SEARCH FOR ANTIBIOSIS BACTERIA IN CUTTING BOARD OF SEA FISH ON

MDR-Salmonella sp. AND Shigella dysentriae

Aprimawita, Periadnadi Periadnadi *, Nurmiati Nurmiati
Biology Depertment, Faculty of Mathematics and Natural Science,
Andalas University, Padang

ABSTRAC
Strong pathogenic bacteria and irregular handling will give rise to the ability of bacterial defenses against
antibiotics which are often referred to as resistance. Resistance to antibiotics is increased due to the
inappropriate use of antibiotics in humans for medicinal purposes, however there are factors that originate from
animal origin foods such as fish having the largest contribution. This study uses survey and experimental
methods and the data are analyzed descriptively. This study aims to find antibiosis by screening and in vitro
testing of bacteria found in cutting fish cutting board which has the potential to inhibit the growth of MDR
Salmonella sp. and Shigella dysentriae. There were 8 isolates of cutting fish cutting bacteria which had the
potential to fight test bacteria and potentially antibiosis. The highest potential for antibiosis was obtained from
sp. Isolates. 3 (18.47 mm), followed by isolate sp. 8 (16.54 mm), isolates sp. 6 (15.86 mm), isolates sp. 7 (15.51
mm), isolate sp. 5 (15.50 mm), isolate sp. 8 (14.95 mm), isolate sp. 9 (14.56 mm) and isolates sp. 5 (14.22 mm).
Therefore it is stated that the microbial free area formed can be categorized as Strong (K).

Key Words : Antibiosis, MDR Salmonella sp, Shigella dysentriae, Resistance.

BACKGROUND
Antibiotics are one of the secondary metabolites produced by microorganisms, both fungi or
bacteria that are used for various purposes in the health sector. Most antibiotics are currently
produced from bacteria because they are easily isolated, cultured, and can be stored for a long
time without losing viability. Antibiotics are one of the important pillars of modern medicine,
but early generation antibiotics have lost their effectiveness and have always been replaced
with new ones for handling various species of pathogenic bacteria. Strong pathogenic
bacteria and irregular handling will give rise to the ability of bacterial defenses against
antibiotics which are often referred to as resistance (Kourkouta et al., 2017).
Resistance to antibiotics is increased due to the pattern of antibiotic use which is not
appropriate for humans for medicinal purposes, however, there are factors that originate from
food of animal origin such as fish having the greatest contribution. The decreasing
development of new antibiotics and the absence of a global control system for antibiotic use,
estimated that by 2050, microbial resistance will be the highest killer in the world with an
estimated mortality rate of 10 million people/year, where one bacterium that increases
resistance to antibiotics is Salmonella sp and S. dysentriae have been reported in several
countries. Increased bacterial resistance from several types of antibiotics triggers the
emergence of Multi Drug Resistence (MDR). This is indicated by the resistance of
Salmonella sp. on macrolide antibiotics, tetracyclin, quinolonserta, combined amoxicillin and
clavulanic acid (Yenie, Aulia. and Handayani, 2017). While Shigella dysentriae is reported to
be resistant to ampicillin, tetracyclin, streptomycin, and chloramphenicol (Dewi, Joharman
and Lia, 2013).
Various studies have shown that microorganisms isolated from waste or dirty places
can produce antimicrobial substances. Research by Kaur et al. (2014) has successfully
isolated soil bacteria that are capable of producing antimicrobial substances and have been
tested against E. coli, Pseudomonas aeruginosa, Staphylococcus aureus,
Mycobacteriumsmegmatis, Proteus vulgaris and Bacillus subtilis. In waste or dirty places
such as cutting fish cutting board which is a source of pathogenic bacteria, it is estimated that
it is also a source for the type of antibiosis bacteria which if isolated and tested on pathogenic
bacteria will appear clear (halo) in the culture medium, which indicates that antibiotic
bacteria are capable suppress the growth of pathogenic bacteria and can be used as a source
of antibiotics.
Based on the background above, it can be seen that there are very few reports about
the types of antibiotics that can be produced by bacteria from waste and dirty places.
Therefore, this research needs to be done for screening and in vitro testing of bacteria found
in fish cutting cutting board which has the potential to inhibit growth. MDR-Salmonella sp.
and Shigella dysentriae.

RESEARCH IMPLEMENTATION
Methods
This research was conducted using survey methods for the location of sampling and sampling
using purposive sampling technique. Furthermore, the samples were carried out at the
Microbiology Laboratory, Biology Department, Faculty of Mathematics and Natural
Sciences, Andalas University, Padang using experimental methods and the data obtained were
analyzed descriptively.
Tools and Materials
The tools used consist of autoclave, centrifugator, petri dish, erlenmeyer, testube, vortex, osse
needle, test tube rack, measuring cup, object glass, glass cover, bunsen, funnel, drip pipette,
hot plate, magnetic stirrer, incubator, colony counters, micro pipettes, plastic wrap, label
paper, tissue, cotton, cutton bud, gauze, digital cameras, gloves, masks, permanent markers,
rulers and stationery. The material used in the form of waste samples in the form of crust on
the sea fish cutting board, Salmonella sp. and Shigella dysentriae test bacteria, Mc solution.
Farland's, Chloramphenicol antibiotics, Nutrient Agar (NA) medium, 70% alcohol, spirits and
aquadest.

RESULT AND DISCUSSION

Antibiotic Test of Fish Cut Chopping Board Bacteria
Antibiotic testing of cut fish cutting boards was performed to see antibacterial activity. The
antibacterial activity of cut fish cutting board was carried out using the diffusion method
using paper discs on Nutrient Agar (NA) medium and incubated at 37oC for 24 hours.
Measuring the diameter of each bacterial-free region can be seen in Table 1.
Table 1. Diameter of the Bacteria Free Area Cutting Board of Sea Fish
Bacteria sp. Diameter halo zone (mm)
sp. 5 14.22
Salmonella sp. sp. 8 14.95
sp. 9 14.56
sp. 3 18.47
sp. 5 15.50
Shigella dysentriae sp. 6 15.86
sp. 7 15.51
sp. 8 16.54

Based on the results of bacterial isolation above, the pathogenic bacteria from the cutting
board of fish can inhibit bacterial growth and serve as an antibiotic against infection with
Salmonella sp. and S. dysentriae because around paper discs form each halo zone. According
to Davis and Stout (1971), the microbial free area diameter ≥ 20 mm is categorized as Very
Strong (SK), if the diameter 10-20 mm is categorized as Strong (K), then a diameter of 5-10
mm is categorized as Medium (S) and diameter ≤ 5 mm categorized as Weak (L). Based on
this, it can be stated that microbial-free areas formed from isolation of pathogenic bacteria on
cutting boards of fish cutters as a source of new antibiotics against infection with Salmonella
sp. and S. dysentriae can be categorized as Strong (K)
Samples found in liquid waste or dirty places containing antibiotic-producing
microbes are generally used to produce appropriate antibiotics. These antibiotics can be
bacteriostatic in nature. Microorganisms that have more capacity to produce antibiotics
generally can survive longer than those producing fewer amounts of antibiotics (Kaur et al,
2014). Various studies have shown that microorganisms isolated from waste or dirty places
can produce antimicrobial substances. Research by Kaur et al., 2014 has succeeded in
isolating soil bacteria that are capable of producing antimicrobial substances and have been
tested against E. coli bacteria, Pseudomonas aeruginosa, Staphylococcus aureus,
Mycobacterium smegmatis, Proteus vulgaris and Bacillus subtilis.
Based on the results obtained from the study it was found that the isolation and
potential test in vitro on cutting fish cutting board contained various types of bacteria that
have the potential to inhibit bacterial growth so that it has the potential to be used as a source
of antibiotics against Salmonella sp. and S. Dysentriae. Bacteria that have been obtained have
the potential as bacteria that inhibit bacterial growth because at the time of the potential test
the formation of the halo zone. This can be a new knowledge that can be contained in
scientific articles that explain the presence of bacteria on cutting board fish also has the
potential to be used as a source of antibiotics.
CONCLUSION
Based on the research that has been done, it can be concluded that :
1. 8 isolates of cut fish cutting bacteria were found that could potentially fight test
bacteria and potentially antibiosis.
2. The highest potential for antibiosis is obtained from sp. Isolates. 3 (18.47 mm),
followed by isolate sp. 8 (16.54 mm), isolates sp. 6 (15.86 mm), isolates sp. 7 (15.51
mm), isolate sp. 5 (15.50 mm), isolate sp. 8 (14.95 mm), isolate sp. 9 (14.56 mm) and
isolates sp. 5 (14.22 mm).

REFERENCES
Davis & Stout. (1971). Disc Plate Method Of Microbiological Antibiotic Essay. Journal Of
Microbiology. Vol 22 No 4.
Dewi IK, Joharman, LiaYB. 2013. Perbandingan Daya Hambat Ekstrak Etanol Dengan
Sedian Sirup Herbal Buah Belimbing Wuluh (Averrhoa bilimbi L.) Terhadap
Pertumbuhan Shigella dysenteriae In Vitro. Jurnal Berkala Kedokteran. 9 (2): 191-
198
Kaur. 2014. Isolation and Characterization of Antibiotic ProducingMicroorganisms from Soil
Samples of Certain Area of Punjab Region of India. International Journal of
Pharmaceutical and Clinical Research.6(4): 312-315.
KourkoutaL, Kotsiftopoulos CH, Papageorgiou M, Iliadis CH and Monios A. 2017. The
Rational Use of Antibiotiks Medicine. Journal of Healthcare Communications. 2(3):
27.