C H A P T E R V
D I S C U S S I O N A N D C O N C L U S I O N
5 . 1 T h e m i c r o b i o l o g i c a l q u a l i t y ( i n t e r m o f E . c o l i ) o f
f r e s h b r o i l e r m e a t i n B o g o r
Recovery rate of E. coli
was 87.5% of total samples in this study, resulting in a
high number of isolates. In general, concentration of E. coli
significantly exceeded the
limit as set in the Indonesian National Standard (SNI: 01-6366-2000) (P<0.05). The
Indonesian National Standard (SNI: 01-6366-2000), accepted E. coli
as an indicator
of microbiological quality of poultry meat with a maximum permissible level of
presumptive numbers of E. coli
50 MPN/g in fresh or frozen meats. The recovery
rate of E. coli
from this study was much higher than recovery rate found in a study
conducted by Aryanti et al.
(2007), which reported a recovery rate of 75.68% from
poulty meat. Difference in choice of samples for bacteriological examination might
have resulted into the high recovery rate of E. coli
in our study. This can be further
affirmed by Mead (2007b) who suggested that skin maceration normally had high
recovery rate of microorganisms and was the “gold standard” for detection or
enumeration of organisms present in poultry carcass. Concerning antimicrobial
resistance of E. coli
, Persoons et al.
(2010) stated that skin sample can give a better
indicator for the resistance risk posed to consumers, but it is not a good predictor for
resistance level in primary production.
Many factors during the slaughtering, storage, transport, and processing of
meat can contribute to contamination. E. coli
can occupy multiple niches (Levy,
1997). E. coli
is routinely shed into the environment. The contamination of meats by
occurs most often during slaughter (Bhunia, 2008). These organisms can also
be transmitted between humans, animals, the environment, and other food along the
processing line (Meng and Schroeder, 2007). In this study, we found that most
of the broilers were slaughtered in small scale slaughterhouses. Since these
slaughterhouses used unhygienic practices, it is conceivable that there was a higher
possibility of cross contamination. Aryanti et al.
(2007) reported that the
contamination of food of animal origin in Indonesia by E. coli
unhygienic practices along the processing line. In order to reduce the amount of food
contamination, the improvement of hygienic practices was highly recommended.
5 . 2 A n t i m i c r o b i a l r e s i s t a n c e o f E . c o l i i s o l a t e d f r o m
s e c t o r 3 f r e s h b r o i l e r m e a t i n B o g o r
In total 402 isolates E. coli
were analyzed, and variety of combinations of
antimicrobial resistance pattern among isolates from the same sample has been found.
There are many possible reasons for the variation in antimicrobial resistance pattern
found in the E. coli
samples. It is possible that some isolates of E. coli
sources outside of farms. The transfer of resistant bacteria due to ecological factors
such as the presence of other farm animals, rodents, pets, or personnel/workers/sellers
is also a possible cause of the variation in antimicrobial resistance pattern (Smith et
The highest prevalence of resistance in E. coli
was found for the most
commonly used poultry antimicrobials, which are tetracycline, erythromycin,
ampicillin, nalidixid acid, enrofloxacin, and trimethoprim-sulfamethoxazole. A
similar result was also reported by Poernomo et al.
(1992), who found a high
prevalence of resistance to tetracycline, erythromycin, ampicilin, and trimethoprim-
sulfamethoxazole in E. coli
isolated from poultry farms in the area of West Java. High
accessibility to antimicrobial drugs without the need for prescription, which is
common in West Java, may lead to an increase amount of antimicrobial resistance
(Murdiati and Bahri, 1991). The cheap cost of tetracycline ascompared to other
permitted antimicrobial feed additives (Appendix 4) directs the choice of tetracycline
as feed additives in poultry in Indonesia. This high selection pressure can be the
reason behind the high prevalence of resistance to tetracycline in E. coli
Bahri, 1991). Commercially available feed containing multiple antimicrobials can
also contribute to the increasing amount of resistance (Furtula et al.
, 2010). Brady and
Katz (1992) stated that about 80% of commercial poultry animals have been fed by at
least one antimicrobial substance during their life time. Routine use of antimicrobials
in food animals for growth promotion constitutes a serious public health concern,
especially in the case where the same classes of antimicrobials are being used in
humans (Wegener, 2003). Many studies have mentioned that the use of antimicrobial
feed additives in farms is major factor driving the emergence of antimicrobial
resistance (Cohen, 1992; Silbergeld et al.
, 2009). Based on economic analysis point of
view Silbergeld et al.
(2009) had concluded that, there is only little economic benefit
from using antimicrobials as feed additives, and that equivalent improvements in
growth and feed consumption can be achieved by improved hygiene.
In comparison to another study, the frequency of E. coli
chloramphenicol was much lower in our study. Poernomo et al.
(1992) reported that
26.6% of E. coli
isolated from colibacillosis (colisepticaemia) cases in poultry from
West Java were resistant to chloramphenicol. Resistant against chloramphenicol and
cephalothin was not expected as both antimicrobials are prohibited for use in poultry.
The illegal use of these antimicrobials in poultry may be one of the factors causing the
persistence of resistance in poultry isolates of E. coli.
It might also be possible due to
contamination of the meats by E. coli
from other sources having selection pressure of
chloramphenicol and cephalothin. Smith et al.
(2007) on contrary reported that
antimicrobial resistance pattern in E. coli
isolated from broiler under experimental
condition were not correlated with pattern of antimicrobial usage. Moreover, they also
stated that, majority of multi-resistant phenotypes are obtained by the acquisition of
mobile genetic materials, and it may provide resistance to an entire class of
antimicrobials, such phenomena called as co-selection. These events have been
detected frequently in resistant E. coli
isolated from consumer meat products in
Norway by Sunde & Norstrom (2006). The Australian Joint Expert Advisory
Committee on Antibiotic Resistance (JETACAR) mentioned in their reports in 1999
that, a single mutation in some bacteria can cause resistance to multiple antibiotics
such as tetracycline, chloramphenicol, trimethoprim, and some penicillin groups. The
resistance appears to be due to a complex set of changes that alters the permeability of
the cells, in particular, pumping these antibiotics out of the cell (efflux system).
A similar study in Vietnam has been reported by Van et al.
(2008). The results
demonstrated that high single and multiple resistances to antimicrobials in E. coli
isolated from fresh chicken meats were found. Comparing to our study, their results
were much higher with regard to resistance of antimicrobials in E. coli
tetracycline (84.2%), ampicillin (84.2%), gentamycine (47.4%), enrofloxacin
(63.2%), nalidixic acid (64.4%), and chloramphenicol (57.9%). Higher multiple
resistances of antimicrobial in E. coli
were also found which amount to 89.5% of
isolates resistant to at least three different antimicrobials.
Other studies of antimicrobial resistances in E. coli
isolated from broiler
poultry in Asian region have been reported. In Malaysia, Apun et al.
reported a high degree of resistance of antimicrobials in E. coli
isolated from broiler
chicken. E. coli
isolates were found much more resistance against chloramphenicol
(46.43%), tetracycline (95.86%), cephalotin (14.29%), nalidixic acid (64.29%), and
sulphamethoxazole-trimetrhoprim (82.14%). High multiple resistances in E. coli
isolates were also mentioned, 85.72% resistance was found against more than three
antimicrobial agents in broiler chicken. In India, Sharada et al.
(2010) reported high
resistant of E. coli
isolates against chloramphenicol (16.92%), tetracycline (83.08%),
erythromycin (94.19%), and gentamycin (40.00%). However, the prevalence of
antimicrobial resistance differs widely among countries. These differences might be
explained by differences in the degree of antimicrobial use, geographical differences,
and variations in poultry production systems (Bywater et al.
In the last decade, the frequency and spectrum of antimicrobial resistant
bacteria infections has increased in both hospital and community (Byarugaba, 2004).
Some reports mentioned that antimicrobial resistance among commensal E. coli
isolated from community and hospitalized patients has emerged in Indonesia (Lestari
, 2008; Refdanita et al.
, 2004). A high prevalence of resistance for
chloramphenicol, trimethoprim-sulfamethoxazole, ampicillin, and tetracycline was
observed among E. coli
isolated from hospitalized patients. Lestari et al.
mentioned that the E. coli
resistance rate in healthy persons discharged from the
hospital were significantly higher than patients being admitted in the hospital.
Silbergeld et al.
(2009) mentioned that because of large size of the animal reservoir of
resistance which includes consumer meats in poultry, the antimicrobials used in
agricultural may generated a greater range of resistance, and it may result in a larger
reservoir of non-hospitalized populations carrying antimicrobial resistance in term of
pathogen and non pathogen bacteria, as well as transposable resistance genes.
Concerning antimicrobial resistance, commensal E. coli
has capacity to be a
major threat for human consumers significantly. Commensal E. coli
may serve as a
reservoir of potential antimicrobial resistance genes in the environment, where the
resistance may be transferred to other commensal or pathogenic bacteria (Angulo et
, 2004). Food contaminated with resistant commensal E. coli
, which possibly carry
resistance genes, may transfer the genes to other pathogenic bacteria of human
clinical significance (Bester & Essack, 2010). Bacteria of animal origin possibly
spread to food products during slaughter and further processing, which has been
extensively documented for conventional foodborne pathogens, such as Salmonella
, and E. coli
(Wegener, 2003; Bester & Essack, 2010). However, the
treatment of pathogenic organism on farms may increase the likelihood of selective
pressure in commensal E. coli
(Bester & Essack, 2010).
Intensification and commercialization of poultry production can be associated
with the widespread use of antimicrobial substances in poultry (Soeripto, 1996) which
can lead to high prevalence of resistance in gut commensal in poultry. The high
number of multi drug resistant E. coli
isolates raises questions about the control of
antimicrobials use, management practices in the slaughter house and farm, and
ultimately the safety of food. The indication of multiple antimicrobial substances used
in the farms must not be economically efficient. Practically, the minimal biosecurity
measures would be compensated by using antimicrobial substances to control
bacterial infection. Thus, prudent use of antimicrobials should no way result in
antimicrobials used on farms with poor management practices.
The intervention strategies used for controlling antimicrobials use on farms
with low to minimal biosecurity level should be reconsidered. It is required to set up a
monitoring and surveillance program, and improve farming practices in order to
reduce the transmission of antimicrobial resistance genes and thereby minimize the
likelihood of horizontal gene transfers of these antimicrobial resistance genes to other
microbes in the food chain. Training of consumers and food handlers on safe food
handling and proper cooking are therefore important to reduce or eliminate the risk of
antimicrobial resistance and pathogenic bacteria originating from raw foods.
Additionally, it is recommended that antimicrobial usage regulations in animal feed
must be strongly enforced. Further studies are also required to get more information
as commensal E. coli
can serve as a reservoir of resistance genes, determining
genotypic patterns in E. coli
may help us to illustrate the potential risks posed to
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