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Effects of crude kerosene on testosterone levels, aggression and toxicity in rat

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DOI: 10.1016/j.toxrep.2014.11.017

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 Toxicology Reports 2 (2015) 175–183

Contents lists available at ScienceDirect

Toxicology Reports
journal homepage: www.elsevier.com/locate/toxrep

Effects of crude kerosene on testosterone levels, aggression

and toxicity in rat
Rachel W. Njoroge a,1 , Benson N. Macharia b , Dinah J. Sawe c ,
Geoffrey K. Maiyoh c,∗,1
a
Department of Chemistry and Biochemistry, University of Eldoret, P.O. Box 1125, Eldoret, Kenya
b
Department of Human Pathology and Forensic Medicine, School of Medicine, Moi University, P.O. Box 4606, Eldoret, Kenya
c
Department of Medical Biochemistry, School of Medicine, Moi University, P.O. Box 4606, Eldoret, Kenya

a r t i c l e i n f o a b s t r a c t

Article history: The use of crude kerosene as a dietary supplement in boarding schools has been a common
Received 24 September 2014 practice in east Africa and other countries for many years, with the belief of it reducing the
Received in revised form sex drive (libido) at the pubertal stage. There is however no scientific basis for this belief.
20 November 2014
The present study aimed at using a rat animal model to investigate the effects of crude
Accepted 24 November 2014
kerosene on serum testosterone levels, aggression and its possible toxic effects. Fifteen
Available online 2 December 2014
male albino rats of approximately similar age and average weights were put into three
groups of five animals each; the control group (placebo), low kerosene dose (10 ␮l/day)
Keywords:
group and high kerosene dose (300 ␮l/day) group. ELISA was used to determine the serum
Crude kerosene
Testosterone testosterone levels. During treatment, changes in aggression were observed and noted.
Sex drive Liver toxicity was determined using enzyme assays, total protein and albumin while renal
Aggression toxicity was monitored using serum creatinine levels. A full hemogram was conducted
Toxicity to determine hematological effects. Various tissue biopsies were obtained and examined
Gastritis using histopathological techniques for evidence of toxicity. Contrary to the common belief,
our findings showed an overall increase of serum testosterone levels of up to 66% in the
Chemical compounds studied in this article:
Testosterone (PubChem CID: 6013)
low dose and 75% in the high dose groups, with an increasing trend by the end of the
Picrate (PubChem CID: 62496) study. The high dose group showed significantly increased levels of white blood cells (WBC)
Bromocresol green (PubChem CID: 6451) (p = 0.036), red blood cells (RBC) (p = 0.025), hematocrit (HCT) (p = 0.03), red cell distribution
Creatinine (PubChem CID: 588) width (p = 0.028) and platelets (p = 0.017). The histological results of the stomach indicated
Hematoxylin (PubChem CID: 442514) chronic gastritis.
Formaldehyde (PubChem CID: 712) © 2014 The Authors. Published by Elsevier Ireland Ltd. This is an open access article under
Eosin (PubChem CID: 11048) the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).
Ethylenediaminetetraacetic acid (PubChem
CID: 6049)

1. Introduction

Kerosene is a distillate of crude petroleum that contains

Abbreviations: ALP, alkaline phosphatase; ALT, alanine transaminase;
aliphatic, aromatic and a variety of other branched satu-
AST, aspartate transaminase; EDTA, ethylenediaminetetraacetate; ELISA, rated and unsaturated hydrocarbons [1]. The use of crude
enzyme linked immunosorbent assay; HCT, hematocrit concentration; kerosene has been a common practice in east Africa and
LFT, liver function tests; RBC, red blood cells; RDW, red cell distribu- other countries for many years, with the belief of it reduc-
tion width; RDW, red cell distribution width; RFT, renal function tests;
ing the sex drive (libido) at the pubertal stage. In the course
T, testosterone; WBC, white blood cell.
∗ Corresponding author. Tel.: +254 713 592879. of daily meals consumption students are exposed to doses
E-mail address: [email protected] (G.K. Maiyoh). of kerosene as a dietary supplement, usually without their
1
These authors contributed equally to this work. consent.

http://dx.doi.org/10.1016/j.toxrep.2014.11.017
2214-7500/© 2014 The Authors. Published by Elsevier Ireland Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
176 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183

The process of puberty results in the release of some supplementation is effective in reducing libido has not been
specific hormones which are primarily responsible for the scientifically tested. Further, the dietary use of kerosene in
development of secondary sex characteristics and for the schools to tame sexual drive occurs with little or no care
emergence of reproductive capabilities in boys [2]. During at all on its possible hazardous effects on the health sta-
this stage an increase in testosterone causes an increase tus these students. Although some information is currently
in the sex drive (libido), enlargement of the reproduc- available on the effect of dietary kerosene supplementation
tive organs such as the penis and testes, the production in animals and/or humans [12,19], such studies have failed
of sperm, increase of muscle mass and lowering of the to provide comprehensive information on effects on T lev-
voice, increased frequency of erection, and the growth of els, link to aggression and body tissue toxicity.
facial, chest, nipple and pubic hair among boys [3]. The link The present study was designed to monitor the
between testosterone (T) levels and the sexual drive was effects on serum T levels, hematological, biochemical
demonstrated in a study done using adolescent boys with and histopathological changes in rats exposed to crude
the findings indicating that the adolescent boys who had kerosene as a dietary supplement at doses that are compa-
higher levels T levels also reported higher levels of sexual rable to those commonly used in Kenyan boarding schools.
activity (i.e. coitus) [4–7]. From the studies by Brooks-Gun
and Halpern [5,6] it can be inferred that hormones may 2. Materials and methods
enhance feelings of sexual arousal in adolescents but how
they act on those feelings is very much determined by mul- 2.1. Ethics statement
tiple internal and external variables.
From the study conducted by Olweus et al. [4,8] it was All the animal protocols and experiments were
noted that adolescent boys with higher T levels were more approved by the Institution animal care and use committee
likely to engage in aggressive behavior. Under conditions of the University of Eldoret (Protocol No. UOE/001/14).
of threat or unfair treatment, [9] they were shown to be
aggressive. They further showed a link between higher
2.2. Animals
T level and a lower tolerance for frustration. Further to
these, they also observed that when no provoking situa-
Male Wistar rats (Rattus norvegicus) of approximately
tion occurred, T levels did not predict aggression. Various
the same age (six weeks old) corresponding to early ado-
animal studies conducted on mice demonstrated the link
lescent boys [20] and similar body weights were obtained
between aggressive behavior and increased T levels [10,11].
from the University of Eldoret animal facility. They were
In a study on mice exposed to jet kerosene continuously for
acclimatized and given free access to water and standard
90 days, there was an observed increased incidence in the
rodent chow diet (Unga Farmcare East Africa Limited,
fighting of the test group mice [12].
Nakuru, Kenya) for two weeks prior to initiation of the
There is increasing trend regarding the percentage
experimental diet. The rats were housed and maintained at
of teenagers reporting sexual initiation at younger ages
ambient temperature of 25 ◦ C under a photoperiod of 12 h
[13]. This early sexual initiation (before age 16) is likely
of light and 12 h of darkness. The animals were assorted
to involve sexual risk-taking and expose young people
into three groups of five rats each with all groups having
to unwanted sex, sexually transmitted infections, and
similar average serum testosterone levels.
teenage pregnancy. This may be attributed to exposure to a
highly sexualized media environment that may represent
a primary source of sexual socialization [14,15]. Middle- 2.3. Sample size determination
childhood problems, poor parenting and peer pressure
have also been shown to contribute to early sexual behavior The sample size was determined according to the for-
[16,17]. Due to the dire consequences of early sexual activ- mula by Charan et al. [21].
ity [18], there have been efforts toward finding effective E = total number of animals − total number of groups (if
remedies to tame teenage sexual hyperactivity. In many the value of E is between 10 and 20 then it is considered as
Kenyan boarding schools, especially high schools, one such an adequate, for this study, E = 12).
remedy that has been used traditionally is crude kerosene.
In a recent survey that we conducted using structured ques- 2.4. Animal treatment
tionnaires at a Public University admitting students from
all over the country, (data not shown) we found out that Animal groups consisted of the control (placebo group
68% female and 76% male first year, random respondents – distilled water) low dose (10 ␮l kerosene) and high dose
from 28 of 47 counties in Kenya, reported that at least one (300 ␮l kerosene). All animals were maintained on regular
of their main meals (lunch or dinner) was supplemented rodent chow diet throughout the study. Kerosene (National
with kerosene on daily basis during their high school years. Oil Corporation, Eldoret, Kenya) was delivered orally on a
Interestingly, over 60% of respondents in the above cate- daily basis. Blood samples from animals in all groups (con-
gory gave why they thought kerosene was included in their trol and treatment) were collected from the tail under local
diets as being to reduce their desire for sex. The remainder anesthesia at baseline, day 7 and day 14. Since T levels in
(40%) did not know why it was added. young male rats have been shown to vary with time of
Kerosene is readily available and at fairly low costs the day [22,23], all blood collections were done between
throughout the country. The primary use is for light- 12.00 noon and 1.00 pm at all time points. Animals were
ing and in cooking stoves. Whether or not kerosene also observed for changes in behavior on daily basis during
 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183 177

and after treatment. At the end of study (day 28) blood sam- Imaging America Inc., PA, USA) and images were captured
ples were collected via cardiac puncture under chloroform using a Smartphone digital camera obtained by its autofo-
anesthesia. The stomach and the brain and esophagus tis- cus and automatic exposure control.
sues were dissected and fixed in 10% formalin and used for
histological analysis. Whole blood was collected in EDTA 2.8. Statistical analysis
vacutainers for full hemogram while blood samples for
serum were collected in plain tubes and serum obtained Statistical analysis was conducted using Student’s t
after centrifugation at 3000 × g for 10 min at 4 ◦ C and kept tests. Values are expressed as means ± SD or SEM and
at −20 ◦ C until use for determination of the biochemical p < 0.05 was considered significant.
markers. Animal weights were monitored on weekly basis.
3. Results
2.5. Changes in animal aggression
3.1. Crude kerosene supplementation and associated
To evaluate the effect of kerosene supplementation on
effects among high school students
our experimental animal behavioral changes, an obser-
vational method was used. In brief, rats were monitored
To determine the extent of kerosene dietary supple-
for observable changes in behavior following dietary
mentation in Kenyan high schools; a pre-study survey was
kerosene supplementation and also after bleeding. Aggres-
undertaken prior to our animal studies. Out of a total of
sive behavior was defined as burrowing (mechanical
50 (half from either gender) fresh high school graduates
removal/moving of bedding material by rats within their
who had recently enrolled at a local University taking part
cages) and fighting (chasing after other animals, voluntary
in our pre-study survey, 72% of respondents indicated that
attacks by one animal on another including biting and/or
kerosene was routinely added to their school diets with
scratching) within a period of 20 min post supplementation
slightly higher number of male (76%) than female students
among animals in the same group. Level of aggression
(68%) (Fig. 1A). Most of the respondents in the supple-
was rated in terms of proportion of animals per group
mentation category thought that the reason for kerosene
engaged in burrowing and fighting following kerosene
supplementation was intended to tame sex drive among
supplementation. Comparisons in behavioral changes were
students. Among students where kerosene was added to
made between the various groups to determine the relative
their diets, 46% reported having experienced at least one
aggression.
of the various diet related stomach problems with stom-
ach ulcers, heart burns and stomach ache and/or nausea
2.6. Biochemical/hematological analyses
collectively comprising 47.8% of these problems (Fig. 1B).

Serum testosterone levels were determined by Enzyme

linked immunoassay kit, (Human Diagnostics worldwide, 3.2. Crude kerosene supplementation had no effect on
Wiesbaden, Germany); ALT and AST activity were mea- animal body weights
sured using reagent kits (Human Diagnostics worldwide,
Wiesbaden, Germany; total proteins by biuret methods Body weights were monitored regularly from start to
(Human Diagnostics worldwide, Wiesbaden, Germany); end of study. No differences were seen at all time points in
albumin by bromocresol green reagent (Human Diagnos- the body weights among the three groups (data not shown)
tics worldwide, Wiesbaden, Germany) according to the (p > 0.05).
manufacturer’s instructions. Renal functions were mon-
itored using serum creatinine levels by alkaline picrate 3.3. Crude kerosene increased serum testosterone levels
method (Biosystems, Barcelona, Spain) according to the
manufacturer’s instructions. The average initial T values for the animals in our study
The hematological parameters were determined using were 3.05 ng/ml, 2.98 ng/ml and 2.9 ng/ml for control, low
the ADVIA 120D hematology system (Global Siemens dose and high dose groups, respectively. The levels of T
Healthcare, Henkestrasse, Germany) according to the man- in our study animals are comparable to those obtained
ufacturer’s instructions. by other earlier studies conducted in rat where T levels
were measured [24,25]. Although the T levels in the control
2.7. Histopathological assays group showed a gradual decline in the course of our study
duration, however this decline did not reach statistical sig-
The stomach, brain and esophagus tissues were excised nificance (2.20 ng/ml at day 28).
and fixed in 10% neutral buffered formalin at room The ELISA results at the various time points (Fig. 2)
temperature and the representative tissues were taken indicated that kerosene supplementation caused a marked
from the stomach, brain and esophagus then processed increase in the rat’s serum testosterone levels in the test
using Automated Vacuum Tissue Processor Leica ASP6025 groups relative to the control group. Both the low and high
(Biosystems, Barcelona, Spain). They were embedded using dose groups had an upward trend with an overall increase
paraffin wax and the blocks were sectioned into 5 ␮m of 66% in the low dose and 75% in the high dose group with a
thick slices, and stained with Hematoxylin and Eosin. continuing upward trend at the end of the study duration.
Specimens were examined for morphological changes There was however, no significant change in the T levels
under light microscope (Olympus 6V20WHAL) (Olympus following acute (1st seven days) supplementation.
178 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183

Fig. 1. Kerosene supplementation and associated effects among high school students. Structured questionnaires were used to conduct a pre-study survey.
(A) The graph shows proportion of respondents who indicated that kerosene was included in their high school diet against those who stated otherwise and
their supposed reason for inclusion. (B) Shows data for diet related problems experienced by students where kerosene was included in the diet. K SUPL:
kerosene supplementation; ULC: stomach ulcers; HBN: heart burns; ACNA: stomach ache and or nausea; OTH: other.

3.4. Crude kerosene supplementation resulted in treatment administration and following bleeding at the
increased aggressive behavior various time points. Among the control animals, there were
no observable changes in behavior before and after supple-
Crude kerosene supplementation resulted in increased mentation throughout the study. Incidences of burrowing
incidences of aggression among the test group animals. This and fighting were also minimal (attributable to normal
increased aggression (including fighting and burrowing) behavior in rats). However, among the treatment groups,
was specifically observed during and immediately after a progressive increase in number of animals engaged in
burrowing and fighting was noted during the study period.
It was also noted that the ease of handling during dosing
became increasingly difficult in these groups. Although dif-
ficult to quantify, it was also observed that as the study
progressed an increasingly higher proportion of animals in
the high dose category displayed aggressive tendencies as
compared to the low dose animals.

3.5. Crude kerosene had no effect on the hepatic and

renal functions

We investigated the potential toxic effects of the

kerosene supplementation rat liver. Our results showed
no statistically significant effects on the liver enzymes
Fig. 2. Effect of dietary kerosene supplementation on serum testosterone (AST and ALT) for both doses tested (Fig. 3A). Total pro-
levels. Blood samples were collected from the tail at baseline (day 0), day 7 teins showed a decreasing trend but it did not reach
and day 14. At the end of study (day 28) blood samples were collected via statistical significance (Fig. 3B) (low dose p = 0.064, high
cardiac puncture under chloroform anesthesia and analyzed for T levels dose p = 0.068). Serum albumin levels showed a signifi-
by ELISA. The values represent means ± SEM of five animals per group.
cant decrease (Fig. 3B) (p = 0.038) for the low dose group.
Data that share the same letters indicate values that are not significantly
different (p < 0.05). T: testosterone; ELISA: enzyme linked immunosorbent Kerosene supplementation did not significantly affect the
assay. kidney’s ability to eliminate creatinine from blood (Fig. 3A).
 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183 179

Fig. 3. Effects of dietary kerosene supplementation on hepatic and renal functions. At day 28, blood was collected via cardiac puncture following an
overnight fast and analyzed for hepatic and renal functions. Serum was obtained and analyzed for AST, ALT, total protein and albumin to determine hepatic
functions while creatinine levels were used to determine the renal function. Data show an average of five animals per group ±SEM. Different letters show
significantly different values (p < 0.05); ALT: alanine transaminase; AST: aspartate transaminase; CRET: creatinine.

3.6. Effects of crude kerosene on different blood 3.7. Crude kerosene supplementation resulted in chronic
parameters gastritis

Crude kerosene supplementation increased white blood Kerosene supplementation resulted in an active chronic
cells (WBC), red blood cells (RBC), platelets, hematocrit gastritis in the stomach in both test group animals.
concentration (HCT) and the red cell distribution width This effect was demonstrated by the infiltration of the
(RDW) counts in a dose depended manner (Fig. 4A). eosinophils, lymphocytes and plasma cells present on the
Although there were increases in the counts for low dose gastric mucosa and sub-mucosa. Despite being on simi-
group, the values did not reach statistical significance. lar diets and environmental condition, the control animals
The animals on a high dose kerosene supplementation showed no signs of gastritis (Fig. 5).
had a significant increase in the WBC (p = 0.036, RBC
(p = 0.025), HCT (p = 0.029), RDW (0.029) and platelets 3.8. Crude kerosene supplementation resulted in no
(p = 0.018) as compared to the untreated controls. WBC dif- pathological effects on the brain and esophagus
ferential count showed a significant increase in the levels
of monocytes in the low dose group relative to the con- There were no morphological changes in the brain
trol group (Fig. 4B). Differential counts of the other types (Fig. 6A–C) and the esophagus (Fig. 6D–F) for the ani-
of WBC remained essentially unaltered between all the mals in the various groups including control and treatment.
groups. This is indicative that the kerosene supplementation at our

Fig. 4. Effects of dietary kerosene supplementation on hematological parameters. At day 28, whole blood was collected via cardiac puncture following an
overnight fast. Hematological parameters were determined using the ADVIA 120D hematology system. (A) The graph shows counts of various blood cells
and (B) shows the relative percentages of the different types of white blood cell. Data show ±SEM. Different letters show significantly different values with
p < 0.05. WBC: white blood cells; RBC: red blood cells; HCT: hematocrit concentration; RDW: red cell distribution width.
180 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183

Fig. 5. Light micrograph images of different sections of the rat’s stomach: (A) gastric mucosa from the control group animals (×400), (B) gastric mucosa
from low dose group shows moderate inflammation by the eosinophils, lymphocytes and plasma cells which has extended to the sub mucosa (×100) and
(C) gastric mucosa from high dose group shows severe active chronic gastritis with marked infiltration of both the mucosa and sub mucosa by eosinophils,
lymphocytes and plasma cells.

Fig. 6. Light micrograph images of different sections of the brain and esophagus. (A–C) Brain section showing the absence of parenchymal abnormalities.
Similar findings (lack of pathology) were also observed in the brain stem and cerebellum. (D–F) Esophageal section showing epithelium with no sign of
pathology.

experimental doses had no toxic effects on both the brain lead to an increase in the incidences of STDs (including
and the esophagus. HIV), unsafe and illegal abortion, adolescent pregnancy and
motherhood, single mother child/abandoned child, juve-
4. Discussion nile delinquency and many more [26]. The use of kerosene
for the above purpose has however not been backed sci-
Kenyan boarding schools have for a long time continued entifically. Several studies have shown that accidental
to supplement student meals with crude kerosene with the ingestion of kerosene results in toxic effects [27–30].
belief that this would restrain their sexual desires that pre- Since T is known to regulate libido [6,31], we hypothe-
dispose them to high risk behaviors. Such risky behaviors sized that if kerosene indeed reduces libido, then it might
 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183 181

mediate its effects through modulation of T levels. A reduc- Reported clinical effects of accidental ingestion or suicide
tion in plasma levels has been associated with reduced attempt are quite varied ranging from mild to fatal. The
sexual drive [7]. Further, increase in T has also been asso- severities of the effects appear to be largely dependent on
ciated with aggressive tendencies [32,6,33]. We therefore the quantity ingested, the age and interaction with drugs
investigated the effects of dietary crude kerosene supple- (such as metformin) that the victim might be using at
mentation on the plasma levels of this hormone and the time of ingestion [39,40]. The common effects include
aggression behavior in a rat animal model. cough with difficulty in breathing, vomiting, fever, cen-
Our results indicate that there was no change in the tral nervous system involvement, severe lactic acidosis and
T level following acute (1st seven days) supplementation acute renal failure, pyopneumothorax and deaths [39–41].
(Fig. 2). However, the trend changed drastically follow- It is important to note that effects reported on accidental
ing continued prolonged (chronic) administration. Both the ingestion or intended suicide are acute effects occurring
low dose and high dose groups showed an upwards trend within a short period of time post ingestion and are usually
with an overall increase of serum T levels of up to 66% in the due to ingestion of large quantities. We therefore postu-
low dose and 75% increase in the high dose groups, respec- lated that chronic dietary kerosene supplementation albeit
tively, by the end of the treatment period (Fig. 2). The levels at lower doses than above (accidental or suicide attempt)
were on an upward trajectory even at the end of study sug- may also be harmful to body tissues. We thus investigated
gesting that longer durations of supplementation are more the potential toxic effects of kerosene on the liver, kidney,
likely to result in even higher increases in T levels. It can be blood and the brain, esophagus and stomach lumen.
inferred therefore that initial (acute) dietary supplemen- It was notable from our findings that there was a uni-
tation with kerosene in boarding school has no effects on form steady rate of increase in the body weights from all
blood T levels among students. On the contrary, prolonged the three groups with no significant difference (p > 0.05)
(chronic) use over the extended schooling years may with among the three groups.
time result in elevated levels of T among students with the Regarding potential toxic effects to the liver, relative
concomitant increase in desire for sexual activity. to the control group, kerosene supplementation showed
This result associating kerosene supplementation to little to no effects (Fig. 3A and B). The liver enzymes
increase levels of serum T may in part explain the ris- remained unchanged (ALT, p = 0.97 and p = 0.35, AST,
ing cases of premature sexual activity leading to high p = 0.11 and p = 0.34 for low and high dose groups, respec-
cases of sexually transmitted infections, unwanted sex tively. Similarly, kerosene supplementation did not have
and teenage pregnancy [14,15]. As indicated earlier, evi- a significant effect on the serum total proteins. Although
dence has shown that high levels of T are also associated results depicted a decreasing trend, it did not reach statis-
with aggressive tendencies [32,6,33]. It was interestingly tical significance (low dose p = 0.064, high dose p = 0.068).
observed that animals on kerosene supplementation dis- Serum albumin levels showed a significant decrease of
played increasing aggression over the study period. The p = 0.038 for the low dose however for the high dose there
higher kerosene dose group displayed even higher levels of was no significant effect (p = 0.77). This finding may require
aggression during and immediately after either kerosene further investigations. Overall, for the period of study,
supplementation or bleeding. This corroborates the find- kerosene supplementation resulted in minimal signs of
ings by Olweus et al. [34,35] in a study where it was liver toxicity.
noted that adolescent boys with higher T levels were not Further, no toxic effects were observed with respect to
only more likely to engage in aggressive behavior but kidneys. Kerosene supplementation did not significantly
under conditions of threat or unfair treatment (provoca- affect the kidneys ability to eliminate creatinine (Fig. 3A).
tion), they were shown to be more likely to be aggressive. It was interestingly observed that on the contrary to our
These results may provide a partial link between kerosene expectation, the kidneys in the treated groups relative to
supplementation in boarding schools and the ever increas- the control group appeared to be eliminating creatinine
ing teenage sexual hyperactivity, teenage pregnancy and from the blood more efficiently as shown by their lower
aggressive behavior such as riots. For our animals, aggres- serum creatinine levels (Fig. 3A). In their earlier studies,
sive behavior was observed during and after treatment and Starek et al. observed signs of liver and kidney respiratory
also during and after blood collection from the tails. These toxicity by kerosene in rats, however effects were noted
events may mimic provocative conditions that have often mainly in rats acutely poisoned, while in sub-chronic poi-
led to student unrests. Studies conducted earlier however soning they were less pronounced [10]. This may suggest a
showed that given acutely, over a few hours, no abnormal possible adaptation over time as minimal toxic effects were
neurologic signs or behavior were notable in baboons [36]. also seen in our chronic study.
This is in agreement with our findings where we noted Unlike the other effects noted so far, kerosene supple-
no significant alteration in testosterone levels for the first mentation appeared to have a possible dose related effects
week of supplementation. This means that it may require with respect to the WBC, RBC, platelets, HCT and the
chronic kerosene supplementation to see both increase in RDW. Relative to the control group there was an increas-
T levels in blood and the T mediated effects on behavior ing trend in these cell counts (Fig. 4A) which appeared to
such as increased aggressive tendencies. The mechanism be dose-dependent. Although there were increases in the
through which the kerosene results in the increase of T counts for low dose group, the values did not reach sta-
remains to be elucidated. tistical significance (Fig. 4A). The animals on a high dose
Various studies have shown that ingestion or inhalation kerosene supplementation had a significant increase in the
of kerosene could lead to various toxic effects [27,37,38]. WBC (p = 0.036) corroborating findings by Dede et al. [37],
182 R.W. Njoroge et al. / Toxicology Reports 2 (2015) 175–183

RBC (p = 0.025), HCT (p = 0.029), RDW (0.029) and platelets sexual hyperactivity in boarding schools. Our findings also
(p = 0.018). This RBC and HCT increase may be beneficial demonstrate the relationship between increased serum
since it may lead to increased oxygen carrying capacity T levels with increased aggression. Kerosene supplemen-
of the blood. The initial increase in the platelets may be tation in boarding schools may result to similar effects.
beneficial but continued increase could be toxic if it goes These findings may explain the increase in the numbers
beyond the limit of the normal ranges as then it could lead of teenage pregnancies, rebellion to authority and violence
to increased incidences of clotting disorders such as stroke. as seen in school going teenage children. The findings from
RDW is used as a measurement of the red blood cells varia- the present study further show that crude kerosene supple-
tion in size and an increase is commonly used in humans as mentation caused gastritis in our animal model. Kerosene
a prognostic marker of either a cardiovascular event, or a supplementation in schools thus may be a contributing fac-
metabolic inflammatory event [42–45]. What was interest- tor in the increasing cases of gastritis and ulcers among
ing to note is that kerosene supplementation at the doses students.
used in our study did not cause anemia as is commonly We recommend that alternative, effective and safe
observed in petroleum products toxicity reported earlier ways to control sexual hyperactivity that are scientifi-
[46–48]. This might be explained by the relatively high cally proven need be sought as a replacement to kerosene
doses used in these studies, i.e. 6 ml/kg which are over four dietary supplementation.
times higher than the high doses used in our study (low
dose = 0.05 ml/kg, high dose = 1.3 ml/kg). Funding
As noted earlier, there was an overall increase in the
WBC counts in test groups (Fig. 4A), the reason for this This research did not receive any specific grant from any
observed increase was suggested by Krishan Veena [49] funding agency in the public, commercial or not-for profit
to be due to a defensive mechanism triggered by the sector
immune system. The low dose group showed a marginal
(6%) increase while the high dose group had a signifi-
Conflict of interest
cant increase of 61%. Similar findings were obtained by
Dede [37] and Krishan Veena [49]. WBC differential count
The authors declare that there is no conflict of interest
showed a significant increase in the levels of serum mono-
that could be perceived as prejudicing the impartiality of
cytes in the low dose group relative to the control group
the research findings reported.
(p = 0.023), (Fig. 4B).
Kerosene supplementation had an inflammatory effect
on the stomach lumen in all the test groups. This effect Transparency document
was demonstrated by the active and chronic inflammation
observed histologically (Fig. 5A–C). From these findings, it The Transparency document associated with this article
can be concluded that kerosene supplementation causes can be found in the online version.
gastritis. The inflammation was observed to be more pro-
nounced at the gastro duodenal junction of the stomach. Acknowledgments
Although studies have shown that Helicobacter pylori is the
chief cause of gastritis in Kenya [50], there may be need to The authors thank Edwin Kirwa for helping with animal
re-examine the contribution of dietary kerosene supple- care and treatment and Winfridah Cheriro for assistance
mentation especially among school going children. From during the running of hormone assays.
data obtained during an earlier pre-animal study survey
(Fig. 1B), 47.8% of respondents with kerosene supplemen-
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