Download Screening resistant gram positif bacteria from meat products

Screening bacteria from meat products and
its antagonist effects to antibiotic
Introduction
Food can be important vehicle in the transmission of diseases. This has been
documented for along time specially in the developing countries. Where hygienic
standards are not strictly followed and enforced. These disease comonly are caused by
microbial infection. Further more the wide aplplication of antimicrobial has led to large
scale dissemination of bacteria resistant to antimicrobial in the environment (Harakeh S.
et. al. 2005).
Some studies have evidence the spreading of resistant bacteria such as
staphylococci, enterococci, and lactic acid bacteria during meat processing (Simeoni et.
al .2007). Staphylococcal food poisoning is considered to one of the leading causes of all
food-borne disease (Normanno G. et. al. 2006).The aims of this study was to screen
bacteria in meat products that might have spectrum in antagonist effect to antibiotic.
Materials and Methods
Samples
Two meat products were obtained from different manufactures. They included
sausage and chicken nugget. The sample were stored in the refrigerator.
Isolation of bacteria
A 1-g portion of each meats sample was aseptically squasshed. As follows the
squashed sample were transferred to a tubes filled with 9 ml steril destilate water. Serial
dillution were carried and than dillution were inoculated onto Nutrient Agar (NA) and
incubated for 24 hours in incubator. Plates that contains with bacteria were purrified. The
purified isolates were examined with gram staining.
Detection of antagonistic activity
Antagonistic activity were assayed with antibiotics discs.on Luria Bertani agar.
Four antibiotics disc were used : CC 2 (Clindamycin, 2 g), F/M 300 (Nitrofurantoin,
300 g), GM 10 (Gentamicin, 10 g), VA 30 (Vancomycin, 30) g. Four discs spread in
one plate contain single strain of bacteria in a clear distance. The plate were incubated for
24 hours in incubator.
Result and Discussion
Of the 2 samples were analysed all of them contaminated with bacteria. The
bacteria namely K1 and K2 from nugget and K3 and K4 from sausage. Almost all of the
isolates were gram positive except K4 (Table 1). Cell shape varrys from cocci to rod
shape. Common bacteria that have been succeded in screening are Staphylococci
(Simeoni D. et. al. 2007), Staphylococcus aureus (Normanno et. al 2006), Bacteriocinproducting lactic acid bacteria (Bromberg R. et. al. 2004), Salmonella sp. and E.coli
(Harakeh S. et. al. 2005).
Antibiotics assay showed antimicrobial resistance in K3 and K4 to antibiotic disc
CC 2 and VA 30 (Table 2). The highest inhibition zone were occurred in CC 2 in
treatement with K1, and the second one was F/M 300 in treatement with K2 (Figure 1).
Colonu forming unit were more abundant in nugget sample rather than in sausage sample
(Table 3).
Table 1. Gram staining result and cell shape
Bacteria
Cell shape
Gram
K1
Cocci
Positive
K2
Cocci
Positive
K3
Rod
Posiitve
K4
Rod
Negative
Source
Nugget
Nugget
Sausage
Sausage
Table 2. Inhibition zone of antibiotics
Bacteria
K1
K2
K3
K4
CC 2
2,33 cm
2,1 cm
-
Antibiotics discs
GM 10
F/M 300
1,45 cm
1,7 cm
1,85 cm
2,15 cm
1,7 cm
1,9 cm
1,75 cm
2,1 cm
VA 30
1,3 cm
1,55 cm
-
Table 3.Colony forming unit (CFU) for each sample
Sample
colony
Nugget
133
Sausage
9
dillution
10-1
10-3
Volume
inoculated
CFU
0,5
2,66 x 105 cfu
0,1
0,9 x 103 cfu
a
b
VA 30
GM 10
F/M 300
CC 2
F/M 300
VA 30
GM 10
CC 2
d
c
CC 2
VA 30
GM 10
F/M 300
CC 2
VA 30
GM 10
F/M 300
Figure 1. Antimicrobial activity of CC 2, F/M 300, GM 10, and VA 30
againts K1 (a), K2 (b), K3 (c), and K4 (d).
Antibiotic discs were used CC 2, F/M 300, GM 10, and VA 30. CC also called
with clyndamycin, this antibiotic ussually used to treat infections with anaerobic bacteria
but can also be used to treat some protozoal disease such as malaria. It is a common
topical treatment for acne, and can be useful againts some methicillin resistant
Staphylococcus aureus (MRSA) infection (Wikipedia 2008a). Clyndamycin most
effective againts infections involving the following types of organisms: Aerobic grampositive cocci, including some members of the Staphylococcus and Streptococcus (e.g.
pneumococcus) genera; Anaerobic, gram-negative rod-shaped, including some species of
Bacteroides and Fusobacterium. As the result showed all the bacteria were inhibited
gram-positive and that resistant were gram negative (rod shape) K4 and gram positive
(cocci) K3.Simillar results for gram negative bacteria E. coli and were isolated from meat
based fast food showed resistant result in antimicrobial test with clyndamycin (Harakeh
S. et. al. 2005). GM or so called gentamicin is an aminoglycoside, an antibiotics that
works by binding the 30S, ribosome unit of the bacterial. It is interrupting protein
synthesis. Gentamicin is of the few heat-stable antibiotics that remain active even after
autoclaving., which makes it particulary usefull in the preparation of certain
microbiological growth media (Wikipedia 2008b). Gentamicin reported susceptible to
E.coli that were isolated from meat based food from Libanon by Harakeh et. al. (2005).
This antibiotics susceptible in treatment normally gram-negative bacteria including
Pseudomons, Proteus, and Serratia and gram positive Staphylococcus.
F/M also know as nitrofurantoin, this antibotics works by damaging bacterial
DNA, since its reduce from is highly reactive. Nitrofurantoin and the quinolone
antibiotics are mutually antagonistic in vitro. It is not known whether this is of clinical
significance, but the combinaton should be avoided (wikipedia 2008c). Nitrofurantoin is
only clinically proven for use againts E. coli, Staphylococcus saprophyticus, It may also
have in vitro activity againts: Enterococus faecalis, Staphylococcus aureus,
Streptococcus agalactiae, Citrobacter sp., Klebsiella sp., and coagulase-negative
staphylococci. As the result showed that there were no resistant occurred. Thus can be
suggested one of the following bacteria were mentioned susceptible with nitrofurantoin
one of the one observed in this experiment.
VA or vancomycin speciffically, vancomycin prevents incorporation of Nacetylmuramic acid (NAM)-matrix; peptidoglycan and N-acetylglucosamine (NAG)peptide subunits into the which forms the major structural component of gram-positive
cell walls (Wikipedia 2008d). The results in the experiment showed that gram positive,
cocci, K3 tends to resistant to this antibiotics. The same with Staphylococcus epidermidis
(gram positive) collected from the production chain of swine meat comodities showed
resistance with a low level vancomycin (6 g ml-1) reported by Simeoni D. et. al. (2007).
Other reistant bacteria is K4, gram negative rod shaped bacteria. This results the same as
reported by Harakeh S. et. al. (2005) that most of E. coli (gram negative) collected from
meat-based fast food tends to resistant with vancomycin.
Conclussion
Finally, the data obtained that meat products that is nugget and sausage are
potential of carrying bacteria posses resistance ability to antibiotics.
Reference
Bromberg R. et. al. 2004. Isolation of Bacteriocin-producing lactic acid bacteria from
meat and meat products andits spectrum of inhibtory activity. Brazillian J. of
Microbiol 35:137-144.
Harakeh S. et. al. 2005. Isolation, molecular characterization and antimicrobial resistance
patterns of Salmonella and Eschericia coli isolates from meat-based fast food in
Lebanon. Science of the total environment 341: 33-44.
Normanno G. et. al. 2006. Occurrence, characterizations and antimicrobial resistance od
enterotoxigenic Staphylococcus aureus isolated from meat and airy products. Int J
of food microbiol 115: 290-296.
Simeoni D. et. al. 2007. Antibiotic resistance genes and identification of Staphylococci
collected from the production chain of swine meat comodities.Food Microbiology
(25): 196-201.
[wikipedia]. 2008a. Clindamycin. http//:en.wikipedia.org/wiki/Clyndamycin. [13 jan
2008]
[wikipedia]. 2008b. Gentamicin. http//:en.wikipedia.org/wiki/Gentamicin. [13 jan 2008]
[wikipedia]. 2008c. Nitrofurantoin. http//:en.wikipedia.org/wiki/Nitrofurantoin. [13 jan
2008]
[wikipedia]. 2008d. Vancomycin. http://en.wikipedia.org/wiki/Vancomycin. [13 jan
2008]