CHAPTER IV

DISCUSSION

From the bacterial resistant to antibiotics and marine microorganisms for future natural
antibiotics, the researchers would like to isolate microorganisms from sea sand and sea water. On
this chapter, the statement of the problem in the study are answered and thereby explained how
the results are related in improving future natural antibiotics.

This study adds to the growing evidence that the E.coli is highest than staphylococcus
aureus is the lowest based to the measurement of the turbidity.

According to De Oliveira and Pinhata (2008), although there have been recent studies
aiming to describe the pattern of distribution of antibiotic-resistant bacteria in freshwater,
estuarine water, water distribution systems and sewage, there are few studies in the marine
environment and coastal zone. And in a study conducted by these authors in the city of So
Vicente, located in Santos Lowland (Baixada Santista), So Paulo, Enterococcus spp. were
isolated in 94.2% of the samples of sea water and sand. Of these, 51.9% were resistant to some
antimicrobials, predominantly erythromycin and streptomycin.

This same study shows that other studies have demonstrated that beach sand can act as a
reservoir and/or vector for a variety of diseases and that, although it is a fundamental part of
recreational activities in the coastal environment, it has been largely neglected from the point of
view of public health (De Oliveira and Pinhata, 2008).

The seas and oceans cover more than 70% of the Earth's surface and considered a
valuable source for microorganisms and bioactive secondary metabolites
(T.I. Orlova, V.G. Bulgakova, A.N. Polin, 2015)
 The main source of antibiotics in aquatic environments comes from the discharge of
wastewater, disposal of waste and aquaculture (Ding and He, 2010), and the increased levels of
antimicrobial resistance among bacterial strains isolated from polluted aquatic environments,
particularly in developing countries, which may be an indicative of overuse or misuse of
antimicrobial agents in these environments (Souza et al., 2000).

References
DING, C.; HE, J. Effect of antibiotics in the environment on microbial populations. Applied
microbiology and biotechnology, v. 87, n. 3, p. 925-41, jul. 2010.

DE OLIVEIRA, A. J. F. C.; PINHATA, J. M. W. Antimicrobial resistance and species

composition of Enterococcus spp. isolated from waters and sands of marine recreational beaches
in Southeastern Brazil. Water research, v. 42, n. 8-9, p. 2242-50, abr. 2008.

T.I. Orlova, V.G. Bulgakova, A.N. Polin [Secondary metabolites from marine
microorganisms. I. Secondary metabolites from marine actinomycetes]

SOUZA, W. G. S. et al. Resistance profile of Bacteroides fragilis isolated in Brazil. Do they

shelter the cfiA gene? The Journal of antimicrobial chemotherapy, v. 45, n. 4, p. 475-81, abr.
2000.
 APPENDIX A

TURBIDITY COMPARISON OF E. COLI AND STAPHYLOCOCCUS

E. coli Staphylococcus
Control 1 1.282 0.632
Ave Trial 1 1.038 0.768
Control2 0.652 0.666
Ave Trial 2 0.961 1.238
 APPENDIX B

TURBIDITY OF STAPHYLOCCOCUS AND E. COLI

 Turbidity - Staphy
1.755molL^(-1) 1.628molL^(-1) 1.510molL^(-1)
Control-1 0.815 0.657 0.425
Trial 1 1.0080 0.7666 0.5300
Control -2 1.48 1.24 0.993
Trial 2 0.9153 0.6594 0.4243

Turbidity - Ecoli
1.755molL^(-1) 1.628molL^(-1) 1.510molL^(-1)
Control1 1.462 1.288 1.095
Trial 1 1.2557 1.0503 0.8089
Control2 0.902 0.648 0.407
Trial 2 1.2201 0.9601 0.7019
 APPENDIXC

MORPHOLOGICAL CHARACTERISTICS OF SEA SAND

 APPENDIX D

MOPRPHOLOGICAL CHARACTERICS OF SEA WATER

 APPENDIX E

BIOCHEMICAL TEST RESULT OD SEA SAND AND SEA WATER

 APPENDIX F

SPECTROPHOTOMETER RESULTS OF E.COLI AND STAPHYLOCCOCUS

Note: Higher absorbance (A) = more light absorbed.
APPENDIX G

ISOLATION OF SEA SAND

 APPENDIX H

ISOLATION OF SEA WATER

 APPENDIX I

STATISTICAL RESULTS

A. ANOVA

Source of Variation SS df MS F P-value F crit

Between Groups 0.417374 2 0.208687 3.062801 0.096691 4.256495
Within Groups 0.613224 9 0.068136
Total 1.030598 11

B. ANOVA

Source of
Variation SS df MS F P-value F crit
Between Groups 0.426093 2 0.213046 2.693926 0.12109 4.256495
Within Groups 0.711755 9 0.079084
Total 1.137848 11

C. T-Test: Two-Sample Assuming Unequal Variances

E coli Staphy
Mean 1.072893 0.826131
Variance 0.200956 0.078604
Observations 4 4
Hypothesized
Mean Difference 0
df 5
t Stat 0.933406
P(T<=t) one-tail 0.196728
t Critical one-tail 2.015048
P(T<=t) two-tail 0.393457
t Critical two-tail 2.570582