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DOI 10.1007/s10661-009-1180-6
Abstract Antimicrobial resistance of marine het- The positive correlations obtained between the
erotrophic bacteria to different antimicrobials degree of seawater contamination and frequency
agents were evaluated in seawater, dry and wet and variability of bacterial resistance indicate that
sands from three marine recreational beaches with polluted marine recreational waters and sands are
different pollution levels. In all studied beaches, sources of resistant bacteria contributing thus, to
the greatest frequencies of resistance were found the dissemination of bacterial resistance.
in relation to penicillin. On Gonzaguinha, the
most polluted beach, 72.3% of all isolated strains Keywords Marine heterotrophic bacteria ·
showed simple resistance, whilst 8.33% had mul- Resistance · Seawater · Sand beach ·
tiple resistance. The values found on Ilha Porchat Contamination
beach, were 70.8% and 6.9% for simple and mul-
tiple resistances, respectively. On Guaraú, the less
polluted beach, only 35.3% of isolated strains
had simple resistance. Multiple resistance was not Introduction
observed. While samples from Gonzaguinha and
Ilha Porchat beach showed isolated strains re- Coastal areas are complex formations including
sistant to seven and six different antimicrobial bays, estuaries, beaches and large semi-closed
agents, respectively, samples from Guaraú beach areas, where there is a high concentration of popu-
were resistant only to penicillin and erytromicin. lation development. The relation between popula-
tion increases in coastal regions and changes in the
environment has been known since many years.
However, attention has been given only recently
to environmental impacts on marine ecosystem
and on population health caused by the develop-
ment of these areas.
A. J. Fernandes Cardoso de Oliveira (B) ·
P. T. Ranzani de França · A. B. Pinto Besides the marine microbiota, seawater and
Laboratory of Marine Microbiology, sediments can contain a significant non-
Campus do Litoral Paulista, indigenous microbiota composed by bacteria,
Universidade Estadual Paulista,
virus and protozoan (some of them are patho-
Praça Infante Dom Henrique, s/n, 11330–900,
São Vicente, SP, Brazil gens), that are discharged to the environment
e-mail: ajuliaf@clp.unesp.br in domestic sewage and urban drainage water
Environ Monit Assess
(Austin 1988; Sato et al. 2005; Oliveira et al. Materials and methods
2007; Oliveira and Pinhata 2008). The presence
of such microorganisms in recreational seawater, Study areas
sediments and beach sands affect the quality of
these habitats, leading to high risk of beachgoer’s São Vicente (23◦ 57 S and 46◦ 23 W) and Peruíbe
health due to waterborne and other illness, as (24◦ 19 S and 46◦ 59 W) are located on the Baixada
well as to the high resistance that can be showed Santista (“Santos Lowland”), south coast of São
by these microorganisms to several antimicrobial Paulo State (Fig. 1). The Baixada Santista is so
agents such as antibiotics (Erdem-Kimiran et al. named due to its geography, since it borders the
2007; Goni-Urriza et al. 2000; Irvensen et al. 2004; Serra do Mar (“Sea Highlands”). It occupies an
Meirelles-Pereira et al. 2002; Oliveira and Pinhata area of 2,373 km2 , with 161 km2 of beaches. Due to
2008; Schwartz et al. 2003; Tejedor et al. 2001). the heterogeneity of its beaches and the proximity
The bacterial resistance to antimicrobial agents to urban centres, such as São Paulo city, one of
can be intrinsic, associated to the incapacity of the most important economical activities in São
these agents to reach the sites of action in the Vicente and Peruíbe is tourism.
cell. In addition, it can be extrinsic, related to Peruíbe is located within four conservation
dynamic processes such as changes in antibiotics units: Juréia Itatins (considered by UNESCO as
transport within the cell, molecular modifications Natural Site on World Heritage), Tupiniquins,
or enzymatic production that modify and inac- Cananéia-Iguape-Peruíbe and Serra do Mar. It
tivate the antibiotic (Hermansson et al. 1987; contains 18 beaches, from which six have their
Davies 1994). This resistance can be determined water quality monitored by São Paulo State Com-
by both genes from chromosomes as well as by pany for Environmental Sanitation Technology
extra-chromosome genetic elements (R-plasmid; (CETESB). Since it has a great part of its area
Gomes-Lus 1998; Hirsch et al. 1999). under legal protection and also due to its longer
Besides their resistance to several antimicro- distance to large cities (e.g. São Paulo), Peruíbe is
bials, bacteria can also, through conjugation with still well preserved. Guaraú beach is considered to
bacteria from the same or different species, have the best sanitary quality among all evaluated
acquire resistance factors present in plasmid beaches and its water quality is considered to be
(Huycke et al. 1998; Kühn et al. 2005). There- excellent over 90% of the time, which is attributed
fore, faecal bacteria in domestic sewage with a to its geographical configuration, distance from
high level of antimicrobial resistance can dissem- city downtown and low rates of urban occupation.
inate this resistance to indigenous bacteria. The Domestic sewage is collected and directed to a
frequency and variability of resistant strains of sewage treatment plain, where it is preconditioned
bacteria to different antimicrobial has increased and submitted to the reduction of organic load
in the last time, becoming a great challenge for in stabilisation lagoons. After organic load reduc-
the treatment of infectious diseases, which should tion, effluents are discharged into Preto River,
be carefully considered (Arvanitidou et al. 2001; which runs directly to the sea, in the southern part
Halling-Sorensen et al. 1998). Therefore, this of Peruíbe (CETESB 2006).
study aims to evaluate the resistance of het- Five beaches belong to São Vicente:
erotrophic bacteria to antimicrobial agents, which Itararé, Milionários, Ilha Porchat, Divisa and
were isolated from seawater and sand samples in Gonzaguinha beach. The last ones are the most
three recreational beaches under different levels affected by organic contamination, whilst Ilha
of pollution. In addition, environment potentials Porchat shows the lowest levels of pollution
as sources of dissemination of bacterial resistance (CETESB 2008).
were evaluated based on the relationship between The worse bathing water quality at
quality of marine recreational waters, bacterial Gonzaguinha beach is associated with its
habitat, frequency and type of resistance. geographical configuration and the high inflow of
Environ Monit Assess
BRAZIL
SÃO
PAULO
Fig. 1 Detail of studied areas on Gonzaguinha and Ilha Porchat beaches, located in São Vicente, Baixada Santista (São
Paulo State, Brazil)
organic load brought by the effluents discharged of Santos city (CETESB 2006). After passing by
at Mar Pequeno, São Vicente Channel and a bar screen to remove solid residuals and after
the adjacent hydrographical system, which ulti- chlorination, it is released 4 km away from the
mately reach this beach (CETESB 2006). Since coast, in the central part of Santos Bay. During
Gonzaguinha Beach is located in a sheltered summer, the treatment of domestic sewage is not
area, dispersal of the microorganisms is limited, sufficient, since population in these areas can dra-
allowing bacteria and other microorganisms to matically increase, reaching twice the number of
be accumulated in areas used by beachgoers. On local residents (CETESB 2006). Therefore, there
the other hand, Ilha Porchat Beach is located in is an increase of domestic sewage load, affecting
a less sheltered area, where circulation processes the quality of marine recreational water in the
and the constant renewal of the water facilitate region (Sato et al. 2005; CETESB 2006).
dispersal of the pollutants. These processes Besides the increase of domestic sewage dis-
favour the dispersal of microorganism seaward charge in seawaters, the urban water drainage
and provide better bathing water quality. reaching beaches, litter brought by beachgoers
In São Vicente, there is a sewage treatment and tides as well as the presence of animals on
plant in operation, which is responsible for treat- the beach can also be pointed out as noteworthy
ing the total collected sewage (56.6%). Part of the problems in the region that are intensified during
collected sewage is sent to the submarine outfall the holiday season.
Environ Monit Assess
Table 1 Percentage of isolated strains resistant to one ceptibilities, on seawater, wet and dry sand samples from
or two antimicrobials (simple resistance), and to at least Gonzaguinha, Ilha Porchat (São Vicente) and Guaraú
three antimicrobials (multiple resistance) and their sus- (Peruíbe) beaches, Brazil
Strains Gonzaguinha (n = 144) Ilha Porchat (n = 72) Guaraú (n = 102)
Number % Number % Number %
One or two antimicrobials 104 72.3 51 70.8 36 35.3
At least three antimicrobials 12 8.3 5 6.9 0 0
Total of resistant 116 80.6 56 77.7 36 35.3
Susceptibilities 28 19.4 16 22.3 66 64.2
Environ Monit Assess
Table 3 Percentage of resistant heterotrophic bacteria iso- Table 4 Percentage of resistant heterotrophic bacteria
lated from seawater and sand samples from Ilha Porchat isolated from seawater and sand samples from Guaraú
beach, in relation to antimicrobials (amoxicillin, ampicillin, beach, in relation to antimicrobials (amoxicillin, ampicillin,
chloramphenicol, ciprofloxacin, erythromicin, gentamicin, chloramphenicol, ciprofloxacin, erythromicin, gentamicin,
penicillin, rifampicin, streptomycin and vancomycin) penicillin, rifampicin, streptomycin and vancomycin)
Antimicrobial agent % of resistant strains (n = 56) Antimicrobial agent % of resistant strains (n = 36)
Ilha Porchat beach Guaraú beach
Water Wet sand Dry sand Water Wet sand Dry sand
(18) (22) (16) (19) (11) (6)
Amoxicillin 20.8 20.8 0 Amoxicillin 0 0 0
Ampicillin 0 8.3 0 Ampicillin 0 0 0
Chloramphenicol 0 0 0 Chloramphenicol 0 0 0
Ciprofloxacin 0 25.0 12.5 Ciprofloxacin 0 0 0
Erytromicin 16.7 0 0 Erytromicin 21.0 18.2 33.3
Gentamicin 0 0 0 Gentamicin 0 0 0
Penicillin 70.8 86.4 58.3 Penicillin 79.0 81.8 66.7
Rifampicin 0 0 0 Rifampicin 0 0 0
Streptomycin 0 0 0 Streptomycin 0 0 0
Vancomycin 20.8 0 0 Vancomycin 0 0 0
Environ Monit Assess
From both the quantitative and qualitative do Estado de São Paulo: Balneabilidade das praias
point of view, results obtained in this study 2005. São Paulo, Brasil: Companhia de Tecnologia de
Saneamento Ambiental.
demonstrated that marine recreational waters and
CETESB, Companhia de Tecnologia e Saneamento Am-
sands may be sources of bacterial resistance. It biental (2008). Reltório de qualidade das águas
may thus, contribute to further dissemination of litorâneas do Estado de São Paulo: Balneabilidade das
resistance to antimicrobials, possibly in high lev- praias 2007. São Paulo, Brasil: Companhia de
Tecnologia de Saneamento Ambiental.
els, which ultimately may render inefficiently the
Chandrasekarn, S., Venkatesh, B., & Laithakumari, D.
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