No está claro cuán grande es el papel de los antibióticos https://antibioticos-wiki.es en las relaciones competitivas entre los microorganismos en condiciones naturales. Zelman Waxman creía que este papel era mínimo, los antibióticos no se forman sino en culturas limpias en entornos ricos. Posteriormente, sin embargo, se descubrió que en muchos productos, la actividad de síntesis de antibióticos aumenta en presencia de otros tipos o productos específicos de su metabolismo.
Microsoft word - mehel et al pdf
African Journal of Biotechnology Vol. 9 (21), pp. 3223-3230, 24 May, 2010
Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 2010 Academic Journals
Effects of metronidazole and probiotics
oligosaccharide on bacterial translocation in protein
Benakriche Ben Mehel1*, Boudali Souad1, Gorine H1, Bekada Ahmed2, Philippe Pochart3,
Saïdi Djamel1 and Kheroua Omar1
1Laboratory of Physiology of Nutrition and Food Safety. Faculty of Science. University of Oran, Algeria.
2Laboratory of Microbiology and Food Security. Faculty of Science. University of Oran, Algeria.
3Laboratory of Biology – EA 3199, Institute of Scientific and Technical Food, CNAM, Paris, France.
The present study aims to evaluate the effects of metronidazole, probiotics oligosaccharide on
indigenous microflora and bacterial translocation (BT) in protein malnourished rats. Thirty male Wistar
rats were divided into three groups: protein malnourished rats PM (group1, n = 10) were fed with maize
only, protein malnourished rats (group 2, n = 10) were received metronidazole and protein malnourished
rats (group 3, n = 10) were received both metronidazole and probiotics-oligosaccharide for fifteen days.
Metronidazole (1000 mg/kg/day) was given via an orogastric feeding tube to the second and third
groups. Lyophilized probiotics-oligosaccharide (0.5 mg/g body weight/day) was given in two doses via
the same route to the third group. All animals were sacrificed after fifteen days of protein malnutrition
and cultures of the mesenteric lymph nodes (MLNs), liver, spleen and cecal contents were done. The
incidence of bacterial translocation (BT) was 30% (3/10) in protein malnourished group 1,60% (06/10) in
group 2 where protein malnutrition was associated with metronidazole and 25% (2.5/10) in group 3
whose animals were subjected to protein malnutrition associated with metronidazole and probiotics
oligosaccharide. A significant increase in the BT incidence was found in group 2 (P < 0.05), while a
significant decrease was found in group 3 when compared to group 1. The total bacterial count of cecal
flora was significantly low in group 3 than in group 1 (P < 0.01). These results suggest that the
incidence of BT in protein malnutrition is increased by using an antibiotic while probiotics-
oligosaccharide decreases this incidence in protein malnutrition induced by antibiotic. Thus, we
conclude that probiotics-oligosaccharide can effectively protect the intestinal mucosa and prevent BT
in protein malnourished infants.
Bacterial translocation, protein malnutrition, probiotics oligosaccharide, metronidazole and
In light of the wide usage of antibiotic drugs in developing
the effects of malnutrition on the disposition of antibiotics.
countries, it becomes extremely important to determine
In the world, severe acute protein malnutrition (PM) affects
approximately 13 mil ion children under the age of 5 and it is
associated with 1 - 2 mil ion preventable child deaths each
year (Col ins, 2007). Use of antibiotics promotes the
*Corresponding author. E-mail: email@example.com.
emergence of resistant organisms and multiple-antibiotic
Tel: + 213773926535. Fax: + 21345301018.
resistance has become a major public health issue.
Nowadays, metronidazole (2-methyl-5-nitroimidazole-1-
lation of the intestinal flora, especial y during an antibiotic
ethanol) is widely used in the treatment of parasitic
in case of diarrhea or intestinal parasites. For al the
diseases and mainly against anaerobes, including Bacte-
positive effects of probiotics, three conditions can be met,
roïdes (Freeman et al., 1997). Metronidazole at the doses
a cocktail of the best micro-organisms in sufficient
used (1 mg/ml drinking water ad libitum
) disrupts the
intestinal bacterial flora resulting in the destruction of
Protein malnutrition disrupts the normal ecology of the
strict anaerobic bacteria and therefore, an overgrowth of
microflora affecting strictly anaerobes (Tannock and
enterobacteria in the cecal (Berg, 1981). This al ows
Savage, 1974; Poxton et al., 1997), impairs host immune
studying the simultaneous influence of malnutrition and
response and antibacterial defenses (Reynolds et al.,
antibiotics. There has been a growing interest in the
1992; Chandra, 1993), enhances the susceptibility to
effects of malnutrition on drug metabolism and pharma-
infection and leads to mucosal atrophy (Reynolds et al.,
1996). Malnutrition is a common problem for critical il
Although malnutrition itself cannot be treated by drug
patients and nutritional support is mandatory.
therapy, it can induce situations where drugs are the
The effect of probiotics oligosaccharide in combination
primary form of treatment. The treatment of infections is
with antibiotics on protein malnutrition has not been
the most common and severe drug-related problem asso-
proposed until now as an alternative to the use of
prophylactic antibiotics. Antibiotic prophylaxis is intended
In addition, probiotics were used in conjunction with
to prevent the potential of bacterial contamination as a
antibiotic therapy to prevent or lessen the severity of
situation risk. It is unknown if modification of the intestinal
antibiotic-associated diarrhea in children (Vanderhoof et
flora with such a multispecies probiotics mixture with
al., 1999), although a meta-analysis of this issue showed
fructo oligosaccharide reduces bacterial overgrowth and
significant problems in study design in several studies
bacterial translocation from the gut and, consequently,
alters the course of disease. Therefore, the present study
A malnourished individual is known to be much more
was as an attempt to assess if modification of intestinal
susceptible to infections. In light of the wide usage of
flora by a specifical y designed, multispecies probiotics
antibiotic drugs in developing countries, it becomes extre-
mixture with fructo oligosaccharide changes disease
mely important to determine the effects of malnutrition on
course using a wel -established rat model of protein
the disposition of antibiotics (Jung and Shahl, 1986).
The gut microflora is an important constituent of the
intestinal mucosa barrier and this has led to the concept
of probiotics therapy, that is, the application of potential y
MATERIALS AND METHODS
beneficial microorganisms (Ful er, 1992). Probiotics is a
microbial dietary adjuvant that beneficial y affects the
Animals and diets
host physiology by modulating mucosal and systemic
Thirty male Wistar rats weighing 60 - 70 g and aged 28 days were
immunity, as wel as improving nutritional and microbial
used. Al animals were obtained with approval from the Animal
balance in the intestine tract. They are nonpathogenic
Research Center of Bab Ezzour University, Algeria. The rats were
microorganisms that, when administered in adequate
housed in stainless-steel cages in an animal room maintained at 22
amounts, confer a health benefit on the host. Probiotics
± 2°C on a 12-h light cycle. After three days of acclimatization, they
bacteria are noninvasive, yet they need to interact with
had free access to water ad libitum
and conventional y, pel et food
(UAR, Vil emoisson-sur- Orge, France). The conventional y diet is
gastrointestinal (GI) epithelial cel s to elicit their
containing proteins, fat, carbohydrate, vitamin and minerals (Table
immunomodulatory effects. Probiotics have been shown
to induce various epithelial cel responses by competing
We propose here a new experimental model of malnutrition,
with pathogenic bacteria for host adhesion binding sites,
based on exclusive use of maize. This diet, similar to that
improving epithelial cel barrier function (Myl yluoma et
consumed by the severely malnourished children in poor countries,
is severely deficient in essential acid amine and vitamins (Ribeiro et
Probiotics and prebiotics can be used to either prevent
During fifteen days of experimental feeding period, animals were
or reduce the severity of microbe-induced gut
divided into three groups of 10 rats each. A protein malnourished
inflammation. The best known prebiotics are fructo-
rats (group 1, n = 10) were fed only with maize, a protein
oligosaccharides, sugars of plant origin. It has interesting
malnourished rats (group 2, n = 10) were received an antibiotic
properties such as stimulating the growth of probiotics
(Metronidazole) and a protein malnourished animals of group 3 (n =
and renewal of cel s in the intestinal mucosa.
10) received both Metronidazole and Probiotics Oligosaccharide.
The malnourished rats of the three groups were fed with maize (diet
PROBIONAT® is a product of both a probiotics and
poly-deficient in essential amino acids) with 10 g/kg of rat/day (EPE
Fructo-oligosaccharide (FOS); it is advisable for its regu-
Grope Avicole de l’Ouest, Mostaganem, Algeria); the composition of
Standard diet for rats (UAR, Vil emoisson-sur- Orge, France).
Co-products of the cereals transformation
Oil cakes and other nitrogenized products of vegetable origin Nitrogenized product
Caloric value = 2900 kcal /kg. Ration day laborer = 18 – 25 g.
Composition of maize (EPE Groupe Avicole de l’Ouest, Mostaganem, Algeria).
maize was given in (Table 2). Food intake was measured daily at
The MLNs, spleen and liver were removed and al organs were
17 h. Induction of malnutrition after 15 days with this diet was
weighed separately. The MLN complex was placed in a sterile
published earlier (Dock et al., 2003; Dock-Nascimento et al., 2007).
grinding tube and homogenized with 9 volume of brain– heart
Metronidazole (Hikma Pharmaceuticals, Jordan) was given via
infusion using sterile ground-glass stoppers (Heimo et al.,
orogastric feeding tube to the rats of groups 2 and 3 for 15 days
determine the bacterial concentrations of homogenates, each organ
with the dose of 1 g/kg of animal/day). A complex of lyophilized
was diluted in decimal steps up to 1.105 in a sterile solution. After
probiotics (Lactobacil us acidophilus
, Lactobacil us rhamnosus,
manual grinding, 1 ml of the homogenate was transferred into a
, Bifidoboctenium longum
tube containing 9 ml of physiologic serum; from this dilution 100 µl
, Steptococcus thermophilus
) and Fructo-Oligosaccharides
aliquots were plated on DRIGALSKI agar plates for enterobacteria
(PROBIONAT® Safetynat Limited Epps Building Bridge Road,
culture (Sanofi, Diagnostic Pasteur; France). Spleen and liver was
France) in the form of a capsule contained about 109 germs in
analyzed in the same way as the MLNs. Al agar plates for aerobic
lyophilisate of 390 mg. It was given as three doses per day via
culture were incubated at 37°C under aerobic conditions for 1 day
orogastric feeding tube (1 mg/g of body per day) to the rats of group
and then interpreted. The gram-negative enteric was identified
3 (10 mg/ 10 ml). On the day 15 of this treatment, the animals were
using the API 20 E system (Analytab Products, Plainview, New
sacrificed by cervical dislocation and a 0.5 ml blood sample was
col ected from the inferior vena cava for blood cultures.
Quantitative culture results were determined by the number of
Using sterile procedures, the chest and abdominal cavities were
Colony Forming Units per gram of tissue, calculated from the
reflected with sterile forceps and the exposed viscera were
dilutions of organ homogenate and positive tissue cultures. We did
swabbed with a sterile, cotton-topped applicator stick, which was
not study obligate anaerobic because these organisms are rare
then placed in a tube of brain-heart infusion. The tube was
members of the intestinal flora of rodents early in life (Raibaud P,
incubated aerobical y at 37°C for 24 h to test the bacterial
1988) and because they have a low tendency to translocate to extra
intestinal sites (Stefen EK et al., 1988; Stefen EK et al., 1983). The
Incidence and sites of bacterial translocation (BT) in each group after 15 days of treatment.
PM, protein-malnourished group; BT, bacterial translocation; MLN, mesenteric lymph nodes.
*P < 0.01 compared to group 1, †P < 0.05 compared to groups 1 and 2, ‡P < 0.05 compared to group 1 and ¶ P < 0.05 compared to
Mesenteric Lymph Nodes (MLNs), spleen, liver and cecal contents
Number of translocating bacteria in terms of FCU per
were removed and homogenized for quantitative cultures. The
gram of mesenteric lymph nodes in each group after 15 days
colony-forming units (CFU) of bacteria per gram of tissue were
Data were expressed as means ± standard error of the mean. The
Mean ± standard error of mean × 103 colony-forming units per
differences between the different groups were evaluated by chi
square analysis with the Yates correction. Continuous data are
expressed as mean 6 SEM and analyzed with analysis of variance
(ANOVA) and the Student unpaired t test. A P-value < 0.05 was
The results of quantitative MLNs cultures are shown in
Table 4. Although the number of translocating bacteria
per gram of MLNs was decreased by the administration
The incidence of bacterial translocation was 30% (3/10)
of probiotics oligosaccharide, there was a significant high
in protein-malnourished group 1 (PM), 60% (6/10) in rats
difference between groups 2 and 1 (P < 0.001) and between
of group 2 (where PM was associated with
groups 3 and 1 (P < 0.05). In addition, the difference bet-
metronidazole) and 25% (2.5/10) in group 3 (where
ween groups 2 and 3 was significantly high (P < 0.001).
protein malnutrition was associated with metronidazole
There was a significant difference between groups 1 and
and probiotics oligosaccharide) (Table 3). The incidence
2 (400 ± 95 UFC/g Vs 160 ± 43 UFC/g, P < 0.05) as
of BT in group 3 was not significantly different from that of
shown in Table 5. The total bacterial counts, Gram-
group 1 (P > 0.05). The same result was obtained when
negative and Gram-positive, of cecal flora were signi-
comparing group 2 to group 1 (P > 0.05). The difference
ficantly low in group 1 than that in group 3 (400 ± 95
in the incidence of BT between groups 2 and 3 was
UFC/g versus 36 ± 4 UFC/g, P < 0.001). 42% of the total
significant (P < 0.05), 60% (6/10) in group 2 and 25%
bacterial count was Gram-negative in group 1, 51% in
group 2 and 13% in group 3. Metronidazole caused a
Bacterial translocation was detected only in the MLNs
slight overgrowth of Gram-negative bacteria in group 2.
(3/10, 30%) of the rats of group 3 which were treated with
Otherwise, this decrease in the Gram-negative population
probiotics oligosaccharide. However, it was detected in
of cecal flora was significant (P < 0.01). No micro-
the MLNs (4/10, 40%), spleen (2/10, 20%) and liver
organisms were isolated from the blood samples.
(3/10, 30%) of group 2. The incidence of BT was
significantly high in the MLNs of group 2 than that of
group 1 (P < 0.05). Regarding the liver, the same result
was obtained in group 2 comparing to groups 1 and 3 (P
< 0.05). The incidence of BT in the MLNs was not
These results suggest that protein-malnutrition induced
significantly different between groups 2 and 3 (P > 0.05),
translocation of enterobacteria. Metronidazole acted
or between groups 1 and 3 (P > 0.05). These data
specifical y on the strict anaerobic bacteria by reducing
indicated that BT did not spread beyond the MLNs in
their numbers in cecal flora which explains the high rates
protein malnourished rats treated with probiotics
of overgrowth and translocation of enteric bacteria in the
Quantitative results of Gram-negative, Gram-positive and total bacteria in cecal cultures for each group after
3. PM + antibiotic + Probiotics-Oligosaccharide
Mean ± SEM X 102 colony-forming units per gram cecal content.
*P < 0.05 and **P < 0.01 compared to group 1.
malnourished group received metronidazole.
(1985) found that the cecal overgrowth of enteric
In this context, the current observation is that the PM
bacilli caused an enhanced BT after the admini-
and the PM particularly associated with metronidazole
stration of Metronidazole (500 Units/ml) in PM rats,
results in overgrowth cecal bacteria. This bacterial
but the mean of enteric cecal population was
overgrowth is most marked in the Gram negative. These
significantly decreased by the administration of high
levels of intestinal bacterial population help to explain the
dose of penicil in (1500 Units/ml). Al of these
col apse of the strictly anaerobic bacterial flora by the use
studies demonstrate that oral antibiotics given in a
of metronidazole and the consequent apparition of
dose dependent manner may inhibit the growth of
bacterial translocation in the GLM, spleen and liver.
anaerobic bacteria and allow intestinal overgrowth
In previous studies, we found that protein malnutrition
of facultative anaerobic Gram-negative bacil i.
and malnutrition associated with metronidazole pro-
Therefore, the administration of oral antibiotics may
mote bacterial overgrowth in cecal and translocation
enhance BT to the MLNs and other organs after PM,
of enterobacteria to MLNs (Benakriche et al., 2008).
which impairs both humoral and cel ular immunity in
Berg and Garglinton (1979) demonstrated that
addition to the damage of local barriers. In the pre-
penicil in, clindamycin, or metronidazole given orally
sent study, we found that oral antibiotic treatment
to specific pathogen-free mice for 4 days decreased
enhances BT and promotes the spread of trans-
the cecal population of indigenous bacteria, espe-
locating bacteria to the liver in PM rats. However,
cially anaerobic bacteria. A decreasing anaerobic
antibiotic treatment is usual y required in the treat-
population allows the cecal overgrowth of indi-
ment of the inflammatory victims. Bacterial overgrowth
genous, Gram-negative facultative anaerobic enteric
in the smal intestine has been documented in children
bacilli and enhances BT to the MLNs. Berg et al.
with severe malnutrition and might contribute directly to
(1988) demonstrated that a combination of anti-
ineffective solubilisation, digestion and absorption of lipid
biotics and immunosuppressive drugs promotes the
systemic spread of translocating bacteria, resulting
Despite knowledge in the area of overgrowth and
in lethal sepsis. General y, metronidazole is considered
intestinal bacterial translocation, probiotics and prebiotics
as a safe drug because of its relative high therapeutic
remain a hot topic. They are the subject of several
index and limited duration of treatment. Deitch et al.
studies which have shown in mice and rats that some
(1985) reported that the administration of antibiotics
probiotics like S. boulardi , B. longum, P. acnes
reduces the levels of indigenous GI tract flora and
, L. rhamnosus
and oligosaccharide have a
delays colonization by indigenous bacteria in the
protective effect against bacterial translocation. In
addition, it has been shown that fructo-oligosaccharide
However, Beaugerie and Petit (2004) postulated that
not only have a protective effect against bacterial trans-
giving oral antibiotic treatment to protein malnou-
location but also stimulates the proliferation of Bifido-
rished victims disrupts the ecological balance of
indigenous bacterial flora and causes the intestinal
It was reported that protein malnutrition decreases the
overgrowth or colonization of the intestines by exo-
number of lactobacil i and strict anaerobic bacteria of the
endogenous flora resulting in enterobacteria overgrowth
, or Staphylococcus
Based on this theoretical suggestion, the same
Recent studies have shown that probiotics, which
researchers investigated the effects of oral antibi-
have been used in the treatment of intestinal
otics on bacterial overgrowth and bacterial translo-
cation in protein malnourished rats. Deitch et al.
colitis, are effective for maintaining
intestinal equilibrium and reducing BT. Probiotics
promote an increasing anaerobic population of
translocation, but it enhanced the functional
gastrointestinal tract flora (Duffy, 2000; Mattar et al.,
adaptation of the remaining intestinal segments.
2001) but anaerobic bacterial overgrowth may not be
Another study by Berg et al. (1993) investigating the
important for BT because anaerobic bacteria rarely
effects of S. boulardi
on the translocation of C.
translocate to the MLNs. On the other hand, anae-
in antibiotic-decontaminated specific patho-
robic bacterial overgrowth have many beneficial
gen free mice revealed that it decreased the
effects such as: strengthening gut mucosal barrier
incidence of C. albicans
translocating to the MLNs,
function, balancing microbial ecology, adhering to
liver, kidneys and also the number of translocating
the intestinal mucosa, impeding invasive pathogens,
per gram MLNs, spleen, and kidneys.
metabolizing dietary proteins and enzymes by intes-
The present study, which was conducted to
tinal microflora and promoting resilience of the
evaluate the effects of probiotics oligosaccharide on
epithelium to gut mucosal permeability. Col ins and
BT in protein malnutrition associated with metro-
Gibson (1999) reported that smal -bowel colonization
nidazole treatment, showed that probiotics oligo-
by Escherichia coli
K1A and BT was decreased by
saccharide significantly decreased the population of
the administration of the probiotics oligosaccharide
Gram-negative bacteria in cecal flora. Although we
and suggested that probiotics oligosaccharide may be
could not examine the anaerobic flora, these data
used for the treatment of BT and sepsis.
correlate with that of other reports in which
It has been recently shown that impaired gut barrier
probiotics increase the anaerobic bacterial count
and mucosal immune function by malnutrition can be
and decrease the Gram-negative facultative anae-
reversed by L. casei
used as an oral adjuvant of
robic and aerobic bacteria count in the GI tract
renutrition diet. The clinical significance of these findings
(Mattar et al., 2001; Ishibashi and Yamazaki, 2001).
wil be important, as wel as improving mucosal immunity,
Anaerobic bacteria are generally decreased by
may also induce protection against enteropathogens
antibiotic treatment and this ecological imbalance
(Gauffin et al., 2004). Probiotics can protect the intestine
allows Gram negative bacteria to proliferate and
by competing with pathogens for attachment, strength-
enhances BT (Berg, 1981). The results of our study
ening tight junctions between enterocytes and enhancing
suggest that probiotics oligosaccharide may coun-
the mucosal immune response to pathogens (Lei and
teract this undesirable side effect of antibiotics.
Interestingly, the incidence of BT in the rats treated
Hirofumi et al. (1999) demonstrated that two kinds of
with probiotics oligosaccharide (group 3) was not
probiotics derived from different bacterial genera
significantly higher than that of the group 1, but was
enhanced epithelial cel proliferation of the gut without
significantly higher than that of the rats given the
altering the gross population levels of cecal microflora. In
antibiotic alone (group 2). Although there was no
humans in vivo
, lack of protective effect of probiotics (L.
significant difference in the incidence of BT and the
299V) on bacterial translocation to lymph
number of translocating bacteria per gram of MLN
nodes was confirmed by the same team and same
between groups 2 and 3, we found that probiotics
methodology for combining oligosaccharide with L.
oligosaccharide decreased the incidence of BT from
acidophilus, B. lactis Bb12, S. thermophilus
that which occurred in group 1 and also decreased
(Anderson et al., 2004).
the number of translocating bacteria. Furthermore,
the administration of probiotics oligosaccharide
salmonel a numbers in cecal contents and mucosa and
ensured that BT was limited to the MLNs, whereas it
caused a major increase in infection-induced diarrhea. In
extended to the liver in rats not given probiotics
addition, FOS enhanced translocation of salmonel a.
oligosaccharide. Selection of strains was based on their
Thus, in contrast to most expectations, FOS dose-
antibacterial and immunomodulatory properties. L.
dependently impairs the resistance to salmonel a
, L. rhamnosus
, B. lactis
, B. longum
infection in rats (Sandra et al.,
, S. thermophilus
L. fermentum KLD
has previously been used as both a
were selected for their ability to suppress the growth of
prophylactic and therapeutic agent in the treatment of
Gram- negative bacteria
and their antimicrobial effects.
gastrointestinal disturbances and is an interesting
The lower incidence of BT in the rats given oral
candidate probiotic strain (Marteau et al., 2001).
probiotics oligosaccharide may not only related to
Zaouche et al. (2000) investigated the effects of S.
improved intestinal ecological balance, but also to
on bacterial overgrowth and translocation in
the effects of probiotics oligosaccharide on host
rats with resected smal intestine and found that it
immune responses, including stimulation of the
production and secretion of intestinal s-IgA, enhan-
cing both phagocytic activity and the maturation of
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DWELLINGS Source: Housing Construction, Statistics Sweden Reconditioned dwellings are now reported under the heading converted dwellings. Conversion refers to more extensive measures where the disposition of dwellings is changed. Municipal facts 2009 will only be presenting the net change in the number of dwellings in renova- ted multi-dwelling buildings, as opposed to previous years when t
Bewertung der Sartane im Vergleich zu ACE-Hemmstoffen Prof. Dr. med. Wolfgang Motz Klinikum Karlsburg Herz- und Diabeteszentrum Mecklenburg-Vorpommern Greifswalder Str. 11 17495 Karlsburg In den letzten 20 Jahren konnte gezeigt werden, dass das Enzym Angiotensin II eine Schlüsselrol e bei der arteriel en Hypertonie und der Herzinsuffizienz spielt. Eine Hemmung der Konversion von Angiotensin I