not interfere
with the accuracy of the tests.
The heating should at first be gentle and the temperature
afterward elevated little by little, and finally the heat should be
sufficiently great to distill about one and one-half cubic centimeters
of sulfuric acid. The operation lasts from twelve to thirteen hours. As
the reaction is terminated the cooled mass is taken up with water
absolutely free from ammonia. It is filtered into a flask, and washed
upon the filter until the volume of the filtered liquid is about 350
cubic centimeters. Afterward an excess of soda-lye, at 50° baumé is
added, then a few pieces of quartz to facilitate boiling. The flask is
then connected with a condenser, the liquid distilled and received in
a conical flask closed by a cork having two holes, of which one
permits the entrance of the end of the condenser, and the other a
glass tube which is connected with a small flask containing water, the
neck of the receiving flask being inclined toward the condenser to
avoid the entrainment of any of the alkaline liquid which may be
distilled. The receiving flask rests upon two or three pieces of sheet
iron and is heated with an ordinary burner, and ebullition is perfectly
regular. From 170 to 180 cubic centimeters of the liquid are distilled
in from three and one-half to four hours. The distilled liquid, treated
with a few drops of litmus, is titrated by a solution of sulfuric or
hydrochloric acid, of which one cubic centimeter corresponds to
0.001 cubic centimeter of nitrogen.
399. Modification of the Kjeldahl Method by Arnold and
Wedemeyer.[264]—For the oxidizing liquid a mixture of three grams
of benzoic acid with forty cubic centimeters of H₂SO₄ is employed.
After placing in the digestion flask with the nitrogenous body the
whole is gently shaken for a few minutes to prevent clotting. The
temperature is then raised until acid vapors begin to come off, when
one gram of copper sulfate and one gram of mercuric oxid are added;
and after ten to fifteen minutes, to avoid foaming, ten to twenty
grams of potassium sulfate. The sublimate noticed on the walls of the
flask is benzoic acid and does not interfere with the accuracy of the
determination.
This method has given good results with the alkaline nitrates, the
nitrates of barium, mercury, silver, lead, and with strychnia,
ammonia, pyridin, azobenzol, dinitrobenzol, and picric acid.
400. Prevention of Bumping During Distillation.—Daffert
has employed the modified kjeldahl method, but found considerable
difficulty in using the same owing to the violent bumping of the
liquid in the distillation. This was especially the case where the
sample contained a large proportion of sand. To overcome this
annoyance and danger he devised the following process:[265]
Fit into the mouth of a large-mouthed distillation flask a stopper
having two perforations. Through one of the perforations pass the
usual distillation tube, through the other a similar tube connected
with a supply of steam. Bring the contents to a brisk boil, after which
a small current of steam is turned on, allowing the same to pass in a
small stream throughout the distillation. By this means, not only is
all danger from bumping avoided, but the time required for the
distillation shortened. By the old method it usually requires from
fifteen to twenty minutes, whereas the former requires from six to
ten minutes.
It is advisable to filter all samples of soils having a large proportion
of sand.
401. Determination of Organic Nitrogen by the Soda-
Lime Method.—In the description of the method following, the
directions of the French Commission of Agricultural Chemists have
been taken as the basis of the analytical process.[266] This method is,
in this country, almost superseded by the moist combustion process
with sulfuric acid. By reason of its long use, however, and because it
is still regarded as the best method by the agricultural chemists of
France, Italy, and England, it merits a full description. It is
recommended also by Berthelot and André,[267] by the International
Congress of Chemists, held in Paris in 1889, by the Italian chemists,
and by the official Belgian method,[268] in all cases where nitrates are
not present in notable quantities.
The nitrogen which is found in soils in the organic state is
transformed into ammonia when it is heated with soda-lime. This
reaction is the base of the process of analysis which has so long been
used for this class of bodies. The analytical process is conducted as
follows:
A well-cleaned glass combustion tube, closed at one end, is used.
The length of the tube is from thirty-five to forty centimeters. It is
filled first to a depth of two centimeters with calcium oxalate;
afterwards to a depth of five centimeters with soda-lime in small
fragments; afterwards with the mixture to be analyzed; viz., of ten
grams of the sample of soil, or twenty grams if poor in nitrogen and
organic matters, with soda-lime reduced to a coarse powder. This
mixture should occupy a length of about twenty centimeters in the
tube. The soil and soda-lime are mixed in a mortar. Afterwards the
mortar is rubbed with small quantities of soda-lime, and this,
together with the copper boat which has been used in introducing the
mixture, is thoroughly washed with the soda-lime, which is poured
into the tube until it is filled to within four centimeters of its open
extremity. The open end of the tube is then closed with a wad of
asbestos packed sufficiently tight to prevent the carrying off of the
soda-lime by the gas which may be generated during the combustion.
The combustion should be commenced by heating the tube near the
open extremity until it is red and carrying the heat progressively
towards the part containing the soil mixed with the soda-lime. An
ordinary gas combustion furnace should be used and the heat
graduated in such a way that the bubbles of gas pass off regularly and
not too rapidly. The gas is conducted into a bulb tube containing a
decinormal standard sulfuric acid colored with litmus. The
combustion is continued until the whole of the organic material is
decomposed, care being taken not to raise the combustion tube
above a low redness in order to avoid its softening. At the end,
however, the temperature of the combustion tube should be raised to
a bright red, and the part containing the calcium oxalate should be
heated little by little for the purpose of evolving hydrogen, which is
used to drive out the last traces of ammonia. After the combustion is
completed, and the last traces of ammonia driven out, the standard
acid which has received the evolved ammonia is removed, the tube
leading to it washed, the wash-water collected with the rest of the
liquid and titrated with a standard solution of lime-water, the
strength of which has previously been determined against standard
sulfuric acid.
402. Preparation of the Standard Sulfuric Acid.—The
sulfuric acid to be used in making the standard solutions should be
previously boiled for half an hour in a platinum dish and allowed to
cool in a desiccator. It should contain 61.25 grams of sulfuric acid in
one liter. It is recommended that the flask which holds the sulfuric
acid should be one which has been used for a long time for holding
concentrated sulfuric acid, in order to avoid any action of the alkali
in the glass upon the acid after its strength has been determined. The
solution before described is of such strength as to have each cubic
centimeter equivalent to one milligram of nitrogen.
For the estimation of the nitrogen in the soil a tenth normal
solution should be used, which is prepared by taking 100 cubic
centimeters of the normal solution, described above, and diluting to
one liter.
Preparation of the Lime-Water.—From 200 to 300 grams of
slaked lime are placed in a closed flask of about five liters capacity.
This is filled with water and shaken frequently, and left to deposit the
matter in suspension. The water which contains the saline particles
which may have been present in the lime is then poured off. Fresh
water is then poured on and the flask shaken from time to time. To
use this lime-water the clear part of it is decanted into a flask,
avoiding, as much as possible, access to the air. The flask is closed
with a cork carrying two tubes drawn out and bent at a right angle.
One of these serves for pouring off the water and the other serves for
the entrance of the air. These two tubes are themselves closed by
means of a rubber tube carrying a pinch-cock. The strength of the
lime-water is fixed by titration with the decinormal standard sulfuric
acid.
Preparation of the Soda-Lime.—Six hundred grams of slaked lime
in fine powder are saturated with 300 grams of caustic soda
dissolved in 300 cubic centimeters of water. The whole is rubbed
into a paste and introduced into a crucible which is heated to
redness. The contents of the crucible, still hot, are poured out, and
rapidly reduced to fragments in a copper mortar in such a manner as
to have the pieces about the size of a pea, and without having too
much finely powdered soda-lime mixed with it. While the matter is
still hot it is placed in a flask and well-stoppered. In order that this
reagent should contain no nitrogen it is indispensable to use in its
preparation materials which contain no trace of nitrates.
Preparation of the Calcium Oxalate.—In a small copper vessel
place 100 grams of oxalic acid and add gradually, bringing it to
boiling, enough water to dissolve it. Afterwards place in the solution
small portions of slaked lime in a state of powder, constantly testing
it until turmeric paper indicates that there is a little lime in excess. It
is then evaporated, stirring vigorously on the open fire, and the
evaporation is finally finished on a steam-bath. The dried material is
placed in a flask and well-stoppered. The oxalic acid which is used in
this preparation should be free from every trace of nitrogen.
Preparation of the Litmus Solution.—Five grams of litmus are
placed in a flask with a flat bottom. Afterwards a few cubic
centimeters of ammonia are added, twenty five grams of crystallized
sodium carbonate, and ten cubic centimeters of water. This mixture
is left to digest for sometime, with frequent stirring, at a temperature
of from 60°–80°. The digestion is finished in about four or five days,
during which time, at intervals, a few drops of ammonia are added,
sufficient to maintain always the ammoniacal odor. At the end of this
time 200 cubic centimeters of water are added and the digestion
allowed to continue several days more, still maintaining the solution
alkaline with ammonia. A slight excess of hydrochloric acid is added,
and the matter which is precipitated is received upon a filter where it
is washed several times with cold water and allowed to dry at a low
temperature.
For use, from one to two grams of this dry precipitate are dissolved
in 100 cubic centimeters of alcohol, and there is thus obtained a
litmus solution of extreme sensibility.
403. Treatment of Soil Containing Nitrates.—Nitrates exist
in small quantities in all arable soils. When treated for nitrogen by
the soda-lime method above described, a part of the nitric nitrogen is
changed to the state of ammonia, while another part escapes
estimation altogether, causing an error which it is important to point
out. When the soils contain only small quantities of nitrates this
error is insignificant and does not affect sensibly the results, but in
the case of earths rich in nitrates it is necessary first to eliminate
them before the determination of the nitrogen by the soda-lime
method. The operation is carried on as follows:
Twenty grams of the soil are washed on a small funnel, furnished
with a plug of asbestos, with small quantities of pure water, in such a
way as to cause thirty to forty cubic centimeters of water to pass
through. The whole of the nitrate is thus removed. The soil is now
dried and submitted to analysis by the soda-lime method as just
described. There are removed with the nitrate only small traces of
organic nitrogen, too small to influence the results of the analysis. If,
however, it is desired to remove altogether this slight cause of error,
evaporate the wash-waters, above described, to two or three cubic
centimeters; add a few drops of a concentrated solution of ferrous
chlorid and as much hydrochloric acid, and boil some minutes in
order to drive off, in the state of nitrogen dioxid, all the nitric acid.
The residue is evaporated to dryness and contains the traces of
organic nitrogen. This is added to the soil which is to be treated by
the soda-lime method.
404. Müller’s Method.—The determination of nitrogen in the
soil by soda-lime is carried on as follows by Müller:[269]
Fifteen grams of fine earth, dried and mixed with a little sugar, are
mixed with thirty grams of soda-lime in powder. The bottom of the
combustion tube contains a little moist soda-lime, which is heated at
the end of the operation at the same time that a current of pure
hydrogen is made to pass through it, and the temperature of the tube
is raised, little by little, to a distinct redness. The contents of the
receiving bulbs are distilled, after the addition of water and soda, in
the same apparatus which served in the estimation of nitrogen, by
the kjeldahl method; the determinations and titrations are made also
under the same conditions.
Blank determinations are also made under the same conditions to
determine the amount of correction to be made by the two methods.
Soda-lime, heated with pure sugar, gave 0.0002 gram of nitrogen for
a total weight of fifty-five grams of the soda-lime contained in the
tube. The fuming sulfuric acid gave 0.0011 cubic centimeters of
ammoniacal nitrogen for the volume of thirty cubic centimeters.
The numbers obtained by the kjeldahl method in general, are
lower than those obtained by the soda-lime method when no stearic
or benzoic acid is used. The numbers obtained when stearic acid
alone was used were sometimes inferior to those obtained by the
soda-lime method. The numbers obtained when benzoic acid is used
are, in general, about the same as those obtained by the soda-lime
method.
It would seem that the double distillation, outlined above, for the
kjeldahl method, would not be necessary if due care were exercised
in the first distillation. This variation, therefore, seems to be
unnecessary.
In the soda-lime method, time would be saved by the reception of
the ammonia in standard acid, and its titration in the usual way,
unless a further purification of the nitrogenous products of the
combustion by the final distillation be desired.
405. Volumetric Determination of the Nitrogen.—Instead
of separating the nitrates, the total nitrogen in the soil can be
determined directly by the classic method of Dumas, which consists
in bringing the whole of the nitrogen into a gaseous state and
afterwards measuring its volume.
The following method illustrates the general principles of the
determination:
 A glass combustion tube closed at one end, about one meter in
length, is selected. In the bottom of this tube is placed some
potassium bicarbonate in a crystalline form, in small pieces, filling
the tube to a distance of about twenty centimeters. Afterwards
copper oxid is placed to the depth of ten centimeters and finally a
mixture of from twenty to thirty grams of the earth with thirty to
forty grams of copper oxid in a fine state of subdivision, and about
ten grams of metallic copper obtained by reducing the copper oxid by
hydrogen. Next the tube is filled with copper oxid to a depth of from
twenty to twenty-five centimeters, and afterwards with reduced
copper to the depth of at least twenty-five centimeters, and after this
another layer of copper oxid of about five centimeters, and finally a
plug of asbestos. The combustion tube is closed with a stopper
carrying a glass tube of about ninety centimeters in length, of which
the extremity, bent into the form of a ᥩ, extends to a mercury
trough. The glass combustion tube is surrounded with brass gauze,
except that part which contains the potassium bicarbonate. The
beginning of the operation consists in heating the tube to decompose
a part of the potassium bicarbonate, until the whole of the apparatus
is filled with carbon dioxid. In order to determine that the whole of
the air has been expelled and that the apparatus is entirely filled with
carbon dioxid, a part of the gas which is disengaged, is received into
a jar filled with mercury, in which a little potash-lye has been placed.
If the gas is entirely absorbed by the potash, so that there remain
only unappreciable particles, the tube can be regarded as completely
free of air. When assurance is given that the air is all out of the
apparatus, a jar of about 300 cubic centimeters capacity, filled with
mercury and containing from thirty to forty cubic centimeters of a
solution of potash of a density of 42° baumé, is placed over the outlet
tube. The combustion is commenced by heating the anterior part of
the tube, avoiding the heating of the part containing the earth. When
the first part of the tube has reached the red stage the part containing
the earth is gradually heated in order to obtain a gentle evolution of
gas. The temperature of the tube is carried to redness and the
heating gradually carried back toward the closed extremity, but
avoiding raising the temperature of the part containing the
potassium bicarbonate. The red heat is continued as long as bubbles
of gas are discharged into the reservoir. When the evolution of gas
has ceased the apparatus is again filled with carbon dioxid for the
purpose of driving out the last traces of nitrogen, by heating again
the part of the tube containing the potassium bicarbonate. The
evolution of the carbon dioxid should be maintained for about fifteen
minutes. At the end of this time all the nitrogen will be found in the
receiving jar. Sometimes a small quantity of nitrogen dioxid is
formed incidentally in the operation. After waiting for a quarter of an
hour, in order to permit all the carbon dioxid which may have
escaped into the reservoir to be completely absorbed, the receiving
jar is carried to a water-basin and the mercury allowed gradually to
escape; its place being taken by the water. The gas is then transferred
into an azotometer where its volume and temperature are read in the
usual way.
In order to absorb any nitrogen dioxid which may be admixed with
the nitrogen itself, a little crystal of ferrous sulfate is introduced. The
reservoir containing the nitrogen is carried to the mercury trough,
and the water which it contains is nearly all run out in such a way as
to be replaced with mercury, great care being exercised to avoid any
escape of gas. Afterwards there is introduced over the mercury a
crystal of ferrous sulfate and the azotometer is shaken until this
crystal is dissolved by the water which it still contains. It is then
allowed to remain for twenty hours. At the end of this time the
nitrogen dioxid is absorbed and the volume of the gas is again read
as before. One-half only of the total loss should be subtracted, since
the volume of the nitrogen dioxid is twice the volume of the nitrogen
itself. For the practice of this method, in connection with the use of a
mercury pump, the directions which will be given under fertilizers
may be consulted.
406. Estimation of Ammonia.—Ammonia exists ordinarily
only in very small quantities in the soil, since it is incessantly
transformed into nitrate or diffused in the air. Nevertheless, it is
sometimes interesting to determine its quantity.
The method of determining the ammonia in soils is one of extreme
delicacy on account of the small proportion therein, and the difficulty
of expelling it without at the same time converting some of the
organic nitrogen into ammoniacal compounds. The various methods
employed for this purpose may be classified as follows:
 1. Treatment of the soil with soda-lye in the cold, and the
absorption of the ammonia given off by standard sulfuric acid.
2. The method of Boussingault, which consists in replacing the
soda-lye with magnesia and distilling the ammonia at a boiling
temperature, absorbing the distillate in a standard acid.
3. A modification of the above method, due to Schloesing, which
consists first in extracting the ammonia by hydrochloric acid and
subjecting the extract to distillation with magnesia.
4. The method of Knop consists in treating the soil in a closed
cylinder with soda-lye containing bromin. The ammonia set free by
the lye is decomposed in the presence of bromin into free nitrogen
and hydrochloric acid. The nitrogen is collected and measured in an
azotometer. The brom-soda-lye is prepared by dissolving 100 grams
of sodium hydroxid in 1,200 cubic centimeters of water and adding
twenty-five cubic centimeters of bromin.
5. The process described under 4, as shown by Baumann,[270] does
not give accurate results and it has been modified by him as follows:
Two hundred grams of soil are treated with 100 cubic centimeters of
dilute hydrochloric acid (one part acid and four of water) free of
ammonia; 300 cubic centimeters of ammonia-free distilled water are
added and the whole digested for two hours with frequent stirring. If
a soil contain much calcium carbonate larger quantities of acid must
be used. Two hundred cubic centimeters of the filtrate are placed in
an evolution flask, connected with an azotometer, with five grams of
freshly burned magnesia. The mixture is then oxidized as follows:
Ozone is generated by adding three parts by weight of sulfuric acid to
one part of dry and powdered potassium permanganate. A stream of
air is drawn through the ozone generator by an aspirator, and the
ozone is conducted into a flask containing the hydrochloric acid
extract of the soil and magnesia. The oxidation is completed in about
ten minutes. The mixture is then brought into the evolution flask of
the azotometer and the nitrogen set free and measured in the usual
way.
It has been shown that if asparagin or glutamin be present in the
soil they are decomposed by the soda-lye and the results obtained are
too high. It has been further proved that soils which contain a
notable quantity of humus give, with soda-lye in the cold, a
practically continuous evolution of ammonia. Moreover, soils which
are rich in humus and which have been treated by distillation with
magnesia give, on subsequent treatment with soda-lye, considerable
additional quantities of ammonia.
Comparison of Methods of Estimating Ammonia.—Baumann has
determined the ammonia-nitrogen in various soils by the soda-lime
method; distillation of the hydrochloric acid extract with magnesia,
and the azotometric method modified as indicated above. These
methods will be designated as 1, 2, 3, respectively in the following
table.

Method.
1. 2. 3.
Ammonia-nitrogen in one kilogram of soil.

No. of sample. Gram. Gram. Gram.

1 0.0448 0.02227 0.02781
2 0.0168 0.01105 0.01326
3 0.0336 0.01771 0.02214
4 0.0056 0.00443 0.00443
5 0.0280 0.02337 0.02894
6 0.0196 0.01243 0.01672

From the above figures it is seen that the method usually

attributed to Schloesing gives uniformly higher numbers than either
of the other processes, while the third gives slightly higher values
than the second.
407. The Magnesia Distillation Process.—If a sample of soil
be distilled directly with magnesia and water, there is danger on the
one side of not extracting all the ammonia, by reason of the
absorbing power of these bodies, and on the other, of transforming
into ammonia the nitrogen of the organic matters. It is therefore
preferable to separate the ammonia from the soil in the form of
chlorid, and to subject this extract to distillation.
In fifty grams of the soil the humidity is determined by drying at
100° until there is no further loss of weight. The quantity of moisture
being known, 200 grams of soil are taken and moistened with water,
and then there is added, in small portions, some dilute hydrochloric
acid, shaking frequently until the whole of the calcium carbonate
present is decomposed. The liquor should remain acid at the end of
the operation, but without containing a notable excess of acidity.
Knowing beforehand the quantity of moisture contained in the 200
grams, water is added until the total quantity shall be equal to 500
cubic centimeters. The whole is then shaken and allowed to repose,
and filtered rapidly, covering the funnel with a glass vessel and
receiving the liquid which runs through in a flask with a narrow
opening. Two hundred and fifty cubic centimeters of this liquor, or
mixture, represent 100 grams of earth of known humidity. This
quantity is introduced into a flask for determining the ammonia and
five grams of calcined magnesia added. Before commencing the
distillation, assurance should be had that the magnesia has
completely saturated the acid in excess, and that the liquor is
alkaline.
If, by chance, the liquor should be still acid it would be necessary
to add sufficient magnesia in order that the reaction should be
manifestly alkaline. Afterwards the distillation is begun and the
ammonia is received in an appropriate vessel containing one-tenth
normal sulfuric acid and titrated in the usual way, or nesslerized.
Inasmuch as the quantities of ammonia contained in the earth are
generally very small it is necessary to be very particular in order to
avoid errors. The distilled water which is employed should be
deprived of all traces of ammonia by prolonged ebullition, and the
hydrochloric acid should be distilled in the presence of a little
sulfuric acid. The treatment with hydrochloric acid is for the purpose
of destroying the absorbing properties of the soil for ammonia, and
to permit this last to enter into solution as chlorid. When there is
need of very great precision it is convenient to make a blank
operation with the hydrochloric acid and water which are employed,
in order to make a correction for the traces of ammonia which these
reagents may contain.
408. Estimation of Ammoniacal and Amid Nitrogen by
the Method of Berthelot and André.[271]—Heat one hundred
grams of earth for thirty hours on a steam-bath with about .500
cubic centimeters of dilute hydrochloric acid (fifteen grams of
hydrochloric acid to 500 cubic centimeters of water). At the end of
this time throw the contents of the flask on a filter and wash with hot
water until acid reaction has ceased. Determine both the ammoniacal
and amid nitrogen in the soluble, and the total nitrogen in the
insoluble portion, the ammoniacal by distillation with magnesia, and
the amid and total with soda-lime.
Example. A soil contained 0.1669 per cent total nitrogen. Of this
there were obtained:

As ammoniacal nitrogen 13.7 per cent

In the soluble part as amid nitrogen 56.2 „ „
In the insoluble part, total nitrogen 29.7 „ „

Sum 99.6 „ „

Treatment of the Insoluble Portion.—Treat the part insoluble in

hydrochloric acid with a three per cent solution of potash on a steam-
bath for thirty hours. Estimate the nitrogen remaining insoluble,
from which the part dissolved can be determined by difference. The
potash will dissolve usually about two-thirds of the remaining
nitrogen.
About ninety per cent of the total nitrogen present in an arable soil
will be rendered soluble by successive treatment with acid and alkali.
The reverse treatment will give practically the same result. It is
therefore immaterial, from an analytical standpoint, whether the
acid or alkali be used first.
409. Estimation of Volatile Nitrogenous Compounds
Emitted by Arable Soil.—The following method, due to Berthelot
and André,[272] may be practiced:
Porcelain pots, containing one kilogram of soil, are placed under
bell-jars of fifty liters capacity adjusted to glass dishes designed to
receive the waters of condensation.
During the first period the pots are to be sprinkled from time to
time, during the duration of the experiment, through the upper
tubulature, so as to prevent the soil from becoming dry. The water is
partly condensed on the sides of the bell-jar. It is removed each week
through the inferior tubulature, treated with a little dilute sulfuric
acid, and preserved for further study. A small vessel containing
dilute sulfuric acid is placed near the porcelain pot for the purpose of
collecting, as far as possible, the evolved ammonia.
During the second period the pots are not sprinkled, the soil
becomes dry and there is no longer any condensation of water on the
walls of the bell-jar. The two periods should include about five
months, from May to October.
At the end of the second period the following determinations are to
be made:
1. The ammonia absorbed by the dilute sulfuric acid.
2. The ammonia set free by distillation with magnesia, such as may
have accumulated in the condensed water.
3. The organic nitrogen contained in the latter after elimination of
the ammonia. This is determined by adding a slight excess of acid,
evaporation to dryness, and combustion with soda-lime, or by moist
combustion with sulfuric acid.
Example:
Earth Employed.—One kilogram of sandy clay containing total
nitrogen, 0.09 gram. Nitrogen in sprinkling water, 0.000048 gram.
Nitrogen in Exhaled Products.—

First Period. Sprinkling.

Ammoniacal nitrogen collected in the dilute sulfuric acid 0.00012 gram.
Ammoniacal nitrogen collected in the condensation waters 0.00012 „
Organic nitrogen in condensation waters 0.00220 „

Sum 0.00244 „

Second Period. No Sprinkling.

Ammoniacal nitrogen in dilute sulfuric acid 0.000007 gram.
„ „ „ condensed water 0.000007 „
Organic „ „ „ „ 0.000040 „

Sum 0.000054 „

Conclusions.—The exhalation of nitrogenous compounds takes

place with a certain relative activity, about two milligrams in two
months and a half, as long as the soil is kept moist by sprinkling.
In the second period, without sprinkling, the exhalation is reduced
to a mere trace.
The vessel containing the dilute sulfuric acid placed near the
porcelain pot absorbs only about one-half of the ammoniacal
nitrogen set free. The nitrogen emitted under other forms than
ammonia is, in every instance, greatly superior in quantity, and this
is the most important of the observed phenomena. This is true at
least with the kind of soil with which the experiment was made. With
arable soil containing twenty times as much nitrogen as the soil
described above this order is reversed,[273] the ammoniacal prevailing
over the non-ammoniacal nitrogen volatilized.
These phenomena are doubtless greatly influenced in soil under
culture by microbes, and the lowest orders of vegetation to which are
doubtless due the traces of non-ammoniacal volatile nitrogenous
compounds, a sort of vegetable ptomaines.
410. General Conclusions.—In the light of our present
knowledge concerning the methods of nitrogen determination in the
soil in the form of organic compounds and ammonia, moist
combustion with sulfuric acid is to be preferred to the older soda-
lime process. For the nitrogen combined as ammonia, the extraction
of the sample with hydrochloric acid and subsequent distillation with
an excess of freshly calcined magnesia, are recommended. For the
study of the progressive decomposition of the nitrogenous
compounds, the various processes devised by Berthelot and André
are the best.
The origin of the nitric acid in the soil, the methods of studying the
various nitrifying organisms, and of estimating the nitric acid
produced, will form the subject of the next part.
Note.—At the Eleventh Annual Convention of the Association of
Official Agricultural Chemists, held in Washington, August 23, 24
and 25, 1894, the following process of soil extraction was adopted as
the official method:
Preparation of the Sample.—500 grams or more, of the air-dried
soil, which may be either the original soil or that which has been
passed through a sieve of coarser mesh, are sifted upon a sieve with
circular openings one-half millimeter in diameter, rubbing, if
necessary, with a rubber pestle in a mortar, until the fine earth has
been separated as completely as possible from the particles that are
too coarse to pass through the sieve. The fine earths thoroughly
mixed and preserved in a tightly stoppered bottle from which the
portions for analysis are weighed out.
The coarse part is weighed and may be subjected to further
examination, (as in Bulletin 38, Div. of Chem., pp. 65, 75 and 200.) It
may sometimes be necessary to wash the soil through the one-half
millimeter sieve with water, in which case proceed as directed on pp.
65 and 75 of the above Bulletin. The use of water is to be avoided
whenever possible.
Determination of Moisture.—Heat two to five grams of the air-
dried soil in a flat-bottomed, tared platinum dish; heat for five hours
in a water-oven kept briskly boiling; cover the dish, cool in a
desiccator, and weigh.
Repeat the heating, cooling, and weighing at intervals of two hours
till constant weight is found, and estimate the moisture by the loss of
weight. Weigh rapidly to avoid absorption of moisture from the air.
An air-bath must not be used in this determination.
Determination of Volatile Matter.—The platinum dish and soil
used to determine moisture are used also to determine volatile
matter. Heat the dish and dried soil to full redness until all organic
matter is burned away. If the soil contain appreciable quantities of
carbonates, the contents of the dish, after cooling, are to be
moistened with a few drops of a saturated solution of ammonium
carbonate, dried and heated to dull redness to expel ammonium
salts, cooled in the desiccator and weighed.
The loss in weight represents the organic matter, water of
combination, ammonium salts, etc.
Extraction of Acid-Soluble Materials.—In the following scheme for
soil analysis it is intended to use the air-dried soil from the sample
bottle for each separate investigation. The determination of
moisture, made once for all on a separate portion of air-dried soil,
will afford the datum for calculating the results of analysis upon the
soil dried at the temperature of boiling water. It is not desirable to
ignite the soil before analysis or to heat it so as to change its chemical
properties.
 The acid digestion is to be performed in a flask so arranged that
the evaporation of acid shall be reduced to a minimum, but to take
place under atmospheric pressure and at the temperature of boiling
water. Any flask resistant to acids is suitable, but it is not necessary
to use a condenser, as a simple bohemian glass tube eighteen inches
in length will answer the purpose of preventing loss of acid. Where it
is not desired to determine sulfur trioxid, an erlenmeyer fitted with a
rubber stopper and hard glass tube will answer. The flask must be
immersed in the water-bath up to the neck or at least to the level of
the acid and the water must be kept boiling continuously during the
digestion.
In the following scheme, ten grams of soil are taken, this being a
convenient quantity in most soils, in which the insoluble matter is
about eighty per cent. If desired, a larger quantity of such soil may be
taken, using a proportionately larger quantity of acid and making up
the soil solution to a proportionately larger volume. In very sandy
soils, where the proportion of insoluble matter is ninety per cent or
more, twenty grams of soil are to be digested with 100 cubic
centimeters of acid and the solution made up to 500 cubic
centimeters or a larger quantity may be used, preserving the same
proportions. It is very important that the analyst assure himself of
the purity of all the reagents to be used in the analysis of soils before
beginning the work.
Acid Digestion of the Soil.—Place ten grams of the air-dried soil in
a 150 to 200 cubic centimeter bohemian flask, add 100 cubic
centimeters of pure hydrochloric acid of specific gravity 1.115, insert
the stopper with condensing tube, place in a water or steam-bath and
digest for ten hours continuously at the temperature of boiling water,
shaking once each hour. Pour the clear liquid from the flask into a
small beaker, wash the residue out of the flask with distilled water on
a filter adding the washings to the contents of the beaker. The
residue after washing until free of acid, is to be dried and ignited as
directed below. Add one or two cubic centimeters of nitric acid to the
filtrate, and evaporate to dryness on the water-bath, finishing on a
sand or air-bath to complete dryness; take up with hot water and a
few cubic centimeters of hydrochloric acid, and again evaporate to
complete dryness. Take up as before, filter and wash thoroughly with
cold water or with hot water slightly acidified at first with
hydrochloric acid. Cool and make up to 500 cubic centimeters. This
is solution “A.” The residue is to be added to the main residue and
the whole ignited and weighed, giving the insoluble matter.
The determination of the various components of the solution
remains essentially as described in the provisional methods of the
Association which have already been given.
It is directed that all results of soil analysis be calculated on the
basis of the sample dried to constant weight at the temperature of
boiling water.
AUTHORITIES CITED IN PART SIXTH.