Animal, page 1 of 9 © The Animal Consortium 2019 animal

doi:10.1017/S1751731119002593

Effects of ochratoxin A on membrane phospholipids of the

intestine of broiler chickens, practical consequences
I. El Cafsi1,2† , S. Bjeoui2, I. Rabeh2, S. Nechi3, E. Chelbi3, M. El Cafsi2 and A. Ghram1
1
Institute Pasteur of Tunis – Laboratory of Veterinary Epidemiology and Microbiology, University Tunis-El Manar, 1002 Tunis, Tunisia; 2Faculty of Sciences of Tunis, Unit of
Physiology and Aquatic Enviroment, University Tunis-El Manar, 2092 Tunis, Tunisia; 3Mohamed Tahar Maamouri Hospital, Anatomy and Cytology Service, Road Mrezka
8000, Nabeul, Tunisia

(Received 1 March 2019; Accepted 24 September 2019)

Ochratoxin A (OTA) is a mycotoxin produced by various species of Aspergillus and Penicillium. Ochratoxin A was classified as a
group 2B carcinogen and is one of the major intestinal pathogenic mycotoxins. One of the most frequent modes of intoxication is
consumption of contaminated food with mycotoxins. Feed represents the major cost and has a direct impact on the economical
viability of broiler’s production system, since it must contain the necessary elements that allow the animal to express the
maximum genetic potential while providing its nutritional requirements. Thus, the animal has to digest the feed and absorb its
nutrients, which is in direct correlation with the gastrointestinal tract, especially the small intestine and the development of the
mucosal surface area. Once ingested, OTA is absorbed by passive diffusion, mainly the jejunum. Ochratoxin A’s presence affects
lipid membranes and could lead to the degradation of their normal structure and functionality. All of these effects contribute to
the development of malabsorption. It was very interesting to study the effect of OTA on the layer of phospholipids of the bowel.
The experimental group received OTA (0.05 to mg/kg BW) through an intra-peritoneal injection, every other day for 21 days. We
noted that feed conversion ratio and average daily gain were reduced. Histological studies showed important alterations at the
level of the mucosal membrane of the intestine (villosities, crypts) following intra-peritoneal administration of the mycotoxin.
Thinning and enlargement at the base of the villosities, hyperplasia and crypts in irregular forms, blunting and denudation were
observed through the examination of intestinal morphology. Biochemical studies, such as total lipid and phospholipid
compositions, allowed us to have more detailed results. All identified mucosal phospholipids were modified, particularly the
phosphatidylcholine (PC) and the phosphatidylethanolamine (PE) in the jejunum mucosa. In fact, there was a decrease by
55.81% for PC, 56.66% for PE, while a significant increase by 32.91% was noted for phosphatidylserine in the jejunum. It was
very interesting to study the effect of OTA on the phospholipids layer of the bowel, as the mucous membrane of the small
intestine represents the main site of absorption and transformation of nutriments. To avoid such disturbances and prevent the
effects of the OTA, precautions must be taken to inhibit mold growth at the level of the feed manufactory units.
Phosphatidylcholine and PE administrations may represent an option that could allow reestablishment of phospholipid equilibrium
in the intestine.

Keywords: mycotoxin, membrane border brush, jejunum, phosphatidylcholine, phosphatidylethanolamine

Implications the phospholipid bilayers, it is recommended to make a

dietary addition of phospholipids, especially phospatidylcho-
Ochratoxins are carcinogenic mycotoxins that contaminate lines and phosphatidylethanolamines.
food and thus induce a fall in technical performances, in
addition to contamination of human’s food. In practice, no
poultry feed is completely free of mycotoxins. Once ingested,
ochratoxin A is partially absorbed through the digestive tract, Introduction
mainly in the animal’s intestine, which engenders degrada-
tion of normal structures and functionalities. To prevent and There is a common perception that ‘human made’ chemicals are
mend the damaged structures of the intestine, particularly more dangerous than natural substances. Interestingly, the most
toxic compounds such as mycotoxins are natural. Mycotoxins,
†
E-mail: imneelcafsi@yahoo.fr from the Greek «mycos» for ‘mold’ (Fink-Gremmels, 2008), are

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El Cafsi, Bjeoui, Rabeh, Nechi, Chelbi, El Cafsi and Ghram

secondary metabolites produced by fungi (Marquardt, 1996), any effect on nutrient absorption would induce a deficit in the
mainly Aspergillus, Penicillium, Alternaria, Fusarium and growth of the animal, the activity of the immune system, the
Claviceps, which may be dangerous to vertebrates upon lymphatic circulation and the level of hormonal synthesis.
ingestion (Stoloff, 1979), inhalation or skin contact. Food
contamination occurs after being contaminated by spore,
conidia or mycelia fragments of fungi producing mycotoxins,
which represents a great concern for humans and animals Material and methods
(Food and Agriculture Organization, 2004). Consumption Animals and sampling
of such contaminated food by humans or animals leads to Thirty-one-day-old male broiler chicks (Hubbard JV, The Foeil -
mycotoxicoses (Hanif, 2016). Despite the ever-increasing Quintin France) were obtained from a local supplier. All exper-
understanding of mycotoxins and their effects, they have imental animals were maintained and used in accordance
continuous and severe economic impact worldwide. These with the Guidelines of the Committee for the Care and Use
losses are associated with lower productivity, reduced gain of Experimental Animal Resources of the Institute Pasteur of
weight, decreased feed efficiency, diminished meat and Tunis, Tunisia (F16-00170(A5743-01)). Animals were ran-
egg production, greater disease incidence due to immune domly divided into one control group (n = 15) and one exper-
suppression, subtle damage to vital body organs and inter- imental group (n = 15), housed in an experimental isolators.
ference with the reproduction system, all of which have a All birds received feed (formulated to meet the nutritional
much more immediate impact on morbidity and mortality requirements for starter and grower broilers, as shown in
(Marquardt, 1996). In the field, changes in animal perfor- Table 1) and water ad libitum. After a period of acclimatisa-
mances or behaviour along with increased susceptibility to tion to environmental conditions (1 day), the experimental
infectious agents represent possible subtle and non-specific group received OTA (0.05-mg/kg BW) through an intra-
signs of mycotoxin exposure and make it difficult to establish, peritoneal injection, every other day for 21 days (Prior et al.,
with certainty, a cause–effect relationship with ‘contami- 1976; Chang et al., 1981) and the control group received
nated’ feedstuffs, thus complicating clinical diagnosis and an equivalent volume of monosodium carbonate (NaHCO3
contributing to causality difficulties. Furthermore, food con- 0.1 M). Bird BWs and feed intake were determined daily until
tamination is more relevant considering the fact that there is 21 days of age, while feed conversion ratio (FCR, feed intake/
no way of efficiently predicting and avoiding mycotoxin con- weight gain) was calculated accordingly. At the end of the
taminations (Duarte et al., 2011). In addition, while scientists experimental period, chicks were slaughtered by bisection
have previously looked at various mycotoxin polluting agri- of the cervical vessels, and intestines from all animals were
cultural resources causing mycotoxicosis; nowadays, their collected and kept frozen (−20°C) until analysed. A portion
focus is been shifted towards ochratoxin A (OTA) contamina- of the intestine was placed in 10% formal in buffered solution
tion (Hanif, 2016). In fact, OTA represents one of the most to be used for histopathological studies.
important causes of economic losses for the poultry’s indus-
try (Hanif, 2016). Therefore, the presence of OTA in the live-
stock feed chain negatively impacts the animal’s health and
production (Hanif, 2016). It is well known that the mucous Table 1 Ingredient composition and nutrient content of the basal diets
membrane of the small intestine represents the main site of broiler chickens (%, as fed basis)
of absorption and transformation of nutriments and is con- Chemical composition Starter Grower
stituted by several cell types including the secretory units,
the endocrine cells and the enterocytes or absorbent cells Ingredients (%)
(Alessandri et al., 1990). Phospholipids represent, with gly- Corn 63 67
colipids and cholesterol, the main lipid constituents of the Soybean meal 32 28
membrane border brush (Vance, 2018). However, the major Vitamin-mineral complex1 5 5
Maximum humidity 12 12
part of the global lipid composition of the intestinal border
Crude fibre (max) 5 5
(Sherman strain) is represented by 34.1% phospholipids CPs (min) 21 18.5
(Thomas and David, 1980). It is thus very important to Fatty substances (min) 3 5
know how the cellular membrane reacts to poisoning by Converted energy (kilo silks/kg) 2800 2800
OTA, especially those constituents that allow absorption of Calcium Ca 1 1
nutriments. In fact, after ingestion, OTA is absorbed by pas- Phosphorus P 0.45 0.42
sive diffusion mainly in the proximal third of the jejunum. Methionine 0.42 0.4
Therefore, its presence affects lipid membranes and could Tryonine 0.8 0.9
lead to the degradation of their normal structure and func- Metabolisable energy (mJ/kg) 0.75 0.7
tionality. Ochratoxin A promotes free radical formations ME = Metabolisable energy.
in the intestines leading to oxidative stress, antioxidant 1
The premix provided per kilogram diet: vitamin A 350 000 IU, vitamin D3 75 000
depletion and apoptosis; all of which contribute to the devel- IU, vitamin E 2000 IU, vitamin K3 75 mg, vitamin B1 70 mg, vitamin B2 190 mg,
vitamin B5 300 mg, vitamin B6 100 mg, vitamin B12 0.50 mg, vitamin K3 75 mg,
opment of malabsorption. We are interested in the effect of choline chloride 500 mg, folic acid 35 mg, biotin 5 mg, niacin 1300 mg, Fe
such toxins on the layer of phospholipids of the bowel, since 1400 mg, Cu 250 mg, Mn 2000 mg, Zn 2000 mg, I 25 mg, Se 5 mg, Co 8 mg.

2

Extraction of total lipids and separation of lipid classes
Before lipid extraction and immediately after the chickens
were slaughtered, chick intestines were kept for 10 min
in boiling water to inactivate any active enzymes, such
as phospholipases (Shewfelt et al., 1981). Total lipids were
extracted according to the method of Folch et al. (1957).
The used solvent was a mixture of chloroform : methanol
(2 : 1, v/v), containing 0.01% butylated-hydroxyl-toluene
as an antioxidant. Lipid classes were separated using
thin-layer chromatography with one-dimensional double
development (Folch et al., 1957). A 200 μl of lipid extract
was separated on ‘silica gel 60’ plates (20 × 20 cm; Merck,
Darmstadt, Germany), using methyl acetate : isopropanol :
chloroform : methanol : 0.25% KCl (25 : 25 : 25 : 10 : 9, v/v),
a developing medium for polar lipids (PL). Lipid classes
were visualised under UV light after spraying a 0.1%
20-7 0 dichloro-fluoresceinin absolute methanol. The phos-
pholipids classes were identified using standards deposed
in the same conditions (Olsen and Henderson, 1989).
Quantification of the lipid categories
After migration on a silica gel plate, each spot corresponding
to an identified lipid category was scratched and recovered
with silica. Then, the fatty acids were extracted, methylated
and finally injected into a gas chromatograph. For fatty acid
analysis, PL fractions were esterified to obtain fatty acid
methyl esters (FAMEs), according to the method reported
by Cecchi et al. (1985). Nonadecanoic acid (C19:0), undetect-
able in our samples, was used as an internal standard to
be able to quantify fatty acids of each lipid category. The sep-
aration of FAMEs was carried out using HP6890 gas chro-
matograph (Agilent Technologies, Santa Clara, CA, USA)
with a split/splitless injector equipped with a flame ionisation
detector at 275°C and a 30-m HP Innowax capillary column
(250-μm internal diameter and 0.25-μm film thickness). The
injector temperature was held at 250°C, rising from 50°C
to 180°C at a rate of 40°C/min, from 180°C to 220°C at
1.33°C/min, and stabilised at 220°C for 7 min. The used gas car-
rier was the nitrogen. To identify the FAMEs, a mixture of methyl
esters (Supelco Polyunsaturated Fatty Acids n-3 or omega 3
(PUFA-3)) was used and administered in the same conditions.
Fatty acid peaks were integrated and analysed using HP
Chem Station software (CPG Agilent 7890A; Agilent
Technologies). The results represented the average of six repli-
cates (n = 6). Quality control of replicates of results obtained is
confirmed in Supplementary Material Table S1.
Analysis of the effects of toxins on the morphology of
intestinal epithelial cells
Pieces of 0.5 to 1 cm of each segment of the small intestine,
far away from the mesentery, were removed and cut longi-
tudinally. Then, each sample was rinsed in three successive
baths of physiological saline solution (NaCl 9 g/l), according
to the method described by Goodlad et al. (1991). The
samples were slightly buffered on absorbent paper before
being placed in 10% formalin solution and then embedded
in paraffin. Finally, the paraffin blocks were cut into
Mycotoxins and intestinal modifications
sections (2 to 4 μm) and stained with haematoxylin–eosin
(H&E). Obtained sections were examined under a light micro-
scope (Leica DM 2500 equipped with a camera Leica D-lux 3
and a Software AxioVision Rel.4.8.2.SP2, Wezlar, Germany);
a histological score was given according to morphological and
lesional criteria, as previously described. Decrease in the total
score revealed a deterioration of the epithelium.
Statistical analyses
The results were presented as the mean of six replicates and
expressed as the mean ± SD. Normal distribution and homo-
geneity of variances of standardised lipid quantities and
phospholipid categories were verified using Shapiro–Wilk’s test
(Shapiro and Wilk, 1965) and Levene’s test (Levene, 1960),
respectively. Analysis of variance/Multivariate Analysis of
Variance (MANOVA) procedure was used for mean compari-
sons, and Dunken’s method for discriminating between the
means (Supplementary Material S1). Differences between the
samples were considered significant at P < 0.05. In case of a
significant deviation from normality and/or non-homogenous
data, the non-parametric test of Kruskal–Wallis (Kruskal and
Wallis, 1952) was conducted. Statistical analyses were per-
formed using R (R i386 3.4.4) for windows.
Results
Feed consumption and feed efficiency
The results of broiler performance are summarised in Figure 1.
From the first day of experiment, we have noticed that the
OTA group presents a lower feed intake, affecting the FCR
and consequently the average daily gain (ADG). The FCR dif-
ference between the control and the OTA groups increased
everyday reaching 13.84% by day 5. After 1 week, a differ-
ence of 16.61% was noticed and this percentage continued
to increase, to reach its maximum of 18.82% on day 13 and
stabilise at 16.69% by day 19 (Figure 1). The ADG showed a
decrease from day 1 to day 6 in the control group and day 7 in
the OTA group. Then, ADG started to increase progressively
up to day 20, the OTA group showing lower ADG during the
entire observation period (Figure 2).
Figure 1 (Colour online) Evolution of feed conversion ratio of broiler chicks
in control and experimental ochratoxin A groups (n = 15).
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El Cafsi, Bjeoui, Rabeh, Nechi, Chelbi, El Cafsi and Ghram
Intestinal lesions and morphology
Different types of lesions were observed along the different
portions of the OTA group. The morphology of the duodenum
of the control group was characterised by developed villos-
ities in the shape of a tongue (Figure 3a). The crypts have
regular form and normal width, different from the OTA group,
which demonstrated crypts of irregular forms and presented
an enlargement on their basis. A hyperplasia and an increase
in the width of the crypts, with shortened and blunt veloc-
ities, were also noticed (Figure 3b).
For the jejunum, the villi of the middle segment of the con-
trol group were distinguished by a long and a rectilinear
shape with crypts showing a regular form (Figure 4a). In con-
trast, the effect of mycotoxin administration on the structure
of the crypts and the villosities of the jejunum demonstrated
villous blunting and epithelial denudation, along with crypts
Figure 2 (Colour online) Evolution of the average daily weight gain of
broiler chicks in control and experimental ochratoxin A groups expressed
as food ingested.
Figure 3 (Colour online) Representative microstructure of duodenum of broiler chicks (a) control group and (b) ochratoxin A group at 21 days of age (scale bar,
100 μm).
Thinning
Enlargement at the basis of the villosities
Hyperplasia of crypts
Crypts in irregular forms
4
showing irregular forms and hyperplasia (Figure 4b). In the
ileum, which is the last segment of the small intestine, the
villosities were shorter and marked by their pyramidal form
with crypts having a normal shape, in the control group
(Figure 5a). However, the treated group showed blunting
villosities and crypts with irregular shapes, as it was shown
for other intestine segments of the OTA group (Figure 5b).
Total lipid composition
Lipid composition of the three segments of the small intes-
tine, the duodenum, the jejunum and the ileum, was exam-
ined (Figure 6). At a first glance, significant differences
(P < 0.05) were detected in the composition of the total lipid
in the three tested segments, in the presence or absence of
OTA. The jejunum showed the highest value of lipid compo-
sition in the absence of OTA, in comparison with the duo-
denum and the ileum contents (P < 0.05). On the contrary,
the total lipid composition, in the presence of OTA, showed
a significant increase (P < 0.05) in the duodenum and the
ileum, while there was a significant decrease (P < 0.05) in
the jejunum. The duodenum was characterised by the highest
amount of total lipid content as compared to the two other
segments, in the presence of OTA.
Phospholipid composition
The phospholipid compositions of the three segments of the
small intestine, expressed as their content of phosphatidyl-
choline (PC), phosphatidylserine (PS), phosphatidylethanol-
amine (PE) and phosphatidylinositol (PI), were examined
through this experiment. The control group showed that
the amount of PC in the jejunum is the most important with
170.76-μg/g fresh matter (FM), followed by the ileum with
128.77-μg/g FM, while the duodenum has the lowest amount
with 120.60-μg/g FM. Concerning the amount of PE, it was
shown that the jejunum presents the highest amount with
 Mycotoxins and intestinal modifications

Figure 4 (Colour online) Representative microstructure of jejunum of broiler chicks (a) control group and (b) ochratoxin A group at 21 days of age (scale bar,
100 μm).
Blunting villi
Denudation
Hyperplasia of crypts
Crypts in irregular forms

Figure 5 (Colour online) Representative microstructure of ileum of broiler chicks (a) control group and (b) ochratoxin A group at 21 days of age (scale bar,
100 μm).
Blunting villi
Denudation
Crypts in irregular forms

314.58-μg/g FM, followed by the duodenum with 105.57-
μg/g FM; the lowest amount being detected in the ileum with
96.79-μg/g FM. Similarly, the jejunum presented the highest
amount of PI with 68.50-μg/g FM, the duodenum showed
62.35-μg/g FM and the ileum had the lowest amount of
25.57-μg/g FM. On the contrary, in the jejunum, the PS pre-
sented the most significant amount with 222.46-μg/g FM,
while the duodenum and the ileum have shown the amounts
of PS equal to 103.53 and 101.62-μg/g FM, respectively.
Overall, the results showed that the impact of OTA on the
various phospholipid contents in the small intestine was
especially perceived in the ileum segment (Table 2). In fact,
there was an increase in the amount of PC by 98.91%, PI by
304.06% and PS by 84.49%; while a significant decrease was
noted in the amount of PE by 54.72% in the ileum. For the
duodenum, there was a significant decrease in the amount of
PC by 78.07%, PE by 43.13%, while an increase was noted in
the amount of PI by 17.92% and PS by 61.78%. For the jeju-
num, a decrease in the amounts of PC by 55.81% and of PE
by 56.66% was observed. On the other hand, an increase in
the amount of PS by 32.91% was noticed.
Discussion
Mycotoxicoses are considered as the second highest cause of
poultry farm losses, behind digestive bacterial infections
(Liew and Mohd-Redzwan, 2018). Sklan et al. (2003) have
found that mycotoxins alter digestive and absorptive func-
tions. Such information confirms our first results, as we have

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El Cafsi, Bjeoui, Rabeh, Nechi, Chelbi, El Cafsi and Ghram

Table 2 Effect of ochratoxin A on lipid and phospholipid composition

in the intestinal segments of broiler chicks
Variable Control RSD OTA RSD P-value

Lipid content (g/100g FM)

Duodenum 8.54 0.006 9.88 0.005 0.013
Jejunum 10.84 0.008 9.19 0.009 0.013
Ileum 8.20 0.009 9.47 0.007 0.07
Phosphatidylcholine
Duodenum 120.60 5.01 26.07 3.54 <0.001
Jejunum 170.76 8.07 75.46 5.21 <0.001
Ileum 128.77 0.85 256.13 19.40 <0.001
Phosphatidylethanolamine
Duodenum 105.57 19.04 60.03 4.36 0.015
Jejunum 314.58 1088 136.34 3.15 <0.001
Ileum 96.79 9.29 43.82 3.74 0.001
Phosphatidylinositol
Duodenum 62.35 10.24 73.53 6.76 0.12
Jejunum 68.50 4.25 69.01 2.68 0.87
Ileum 26.57 2.94 107.37 4.97 <0.001
Phosphatidylserine
Duodenum 10.24 10.24 167.49 19.14 0.007
Jejunum 10.48 10.48 295.68 11.40 0.001
Ileum 11.26 11.26 187.48 15.59 0.0017
RSD = residual standard deviation; OTA = ochratoxin A; FM = fresh matter.

Figure 6 Lipid quantity in the intestinal segments (duodenum, jejunum
and ileum) of broiler chicks (a) control group and (b) ochratoxin A group.
The box portion of the plot represents the interquartile range and the
line drawn through the box represents the median. The vertical whiskers
represent the highest and the lowest values in the data set. GFM = gram
fresh matter.
shown that not only the FCR decreases but also the ADG.
These reductions started on day 7 with a decrease of
16.61% in the conversion ratio/feed efficiency and 16.88%
in the ADG. On days 13 and 19, the FCR showed a decrease
of 18.82% and 16.69%, respectively. Gentles et al. (1999)
reported similar results with a 19% reduction in BW gain
on the second and third weeks following OTA and cyclopia-
zonic acid administrations.
These results could be explained by the fact that one of the
known effects of mycotoxins is induction of an uncontrolled
and extravagant infiltration in the intestinal barrier, leading
to an increased permeability (Maresca et al., 2001). This
might indicate a negative effect on the local immunity
(Piotrowska et al., 2014) and a decrease in the resistance
or an increase in the mucosa permeability, leading to an
impairment of nutrient digestion and absorption.
The villi represent the absorbing functional units, charac-
terised by their numerous capillaries that promote nutrient
absorption and their transport to the cells (Institut National
de la Recherche Agronomique, 1992). Each segment of the
small intestine is also characterised by a typical morphology
of its villosities. Thus, the duodenum, where the villosities in
the shape of a tongue are very developed (Figure 3), filters
the chime and allows only small particles to pass away.
Ochratoxin A induced hyperplasia of the crypts with an
enlargement at their basis, increased the width of the crypts
that are already in irregular forms and finally shortened and
blunted the villosities (Figure 4).
The jejunum, the most important segment and the longest
one, as it occupies half of the entire bowel of the chicken, is
distinguished by a long and a rectilinear shape. Figure 5
shows villous blunting and epithelial denudation, and
hyperplasia of crypts in irregular forms as a consequence
of the OTA’s presence. Ending with the ileum, where the
finalisation of intestinal absorption takes place, the villos-
ities are shorter and marked by their pyramidal form, with
the crypts having irregular shapes different from that of the
control group. In this way, it can be inferred that the effect
of OTA in these three intestine segments induced modifi-
cations in the so-called brush border of the small intestine.
Thus, not only the villosities but also the crypts, where cell
renewal of intestinal mucous membrane takes place, are
impugned.
Other authors have found similar results when studying
several mycotoxins (OTA, deoxynivalenol, T-2 toxin and
diacetoxyscripanol) and showed almost the same histopa-
thological changes in duck and broiler intestines with
shortened, atrophied and thin villi, and elongated crypts
with irregular forms (Ruan et al., 2018), observed in the
jejunum’s villosities of turkeys. Solcan et al. (2015) indi-
cated that high amounts of OTA cause destruction/injury
of the intestinal mucosa of chickens, and crypt hyperplasia
is related to villous atrophy, additionally, crypt elongation
is a remediation feature to villous shortening. As a result,
the immune responses such as the inflammatory reactions

6

were observed and corresponded to the intestinal lesions
and morphology changes (McLaughlin et al., 2004).
Moura et al. (2004) reported that exposing 1-day-old
broiler chicks to low doses of OTA causes a decrease in
the average numbers of monocytes and heterophils, weaken-
ing the animal immune responses against pathogenic agents.
This could be related to the sensitivity of several immune
cells, such as macrophages, lymphocytes B and T, to myco-
toxins but also to the alteration of cytokine’s secretion and
the decrease of antibody’s response (Al-Anati and Petzinger
2006). Obviously, to struggle against these pathogenic
agents, the animal body needs to reassign energy away from
feed efficiency and growth towards immunity and homeosta-
sis (Cleynen et al., 2014).
The intestine represents the most important tie between
the ingested feed and the performance and the health of the
animal. The border membrane in brush is the main site of
absorption and the transformation of nutrients. It is therefore
important to establish and study the effects of mycotoxins on
the structure of the small intestine mucosa. Some authors
theorised that during unfavourable times, fat is used as an
energy source (Blem, 1976). The animal could be facing this
kind of imbalanced situation during mycotoxicosis as they
engender a weak consumption index, and consequently, the
bird’s organism uses its lipid reserves. Thus, the analysis of
the small intestine total lipid composition can corroborate such
a theory since it can be seen that lipid content in the jejunum
has decreased (Te: 10.84 ± 0.8 and OTA: 9.19 ± 0.9), and the
jejunum represents 50% of the whole small intestine, implying
the need for more energy. Concerning the duodenum and the
ileum, it was noticed that, unlike the jejunum, the supply of
total lipids increased (duodenum: Te: 8.54 ± 0.6, OTA:
9.88 ± 0.5; ileum: Te: 8.2 ± 0.9, OTA: 9.47 ± 0.7).
As phospholipids are the principal component of the
erythrocyte membrane, it is important to know how this lipid
profile normally evolves through this disequilibrium, espe-
cially where PC stands out because of its important protective
role of the intestinal mucosa (Vance, 2018). In fact, the PC
decreased by 78.07% in the duodenum and 55.81% in the
jejunum, while it increased by 98.91% in the ileum. The
PE decreased by 43.13% in the duodenum, 56.66% in the
jejunum and 54.72% in the ileum. The PI increased by
17.92% in the duodenum, 0.73% in the jejunum and espe-
cially by 304.06% in the ileum. Finally, the PS increased in the
duodenum by 61.78%, 32.91% in the jejunum and by
84.49% in the ileum.
Concerning the ileum, PC increased by 98.91% in the
presence of mycotoxin and might have been synthesised
either directly through cytidine-diphosphate(CDP)-choline
(Kennedy and Weiss, 1956; Weiss et al., 1958) or through
successive methylations of PE (Bremer and Greenberg,
1960) as shown in Figure 7.
As PE also decreased by 54.72%, this could lead one to
think of two synthetic pathways that may take place; the first
one is directly related to ethanolamine (Kennedy and Weiss,
1956) and the second is in relation to synthesis from PS
(Dennis and Kennedy, 1972) as shown in Figure 7.
Mycotoxins and intestinal modifications
(a)
Diglyceride Phosphatidylcholine
CDP-Choline CMP
(b)
Diglyceride Phosphatidyl ethanolamine
CDP-ethanolamine CMP
(c)
CDP-Diglyceride Phosphatidylserine
Serine CMP
(d)
CDP-Diglyceride Phosphatidylinositole
Inositol CMP
Figure 7 Pathways of phospholipid biosynthesis in the intestinal segments
(duodenum, jejunum and ileum) of broiler chicks: (a) biosynthesis of phos-
phatidylcholine, (b) biosynthesis of phosphatidylethanolamine, (c) biosyn-
thesis of phosphatidylserine, (d) biosynthesis of phospatidylinositole.
These results could be confirmed by the significant
increase in PS content by 84.49%, suggesting that PS
was synthesised through decarboxylation of serine (Kuge
et al., 1986).
In the same way, Folch et al. (1957) have shown that the
CDP-glycerin is used for the synthesis of both PS and PI (Kuge
et al., 1986). As the content of PI increased by 304.06%, this
would suggest that most of the amount of CDP-diglyceride is
used to synthesise PI from inositol (Figure 7).
In the jejunum and in the presence of OTA, PS increased by
32.91% but PE and PC decreased by 55.81% and 56.66%,
respectively. In the duodenum, the changes were also similar,
as we have noted an increase in the amount of PS by 61.78%
but a decrease of PE and PC by 43.13% and 78.07%, respec-
tively. In this way, it can be inferred that both ways of PC
synthesis may be CDP-choline; or by successive methylations,
respectively (Weiss et al., 1958; Bremer and Greenberg,
1960), PC synthesis did not take place. Moreover, as the
amount of PE decreased in both intestine segments, it might
be suggested that the synthesis of PE from ethanolamine is
not activated (Kennedy and Weiss, 1956). On the other hand,
as PI remained virtually unchanged (0.73% in the jejunum
and 17.92% in the duodenum), this would imply that all
the amounts of CDP-diglyceride are used to produce PS from
serine (Kuge et al., 1986), which has effectively increased by
32.91% in the jejunum and 61.78% in the duodenum
(Friedman et al., 2018).
The aim of the present work is to study the effects of OTA
on broiler chicks following intra-peritoneal injection and a
7
El Cafsi, Bjeoui, Rabeh, Nechi, Chelbi, El Cafsi and Ghram
3-week period of experimental observations. An important
decrease of chick BW was noticed, which could be related
to the large lesions caused by this mycotoxin on the small
intestine. The phospholipid bilayer of the intestinal mem-
brane showed modifications that have concerned all phos-
pholipids, especially a deficit in PC and PE at the level of
the jejunum. To prevent these effects of OTA, PC and PE,
administrations may represent an option allowing for the
reestablishment of phospholipid equilibrium in the intestine
(Van Hoogevest, 2017).
Acknowledgements
This work was financially supported by the Ministry of Higher
Education of Tunisia. The authors sincerely thank Mr. Nicolas
Dettman for his English proofreading.
I. El Cafsi 0000-0001-8320-265X
Declaration of interest
The authors declare that there are no conflicts of interest.
Ethic statement
The experimental animal protocol for this study was approved
by the Guidelines of the Committee for the Care and Use of
Experimental Animal Resources of the Institute Pasteur of
Tunis, Tunisia (F16-00170(A5743-01)).
Software and data repository resources
The authors declare that the data of this research are not
deposited in any official repository.
Supplementary material
To view supplementary material for this article, please visit
https://doi.org/10.1017/S1751731119002593
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