(Gato Por Liebre)'': the ''Mota'

EcoHealth DOI: 10.1007/s10393-013-0893-8

Ó 2013 International Association for Ecology and Health

Original Contribution

‘‘Pig in a poke (gato por liebre)’’: The ‘‘mota’’ (Calophysus macropterus) Fishery, Molecular Evidence of Commercialization in Colombia and Toxicological Analyses

Cristian Salinas,1 Juan Camilo Cubillos,1 Rigoberto Go´mez,2 Fernando Trujillo,3 and Susana Caballero1

1Laboratorio de Ecologı´a Molecular de Vertebrados Acua´ticos-LEMVA, Biological Sciences Department, Universidad de los Andes, Carrera 1 No. 18A-10, Bogota´, Colombia 2Chemistry Department, Universidad de los Andes, Carrera 1 No. 18A-10, Bogota´, Colombia 3Fundacio´n Omacha, Calle 86A No. 23-38, Bogota´, Colombia

Abstract: Overfishing has affected the population abundance trends of many commercial fish species. In the Amazon, the fishery of a catfish commonly known as ‘‘mota’’ or ‘‘piracatinga’’ (Calophysus macropterus) has become an important economic activity in the region as this species has replaced a number of other overexploited great catfish species in the markets. Due to this high exploitation, ways in which to increase captures have been identified. One strategy is to use decomposing animal carcasses as bait. Such strategy has increased the hunting pressure on endangered species such as caimans and river dolphins. We investigated which catfish species are currently commercialized in Colombian fish markets using DNA barcoding, and measured mercury concentration in the tissues of fish molecularly identified as C. macropterus. We collected 86 fish samples in markets of four Colombian cities. Sixty-eight of these were identified molecularly as C.macropterus. The mercury concentration of 29 such samples was analyzed. Samples presented total Hg concentrations higher than the limit for human consumption established by the WHO (0.5 lg/g). These results are worrisome and suggest that (1) C. macropterus is a widely used fish species for human consumption in Colombia and (2) C. macropterus has high concentrations of total Hg, making its consumption a public health risk. Results presented here suggest that C. macropterus has replaced capaz in most Colombian markets. This fishery threatens wild species of river dolphins and caimans, and is also a public health risk given the high mercury levels we found in a subsample of these fishes.

Keywords: Calophysus macropterus, DNA barcoding, total mercury, Colombia, Inia geoffrensis

INTRODUCTION

Electronic supplementary material: The online version of this article (doi: The worldwide problem of overfishing has affected the 10.1007/s10393-013-0893-8) contains supplementary material, which is available to population abundance trends of many commercial fish authorized users. (Pauly et al. 2002). For this reason, there is a constant need Cristian Salinas and Juan Camilo Cubillos share first authorship on this manuscript. to find new species to replace commercially important

Correspondence to: Susana Caballero, e-mail: [email protected] species in the market which are now scarce and sometimes Cristian Salinas et al. endangered (Myers and Worm 2003). This problem is y Sotalia fluviatilis), because its flesh is used as bait for C. common in the Amazon, where the fishery of a catfish macropterus (Estupinãn et al. 2003; Silveira and Viana 2003; commonly known as ‘‘mota’’ (Colombia), ‘‘simi’’ (Peru) or Trujillo et al. 2007). Hunting these animals is relatively easy ‘‘piracatinga’’ (Brazil) (Calophysus macropterus, Lichten- and there is a market for this bait in some Amazonian stein, 1819) has become an important economic activity in regions (Hernańdez-Rangel 2010). This hunting pressure the region and this species has replaced a number of other increases exploitation of these endangered species. It has overexploited large catfish species from the Amazon and been estimated that around 1,500 dolphins are killed each other rivers, including the Magdalena River in Colombia year to support this fishing practice (da Silva et al. 2008). (Petrere et al. 2004). Currently, there are no available abundance estimates for The use of this particular species to fill the market river dolphins in their whole distribution in the Amazon niche previously occupied by the large catfish species from and Orinoco basins, so it is unclear what percentage of the the Amazon and the Magdalena is partly due to the fact total river dolphin population is being affected by this that this species is included in the Pimelodidae family, fishing practice. Surveys along the main Amazonian and where the large catfish species for human consumption Orinoco tributaries have been developed since 2006 in have also been classified, with around 40 species (Bogota´- order to determine their current abundance in most of Gregory and Maldonado-Ocampo 2006). This is due to the their distribution (Trujillo et al. 2010). For caimans, fact that catfish from this family may be similar in size and available information suggests that around 150 tons of bait they may look alike. Also, the widespread distribution of C. were used in this fishery in 2001 (Silveira and Viana 2003). macropterus in the Amazon region (Salinas and Agudelo In general, dolphin meat has been used for bait in 2000) increases the chance for its capture in ‘‘black’’ and several places around the world including Chile (Goodall ‘‘white’’ water rivers (these river names depend on the and Jordan 1986) and Peru, where dolphin and porpoise amount of sediments, tannins, and pH characteristics of meat is used as bait in shark fisheries (Mangel et al. 2010). rivers in the Amazon region) (Saint-Paul et al. 2000). This meat has two desirable characteristics for use as bait, This widespread distribution may be partly the result increasing its preference over other types of bait such as of its ecological role, as it is considered a scavenger Pe´rez caiman meat or cattle viscera for C. macropterus fishery. and Fabre´ (2009). For this reason, it is frequently found First, the meat is very rich in fat, something that appears to close to human settlements, feeding on animal remains attract this fish and second, the strong odor that this meat (Salinas and Agudelo 2000). C. macropterus has an average emits when decomposing attracts fish to a particular fishing size of 20.5 cm in length and an average weight of 300 g location in a short time (Estupinãn et al. 2003). Fishermen (Carvalho et al. 2009). Because of its size, it is an ‘‘ideal’’ targeting other fish species perceive dolphins as competi- species to exploit for high demand for fish. The fisheries tors; therefore, they hunt them and sell them to C. macr- effort for this species is around 627.43 tons per month in opterus fishermen to use their meat as bait (Loch et al. Southern Amazonia (mostly in Brazil), making it one of the 2009). most exploited catfish of the whole Amazonian region The commercialization chain of C.macropterus has (Alonso et al. 2007). Due to this high exploitation, several been identified as follows: the fish is obtained along the methods to increase captures have been identified. One of Amazon River and some of its main tributaries, including these strategies has been expanding the capture range to the Purus, Javari, and Ucayali Rivers. Then, from particular include the Peruvian (Ortega et al. 2001), Brazilian localities or cities along the river, it is shipped to main (Carvalho et al. 2009), and Colombian Amazon (Ortiz- cities. For example, in Brazil, Manaus has been identified as Zuluaga et al. 2006), including rivers such as the Putumayo the main provider to markets in Sao Paulo and Rio de (Colombia) (Gonza´lez et al. 2006) and also the low ‘‘Lla- Janeiro. In Colombia, fishermen bring their catch to a nos’’ of Venezuela (Castillo 1996). A second strategy, centralized location in Leticia, from there it is shipped and considering low capture rates in some regions, is to use distributed mainly to Bogota´ and other main cities (Pe´rez decomposing animal carcasses as bait to attract this fish and Fabre´ 2009). Commercialization in cities far away from (Trujillo et al. 2007). the Amazon region can be attributed to the fact that C. The latest strategy has now increased the hunting macropterus is considered a low-valued species in Amazo- pressure on species such as caimans (Melanosuchus niger nian villages and cities due to its scavenger feeding habits and Caiman crocodilus) and river dolphins (Inia geoffrensis (Petrere et al. 2004). Pig in a poke (gato por liebre)

Regarding its position in the food chain, C.macropterus species sold as snapper in a number of markets in the USA can be considered an interesting model organism to study and also to identify tuna and other fish species used in sushi the levels of mercury exposure at the upper levels of preparations (Stokstad 2010; Wong et al. 2011). In addi- Amazonian food chains or scavenger organisms that feed tion, the use of DNA barcodes has allowed an under- on top predator remains in the region. Also, since C. standing of the trading tendencies of shark fins in Asian macropterus is found scavenging close to human settle- markets, as well as providing a tool for species trade ments, it may be more exposed to runoff and pollutants. monitoring (Abercrombie et al. 2005; Clarke et al. 2006). The Amazon Basin has high natural sources of mercury due DNA barcoding may help to establish the geographical to natural biogeochemical processes (Lechler et al. 2000). origin of a particular sample, and also commercialization However, due to increasing mining activities (Uryu et al. or trade routes, in addition to being useful in under- 2001), particularly artisanal mining, and the change of land standing how a particular market or fishery works. use in the Amazonian region (Lacerda et al. 2004), high amounts of mercury have leached into the river. This free inorganic mercury is then converted into organic mercury METHODOLOGY forms, such as methyl-mercury that is then bioaccumulated (accumulates in particular tissues) and biomagnified (in- Fish Sampling creases its concentration) along Amazonian food webs. Eighty-six fish specimens were obtained from different sources These processes represent a toxicological risk for full eco- (fishermen, distributors, and markets) in eight locations in systems (Dudgeon et al. 2006; Dorea et al. 2006; Porto et al. Colombia between June and October 2010. Forty-six of these 2005; Lailson-Brito Jr. et al. 2008; Cubillos 2009). It is to be samples where obtained from ‘‘source’’ localities (a source, expected that this fish bioaccumulates mercury in high defined as places where the fishing activity is done or where levels, particularly if it feed on the remains of top predators fishermen bring their catch to be commercialized), and these such as caimans and river dolphins, animals that have been samples were morphologically identified using identification shown to concentrate mercury in high levels in their tissues guides and species identification was confirmed by taxonomic (Hernańdez-Rangel 2010; Silveira and Viana 2003; Gutleb experts in the field. These specimens were also molecularly et al. 1997; Dove 2009) and in their milk (Rosas and Lehti identified in order to use them for comparison in further 1996). For this reason, measuring and monitoring mercury analyses. These specimens were obtained in the Colombian concentrations in this fish are important considering the Amazon (Puerto Narinõ and Leticia), the Colombian Orinoco pervasive effects mercury has on human and animal health, (Inirida), Meta Province (Puerto Lo´pez), and Putumayo including its neurotoxicity (Malm et al. 1995; Castilhos Province (Puerto Ası´s) (Fig. 1). et al. 1998; Dove 2009). Forty specimens were obtained from markets in Bo- In Colombia, there was some information suggesting gota´, Girardot, and Melgar, where they were sold as ‘‘ca- that C. macropterus was being commercialized as another paz’’ from the Magdalena River. The latest two cities are species, the ‘‘capaz’’ from the Magdalena River (Pimelodus located on the Magdalena River, about 98 km southwest grosskopfii) in the big supermarket chains and also in the from Bogota´. They all were sold at different prices and all main central markets in Bogota´ and other cities. For this were sold under the name ‘‘capaz.’’ A small tissue sample reason, the aims of this study were: (1) to indentify via from each specimen was cut and transferred to 70% etha- ‘‘DNA barcoding,’’ which species of freshwater catfish were nol and refrigerated at 4°C. commercialized in Colombian markets, particularly in Bogota´ and (2) to determine the levels of mercury con- DNA Extraction, Amplification, and Phylogenetic centrations in commercialized C. macropterus. Analyses DNA barcoding, a molecular technique based on sequencing a portion of the mitochondrial DNA cyto- DNA was extracted from tissue samples following a phe- chrome oxidase I (COI), has been widely used to identify nol–chloroform protocol from (Sambrook et al. 1989) species and parts of species that may be legally or illegally modified for small samples by (Baker et al. 1994). traded, including fish species being traded as a different one Approximately 558 base pairs (bp) of the mitochondrial in order to increase its price and availability in the market COI gene were amplified via PCR, using the primers (Baker 2008). DNA barcodes have allowed identification of COXFISH F1-R1 (Ward et al. 2005). Bioline Taq Poly- Cristian Salinas et al.

Figure 1. Sample localities: Putu- mayo River Basin—1 Puerto Ası´s, Putumayo Province (four samples). Magdalena River Basin—2 Girardot, Cundinamarca Province (four samples), 3 Melgar, Tolima Province (five Samples), 4 Bogota´, Cundinamarca Province (31 samples). Meta River Basin—5 Puerto Lo´pez, Meta Province. Amazon River Basin 6 Puerto Narinõ, 7 Leticia, Amazonas Province (33 samples) and Orinoco River Re- gion—8 Puerto Inı´rida, Guainıá Province (seven samples). merase was used and the temperature profile was as follows: COI sequences were manually edited and aligned with an initial denaturation for 2 min at 94°C, followed by 35 previously published COI sequences from different species cycles at 94°C for 30 s, annealing at 59°C for 45 s, elon- from the family Pimelodidae available from GenBank gation at 72°C for 1 min 20 s, and a final extension 72°C (www.ncbi.nlm.nih.gov/) (for accession numbers see Sup- for 15 min. PCR products were run and visualized in a plementary Material Table 1) using the software Geneious 1.5% agarose gel stained with ethidium bromide. Successful version 5.3.3 (Biomatters Ltd). This dataset was further amplification products were purified via a protocol using a used for phylogenetic reconstructions using Neighbor- polyethilenglycol resin and a final wash with 96% ethanol Joining (NJ) in PAUP (Swofford 2011). These reconstruc- and sequenced, following the Sanger methodology, on an tions are considered the standard for DNA barcoding ABI3100 at Macrogen Korea. methodology and species identification in a wide variety of Pig in a poke (gato por liebre)

Table 1. Summary of specimens analyzed in this study and the percentage of species representation in the sample of 86 specimens analyzed in total (in bold).

Commercial name Local name Scientiﬁc name Number of specimens Capaz (B) Mota, Piracatinga, Zamurito, Simi, Picalon, Calophysus macropterus 78 (90.7%) Capaceta (B) (M) (G) Capaceta, Barbuchas, mapuro Capaz (B) Pirabuton, Blancopobre, Mapuro Brachyplatystoma vaillantii 3(3.5%) Capaz (B) (M) (G) Capaz Pimelodus grosskopﬁ 3(3.5%) Capaz (B) Barbancho, Barbichato, Piramutaba Pinirampus pinirampus 1(1.15%) Capaz(B) Mapara´ Hypophthalmus marginatus 1(1.15%)

Commercial name refers to the name used for a particular species in the markets of Bogota´ (B), Girardot (G), and Melgar (M). Local name refers to the name used for a particular species in the Colombian Amazon (Leticia, Puerto Narinõ, and Puerto Ası´s), the Brazilian Amazon (Tabatinga), The Colombian Orinoco (Puerto Lo´pez and Puerto Inı´rida). taxa (Hebert et al. 2004; Hellberg and Morrissey 2011; Ross COI gene. These five species were identified as belonging to et al. 2003). the family Pimelodidae with high similarity percentages (85–95%) (Table 1). Only C. macropterus showed more Total Mercury Analyses than one COI haplotype and the clade for this species was supported with a bootstrap value of 100 (Fig. 2). COI se- From the total of 86 specimens collected in this study, a subset quences were also useful to identify the origin of specimens, of 29 frozen samples from specimens molecularly identified as taking into consideration sequences from specimens col- C. macropterus were included in this analyses. Antero-dorsal lected in field locations in the Colombian Amazon, Bra- muscle samples were collected and acid-digested following the zilian Amazon, and Colombian Orinoco. In the NJ protocols described by Sadiq et al. (1991). Total mercury reconstruction (Fig. 2), a clade-joining specimen from the (HgTOT) concentrations were determined by cold vapor Amazon was clearly separated from specimens from the atomic absorption spectroscopy in a CV-AAS (Perkin-Elmer Orinoco. C. macropterus is the species which is sold in AAnalyst 300). Analyses were carried out at the spectroscopy higher proportion in the Colombian markets and is studied laboratory of Universidad de los Andes, Bogota´, Colombia. as if it was the capaz from the Magdalena River (P. gross- Three replicates per sample (0.5 g of skinless and boneless kopfii), but there is evidence of other species also sold as if flesh) were considered to determine the precision and accu- they were capaz. Only three specimens obtained in the racy of Hg concentrations. A standard biological reference markets in Melgar were identified as the real capaz from the material (DOLT-2 from NRCC Ottawa, Canada) was mea- Magdalena (P. grosskopfii) (Table 1). sured for calibration purposes and comparisons. Once the values for mercury concentration were ob- Total Mercury Analyses tained, the mean concentration of mercury was estimated for each sample and then used for the estimation of the All samples included in the mercury analyses were inden- bioaccumulation curves presented in Figure 3a, b . A two- tified as C. macropterus. These samples had HgTOT con- way ANOVA was performed in order to determine the centrations higher than the maximum permissible limit of differences in mercury concentration among the localities 0.5 lg/g (WHO 2007) established for human consumption. presented in this study (Fig. 4). The concentration values ranged from 1.33 to 2.28 lg/g.

HgTOT concentrations compared to fish length and to fish weight for all sampling sites (including locations of origin RESULTS and one market location) are shown in Figure 3a, b. Analyses for Hg by length showed that even small fish DNA Barcoding TOT (30 cm in length) presented values higher than values A total of five different species were identified from the 86 recommended by the WHO. No clear correlation was specimens analyzed. Morphological identification made in found between HgTOT concentration and fish weight, with the field was confirmed by genetic analyses of 558 bp of the some lighter fish having higher concentration than some of Cristian Salinas et al.

inability to clearly identify which species they are purchasing. An example is found in Colombia, where the final consumer thinks they are purchasing capaz but instead they are being sold C. macropterus or other species. This hap- pens when the final consumer thinks they are buying capaz from the Magdalena River but instead they are being sold C. macropterus or other species. This is of concern for several reasons. First, the C. macropterus fishery poses a risk to the conservation of endangered river dolphins and caimans, since fishermen use these species as bait. Second, as we show here, the high concentrations of mercury in the tissues of C. macropterus result in a serious health concern, particularly in large cities where this fish is consumed in large quantities. The problem of misleading the public into consumption of particular species based on deceiving their decision-making capacity is not unique for this case but has been widely researched in a number of fisheries around the world (Filonzi et al. 2010; Wong et al. 2011). For example, in Brazil, researchers found that 80% of fish filets labeled as Figure 2. Neighbor-joining reconstruction based on COI gene surubim (Pseudoplatystoma spp.) were in reality other than sequences. Bootstrap support values are on top of branches.Sequences catfish species (Carvalho et al. 2011). Also, researchers in labeled with (asterisk)correspondtopreviouslypublishedsequences New York found that red snapper (Lutjanus campechanus) available from GenBank. was the most commonly mislabeled fish in the market (Wong and Hanner 2008). the heavier fish analyzed. The bioaccumulation tendency Also, it is common to present the ‘‘substituted’’ fish as was low according to the correlation coefficients. The fillets, in which case it is almost impossible to visually identify the species of origin. Some vendors may also variation in HgTOT concentration among locations is show in Figure 4. For instance, Putumayo showed a higher scratch the side of the fish to remove the spots that allow differentiating mota from capaz. For this reason, the use of average of HgTOT concentration (2.35 lg/g) compared to other sampling locations such as Bogota´, showing the DNA barcodes to investigate what is sold in the market provides a precise, fast, and powerful technique for lowest average of HgTOT concentration (1.73 lg/g). How- ever, there were no significant differences among the detecting this kind of fraudulent sale behavior (Galimberti localities with regards to mercury concentrations (ANOVA et al. 2013). Its application in market surveys or monitoring P = 0.247, Fig. 4). to ensure quality of the product being sold can be greatly increased (Holmes et al. 2009). Also, it can detect if endangered species are commercialized illegally, as was the DISCUSSION recent case when sushi made with whale meat was detected in a restaurant in the United States (Baker et al. 2010). In In this study, we used multidisciplinary techniques to our case, it confirmed the suspicions of replacement of evaluate which species of fish are currently used in the Pimelodus grosskopfii, capaz from the Magdalena River, Colombian market to replace the commonly known but mostly by C. macropterus in markets of Colombia. Three currently scarce and endangered capaz from the Magdalena other fish species from the Pimelodidae family (Pinirampus River. In addition, we measured mercury levels in a subset pinirampus, Brachyplatystoma vaillantii, and Hypophthal- of samples to determine if the consumption of C. macr- mus marginatus) were identified in this study, all being opterus, commonly known as mota or capacete and found commercialized, but in an apparently much smaller pro- in the Amazon and Orinoco, poses a risk to human health. portion than C. macropterus. The use of other species to Cryptic species of fish, or those morphologically sim- replace the capaz may be due to fluctuations in its capture ilar, pose a challenge for the final consumer given their at different times of the year (Go´mez et al. 2008). All these Pig in a poke (gato por liebre)

Figure 3. a Bioaccumulation curve (scatter-plot)ofmuscle-Hgcon- centrations as a function of body length presented for all sampling locations. b Bioaccumulation curve (scatter-plot)ofmuscle-Hgconcen- trations as a function of body weight presented for all locations.

Figure 4. Box-plot of the variation in HgTOT concentration from specimens among sampling locations. species are found in the Amazon and Orinoco basins and capaz from the Magdalena River (Pimelodus grosskopfii) is not in the Magdalena River. now classified as a vulnerable species in Colombia (Mojica Our results support recently published information by et al. 2012). The low availability of capaz from the the Colombian government (Ministerio de Agricultura y Magdalena has increased its market price. Currently, one Desarrollo Rural—MADR 2010), evidencing higher ex- (1) pound of capaz costs about $6.25 USD, while the cost ports from C. macropterus originating from Colombia and of C. macropterus is about $2.5 USD per pound. The its increase in the internal market as well as decreased considerable difference in price between mota and capaz numbers on capaz being commercialized in the country, also resulted in markets preferring to sell the cheaper fish demonstrating population reductions for this species. The at a higher cost. Cristian Salinas et al.

The final consumer is not capable of telling the two Second, all the regions where samples were collected have species apart and is deceived when not informed properly high mercury levels due to geological characteristics, arti- of what species they are buying. We conducted a small sanal mining, or to the change in land use as has been number of surveys (15 to fish dealers and 20 to final con- previously suggested by Lechler et al. (2000) and Lacerda sumers) in the three cities included as the ‘‘market’’ in this et al. (2004). study (Bogota´, Melgar, and Girardot). Results suggested In this study, we demonstrated the reliability of DNA that fish dealers (fishermen, restaurant owners, and fish barcoding for species identification and presented evidence dealers) were aware of the replacement of capaz by mota of commercialization of mercury-contaminated fish in and they successfully identified mota as capacete (see Ta- Colombian markets. We believe that regulation of this ble 1). Final consumers were not aware of the replacement. fishery, including strong legislation making the use of river Also, it was clear that almost none selling or buying C. dolphins and caimans as bait illegal, must be implemented macropterus was aware of the way in which these fish are in all nations from the Amazon region, particularly Brazil, captured through the use of river dolphins and caimans as Colombia, and Peru. It is necessary to track the origin and bait (Alves et al. 2012; Go´mez et al. 2008; Mintzer et al. fishing technique used in the capture of C. macropterus. 2013; da Silva et al. 2011). An unaware market may be the The use of cattle viscera represents fish bait alternatives and cause of approximately 1,150 tons of C. macropterus being it is being currently used in some areas of Colombia (Go´- obtained in the Brazilian Amazon each year (Trujillo et al. mez et al. 2008). For this reason, a chain of custody, 2010), overexploiting this fish and causing a significant making clear which bait was used to capture C. macropterus decrease in numbers in the populations of the animals used is necessary in order to allow the final consumer to make an as bait. This was recently demonstrated for Inia geoffrensis informed decision. A chain of custody in this context refers in the central Brazilian Amazon. In this study, researchers to the tracking of a product from its origin and in every in the Japura´ and Solimo˜es Rivers used capture/recapture– stage of the supply chain to ensure the end consumers that resighting data from marked Inia geoffrensis for a 17-year the product was obtained from a sustainable source or in period. They estimated that apparent survival of these an environmentally sustainable way. Such chains of custody dolphins was significantly lower in the years following the have been developed for fisheries, as is the case for the start, about 10 years ago, of the harvesting of these animals Marine Stewardship Council eco-labeled products (Gul- for bait (Mintzer et al. 2013). There are no clear estimates brandsen 2009). Also, increased monitoring of fishing of how many caimans are killed each year in the Amazon practices and follow up of testing for mercury content is for bait, but data from 2001 suggested that approximately also needed in order to determine if the trade and con- 150 tons of caimans were used in this fishery, considering sumption should be continued. We suggest that the gov- that for each kg of C. macropterus obtained, 1 kg of caiman ernmental entities working in collaboration with NGOs bait was used (Silveira and Viana 2003). Ganges river and academic institutions should also provide alternatives dolphins (Platanista gangetica) were used as bait (Smith to the fishing communities regarding sustainable manage- et al. 1998) and their oil used as fish attractant (Sinha 2002) ment of aquatic resources and alternatives to the C. in Bangladesh and India in the late 1990s. This caused macropterus fishery. population numbers to crash and to this date remain in very low numbers. The total mercury concentrations in fish muscle found ACKNOWLEDGMENTS in this study were much higher than the ones found in previous studies in the Amazon region (Malm et al. 1995; The authors want to thank all the people who in one way or Cubillos 2009). These concentrations were even higher than another helped in developing this study and in sample the recommended levels allowed for human consumption collection. We would like to thank Luis Fernando Mejia, by the WHO (0.5 lg/g). No significant differences were Veronica, and Melany Villate for their collaboration in the found in mercury concentration among the capture local- total mercury sample analyses. Funds for this project were ities, which may suggest two possible models as a way to made available by Universidad de los Andes (proyecto explain the high concentrations detected in this study. First, Profesor Asistente, Departamento de Ciencias Biolo´gicas migratory organisms may be transporting organic mercury and Departamento de Quı´mica), Cetacean Society Inter- from highly mercury-polluted locations to other regions. national, Whale and Dolphin Conservation Society, Whit- Pig in a poke (gato por liebre) ley Fund for Nature, Global Ocean, WWF and Fundacioń da Silva VMF, Martin AR, Mikesh E (2008) Skin disease and Omacha. lesion of the boto Inia geoffrensis in central Amazon. Report to the Scientific Committee of the International Whaling Com- mission, Santiago de Chile, Chile. Dorea JG, Barbosa AC, Silva GS (2006) Fish mercury bioaccu- REFERENCES mulation as a function to feeding behavior and hydrological cycles of the Rio Negro, Amazon. Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology 142:275–283 Abercrombie D, Clarke S, Shivji M (2005) Global-scale genetic identification of hammerhead sharks: application to assessment Dove V (2009) Mortality investigation of the Mekong Irrawaddy of the international fin trade and law enforcement. Conservation River dolphin (Orcaella brevirostris) in Cambodia based on Genetics 6:775–788 necropsy sample analysis. WWF International. Alonso JC, Camacho K, Usma JS (2007) Economia y usos de la Dudgeon D, Arthington AH, Gessner MO, Kawabata ZI, Knowler biodiversidad. In: Diversidad biolo´gica y cultural del sur de la DJ, Le´veque C, Naiman RJ, Prieur-Richard AH, Soto D, Stiassny Amazonia colombiana, Ruiz SL, Sańchez E, Tabares E, Prieto-C MLJ, Sullivan CA (2006) Freshwater Biodiversity: importance, A, Arias-G JC, Go´mez R, Castellanos D, Garcıá P, Rodrı´guez L threats, status and conservation challenges. Biological Reviews (editors), Bogota´ D. C: Corpoamazonia, Instituto Humboldt, 82:163–182 Instituto Sinchi, UAESPNN, pp 317–321 Estupinãn G, Marmontel M, Queiroz HLd, Souza PRe, Valsecchi Alves LCPdS, Zappes CA, Andriolo A (2012) Conflicts between J, Batista GdS, Pereira SB (2003) A pesca da Piracatinga (Cal- river dolphins (Cetacea: Odontoceti) and fisheries in the Central ophysus macropterus) na Reserva de Desenvolvimento Sustent- Amazon: a path toward tragedy? Zoologia (Curitiba) 29:420–429 avel Mamiraua. M.r.d.C.n.e.T. Technical report, Instituto de Desenvolvimento Sustentavel, Mamiraua. Baker CS (2008) A truer measure of the market: the molecular ecology of fisheries and wildlife trade. Molecular Ecology Filonzi L, Chiesa S, Vaghi M, Nonnis Marzano F (2010) Molecular 17:3985–3998 barcoding reveals mislabelling of commercial fish products in Italy. Food Research International 43:1383–1388 Baker CS, Slade RW, Bannister JL, Abernethy RB, Weinrich MT, Lien J, Urban J, Corkeron P, Calmabokidis J, Vasquez O, Pa- Galimberti A, De Mattia F, Losa A, Bruni I, Federici S, Casiraghi lumbi SR (1994) Hierarchical structure of mitochondrial DNA M, Martellos S, Labra M (2013) DNA barcoding as a new tool gene flow among humpback whales Megaptera novaeangliae, for food traceability. Food Research International 50:55–63 world-wide. Molecular Ecology 3:313–327 Goodall RNP, Jordan J (1986) Use of dolphins for crab bait in Baker CS, Steel D, Choi Y, Lee H, Kim KS, Choi SK, Ma Y-U, Chile, International Union for the Conservation of Nature Hambleton C, Psihoyos L, Brownell RL, Funahashi N (2010) (SSC): Newsletter of the Cetacean Specialist Genetic evidence of illegal trade in protected whales links Japan Go´mez C, Trujillo F, Diazgranados MC, Alonso JC (2008) Cap- with the US and South Korea. Biology Letters 6:647–650 tura dirigida de delfines de rio en la Amazonia para la pesca de Bogota´-Gregory JD, Maldonado-Ocampo JA (2006) Peces de la mota (Calophysus macropterus) una problematica regional de zona hidrogeogra´fica de la Amazonia, Colombia. Revista Biota gran impacto. In: Fauna Acuatica Amenazada en la Amazonia Colombiana 7:55–94 Colombiana, Trujillo FA, Alonso JC, Diazgranados MC, Go´mez C (editors), Bogota´: Fundacion Omacha, pp 39–57. Carvalho DC, Neto DAP, Brasil BSAF, Oliveira DAA (2011) DNA barcoding unveils a high rate of mislabeling in a commercial Gonza´lez JCA, Nun˜ez-Avellaneda M, Co´rdoba EA, Lo´pez LFR, freshwater catfish from Brazil. Mitochondrial DNA 22:97–105 Paéz CLS (2006) Ecosistemas acuaticos de la Amazonia Co- lombiana: Avances y perspectivas. Revista Colombia Amazońica, Carvalho TP, Tang SJ, Fredieu JI, Quispe R, Corahua I, Ortega H, pp 1–18. Albert JS (2009) Fishes from the upper Yurua´ river, Amazon basin, Peru. Check List 5:673–691 Gulbrandsen A (2009) The emergence and effectiveness of the Marine Stewardship Council. Marine Policy 33:1–11 Castilhos ZC, Bidone ED, Lacerda LD (1998) Increase of the background human exposure to mercury through fish con- Gutleb A, Schennck C, Staib E (1997) Giant otter (Pteronura sumption due to gold mining at the Tapajo´s River region, Para´ brasiliensis) at risk? Total mercury and methylmercury levels in State, Amazon. Bulletin of Environmental Contamination and fish and otter scats, Peru Ambio 26:511–514 Toxicology 61:202–209 Hebert PDN, Stoeckle MY, Zemlak TS, Francis CM (2004) Castillo O (1996) La pesca nocturna de bagre zamurito Calophysus Identification of birds through DNA barcodes. PLoS Biology macropterus (Lichtenstein) 1819 (siluriformes, pimelodidae) en 2:e312 rıós del bajo llano, Venezuela. Biollania 12:117–126 Hellberg RSR, Morrissey MT (2011) Advances in DNA-based Clarke SC, Magnussen JE, Abercrombie DL, McAllister MK, Shivji techniques for the detection of seafood species substitution on MS (2006) Identification of shark species composition and the commercial market. Journal of the Association for Laboratory proportion in the Hong Kong shark fin market based on Automation 16:308–321 molecular genetics and trade records. Conservation Biology Hernańdez-Rangel SM (2010) Informe final sobre la evalucioń de 20:201–211 la matanza de delfines de rıó utilizandos como carnada para la Cubillos JC (2009) Determinacioń de mercurio en la cadena pesca de mota (Calophysus macropterus) en el rıó Javarı´. Fun- tro´fica acua´tica en Puerto Narinõ, Amazonas (Colombia): im- dacioń OMACHA—Reserva natural PALMARI—Instituto de plicaciones ecolo´gicas a nivel global. B.Sc. Thesis, Biological desenvolvimiento socioambiental do vale do rio Javari. Sciences Department, Universidad de los Andes, Bogota´, Holmes BH, Steinke D, Ward RD (2009) Identification of shark and Colombia. ray fins using DNA barcoding. Fisheries Research 95:280–288 da Silva VM, Martin AR, Carmo NA (2011) Boto-bait. Amazonian Lacerda LD, de Sousa M, Ribeiro MG (2004) The effects of land fisheries pose threat to elusive dolphin species. In: IUCN use change on mercury distribution in soils of Alta Floresta, Magazine of the species Survival Commision, pp 10–11. Southern Amazon. Environmental Pollution 129:247–255 Cristian Salinas et al.

Lailson-Brito J Jr, Dorneles PR, da Silva VMF, Martin AR, Bastos Ross HA, Lento GM, Dalebout ML, Goode M, Ewing G, McLaren WR, Azevedo-Silva CE, Azevedo AF, Torres JPM, Malm O P, Rodrigo AG, Lavery S, Baker CS (2003) DNA surveillance: (2008) Dolphins as indicators of micropollutant trophic flow in web-based molecular identification of whales, dolphins, and Amazon Basin. Oecologia Brasiliensis 12:531–541 porpoises. Journal of Heredity 94:111–114 Lechler PJ, Miller JR, Lacerda LD, Vinson D, Bonzongo JC, Lyons Sadiq M, Zaidi TH, Al-Mohana H (1991) Sample weight and WB, Warwick JJ (2000) Elevated mercury concentration in soils, digestion temperature as critical factors in mercury determi- sediments, water and fish of the Madeira River basin, Brazilian nation in fish. Bulletin of Environmental Contamination and Amazon: a function of natural enrichments? The Science of Total Toxicology 47:335–341 Environment 260:87–96 Saint-Paul U, Zuanon J, Correa MAV, Garcıá M, Fabre´ NN, Loch C, Marmontel M, Simo˜es-Lopes P (2009) Conflicts with Berger U, Junk WJ (2000) Fish communities in Central Ama- fisheries and intentional killing of freshwater dolphins (Cetacea: zonian white and black water floodplains. Environmental Biology Odontoceti) in the Western Brazilian Amazon. Biodiversity and of Fishes 57:235–250 Conservation 18:3979–3988 Salinas Y, Agudelo E (2000) Calophysus macropterus. In: Peces de Malm O, Branches FJP, Akagi H, Castro MB, Pfeiffer WC, Harada Importancia Economica en la cuenca Amazonica Colombiana, M, Bastos WR, Kato H (1995) Mercury and methylmercury in Bogota´: I.A.d.I.C.-. SINCHI, pp 92–93. fish and human hair from the Tapajo´s river basin, Brazil. The Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a Science of Total Environment 175:141–150 laboratory manual, Cold Spring Harbor, New York: C.S.H.L Press Mangel JC, Alfaro-Shigueto J, Van Waerebeek K, Caćeres C, Silveira RD, Viana JP (2003) Amazonian crocodilians: a keystone Bearhop S, Witt MJ, Godley BJ (2010) Small cetacean captures species for ecology and management…or simply bait? Crocodile in Peruvian artisanal fisheries: high despite protective legisla- Specialist Group Newsletter 22:16–17 tion. Biological Conservation 143:136–143 Sinha RK (2002) An alternative to dolphin oil as a fish attractant Ministerio de Agricultura y Desarrollo Rural—MADR, C.C.I in the Ganges River system: conservation of the Ganges River (2010) Pesca y Acuicultura Colombia 2009. Corporacion Co- dolphin. Biological Conservation 107:253–257 lombiana Internacional, Bogota´, Colombia, pp 70–76. Smith BD, Aminul Haque AKM, Shakhawat Hossain M, Khan A Mintzer VJ, Martin AR, da Silva VMF, Barbour AB, Lorenzen K, (1998) River dolphins in Bangladesh: conservation and the effects Frazer TK (2013) Effect of illegal harvest on apparent survival of of water development. Environmental Management 22:323–335 Amazon River dolphins (Inia geoffrensis). Biological Conserva- tion 158:280–286 Stokstad E (2010) To fight illegal fishing, forensic DNA gets local. Science 330:1468–1469 Mojica JI, Oviedo JSU, Leon RA, Lasso CA (editors), A´ .-L.y.C.A.L. (2012) Libro rojo de peces dulceacuıćolas de Colombia 2012, Swofford DL (2011) PAUP: phylogenetic analysis using parsi- eds I.d.I.d.R. Biolo´gicos, I.d. Alexander von Humboldt, mony, version 4.0b10. C.N.d.l.U. Nacional, W.C.y.U.d. de Colombia, Manizales, p 141. Trujillo F, Crespo E, Damme PV, Usma JS (2010) The Action Plan Myers RA, Worm B (2003) Rapid worldwide depletion of pred- for South American River Dolphins 2010–2020, Bogota´: WWF, atory fish communities. Nature 423:280–283 Fundacion Omacha, WDS, WDCS, Solamac Ortega H, Hidalgo M, Salcedo N, Castro E, Riofrio C (2001) Trujillo F, Go´mez C, Portocarrero M, Diazgranados MC (2007) Diversity and conservation of fish of the lower Urubamba re- Conservacioń de la diversidad biolo´gica y cultural. In: Diversi- gion, Peru. In: Urubamba: The Biodiversity of a Peruvian Rain- dad biolo´gica y cultural del sur de la Amazonia colombiana, forest, Alonso A, Dallmeier F, Campbell P (editors), Ruiz SL, Sańchez E, Tabares E, Prieto-C A, Arias-G JC, Go´mez ´ ´ Washington, USA: Smithsonian Institution, National Zoological R, Castellanos D, Garcıa P, Rodrıguez L (editors), Bogota´, Park, Conservation and Research Center, SI/MAB Biodiversity Colombia: Corpoamazonia, Instituto Humboldt, Instituto Sin- Program, pp 143–150 chi, UAESPNN, p 440. Pauly D, Christensen V, Guenette S, Pitcher TJ, Sumaila UR, Uryu Y, Malm O, Thornton I, Payne I, Cleary D (2001) Mercury Walters CJ, Watson R, Zeller D (2002) Towards sustainability in contamination of fish and its implications for other wildlife of world fisheries. Nature 418:689–695 the Tapajo´s Basin, Brazilian Amazon. Conservation Biology 15:438–446 Pe´rez A, Fabre´ NN (2009) Seasonal growth and life history of the catfish Calophysus macropterus (Lichtenstein, 1819) (Silurifor- Ward RD, Zemlak TS, Innes BH, Last PR, Hebert PDN (2005) mes: Pimelodidae) from the Amazon floodplain. Journal of DNA barcoding Australia’s fish species. Philosophical Transac- Applied Ichthyology 25:343–349 tions of the Royal Society B: Biological Sciences 360:1847–1857 Petrere M, Barthem RB, Co´rdoba EA, Go´mez BC (2004) Review of Wong EHK, Hanner RH (2008) DNA barcoding detects market the large catfish fisheries in the upper Amazon and the stock substitution in North American seafood. Food Research Inter- depletion of piraı´ba (Brachyplatystoma filamentosum, Lichten- national 41:828–837 stein). Reviews in Fish Biology and Fisheries 14:403–414 Wong LL, Peatman E, Lu J, Kucuktas H, He S, Zhou C, Na- Porto JIR, Araujo CSO, Feldberg E (2005) Mutagenic effects of nakorn U, Liu Z (2011) DNA barcoding of catfish: species mercury pollution as revealed by micronucleus test on three authentication and phylogenetic assessment. PLoS One 6:e17812 Amazonian fish species. Environmental Research 97:287–292 Ortiz-Zuluaga GI, Wood AA, Zarate CG (2006) El sector pesquero Rosas FCW, Lehti K (1996) Nutritional and mercury content of en Puerto Narinõ. In: Puerto Narinõ-el pueblo que se mira en el milk of the Amazon River dolphin Inia geoffrensis. Comparative rıó: retos al desarrollo sustentable en los municipios amazońicos, Biochemistry and Physiology, Part A: Physiology 115:117–119 Colombia, Bogota: U.N.d.