JOURNAL OF INTERDISCIPLINARY HISTOPATHOLOGY, 2018

VOL 6, NO. 2, PAGE 33–41

10.5455/jbh.20180605103508

REVIEW ARTICLE Open Access

Impact of gasoline on health. An updated review on gasoline-induced

histopathological effects in laboratory animals
Samar Alsaggaf
Anatomy Department, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia

ABSTRACT
ARTICLE HISTORY
Gasoline was described to induce many biochemical and physiological dysfunctions with
subsequent disturbance of health and development of many diseases. It was reported Received June 05, 2018
Accepted July 18, 2018
to induce health hazards through generating reactive oxygen species with subsequent
Published August 01, 2018
harmful effects on the normal body physiology. This work reviewed the previous experi-
mental studies conducted on animal models and aimed to assess the effect of exposure KEYWORDS
to gasoline through different routes on the different body systems and organs. The histo-
Gasoline; car fuel; health;
pathological impact of gasoline on the nervous, respiratory, reproductive, and immune histology; pathology;
system was reviewed. Adding to that the impact of exposure to gasoline on the skin, trachea; skin; experimental
liver, kidneys, blood, bone marrow, and genes was also reviewed. It is hoped that this animals
review will encourage and support the conduction of more rigorous studies aimed to
assess the impact of gasoline on the health and possibly help in optimizing the exposure
standards for gasoline and other hydrocarbon fuels.

Introduction expanded the creation of petrochemical indus-

Production, transportation storage, and consump-
tion of hundreds of millions of hydrocarbon fuels
gallons occur daily worldwide. Examples of these
fuels are gasoline, jet fuels, diesel fuel, diesel fuel
marine, and kerosene which are all utilized for
the fueling of air, land, and sea vehicles. Adding to
that the action of industrial and commercial equip-
ment and machinery, industrial, commercial, and
residential heating, and for numerous additional
applications. Hydrocarbon fuels are so commonly
distributed and consumed, but unfortunately, there
is no or little obligation for the usage of dermal
or respiratory protection tools with subsequent
frequent exposure to an extensive risk of acute or
repeated human exposure Ritchie et al. [1].
Gasoline is a fractionated product of crude oil
that has volatile components and its emission
induces chemical pollutants in the environment.
“It comprises more than 500 saturated or unsatu-
rated hydrocarbons having 3–12 carbons such as
n-pentane, n-hexane, toluene, and benzene” [2].
The diverse use of gasoline and the increased need
of the growing industrial institutions have greatly
try and petroleum exploration “which has been
reported as one of the main contributors of envi-
ronmental and global problems” [3]. Gasoline is
an example of a xenobiotic which can simply enter
into the organisms through inhalation, ingestion,
dermal contact, and diffusion and alter the normal
physiological status of the organism. It contains
very complex and inflammable substances which
can disrupt the endocrine system [4]. Gasoline was
described to induce many biochemical and physi-
ological dysfunctions through generating reactive
oxygen species. It suppresses the antioxidant effect
of superoxide dismutase, catalase, and reduces the
level of glutathione [5]. Workers exposed to chemi-
cals created and utilized in oil and gas industry were
reported to suffer from occupational diseases like
dermatitis and irritation and inflammation of respi-
ratory system and other organs at levels depending
on the length of exposure time [6]. In a recent study,
Owagboriaye et al. [7] concluded that inhalation of
gasoline fume might be “injurious to the normal
body physiology by increasing serum lipid peroxi-
dation, corticosterone, and aldosterone level”.

Contact  Samar Alsaggaf salsaggaf@gmail.com/ssagaf@kau.edu.sa Associate professor, Anatomy Department, Faculty of

Medicine, King Abdulaziz University, Jeddah, Saudi Arabia.
© 2018 The Authors. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
 Samar Alsaggaf
Although there were many studies conducted on
the effects of exposure to gasoline on different body
organs through different routes, some controversy
existed about these effects in this study. There was a
need to summarize and discuss the results of these
studies in order to be helpful for research studies
aiming to work on this research area. This article
aimed to review the previous experimental studies
conducted on animal models to assess the effect of
exposure to gasoline through different routes on
the different body systems and organs. This review
raised the research question “what were the effects
proved to be induced by gasoline of the different
body organs and systems of the experimental ani-
mal in the previous research studies?”
Effect of Gasoline on the Nervous System
The effect of vapor condensate of baseline gasoline
(BGVC) and Gasoline/methyl tert-butyl ether (G/
MTBE) on the pups of Sprague-Dawley rats exposed
to 10,000 and 20,000 mg/m3 6 hours/day, 7 days/
week was studied by Gray et al. [8]. They reported
no adverse neuropathology in F1 pups and their
study revealed no alterations in F1 offspring-ex-
posed BGVC or G/MTBE in brain length or width
compared to the control. They added that “exposure
to these substances did not cause changes in glial
fibrillary acidic protein levels in any brain region
examined indicated that none of these substances
induced gliosis in the brain regions examined” [8].
Effect of Gasoline on the Respiratory System
Al-Saggaf et al. [9] have studied the effect of car
fuel on histological structure of trachea and lung of
guinea pigs exposed to inhalation of gasoline near
the station tanks for 30 and 90 days. They reported
that trachea of animals exposed to gasoline vapor
for 30 days showed inflammatory cell infiltration,
mainly of lymphocytes and eosinophils, in the
mucosa and submucosa as well as significant reduc-
tion in the number of goblet cells. Scanning electron
microscopic examination showed focal desquama-
tion of epithelial cells together with loss and short-
ening of cilia on some columnar epithelial cells. The
lung alveoli of these animals showed focal inflam-
matory cell infiltrate, intra-alveolar hemorrhage,
and emphysematous changes in some areas while
the bronchi and bronchioles showed sloughing of
their epithelial lining and hypertrophy of their mus-
cle layer. Animals exposed to gasoline vapor for 90
34 J Interdiscip Histopathol • 2018 • Vol 6 • Issue 2
days showed same changes but at a much severe
degree (See Fig. 1).
The same research team was investigating the
tracheal histopathological changes induced in
guinea pigs by inhalation of gasoline vapor in labo-
ratory conditions [10]. The findings were consistent
with those reported by Al-Saggaf et al. [9] following
exposure of animals at the station (See Fig. 2).
Effect of Gasoline on the Skin
It was described that skin is exposed to gasoline
during its use in cleaning, handling contaminated
soil or water, or accidentally if spoiled on it and
finally, exposure of workers at gas stations. Upon
exposure to gasoline, some ingredients of it can
pass through the skin [11].
Some researchers have described that the skin of
guinea pig exposed to car fuel containing gasoline
for 21 days has resulted in erythematous thicken-
ing, ulcerated, firmly adherent to the underlying
tissue, and bled abundantly on removal [12,13].
Microscopically, the thickness of the whole epider-
mal, keratin, and granular layer was significantly
increased compared to the control. Focal elon-
gation of the rete ridges and infected ulcerations
were also observed. Upon examined by the electron
microscope, ill-defined dermo-epidermal junction
and an increase in intercellular spaces among the
basal layer cells due to desmosomal disruption
were observed. The spinous and granular cell lay-
ers showed marked cytoplasmic vacuolation and
the latter showed a marked decrease in fibrillar
contents. The individual layers in the stratum cor-
neum were increased in thickness and the regular
desmosomal junctions disappeared compared with
the control [13] (See Fig. 3).
These findings indicated that gasoline produced
skin irritation as it possessed lipid solving prop-
erties [14]. This irritation could be induced by
“destructing the hydrophobic barrier produced by
the lamellar bodies producing layers and, second,
by a direct effect on cells of the epidermal kerati-
nocytes stimulating the release of inflammatory
cytokines such as interleukin-1α” [15]. Production
of reactive oxygen species free radicals, with the
subsequent cytotoxicity, could be another mecha-
nism behind gasoline-induced skin irritation [16].
Reese and Kimbrough [17] reported that gasoline
increased cellular microsomal enzyme activity
with subsequent production of toxic metabolites of
benzene.
 Histopathological effects of gasoline in laboratory animals

Figure 1. “(A) Light microscopic sections of the trachea from the control group, and the group exposed to gasoline
vapor for 30 days at the station. (B) SM = Submucosa Eosinophils (thin arrow), inflammatory infiltrate (interrupted
arrow), di1ated gland (asterisk) and desquamated epithelium (white arrow). Lost cells (black thick arrows) and
goblet cells (arrow head (stained with Hematoxyline & Eosin). (C and D) Scanning electron microscopic sections
of the trachea from thecontrol group and the group exposed to gasoline vapor at the station. (E and F) Normal cilia
(thick black arrow), lost cilia (interrupted white arrow), short cilia (interrupted black arrow) goblet cells (thick
white arrow)” (cited after permission from Al-Saggaf et al. [13]).

Ahaghotu et al. [18] during their study on hair-
less rats stated that xylene and tetramethyl benzene
isomers (TMB) caused granulocyte infiltration,
swelling of the epidermis, and widespread destruc-
tion of stratum corneum, as well as increased
expression of tumor necrosis factor in the skin
compared to control. They added that “gasoline
led to disruption and separation of keratin layers,
most probably because of delipidization and pro-
tein denaturation with subsequent loss of barrier
function and enhanced transepidermal water loss”
[18]. Gunasekar et al. [19] observed a rise in the
number of mast cells and plasma cells at the epi-
dermal separation from the basement membrane
and in the dermis which suggests immune reaction
induced by TMB in hairless rats.

www.ejmjih.com 35
 Samar Alsaggaf

Figure 2. “Light microscopic sections of the lung of control animals (A–D) and animals exposed to gasoline vapor
for 30 days (E–H). AVL = alveoli, B = intrapulmonary bronchus, Bi = bronchiole, Od = edema. Pneumocyte type I
(white arrow head), Pneumocyte type II (black arrow head). Emphysema (Thick black arrow), lymphoid aggregation
(Hematoxyline & Eosin) (interrupted arrow)” (cited after permission from Al-Saggaf et al. [13]).

Effect of Gasoline on Reproductive System
Gasoline was reported to decrease seminal param-
eters and affect semen quality and induce male
genital anomalies as well as fertility [20]. The sub-
chronic effect of gasoline and gasoline/oxygenate
on the reproductive system and offspring were
assessed [8]. When gasoline blended with oxygenate

36 J Interdiscip Histopathol • 2018 • Vol 6 • Issue 2

 Histopathological effects of gasoline in laboratory animals

Figure 3. “(A) Light micrograph of a section of back skin from control rat showing normal intact keratinocyte layers
as well as hair follicles in the dermis. (B) A section of back skin painted by gasoline for 3 weeks showing thickening
and separation of keratin (star), a marked increase in the granular layer (double head arrow), Hematoxyline & Eosin
×600. (C) Transmission electron microscope (TEM) micrograph of a section of back skin from control showing intact
epidermal cells. (D) TEM micrograph of a section of skin painted with gasoline showing marked alteration of both
basal (GSe) and spinous cell layers [stratum spinosum (SS)]. The nuclei showed irregular outlines (black arrow). The
cytoplasm of some cells contains irregular vacuoles (black interrupted arrow) and dark disrupted filaments (star).
(E) TEM micrograph SS of skin painted with gasoline showing that some cells become distorted and shrunken with
irregular or fragmented nuclei (white arrow). (F) TEM micrograph of skin painted with gasoline showing increased
keratin layers (white arrow), karyolysis of granular cell nuclei, and a coarse electron-dense substance within the
cytoplasm (interrupted white arrow) ×7,500” (cited after permission from Al-Saggaf et al. [9]).

diisopropyl ether (G/DIPE) and ethanol (G/EtOH)
administrated at “10,000 and 20,000 mg/m3 6
hours/day, 7 days/week” to male Sprague-Dawley
rats, they resulted in a rise in absolute weight of
epididymis, prostate, seminal vesicles/coagulating
gland and in their relative weight to body weight,
although there were no histopathological changes
detected in these organs. They added that none
of these substances induced and macroscopic or
microscopic changes in female reproductive organs

www.ejmjih.com 37
 Samar Alsaggaf

[8]. The observed adverse effect level was the Genotoxic Effect of Gasoline
highest level tested (20,000 mg/m3 for gasoline and
The cytogenetic studies of unadditized gasoline and
gasoline/oxygenate) as there are no alternations in
its blending streams did not reveal any changes in
fertility, sperm counts, estrus cycle, or developmen-
chromosome aberrations or positive micronucleus
tal parameters in pups [8]. In another study done by
findings in laboratory animals [28]. In a study, the
Tyl et al. [21], reduced pup weights during lactation
cytogenetic hazard of different gasoline included
was reported in a two-generation study with inha-
oxygenates was assessed by Schreiner et al. [29].
lation exposure of gasoline blended with oxygenate
The vapor condensates of gasoline with oxygen-
t-amyl methyl ether (G/TAME) at concentrations of
ate ethanol (G/EtOH), diisopropyl ether (G/DIPE),
1,040, 2,250, and 12,500 mg/m3.
and t-butyl alcohol (G/TBA) were negative in both
Inhalational exposure of pregnant Sprague-
micronucleus and sister chromatid exchange (SCE)
Dawley rats to gasoline alone, or gasoline blended
tests “indicating an absence of DNA perturbation or
with various ethers or alcohols (on gestation days
cytogenetic hazard from inhalation of these mate-
5–20 for 6 hours/day at different levels) do not
rials” [29]. They reported a significant increase
produce selective developmental toxicity in rats. It
in SCE in rats given BGVC alone or in female rats
is noticed that “A dose responsive maternal effects
given G/MTBE. Gasoline/t-amyl methyl ether (G/
included decreased maternal body weight and/or
TAME) induced increased SCE in both sexes at the
weight change, and reduced food consumption. No
highest dose only. Although DNA perturbation was
significant malformations were described in any
noticed for several samples, DNA damage was not
study” [22].
expressed as increased micronuclei in bone mar-
row cells [29].
Effect of Gasoline on Immune System
In another study, Trevisan et al. [30] conducted
In previous research studies, Jet propulsion fuel-8 a study aimed to assess the occurrences of chro-
was reported to be an immunotoxicant when mosomal abnormalities and SCE in workers of gas
inhaled [23], topically administrated to the skin stations. They could not observe the presence of
[24], or when given orally through gavage [25]. In benzene and its derivatives and did not observed
contrast to these studies, White et al. [26] found chromosomal destruction that may be associated
that exposure of the female rodent species B6C3F1 with the gas station activity in the worker.
mice and Crl:CD rats to jet fuel kerosene through
inhalation for “28 days at levels up to 2,000 mg/m3” Effect of Gasoline on Kidneys
did not suppress innate, humoral, or cell-mediated
The effect of subchronic exposure of Sprague-
immune functions. They added that there was no
Dawley rats to inhalation of BGVC and gasoline
significant effect detected in either species on body
combined with methyl tertiary butyl ether (G/
weights, spleen, or thymus weights and with a few
MTBE) at different concentrations “2,000, 10,000,
exceptions, spleen cell numbers and phenotypes
or 20,000 mg/m3 for 6 hours/day, 5 days/week
were also unaffected.
for 13 weeks” was previously studied by Clark et
In a more recent study conducted by White et
al. [31]. They reported that both male and female
al. [27], they reported that “exposing the female
rats exposed to the BGVC and G/MTBE at all levels
Sprague-Dawley rats to BGVC, G/MTBE, G/TAME,
showed “eosinophilic hyaline granules within the
and gasoline with t-butyl alcohol (G/TBA) 6
cytoplasm of renal proximal convoluted tubular
hours/day, 5 days/week for 4 weeks did not alter
epithelial cells in a dose-dependent manner. Most
the humoral immune response as evaluated in
10,000 and 20,000 mg/m3 exposed males also had
the immunoglobulin M (IgM) antibody-forming
evidence of tubular regeneration. Several animals
cell response to the T-dependent antigen sheep
among each of these exposure groups had corti-
erythrocytes.” Spleen weight, spleen cell number,
comedullary intraluminal tubular granular casts.
or IgM antibody production did not show any sig-
Mallory-Heidenhain staining of renal tissue of
nificant changes when assessed. They added that
male rats showed red staining of granular material
the humoral immune response was significantly
within the proximal tubular epithelium. Minimal
depressed after exposure to the three vapor con-
staining was apparent in control males and slight to
densates: G/EtOH and G/DIPE at the 20,000 mg/m3
moderate staining was present in exposed female
dose and G/ethyl tertiary butyl ether at the 10,000
animals” [31].
and 20,000 mg/m3 doses.

38 J Interdiscip Histopathol • 2018 • Vol 6 • Issue 2

 Histopathological effects of gasoline in laboratory animals

Figure 4. “(A) Photomicrograph of liver exposed to gasoline fume for 3 hours (T3). Hematoxyline & Eosin (H&E)
×400: showing extensive distorted histoarchitecture, shrinkage/degenerating/hepatocytes (yellow arrows), pykno-
tism (purple arrow), and dilated central vein (red arrow). (B) Photomicrograph of liver exposed to gasoline fume for
5 hours (T4). H&E ×100: showing extensive distorted histoarchitecture, degenerated endothelium (green arrow);
dilated sinusoids (blue arrows), and central vein (red arrow); chromatolytic hepatocytes (yellow arrows) and pyk-
notic cells (purple arrows)” (cited after permission from Owagboriaye et al. [7]).

Effect of Gasoline on Liver Effect of Gasoline on the Blood and Bone

Liver is considered the key organ dealing with the
different compounds entering the body and this
denotes that it could be a target organ for environ-
mental pollutants that disrupt the metabolic action
of this organ [32].
Poon et al. [33] reported that oral administration
of gasohol (10% ethanol in gasoline by volume) in
female rats for 4 weeks did not induce any histo-
pathological alternations in the liver, kidneys, and
many other organs although it induces biochemi-
cal changes in the liver. In a recent study, the effect
of gasoline on the liver structural integrity was
assessed by Owagboriaye et al. [32]. The histo-
pathological examination of liver sections from rats
exposed to gasoline for varying periods (1, 3, 5, and
9 hours) daily for 12 weeks revealed “increasing
level of distorted histoarchitecture swelling/degen-
erated hepatocytes, degenerated endothelium,
chromatolytic hepatocytes and pyknotism, patchy
inflammation with remarkable sinusoidal space
and large central vein” (See Fig. 4).
The serum levels of liver enzymes were also
elevated in these rats exposed to gasoline which
might be indicative of liver cell damage and injury.
This indicated that “liver is one of the major target
organs of gasoline fume-induced injury.” These hep-
atotoxic changes induced by gasoline on the liver
were previously attributed to the cumulative oxida-
tive damage as reported by Uboh et al. [34].
Marrow
During the study of the effect of exposure to gaso-
line vapors “17.8 ± 2.6 cm3/h/m3/day, 6 hours/day,
6 days/week, for 20 weeks” on female Wistar albino
rats it was noticed that the rats developed growth
depression and weight loss [35]. In addition, the
rats suffered from a significant reduction in levels
of hemoglobin, hematocrit, red blood cells as well
as significant increase in white blood cells com-
pared to the control rats indicating that exposure to
gasoline vapors may cause hematotoxicity in rats.
In a later study, the bone marrow of the male
Sprague-Dawley rats inhaled petrol fumes at a dose
of 11.13 ± 1.1 cm3/hour for 6 hours daily, 6 days/
week for 11 weeks was extracted by Abubakar et al.
[36] for cytological examination [36]. They noticed
that inhalation of gasoline caused an increase in
the percentage of abnormal megakaryocytes with
detached nuclei, hypolobulation, and/or disintegra-
tion, while it did not induce significant changes in
oxidative markers in the erythrocytes [36].
Conclusions
Gasoline-induced negative impact on the health
was proved in many previous research studies.
The histopathological prospective of this impact on
the nervous, respiratory, reproductive, and immune
systems as well as on skin, liver, kidneys, blood, bone
marrow, and genes was reviewed in this study. It is

www.ejmjih.com 39
 Samar Alsaggaf
hoped that this review will encourage and support
the conduction of more rigorous studies aimed to
assess the impact of gasoline on the health and pos-
sibly help in optimizing the exposure standards for
gasoline and other hydrocarbon fuels. Performing
a future review on the clinical effects of exposure
to gasoline through different routes on the human
body is recommended. Not only that, but studies of
the possible protection against this reported patho-
logical effect of gasoline on different body system
are encouraged in order to help find solutions for
such problem.
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