Environ Sci Pollut Res (2015) 22:17475–17485
DOI 10.1007/s11356-015-4986-5
RESEARCH ARTICLE
Organochlorine pesticides and parasites in Mugil incilis collected
in Cartagena Bay, Colombia
Beatriz E. Jaramillo-Colorado 1 & Bárbara Arroyo-Salgado 2 & Luis Carlos Ruiz-Garcés 1,3
Received: 2 April 2014 / Accepted: 30 June 2015 / Published online: 11 July 2015
# Springer-Verlag Berlin Heidelberg 2015
Abstract Nematode parasites of the Anisakides family are
often found in people living in countries where fish is con-
sumed raw or partially cooked. This research shows the his-
tological changes in the liver and spleen of Mugil incilis, col-
lected in Cartagena Bay. These changes are associated with
pollution by organochlorine pesticides and their possible in-
fluence on the parasite. Organochlorine compounds were ex-
tracted using the headspace-solid-phase microextraction (HS-
SPME) technique. Residual amounts in the muscle of
M. incilis such as β-HCH, γ-HCH, heptachlor, aldrin, endo-
sulfan, 4,4′-DDE, and dieldrin, among others, were identified
by gas chromatography connected to an electron capture de-
tector, indicating that the fauna of Cartagena Bay are exposed
to these pollutants. Histological analysis was carried out on
liver and spleen samples of M. incilis which were fixed, proc-
essed, and embedded in paraffin. The presence of melano-
macrophages, granulomes, and trematodes in the liver was
the most important changes observed. Larval prevalence for
the Anisakis spp. was determined to be 1.6 %; for
Pseudoterranova spp., 25.3 %, and for Contracaecum spp.,
57.8 %. Other parasites such as acanthocephalans were also
reported for a total of 15.3 %. Nevertheless, no significant
Responsible editor: Philippe Garrigues
* Beatriz E. Jaramillo-Colorado
bjaramilloc@unicartagena.edu.co; beatrizjaramilloc@yahoo.com
1
Agrochemical Research Group, Chemistry Program, Department of
Exact and Natural Sciences, University of Cartagena, San Pablo
Campus, Cartagena, Colombia
2
Group GIMUC, School of Medicine, University of Cartagena,
Zaragocilla Campus, Cartagena, Colombia
3
Aquaculture Program, Department of Veterinary Medicine,
University of Córdoba, Montería, Colombia
correlation between parasites and organochlorines was found.
This study is the first to correlate the presence of organochlo-
rine compounds and histological damage in the liver and
spleen of M. incilis, with the presence of parasites in fish from
Cartagena Bay (Colombia).
Keywords Mugil incilis . Zoonosis . Parasites . Anisakis spp .
Pseudoterranova spp . Contracaecum spp . Organochlorine .
HS-SPME . GC-ECD
Introduction
Fish have been used as indicators of environmental quality
because changes in their nervous, endocrine, and immune
systems may indicate an alteration in parameters such as pH,
salinity, temperature, solutes, dissolved oxygen, substrate
composition, or levels of pollution in the ecosystem
(Kuklina et al. 2013; Rašković et al. 2013). Marine pollution
predisposes the appearance of infectious and noninfectious
diseases in fish. These often generate functional biochemical
and physiological alterations in fish blood and tissue, resulting
in pathological changes (Costa et al. 2013; Mhadhbi and
Beiras 2012; Mikryakov et al. 2011).
Toxic and pathological effects of contaminants are present in
aquatic organisms (Matthiessen and Law 2002). These contam-
inants, which bioaccumulate and bioconcentrate, are environ-
mental pollutants, which, over time, may reach levels that cause
physiological impairment in humans (Karouna-Renier et al.
2011; Polsky et al. 2007). The most recognized are mercury
(Sedláčková et al. 2014), polychlorinated dibenzo-p-dioxins,
polychlorinated dibenzofurans (PCDD/F), and polychlorinated
biphenyls (PCBs) (Jiang et al. 2007; Letcher et al. 2010; Smith
and Gangolli 2002), dichlorodiphenyltrichloroethane (DDT),
17476
and dichlorodiphenyldichloroethylene (DDE) (Deshpande et al.
2013; Schiavone et al. 2009).
Actually, coastal waters used for recreation near urban
areas, where untreated wastewater discharges with high con-
tent of pathogens and other pollutants, represent major health
and environmental problems. The highest levels of these con-
taminants have been detected in species of fish in coastal
waters adjacent to industrial parks (Albaigés et al. 1987;
Kannan et al. 1994; Cirillo et al. 2009). The discharges from
industrial, municipal, medical, and agricultural effluents are
often poorly treated or not treated at all (Wolke 1992).
The widespread use of organochlorine pesticides (OCPs)
has created significant environmental concern. The hazardous
nature of OCPs is a result of their toxicity in combination with
high chemical and biological stability and a high degree of
lipophilicity. The two latter characteristics make OCPs prone
to bioaccumulation along the food chain, involving a wide
range of trophic levels. As a consequence, although the use
of most OCPs has been restricted or even banned in many
countries, they continue to be widespread in the environment
especially in biological specimens (Fidalgo-Used et al. 2003).
Though many persistent OCPs are restricted or banned in
developed countries, they are still produced and used to a large
extent in the developing world (Miranda et al. 2008; Sagerup
et al. 2009; Sapozhnikova et al. 2004). However, the informa-
tion about them is scare in Colombia.
Recently, several investigations have demonstrated signif-
icant effects and interactions between parasites, as well as their
contact with contaminants. These are environmental stressors
affecting the immune systems of their intermediate hosts, in-
cluding invertebrates, fish, birds, and mammals (Luong and
Hudson 2011). According toTimi et al. (2011), in several eco-
systems, the fish trophic level is a risk factor for similarity of
parasite assemblages, allowing the identification of the trophic
leve l based on pa rasite data alone. In ad ditio n,
biomagnification is present as a risk factor in the fish
community.
In Colombia, studies conducted by Olivero-Verbel et al.
(2005), Olivero-Verbel et al. (2006a), Olivero-Verbel and
Caballero-Gallardo (2013) show the presence of pollutants
and parasites, in particular Contracaecum spp., in fish species,
especially the Mugilidae family, such as Mugil incilis, which
are localized in the 2.01 trophic level (Olivero-Verbel and
Caballero-Gallardo 2013). M. incilis is a detritivorous fish,
which is the most abundant fish found in the estuaries of
Colombia’s northern coast throughout the year (IUCN
International Union for Conservation of Nature 2014;
Olivero-Verbel and Caballero-Gallardo 2013). It may
bioaccumulate persistent organic pollutants at higher concen-
trations (Aksoy et al. 2012; Karouna-Renier et al. 2011).
Cartagena Bay is located on the Caribbean coast of
Colombia. Although Cartagena Bay is a typical large pouch
bay by geomorphologic definition, today it behaves as an
Environ Sci Pollut Res (2015) 22:17475–17485
estuarine system with a mix of ocean and freshwaters, the
latter coming from the Canal del Dique. It is contaminated
through several sources of pollution: sewage, industrial dis-
charges, sedimentation, ship waste, leaching from garbage,
and solid waste disposal areas. The major impacts are the
dumping of city sewage, the sediment from the Canal del
Dique (part of Magdalena River), the dumping of industrial
waste and oil spills from ships and docks, as well as the in-
dustrial zone (Mamonal), all of which discharge their waste
into the bay. The industries include pharmaceutical com-
plexes, petrochemical factories, a petroleum refinery, food
plants, and electronic plants among others (Restrepo and
Kjerfve 2004; GEF 1998).
Previous studies have reported concentrations of organo-
chlorine (Aldrin, DDT, heptachlor, and dieldrin compounds)
in shrimp (Panameus vannamei) at concentrations between 0,
1–13 ng/g (Jaramillo and Marrugo 2010) in the coastal zone of
Cartagena Bay. DDT, DDD, and DDE were also detected in
fish caught in the Ciénaga de la Virgen (Castro 1998) as well
as other internal waterways in the city, which presented high
concentrations of dissolved oxygen, fecal coliforms, organic
matter, and heavy metals, affecting the water quality and ex-
ceeding the permitted limits acceptable for the preservation of
flora and fauna in marine waters (Tirado et al. 2011).
Approximately 90 % of industrial and domestic waste gener-
ated along of the entire coastline is received in Cartagena Bay,
exceeding the allowable limits for coliforms in recreational
waters in the months of September, October, and November
(PNUMA 2009).
The objective of this research was to determine the histo-
logical changes in the liver and spleen of M. incilis collected in
Cartagena Bay, associated with pollution from organochlorine
pesticides and their possible influence on the parasite.
Materials and methods
Sampling area
This study was carried out in Cartagena Bay, on the northern
coast of Colombia (75° 16′ W and 10° 44 N). Fish samples and
water were collected between June and November 2009, in
three different sites denominated station 1 (E1), station 2 (E2),
and station 3 (E3) as shown in Fig. 1. A representative sample
of 20 (21) M. incilis were taken monthly from each site. Once
collected, fish were placed in polyethylene bags and
transported on ice to the Agrochemical Research Laboratory
(University of Cartagena).
Chemicals
Acetone, methanol, and hexane were purchased from Merck
(Darmstadt, Germany) and Sodium sulfate anhydrous from
Environ Sci Pollut Res (2015) 22:17475–17485
Fig. 1 Map of Cartagena Bay,
showing sampling sites (stations
E1, E2, and E3)
J.T. Baker, Inc (Phillipsburg, NJ, USA). The standard pesti-
cides were all purchased from Chem Service, Inc. (Tower
L a n e We s t C h e s t e r, PA , U S A ) . T h e so l i d - p h a s e
microextraction (SPME) fibers and holder were purchased
from Supelco (Bellefonte, PA, USA). Vials of glass (20 mL)
were bought from Schott (Hofheim, Germany).
Bacteriological characterization of the water in Cartagena
Bay
The levels of pollution in water samples at the collection sta-
tions (E1, E2, and E3) were determined using multiple tube
most probable number in 100 mL (MPN/100 mL), according
to the McCrady table, following the recommendations of the
Food and Drug Administration (Jackson GJ et al. 2001). The
water samples were properly preserved in previously sterilized
bottles and were then taken to the Microbiology Laboratory at
the University of Cartagena at a temperature of about 4 °C in a
styrofoam cooler, for subsequent bacteriological analysis.
Morphometric analysis of M. incilis and identification
of parasites
The total length and weight of the fish were recorded.
Morphometric analysis such as the condition factor (CF)
(Bervoets and Blust 2003), the liver somatic index, and the
spleen somatic index was calculated (Garcia‐Abiado and
Mbahinzireki 2004; Hadidi et al. 2008). Morphological iden-
tification of parasites was carried out using the comparative
features reported by Olivero-Verbel et al. (2005) and the keys
published by Martins et al. (2005). Parameters such as preva-
lence, intensity, and abundance were also determined (Bush
et al. 1997).
17477
Histopathogical analysis
This analysis was carried out according to Takei and Powell
(2007). Liver paraffin microsections (5 μm) were initially
evaluated for histopathological changes. The system was eval-
uated to quantify the extent of the lesions in the liver for the
total sample in each month of study. Recorded lesions includ-
ed nonspecific inflammatory changes (infiltration of inflam-
matory cells and granulomatosis), necrosis, apoptosis, and the
presence of melano-macrophage centers (MMCs) (Martins
et al. 2005; Takei and Powell 2007).
Detection of organochlorine pesticides in the smooth
muscle (M. incilis)
The method of extraction and concentration of organochlorine
compounds in fish through the technique of solid-phase
microextration (SPME) was used, taking into account the rec-
ommendations of Fidalgo-Used et al. (2003), in which the fish
dorsal muscle (10-g wet weight) was ground into a fine powder
with activated sodium sulfate and then was placed in a Soxhlet
extractor, adding 300 mL of hexane/acetone (1:1) for 9 h. The
extract was later cleaned using a Florisil column before GC–
ECD analysis (Fidalgo-Used et al. 2003). The data were
expressed in accordance with maximum level residue-MLR
(0.002–0.1 ng/g) according to FAO/WHO (2013). Because of
the physical and chemical characteristics of organochlorides,
pesticide analysis in water was not undertaken in this study.
SPME equipment and experimental conditions
The fiber selected for this study was a fused silica fiber of
polydimethylsiloxane/divinylbenzene (PDMS/DVB) of
17478
65 μm, held in a SPME device supplied by Supelco (Palo
Alto, USA). Prior to use, the fibers were conditioned by
heating them in the injection port of the gas chromatograph
for 30 min at 250 °C.
The SPME extraction was carried out as follows: 1-mL
aliquots of the fish tissue organic extract from the Soxhlet
were placed into a 10-mL Teflon-lined screw-capped vial
and evaporated to the point of drying using a gentle stream
of nitrogen. The residue was redisolved in 10 mL of methanol/
water (5 % v/v). The vial was placed in a thermostatic water
bath, and a fiber of polydimethylsiloxane/divinylbenzene
(PDMS/DVB) 65 μm was immersed in the solution, remain-
ing there for 90 min, at ambient temperature (25±1 °C).
Previously, the exposure time of the fiber was determined.
During extraction, the solution was vigorously stirred. After
extraction, the fiber was thermally desorbed at 260 °C for
5 min into the chromatograph injection port. Procedure blanks
were run periodically during the analysis to check possible
fiber contamination (Fidalgo-Used et al. 2003).
Gas chromatograph
Sample extracts were analyzed using an Agilent Technologies
4890 D gas chromatograph, equipped with an electron capture
detector (ECD) and a HP-5 fused silica capillary column
(30 m×0.32 mm d.i, 0.25 μm df), with a mobile phase of
helium. The column temperature was programmed at 80 °C,
maintained for 2 min, then increased to 160 °C at 10 °C/min,
ramped to 240 °C at 2 °C/min; and finally 20 °C/min until
280 °C. The injector and detector temperatures were main-
tained at 250 and 310 °C, respectively.
The chromatograms obtained from the samples were com-
pared with the mixture pattern of organochlorine pesticides.
The standard solutions used were as follows: PP-HC5JM
Fig. 2 Measures of water quality
for bacteria, collected at the
sampling stations (E1, E2, and
E3) in Cartagena Bay
Environ Sci Pollut Res (2015) 22:17475–17485
(Chem Service, Inc., Tower Lane West Chester, PA, USA),
consisting of a mixture of 16 organochlorine pesticides in
hexane/toluene (1:1) 2000-μg/mL concentration. The 16 pes-
ticides are listed below: Aldrin, HCH (α-isomer), BCH (β-
isomer), BCH (δ-isomer), 4,4′-DDD, 4,4′-DDE, 4,4′-DDT,
dieldrin, α-endosulfan, β-endosulfan, endosulfan sulfate, en-
drin, endrin aldehyde, heptanoclor, heptanoclor epoxide, lin-
dane. Quality parameters such as linearity, precision, accura-
cy, and limit of detection (LOD) were previously established.
Statistical analysis
All data are presented as mean standard error. For comparison
of means, the morphometric data and prevalence of parasites
were evaluated for normality and homogeneity of variance,
using Kolmogorov-Smirnov and Bartlett, respectively. In case
of failure to provide normal data, they were transformed to
their logarithms. Comparisons between variables for the sta-
tions and sampling campaigns were performed using two-way
ANOVA using the Tukey test as posttest.
The quantitative relationships between variables were de-
termined through correlation and regression analysis. Factor
and cluster analysis were applied to multivariate relationships
of the total data. In all cases, the level of significance is
P<0.05.
Results
Identification of total and fecal coliforms
Bacteria belonging to the group of coliforms were taken as an
indicator of water quality of Cartagena Bay, according to the
World Health Organization (WHO 2004). The presence of
Environ Sci Pollut Res (2015) 22:17475–17485 17479

Table 1 Fish morphometric parameters of the M. incilis of Cartagena Bay

Month of sampling Station n Weight (g) Length (cm) Condiction factor Hepatosomatic index Bazosomatic index

June E1 21 77,5±17,9 21,9±1,7 0,35±0,06 0,011±0,00 0,002±0,00

June E2 21 181±45,8 29,1±2,6 O,61±0,10 0,070±0,00 0,007±0,00
June E3 21 94,1±37,8 22,7±3,1 0,40±0,10 0,013±0,00 0,001±0,00
July E1 21 100,6±10,6 23,1±0,9 0,43±0,03 0,020±0,00 0,001±0,00
July E2 21 97,4±20,5 22,7±1,6 0,42±0,06 0,015±0,00 0,0009±0,00
July E3 21 107,4±18,8 23,7±1,4 0,44±0,06 0,015±0,00 0,0009±0,00
August E1 21 86,4±18,2 20,4±2,9 0,42±0,07 0,013±0,00 0,0019±0,00
August E2 21 102,5±37,8 22,1±3,4 0,43±0,1 0,018±0,00 0,002±0,00
August E3 21 85,1±20,0 22,1±2,0 0,38±0,07 0,010±0,00 0,001±0,00
September E1 21 41,4±11,8 17,0±1,6 0,23±0,05 0,010±0,00 0,001±0,00
September E2 21 39,3±8,9 16,2±1,3 0,22±0,06 0,010±0,00 0,001±0,00
September E3 21 62,1±29,7 19,5±3,0 0,30±0,09 0,012±0,00 0,001±0,00
October E1 21 109,6±13,5 23,7±0,8 0,46±0,06 0,010±0,00 0,001±0,00
Octuber E2 21 40,8±7,7 16,7±1,2 0,21±0,04 0,010±0,00 0,002±0,00
Octuber E3 21 87,1±21,2 22,2±1,7 0,39±0,07 0,010±0,00 0,001±0,00
November E1 21 95,5±21,5 21,8±0,6 0,41±0,06 0,010±0,00 0,001±0,00
November E2 21 55,6±14,8 18,2±0.5 0,23±0,03 0,010±0,00 0,001±0,00
November E3 21 100,7±12,2 22,5±1,3 0,42±0,04 0,010±0,00 0,001±0,00

total coliforms throughout the months of sampling at all sta- a high correlation between fish weight and length (0.903) as
tions was reported, as opposed to fecal coliforms, which were well as condition factor (CF) and weight (0.916) were ob-
not reported as a presence in the month of June at stations 1 served. However, CF has correlation with an average length
(E1) and 2 (E2), in July at stations 1 (E1), 2 (E2), and 3 (E3), (0.707). Whereas station (E2) and station (E3) had a higher
and similarly, in the months of August and September in sta- correlation between weight and size of the fish (0.975, 0952,
tion 2 (E2), assuming their presence with a value of ≤3 MPN/
100 mL microorganisms, according to the McCrady table. Table 2 Quantitative parasitic infection indexes: parasitic infection
The highest values of total coliforms were reported in M. Incilis of the Bay of Cartagena: rate parasitic (RP); prevalence (P);
rate of infestation (I) and parasite intensity (PI)
October at station 3 (E3) and November at station 1 (E1) with
a value of 2100 MPN/100 mL for fecal coliform organisms Month of sampling Station n RP P (%) I PI
with the highest value presented in the months of October with
750 MPN/100 mL microorganisms and November at station 2 June E1 21 627 100 Severe 15
(E2) with a 930 MPN/100 mL (Fig. 2). June E2 21 182 80,9 Moderate 10,7
June E3 21 262 100 Severe 12,5
July E1 21 474 100 Severe 22,6
Morphometric analysis of the fish July E2 21 293 95,2 Severe 14,6
July E3 21 350 100 Severe 16,6
Data from the morphometric parameters of M. incilis during August E1 21 371 100 Severe 19,2
the months of sampling (June–November) at different stations August E2 21 301 100 Severe 15,1
(E1, E2, and E3) are presented in Table 1. The highest and August E3 21 279 90 Severe 13,4
lowest condition factors are observed in station 2 (E2) with an September E1 21 90 100 Light 4,5
average value of 0.61 ± 0.10 in June and 0.21 ± 0.04 in September E2 21 74 85,7 Light 4,1
October; likewise, the longest fish (21.9±1.7) and the heaviest September E3 21 225 100 Severe 10,8
(181±45.8) were reported at station 2 (E2) during the month October E1 21 294 100 Severe 14,2
of June, as well as the lowest values in length (16.2±13) and Octuber E2 21 115 100 Moderate 5,6
weight (39.3±8.9). Octuber E3 21 352 95,2 Severe 16,8
For the evaluation of the correlation and health of fish November E1 21 180 93,2 Severe 8,6
values and relationships, the condition factor, the November E2 21 101 80,5 Moderate 4,8
hepatosomatic index, spleen somatic index, weight, and para- November E3 21 160 82 Moderate 7,6
site abundance in sampling times were analyzed. At station 1,
17480
respectively), while the correlation between weight, length of
the fish, and the parasitic intensity was low (0.952 to 0.237).
Identification of parasites in the M. incilis
Quantitative parasitic infection indexes obtained for all sam-
pled specimens in three sampling stations are shown in
Table 2, as well as three genera of parasites from the family
Anisakidae: Contracaecum spp., Pseudoterranova spp.,
Anisakis spp. The prevalence of parasites was classified for
Anisakis spp. (1.6 %), Pseudoterranova spp. (25.3 %), and
Contracaecum spp. (57.8 %). Other parasites such as acantho-
cephalan (15.3 %) were found. In general, the higher density
Fig. 3 I Contracaecum spp. L3 isolated from Mugil incilis in Cartagena
Bay. A End (CT cuticular tooth, DL dorsal lip, EP excretory pore) ×40. B
End (NR nervous ring, I intestinal cecum, E esophagus, V ventricle) ×10.
C Intermediate part (E esophagus, V ventricle, AV atrial appendage, IN
intestine, I intestinal cecum) ×40. D Rear end (A anus, m mucro, N nodes)
×10. II Pseudoterranova spp. Larval isolated from Mugil incilis in the
Bay of Cartagena of Indias. A End (CT cuticular tooth, PE excretory pore,
DL dorsal lip) ×40. B End (NR nervous ring, I intestinal caecum, E
Environ Sci Pollut Res (2015) 22:17475–17485
of parasites was found in the abdominal cavity (57.9 %), ver-
tebrae (31.3 %) with a few cysts found in the liver and spleen
with the presence of trematodes (10.8 %). The parasites found
in this study are shown in Fig. 3.
Organochlorine pesticide concentrations in M. incilis
muscle
The concentration of organochlorine compounds in the mus-
cle of M. incilis is presented in Table 3. The analysis of the
correlation between morphometric parameters, the parasitic
intensity, and the presence of organochlorine pesticides are
presented in Table 4, accepting the high correlation between
esophagus) ×10. C Intermediate part (E esophagus, V ventricle, I
intestinal cecum) ×10. D Rear end (m mucro) ×40. III Anisakis spp.
Larval isolated from Mugil incilis in the Bay of Cartagena of Indias. A
End (CT cuticular tooth, EP excretory pore, DL dorsal lip) ×40. B End
(NR nervous ring, E esophagus) ×10. C Intermediate part (E esophagus,
In intestine) ×10. D Rear end (m mucro) ×10. IVAB. Trematode found in
the body cavity of M. incilis (A anterior, B back). IV CD.
Acanthocephalans (C anterior, P proboscis, D back) ×10
Environ Sci Pollut Res (2015) 22:17475–17485 17481

Table 3 Concentration of organochlorine compounds in the muscle of the concentration of pesticides and the weight of the fish
M. incilis in different sampling stations (E1, E2, and E3)
(0.824) and the low correlation between concentration of pes-
No station OCP Concentration (ng/g)a LOD MRLb ticide and parasite intensity (0.172).

E1 β-HCH 0,00638±0,0012 0,0005 0,1

Histopathological changes in the liver and spleen
Aldrin 0,00115±0,0008 0,0005 0,2
of M. incilis
4,4’-DDD 0,00452±0,001 0,0002 -*
E2 β-HCH 0,00423±0,001 0,0005 0,1
The histopathological findings of this study showed severe
γ-HCH (Lindane) 0,00851±0,002 0,0005 0,02
damage in the liver and spleen of M. incilis, including high
Heptachlor 0,00725±0,001 0,0005 0,2
MMCs as shown in Fig. 4a, b. Granulomas are shown in
Aldrin 0,00333±0,001 0,0005 0,2
Fig. 4c. A presence of 85 % of MMCs in the liver compared
Endosulfan 0,00415±0,001 0,0005 0,05
with 15 % in the spleen as well as granuloma in the spleen
4,4’-DDE 0,00401±0,001 0,0005 -* (95 %) was found. Ascocotyle spp. was also found in the
Dieldrin 0,00206±0,000 0,0005 0,2 spleen of M. incilis in Cartagena Bay, as is shown Fig. 4d.
E3 β-HCH 0,00185±0,000 0,0005 0,1
Heptachlor 0,00436±0,001 0,0005 0,2
Aldrin 0,00227±0,001 0,0005 0,2
Discussion
4,4’-DDD 0,00404±0,001 0,0002 -*
a
Averages of four independent extractions ± ts/√ n (n = 4, 95 % Persistent organic pollutants, heavy metals, and parasites are
confidence) interrelated (Blanar et al. 2009; Sagerup et al. 2009). In
b
EU pesticide Data Base: FAO/WHO Food Standars (2013) Codex Colombia, few studies have been published on this subject.
Alimentarius However, previous research has reported significant changes
*
MLR for DDT + metabolites=1,0 ng/g in fish health parameters, including elevated tissue levels of
pollutants in M. incilis (Olivero-Verbel et al. 2006b, 2009).

Table 4 Correlation of morphometric parameters, parasitic intensity, and concentration of organochlorine pesticides in the M. incilis collected from
Cartagena Bay

Pesticide W (g) L (cm) Condiction factor (CF) Hepatosomatic index (HI) Bazosomatic index (BI) Parasite intensity

Pesticide
P 1 0.824** 0.383 0.769** 0.027 −0.654** 0.172
P=0.000 P= 0.159 P= 0.001 P= 0.923 P= 0.008 P= 0.540
W (g)
P 0.824** 1 0.540* 0.969** −0.100 −.581* 0.220
P= 0.000 P= 0.038 P= 0.000 P= 0.722 P= 0.023 P= 0.431
L (cm)
P 0.383 0.540* 1 0.451 0.052 -0.063 0.683**
P= 0.159 P= (0.038) P= 0.092 P= 0.854 P= 0.824 P= 0.005
CF(g)
P 0.769** 0.969** 0.451 1 −.023 −0.587* 0.226
P= 0.001 P= 0.000 P= 0.092 P= 0.934 P= 0.021 P= 0.418
HI (g)
P 0.027 -0.100 0.052 -0.023 1 -0.144 0.281
P= 0.923 P= 0.722 P= 0.854 P= 0.934 P= 0.609 P= 0.310
BI (g)
P -0.654** -0.581* -0.063 -0.587* -0.144 1 0.148
P= 0.008 P= 0.023 P= 0.824 P= 0.021 P= 0.609 P= 0.598
Parasite intensity
P 0.172 0.220 0.683** 0.226 0.281 0.148 1
P= 0.540 P= 0.431 P= 0.005 P= 0.418 P= 0.310 P= 0.598
17482
Fig. 4 Histology of the liver and
spleen of M. incilis. a, b
Melanomacrophage (M) in liver
sample; c granulomes, (G) spleen
samples and d Ascocotyle spp. All
samples taken from Cartagena
Bay
The effects of pollution on parasitism may be either positive or
negative for the host. The parasites may reduce pollutant
levels in their hosts below a critical value, and thus, are capa-
ble of reducing negative effects (Sures 2008; Sures et al.
1999). Conversely, when the pollutant affects the host rather
than the parasite, as for example through immunotoxic
chemicals, the result could be a suppressed immune system
and increased parasite load (Höss et al. 2011; Marcogliese and
Cone 1997; Sures 2008).
The main purpose of the research reported here is to sum-
marize and analyze the existence of associations among para-
sites, pollutants, and hosts, with the objective of studying bac-
teriological pollution. We first investigated the community of
the microorganism present in the Bay. Decree 1594 of 1984,
Article 42 of the Colombian Legislation on the use of water
and liquid waste states that the limits for fecal coliform should
not exceed the normal level for total coliform microorganisms
(PNUMA 2009). The results for the present study were in-
creased to compare with the legal level. The coliform group
is closely connected with the increase in human population
living in coastal areas, and similarly with the increase in solid
and liquid domestic, agricultural and industrial waste from
mismanagement and inadequate control, affecting the marine
environment with significant ecological, socioeconomic, and
health implications (Kornacki and Johnson 2001). Variations
throughout the study were probably caused by those sug-
gested by Gray (2008), which have summarized the interpre-
tation of the presence of E. coli due to strong and/or recent
human or animal waste.
On the other hand, the existence of complex interactions
among organisms, pollutants, and hosts linked to trophic
levels has been recognized. In particular, Cartagena Bay is a
Environ Sci Pollut Res (2015) 22:17475–17485
tropical estuary with the presence of contaminants (Jaramillo
and Marrugo 2010) and especially M. incilis, in greatest abun-
dance, with a distribution throughout the year that plays an
important role in this ecosystem. M. incilis reported low levels
of organochlorine compounds in muscle tissues (β-HCH,
Aldrin, 4,4′-DDD, γ-HCH, Heptachlor, Endosulfan, 4,4′-
DDE, Dieldrin, Endosulfan 1, and aldehyde Endrin). They
do not exceed the maximum residue limit (MRL) proposed
by the United Nations Food and Agriculture Organization
(FAO/WHO 2013) for food products.
A relationship between the parasite and the concentration
of organochlorine compounds was not found. However, the
interactions between the host-parasite environments are not
easy to interpret as they depend upon a variety of factors
(Sures et al. 1999). The evidence of the parasites present in
M. incilis found in previous studies (Blanar et al. 2009;
Olivero-Verbel et al. 2005, 2006a; Olivero-Verbel and
Caballero-Gallardo 2013; Roche et al. 2000) suggests a close
relationship between the contamination of ecosystems and the
emergence of the parasite. Additionally, organochlorine com-
pounds can suppress the immune system of fish, degrade the
mucus, affect the endocrine system and reproductive organs,
and induce constant stress making them susceptible to the
onset of disease. One such parasite is that of the Anisakidae
family (Miranda et al. 2008). Furthermore, the correlation be-
tween organochlorine contamination and parasites is low,
possibly due to what was suggested by Lafferty (1997) and
Sagerup et al. (2009), that pollution can cause a positive or
negative effect in parasitosis.
The large number of fish parasites and the degree of infes-
tation indicates a marked environmental deterioration of the
water in Cartagena Bay, probably caused by other types of
Environ Sci Pollut Res (2015) 22:17475–17485
pollutants (Olivero-Verbel and Caballero-Gallardo 2013).
This is consistent with those reported by Marcogliese and
Cone (2001), who studied parasitic infection in fish and or-
ganic sediment enrichment of domestic wastewater.
However, contamination has been linked with other risk
factors such as seasonality, availability/presence of intermedi-
ate hosts, as well as feeding biology and activity of the final
host which must be considered. In this study, the resultsof the
values of the condition factor (CF) indicate the poor nutrition-
al status of the M. incilis from Cartagena Bay; however, this is
somewhat variable in the months of sampling in different sea-
sons. The interpretation of condition indices should be done
carefully because they can depend on several factors (Froese
2006; McPherson et al. 2010) such as food availability
(Bervoets and Blust 2003) or seasonality (Blackwell et al.
2000; Trudel et al. 2005), as well as their interrelation.
As for the parasites, there is an increase in several species
of nematodes that may be parasites for fish in their adult stage,
and many other species that use these hosts as an intermedi-
aries, including the fluke and tapeworm, which increase the
problem. However, and fortunately for man, not all parasites
have an impact on consumer health, and therefore, do not pose
a risk for all food (Mattiucci et al. 2011). Those which present
a potential risk to human health are the nematodes of the
Anisakidae family (Manfredi et al. 2000). In this study, three
species of nematodes of the Anisakides family
(Contracaecum spp., Pseudoterranova spp., and Anisakis
spp.) were found, with high intensity and prevalence.
Additionally, acanthocephalan species were also reported.
The degree of contribution to the life cycle of these nema-
todes by the fish species studied in Cartagena Bay as paratenic
guests is limited by numerous factors: the abundance of the
species in their natural environment (representing greater food
supply), the percentage of parasite prevalence, intensity and
mean abundance of parasites (which are related to the eating
habits of the fish depending on the station), the relationship
between parasitic infection and weight or size of fish (Cruz
et al. 2007). Given the difficulty of establishing all these fac-
tors, in order to approximate the degree of contribution to the
life cycle of nematodes of the Anisakidae family, the parasitic
infection and the degree of correlation between weight and the
average abundance of parasitic species were the only param-
eters taken into account.
The histopathological findings in this study showed severe
damage in the liver and spleen of M. incilis, including a high
incidence of melano-macrophages and granulome in the liver
and an increased presence of granulome in the spleen com-
pared with the liver. Similar injuries have been reported
(Marigomez et al. 2006; Miranda et al. 2008). The presence
of granulome and MMCs found in the liver of M. incilis in
Cartagena Bay can be related to the effects of contaminants,
but cannot prove that organochlorine compounds are respon-
sible for histological damage.
17483
A positive result could be detected if the contaminant is
fatal for the parasite or if it reduces the population of the
intermediate host, thus reducing the infection rate. However,
when the contaminant affects the final host before the parasite,
such as through immunotoxic chemicals, the result could be a
suppressed immune system and increased parasite load
(Lafferty 1997). In this study, a relationship between fish
weight and concentration of organochlorine pesticides was
established because older fish tend to be heavier, and there-
fore, have had longer exposure to chlorinated contaminants.
Furthermore, heavier fish have a higher lipid concentration
and therefore, have greater ability to bioaccumulate chlorinat-
ed compounds (Vives et al. 2005).
Prevalence of Contracaecum spp. nematodes in M. incilis
reported in this study was similar to that found by Fernández
(1987) for Mugil cephalus in Chile, and by Olivero-Verbel
et al. (2005) in Cartagena Bay. Jabbar et al. (2013) and
Lymbery et al. (2002) found heavy infections in M. cephalus
from fish species of southwestern Australia where they report-
ed a high prevalence of Contracaecum type I.
Conclusion
According to morphological analysis, the larvae found in the
species studied belong to the Anisakidae family and the
Anisakis spp., Pseudoterranova spp., Contracaecum spp. gen-
ders. In addition, the incidence of parasites found in the fish
studied is not statistically associated with the concentration of
chlorinated pesticides. The presence of fecal and total coli-
forms in Cartagena Bay could indicate poor quality of water
due to fecal pollution, which could encourage parasite infec-
tions in the fish.
Acknowledgments We would like to thank the University of Cartagena
and its School of Medicine for funding this research project. We would also
like to give thanks to the Agrochemical Research Group, especially Irina
Martelo and Irina Tirado; the Environmental and Computational Chemistry
Group; especially Dr. Jesus Olivero for their cooperation. Orlando de la
Rosa and Margarita Sorock for their collaboration
Conflict of interest The authors declare no conflict of interest.
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