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International Journal for Parasitology: Parasites and Wildlife logoLink to International Journal for Parasitology: Parasites and Wildlife
. 2026 Mar 16;29:101218. doi: 10.1016/j.ijppaw.2026.101218

Morphological and molecular characterization of Contracaecum jorgei (Nematoda: Anisakidae) in five Neotropical freshwater fish species in Colombia

Astrid Rave a,⁎, Manuel Uribe a, Sara López-Osorio a, Carlos Hermosilla b, Jenny J Chaparro-Gutiérrez a
PMCID: PMC13049434  PMID: 41939529

Abstract

Larvae of the family Anisakidae occur in numerous freshwater and marine fish species. Although nematodes of the genus Contracaecum are widely distributed, little is known about their occurrence in fish in South America, particularly in Colombia. In this study, 257 specimens representing five Neotropical freshwater fish species—Andinoacara latifrons, Astyanax sp., Brycon henni, Parachromis friedrichsthalii, and Rhamdia guatemalensis—from the Porce River basin (Colombia) were examined for Contracaecum infection. Infective third-stage larvae (L3) of Contracaecum jorgei were identified using both morphological characteristics and molecular analysis targeting the mitochondrial cytochrome c oxidase subunit II (cox2) gene. Infection frequencies were A. latifrons (28.6%; 2/7), Astyanax sp. (38.3%; 23/60), B. henni (13%; 22/170), P. friedrichsthalii (21.1%; 4/19), and R. guatemalensis (100%; 1/1). To our knowledge, this is the first report of infective C. jorgei L3 in edible, commercially important freshwater fish species from Colombia. These findings contribute to the understanding of the parasitic fauna in Colombia and highlight the potential risks that this nematode may pose to aquatic biodiversity in the Porce River basin, which harbors 238 endemic fish species.

Keywords: Anisakidosis, Contracaecum, Fish parasites, Larvae, Freshwater fish

Graphical abstract

Image 1

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Highlights

  • •

    This is the first morphological and molecular characterization of Contracaecum jorgei in Colombia.

  • •

    We conducted the report of C. jorgei in five Neotropical freshwater fish species in Colombia.

  • •

    This report expands the geographic distribution of the parasite Contracaecum jorgei to Colombia.

1. Introduction

The family Anisakidae includes gastrointestinal zoonotic nematodes with a worldwide distribution and a complex epizootiology (Angeles Hernandez et al., 2020). Several anisakid nematodes have been found as adult stages parasitizing the digestive tract of piscivorous birds (i.e herons, cormorants, seagulls, pelicans, albatrosses, and penguins) (Garbin et al., 2013, 2019, 2023b) and marine mammals (i. e. seals, sea lions, and cetaceans) (Ebmer et al., 2020; Hermosilla et al., 2016; Mattiucci et al., 2018; Moravec et al., 2016) acting as definitive hosts (DH). In contrast to adults, anisakid larval stages parasitize the body cavity, organs and muscle tissue of fish acting as IH and/or paratenic hosts (PH) (Mattiucci et al., 2018). Within the family Anisakidae, the genera Anisakis, Pseudoterranova and Contracaecum are the most widespread (Abdullah et al., 2021). Notably, Contracaecum is among the most common nematode genus transmitted by fish (Ziarati et al., 2022). Infective third-stage larvae (L3) of Contracaecum are typically found worldwide in the coelomic cavity of either freshwater-, euryhaline- or marine fish species (Vuić et al., 2022).

Humans become accidently infected by ingesting either raw or undercooked fish harboring infective Contracaecum L3, however, not contributing to the parasite's epizootiology or geographic dissemination as humans are considered aberrant hosts (AH) (Shamsi, 2019; Shamsi and Butcher, 2011). Given the wide spectrum of IH-, DH- PH- and AH species, Contracaecum has important implications for biodiversity (Jawad et al., 2022). In South America, reports of Contracaecum infections in fish include Argentina, Brazil, Colombia, Chile, British Guiana, Peru, Trinidad and Tobago, and Uruguay (Garbin et al., 2023a). In Colombia, Contracaecum L3 have been identified through conventional morphometric methods as previously reported (Castellanos et al., 2017; Olivero-Verbel et al., 2006; Olivero Verbel et al., 2011; Socarras Campo et al., 2012). These Contracaecum L3 have been recorded in fish inhabiting marine environments along the Caribbean coast, including Mugil incilis (Hancock, 1830; Bustos-Montes et al., 2012) and Hoplias malabaricus (Bloch, 1794) (Olivero-Verbel et al., 2006; Sandra Pardo et al., 2008; Vergara-Flórez and Consuegra, 2021).

In freshwater systems, infections have been reported in H. malabaricus (Olivero-Verbel et al., 2006), Salminus affinis (Steindachner, 1880; Pardo et al., 2007), Trichomycterus caliense (Eigenmann, 1912), Astroblepus cyclopus (Humboldt, 1805; Román-Valencia, 2001), and Sorubim cuspicaudus (Littmann, Burr & Nass, 2000; Sandra Pardo et al., 2009). Notably, in Totumo Marsh, an estuarine ecosystem located in Northern Colombia (Ballesteros Madera and González Porto, 2010), L3 larvae of Contracaecum have been detected in M. incilis (Ballesteros Madera and González Porto, 2010; Olivero et al., 2008). Additionally, adult nematodes identified as Contracaecum bioccai (Mattiucci et al., 2008) were recovered from the stomach of a brown pelican (Pelecanus occidentalis) (Linnaeus, 1766) (Mattiucci et al., 2008). Interestingly, this large-sized piscivorous bird is widely distributed from the Southern Pacific coast to the Galápagos Islands and Southern Chile, and in Colombia it has been documented along both the Caribbean- and Pacific coastlines (Chaparro-Herrera et al., 2022). Migratory behavior of P. occidentalis suggests that it could play a role in the regional dispersal of Contracaecum parasites. Although the life cycle of C. jorgei remains unknown, the general life cycle of Contracaecum spp. sensu lato is heteroxenous (Fig. 1), involving piscivorous birds of the families Pelecanidae, Phalacrocoracidae, Ardeidae (Mattiucci et al., 2010), as well as pinnipeds of the families Otariidae and Phocidae (Johnston and Mawson, 1951; Kuzmina et al., 2018; Linstow, 1906), as DH. These DH release embryonated eggs in their feces, which enter aquatic environments and undergo the first larval molt within the egg. After hatching, the larvae develop into free-swimming second-stage larvae (L2) (Shamsi, 2019). The eggs or larvae are ingested by aquatic invertebrates –the first IH- which include copepod, isopods and amphipods (Moravec, 2009). When consumed by fish, acting as second IH, the larvae migrate into the coelomic cavity and develop into infective Contracaecum L3. Infective Contracaecum L3 are available to DH when infected fish are preyed upon, completing the parasite's life cycle (Garbin et al., 2023b).

Fig. 1.

Fig. 1

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Life cycle of Contracaecum sp. a. Definitive hosts (piscivorous birds and pinnipeds) excrete embryonated eggs through feces, which mature in the water to L2. They are ingested by b. the first intermediate hosts, crustaceans, and aquatic macroinvertebrates, where larvae develop in their coelom. Subsequently, aquatic macroinvertebrates are ingested by c. the second intermediate hosts (fish), where larvae develop into L3. d. Humans serve as accidental hosts when consuming raw or undercooked fish infected with L3 larvae. The cycle is completed when fish are ingested by piscivorous birds or pinnipeds (definitive hosts).

In the current parasitological survey on potential zoonotic anisakid infections, five species of consumable Neotropical freshwater fish species in Colombia were investigated for possible occurrence of either Contracaecum L3 or other anisakid L3. The species investigated were Andinoacara latifrons (Steindachner, 1878), Astyanax sp. (S. F. Baird and Girard, 1854), Brycon henni (Eigenmann, 1913), Parachromis friedrichsthalii (Heckel, 1840), and Rhamdia guatemalensis (Günther, 1864). This study reports for the first time the occurrence of infective C.jorgei (Sardella et al., 2020) L3 in five consumable Neotropical freshwater fish of Colombia. Investigated fish species are popular within Colombian gastronomy and regularly consumed by humans inhabiting riversides of the Porce River basin, Antioquia, Colombia. Accordingly, findings are highly relevant not only on the Porce River basin biodiversity inhabiting 238 endemic fish species (DoNascimiento et al., 2024), but also on fish-as well, given the potential zoonotic relevance of some species of Contracaecum.

2. Methodology

2.1. Study area and sample collection

The Porce River is part of the Magdalena River basin, the largest river system in the Northern Andean region of Colombia. A total of 257 specimens of five Neotropical freshwater fish species were sampled in this study, including four endemic species, i. e. Andinoacara latifrons (n = 7), Astyanax sp. (n = 60), Brycon henni (n = 170), and Rhamdia guatemalensis (n = 1), and one non-native species namely Parachromis friedrichsthalii (n = 19) (Fig. 2). Fish species identification was carried out by ichthyologists from the University of Antioquia, following the identification guidelines as described elsewhere according to Jiménez Segura et al. (2014) and Valencia et al. (2017).

Fig. 2.

Fig. 2

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The fish species analyzed in this study include: a.Andinoacara latifrons (Cichlidae).b.Astyanax sp. (Characidae) c.Brycon henni (Characidae) d.Rhamdia guatemalensis (Heptapteridae).e.Parachromis friedrichsthalii (Cichlidae).

Fish specimens were collected from 20 sampling sites illustrated in Fig. 3, as detailed in (Supplementary Material Table 1). In the flowing channels of this river canyon (streams, small channels, and brooks), three types of cast nets with different mesh sizes (0.5, 1.5, and 3.5 cm) were used. Additionally, 60 min of portable electric fishing sweeps were conducted, using an equipment with a pulsating current of 340 V, 1–2 A, DC, along 100 m stretch of the channels. In reservoirs, capture efforts involved three gill nets, each 100 m long and 3 m deep, with each net having 10 different mesh sizes ranging from 1 to 10 cm.

Fig. 3.

Fig. 3

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Precise geographic location of sampling area.

Captured fish were anesthetized in a eugenol solution (100 mg/L) and euthanized in a concentrated eugenol solution 300 mg/L as described elsewhere (Chacón-Guzmán et al., 2019; Rucinque et al., 2017). During necropsy procedures, a longitudinal incision was made using Metzenbaum scissors, starting from the caudal region of the pectoral fin, ascending towards the midline, and descending towards the anus. This procedure created a necropsy window for observing internal organs (Gelnar et al., 2018). Exposed organs were examined under a Nikon SMZ-1 stereomicroscope in Petri dishes with saline solution.

2.2. Morphological analysis

The parasites recovered during the fish necropsy were carefully extracted from the coelomic cavity of evaluated individuals using toothless Adson forceps. Subsequently, parasites recovered during the necropsy procedure were cleaned in 0.9% sterile saline solution, relaxed in acetic acid, and thereafter fixed in 70% ethanol for morphological identification. Additionally, the helminths collected for molecular studies were preserved in 96% ethanol until further genetic analysis.

Thereafter observations of morphometrical features were conducted. In brief, collected nematodes were clarified in Amman's lactophenol and checked periodically until external and internal structures were completely clarified, mounted on glass slides following the protocol outlined by (Gelnar et al., 2018). Photographs were taken using an Olympus BX53 microscope (Olympus Corporation, Tokyo, Japan) (100x, 400x, and 1,000x), equipped with an Olympus DP74 digital camera (Olympus Corporation, Tokyo, Japan), and the Olympus SZXY stereomicroscope (Olympus Corporation, Tokyo, Japan), equipped with an Olympus DP27 digital camera. These instruments were used with Olympus cellSens Standard software (Olympus Corporation, Tokyo, Japan) for image analysis. Identifications were based on key features and descriptions (R. C. Anderson et al., 1974; R. Anderson, 2000; Martins et al., 2005; Arai and Smith, 2016). The infection frequency, mean intensity and abundance for each species were calculated (Bush et al., 1997).

2.3. Molecular and phylogenetic analyses

Samples from each host species – A. latifrons, Astyanax sp., B. henni, R. guatemalensis – were fixed in molecular grade ethanol at 96-99% and subsequently sequenced. Total genomic DNA was extracted using the GeneJet Genomic DNA Purification Kit® (Thermo Scientific, Waltham, United States). A fragment of the mitochondrial cytochrome c oxidase subunit II (cox2) gene was amplified using primers 211F (5′-TTTTCTAGTTATATAGATTGRTTTYAT-3′) and 210R (5′-CACCAACTCTTAAAATTATC-3′) according to Nadler and Hudspeth (2000). The PCR conditions consisted of initial denaturation 94 °C for 5 min, followed by 35 cycles of 94 °C for 30 s, 55 °C for 60 s, 72 °C for 90 s, and a final extension at 72 °C for 10 min. The PCR products were verified by electrophoresis on a 2% agarose gel.

Afterward, the PCR products were sequenced using the same amplification primers by Macrogen (Seoul, Korea). The Contracaecum cox2 gene sequences obtained corresponded to one nematode from each host species: Andinoacara latifrons (456 bp), Astyanax sp. (597 bp), Brycon henni (602 bp), Rhamdia guatemalensis (611 bp). However, the genetic sample obtained from P. friedrichsthalii was insufficient and showed insufficient quality to be included in further phylogenetic analysis, although the morphometrical traits were identical to those of the larvae recovered from the other hosts.

One sequence from each species was assembled, trimmed, and edited using Geneious Prime 2023.2.1. To explore phylogenetic relationships, the obtained sequences were aligned with previously published cox2 data from Contracaecum sp. specimens and other species within the Anisakidae family (Supplementary Material Table 2). A sequence of Hysterothylacium deardorffoverstreetorum (JF730209) was used as an outgroup taxon, this genus has been used in previous studies for the molecular identification of anisakid nematodes (Martínez-Rojas et al., 2021). Phylogenetic reconstruction was estimated using the maximum likelihood method in IQ-TREE (Nguyen et al., 2015). The best-fit model of nucleotide substitution was TPM2u + F + I + G4. Node supports values were estimated through 10,000 ultrafast bootstrap replicates and 1000 standard nonparametric bootstrap iterations.

2.4. Data accessibility

The parasites isolated from A. latifrons, Astyanax sp., B. henni and R. guatemalensis were deposited in the Parasitology Museum Collection (PMC) at the University of Antioquia, Medellín, Colombia under ID occurrence (178-PA-FCA-UdeA). The nucleotide sequences isolated from Contracaecum reported in this study are available in the GenBank database under accession numbers PP213326, PX127524, PX127523 and PX102573, respectively.

3. Results

3.1. Morphological and morphometric identification

The collected nematodes possessed a slender and robust body tapering towards their ends, with an average length of 19.30 mm ± 7.47. A total of 20 nematodes (four from each host species) were cleared with Amman's lactophenol. All parasites exhibited the typical morphological characteristics of third-stage larvae (L3) of the genus Contracaecum (family Anisakidae), including a longitudinally striated cuticle with fine transverse striations along the body, which were more prominent in the anterior- and posterior regions (Fig. 4). The parasitic frequency was as follows: A. latifrons (28.57%; 2/7), Astyanax sp. (38.33%; 23/60), B. henni (13%; 22/170), P. friedrichsthalii (21.05%; 4/19), and R. guatemalensis (100%; 1/1). The mean intensity of infection was 3 in A. latifrons, 5.34 in Astyanax sp., and 3.59 in B. henni, 1.5 in P. friedrichsthalii, and 3 in R. guatemalensis. The abundance was 0.85 in A. latifrons, 2.5 in Astyanax sp., 0.46 in B. henni, 0.31 in P. friedrichsthalii, and 3 in R. guatemalensis.

Fig. 4.

Fig. 4

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a. Third-stage larvae (L3) of Contracaecum sp. found in Brycon hennib. L3 larvae of Contracaecum sp. located in the coelomic cavity of Astyanax sp.

At the anterior larval end, specimens had three underdeveloped lips with an excretory pore situated below the ventral cephalic tooth, a narrow esophagus measuring 2.18 mm ± 0.40 long and 0.09 mm ± 0.03 wide, ending in a small rounded ventricle measuring 0.12 mm ± 0.04 long and 0.12 mm ± 0.03 wide, with an intestinal caecum end measuring 2.18 mm ± 0.44 long and 0.130 mm ± 0.02 wide, directed towards the anterior part, a ventricular appendage measuring 0.57 mm ± 0.04 long and 0.09 mm ± 0.02 wide, a conical tail with pointed tip measuring 0.12 mm ± 0.02 long, and the presence of 2 - 4 accessory glands (Fig. 5).

Fig. 5.

Fig. 5

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Microphotographs of Contracaecum sp. third-stage larva showing internal structures and characteristic taxonomic traits. a. Nematode anterior end. l: lips, d.t: cuticular tooth, e: esophagus. b. Middle section. i: Intestinal caecum e: esophagus, v: ventricle, v.a: ventricular appendix, c. Posterior end i: intestinal caecum end. m: mucron, a: anus, a.g: accessory glands, i: intestine.

3.2. Molecular identification

The nematodes isolated from A. latifrons, Astyanax sp., B. henni and R. guatemalensis were molecularly characterized. The phylogenetic topology inferred from the analysis of the gene dataset comprising cox2 sequences is presented (Fig. 6). The BLAST comparisons were performed at the National Center for Biotechnology Information (NCBI) with the cox2 gene dataset in GenBank. The BLAST search yielded a 99.41% of identity with the sequence MT304463 isolated from fourth-stage larvae (L4) of C. jorgei, collected from a Neotropical cormorant (Nannopterum brasilianus) in Argentina (Sardella et al., 2020).

Fig. 6.

Fig. 6

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Phylogenetic position of Contracaecum sp. isolated from Brycon henni. Maximum likelihood tree generated by IQ-TREE based on COX2 gene sequences. The best-fitting substitution model was HKY + F + I + G4. Nodal supports were estimated by running 10,000 replicates and standard nonparametric Bootstrap supports (1000 repetitions).

4. Discussion

Infective anisakid L3 found in the coelomic cavity of the consumable Neotropical fish species A. latifrons, Astyanax sp., B. henni, R. guatemalensis, and P. friedrichsthalii presented morphological characteristics that allowed their classification within the genus Contracaecum (Family Anisakidae). Given the challenges in taxonomic classification of larval- and juvenile stages of anisakid nematodes using conventional morphometric methods, genetic tools have become essential to clarify the identification to species level within this genus (Garbin et al., 2023a). The sequence analysis of the mitochondrial cox2 gene has been widely employed for taxonomically identifying species of Contracaecum as reported elsewhere (Davidovich et al., 2022; Garbin et al., 2011, 2023b; Levsen et al., 2022; Mattiucci et al., 2008, 2010; Pekmezci and Yardimci, 2019). Thus, herein the cox2 gene sequence of L3 larvae found in the coelomic cavity of captured freshwater fish were molecularly and phylogenetically analyzed. These further genetic analyses not only confirmed that collected L3 belonged to the genus Contracaecum but also identified at species level as the neglected species C. jorgei. The gene sequences obtained herein (PP213326, PX127524, PX127523, and PX102573) showed a 99.41% identity with the sequence of C. jorgei (MT304463) described previously by (Sardella et al., 2020).

The anisakid C. jorgei is a recently proposed species within the genus Contracaecum described for the first time in 2020 (Sardella et al., 2020). As such, both C. jorgei-adults and fourth-stage larvae (L4) were identified infecting a Neotropical cormorant (N. brasilianus) in Argentina, as well as L3 infecting the freshwater ray-finned fish (Hoplias argentinensis) (Sardella et al., 2020). Thus, the new record of C. jorgei reported in current study expands its geographical distribution range to Colombia. This geographic expansion into previously non-endemic areas might be so possibly due to the migratory capacity of birds of the families Pelecanidae and Phalacrocoracidae (D'Amelio et al., 2012; Peña R and Quirama, 2014). The migratory behavior of the large-sized piscivorous birds such as the brown pelican (P. occidentalis) which is also present in Colombian Atlantic-as well as Pacific coasts (Chaparro-Herrera et al. 2022), might play a potential role in regional dispersal of Contracaecum within South America.

Concerning species identification, the morphology of C. jorgei adult nematodes shows similarities with other Contracaecaeum species previously reported in the Americas, such as C. margolisi, C. fagerholmi, C. mirounga and C. bioccai, due to the number and arrangement of the cephalic papillae of these adults (D’Amelio et al., 2012; Mattiucci et al., 2008; Sardella et al., 2020). Nonetheless molecular- and phylogenetic analyses indicated that C. jorgei belongs to a sister lineage of previously reported Contracaecum species has recently reported (Sardella et al., 2020) were found to be parasitized with infective C. jorgei L3 in this study. As above-mentioned, these species are commonly consumed by inhabitants of regions near rivers, streams, and reservoirs along the Porce River basin (Jimenéz-Segura and Lasso, 2020; Lasso, et al., 2011a,b; López Sánchez et al., 2018). These fish species exhibit omnivorous feeding habits, with a tendency to consume insects (acting as IH) and plants (Blanco-Cervantes and Blanco-Cervantes, 2020; Jiménez et al., 2014). Thus, the fish dietary preferences facilitate the development of C. jorgei by consuming aquatic macroinvertebrates or insects harboring L2 larvae and allowing further development into infective L3.

This is of particularly concern in relation to human anisakidosis which is still poorly understood by the general population and highly underestimated as a public health issue worldwide (Shamsi and Sheorey, 2018). The lack of awareness leads to underdiagnosis of this disease (Castellanos-Garzón et al., 2020). In South America there are some case reports on human anisakidosis from Argentina, Brazil, Chile, Colombia, Ecuador, Perú, Venezuela and Uruguay (Falla-Zuñiga et al., 2021). In 2019, the first human anisakidosis case was reported in an adult female patient from Colombia, when a gastroenteric endoscopy showed the presence of Anisakis L3 (Patiño and Olivera, 2019). Conversely, human anisakidosis due to consumption of Contracaecum L3 is not reported to date in Colombia. Nevertheless, studies have shown that Contracaecum can cause severe and painful gastrointestinal infections in humans after ingesting raw or undercooked fish carrying infected L3 of this genus (D'Amelio et al., 2012; Davidovich et al., 2022; Levsen et al., 2022).

The freshwater fish species A. latifrons (Cichlidae), Astyanax sp. (Characidae), B. henni (Characidae), R. guatemalensis (Heptapteridae), and P. friedrichsthalii (Cichlidae), represent three different families of consumable fish. Based on the molecular data, this is the first report of C. jorgei in the fish species examined here. However, the genus Contracaecum has previously been reported parasitizing Astyanax eigenmanniorum in Argentina (Mancini et al., 2014), Astyanax mexicanus in Mexico (Leal-Olvera et al., 2023), Astyanax lacustris in Brazil (Rocha et al., 2025) and R. guatemalensis in Mexico (León and García-prieto, 1989). Furthermore previous parasitological studies conducted in Colombia on these species have reported the ectoparasite Piscinoodinium sp. in Brycon henni (Londoño-Franco et al., 2016) and the genus Oligogonotylus in Andinoacara latifrons (Vélez-Sampedro et al., 2022). However, parasitological research on these fish species remains limited in Colombia, emphasizing the importance of the present study in advancing the understanding of the parasitic fauna associated with fish distributed throughout the Colombian river basins.

The results of this investigation provide novel information on the occurrence of infective C. jorgei L3 in five Neotropical freshwater fish species. These findings have significant implications for fish health, as anisakid larvae penetrate the fish stomach wall until reaching the peritoneal cavity, organs (liver, kidney, spleen), or musculature (Mehrdana et al., 2014). Migrating anisakid larvaes can trigger a systemic inflammatory reaction and consequently affecting the food absorption and growth of parasitized host fish (Buchmann and Mehrdana, 2016; Haarder et al., 2014). In addition to having implications for fish health, these findings represent a significant potential zoonotic risk for the region. Furthermore, present Contracaeum findings are highly relevant for biodiversity issues within the Porce River basin and the Magdalena river, as this parasite might have unknown implications on fish and fish-consuming birds and mammals [i. e. Amazon river dolphins (Inia geoffrensis) and otters (Lontra longicaudis and Pteronura brasiliensis)] in Colombia. Therefore, it is crucial to conduct further epizootiological research to better understand the adverse effects of C. jorgei on the fish's health, acting as suitable IH, as well as endemic wildlife animals possibly acting as either DH or AH in the parasite life cycle.

Diverse South American countries have culinary preferences including eating raw fish in various traditional preparations, including ‘ceviche’. Given the increasing popularity of raw or undercooked fish consumption, it is imperative to implement monitoring programs for zoonotic fish-borne parasitosis circulating in marine- and freshwater fish, disease prevention and control of potential spillover in local communities residing near rivers, streams, and reservoirs. We also strongly recommend implementing additional diagnostic methods for searching anisakid larvaes in consumable fish species allowing fast and adequate larval detection before human consumption. Techniques that include parasitological inspection of fish abdominal cavity and musculature, trans-illumination method (Muñoz-Caro et al., 2022), enzymatic digestion, and incubation methods are cheap, quick and easy diagnostic techniques that will allow detection of encapsulated anisakid L3 in fish specimens (McGladdery, 1986; Shamsi and Suthar, 2016). Based performing molecular analysis, we showed the presence of C. jorgei in five commonly consumed freshwater fish specimens of Colombia. Overall, these novel data are relevant not only for epidemiological-, fish health considering the annual migration routes of various South American piscivorous birds, but also to fish-eating birds and mammals, underlying the complexity of anisakid life cycles. Particularly for geographic areas with high aquatic biodiversity, as is the case for the Porce River basin in Colombia, a hydrographic basin with the highest levels of endemism among freshwater fish species (i. e. 238 endemic species) (Jiménez et al., 2014). Additionally, it is one of the basins with the highest number of threatened fish species in the country (Jimenéz-Segura and Lasso, 2020) the occurrence of C. jorgei infections calls for future investigations.

CRediT authorship contribution statement

Astrid Rave: Writing – review & editing, Writing – original draft, Visualization, Software, Methodology, Investigation, Formal analysis, Data curation, Conceptualization. Manuel Uribe: Writing – review & editing, Visualization, Supervision, Methodology. Sara López-Osorio: Writing – review & editing, Visualization, Methodology, Conceptualization. Carlos Hermosilla: Writing – review & editing, Visualization. Jenny J. Chaparro-Gutiérrez: Writing – review & editing, Visualization, Resources, Methodology, Funding acquisition.

Ethical statement

This study was conducted with the approval of the Ethics Committee for Animal Experimentation at the Universidad de Antioquia. It received ethical clearance for the project "Respuesta de la Ictiofauna a la formación de embalses en los Andes Colombianos", Act number N°138 dated February 9, 2021. Additionally, present study obtained permission from the National Authority for Environmental Licenses of Colombia [Autoridad Nacional de Licencias Ambientales (ANLA) de Colombia], for the mobilization of biological diversity specimens.

Available of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Consent to publish declaration

Not applicable.

Consent to participate declaration

Not applicable.

Funding

The author(s) declare that financial support was received for the research and/or publication of this article. The project in which the samples were collected was funded by the Convenio BIO. Framework cooperation agreement “Gestión integral del recurso hídrico, la biodiversidad y sus servicios ecosistémicos en áreas de interés de EPM,” CT: 2021-000023-A3, between Empresas Públicas de Medellín and the Universidad de Antioquia. In addition, the CIBAV Research Group—Centro de Investigaciones Básicas y Aplicadas en Veterinaria financed the study through the Strategy of sustainability 2023-2025, Faculty of Agrarian Sciences at the Universidad de Antioquia Medellin, Colombia.

Conflicts of interest

The authors declare no conflict of interest.

Acknowledgments

We would like to thank Luz Fernanda Jiménez-Segura, Daniel Restrepo-Santamaria, and Andrés F. Galeano-Moreno for their assistance with the administrative and sampling procedures. We also extend our gratitude to the ichthyology group, GIUA, at the Universidad de Antioquia for their technical and logistical support in capturing the fish. We would also like to thank Jorge E. García-Melo, and Jose Luis Londoño López Project CavFish, and the GIUA group for the fish photographs, thanks to Edwin F. Rave Espinosa for the illustration design, as well as the CIBAV Research Group—Centro de Investigaciones Básicas y Aplicadas en Veterinaria.

Footnotes

Appendix A

Supplementary data to this article can be found online at https://doi.org/10.1016/j.ijppaw.2026.101218.

Appendix A. Supplementary data

The following is the Supplementary data to this article:

Multimedia component 1
mmc1.docx (33.3KB, docx)

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Associated Data

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Supplementary Materials

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Data Availability Statement

The parasites isolated from A. latifrons, Astyanax sp., B. henni and R. guatemalensis were deposited in the Parasitology Museum Collection (PMC) at the University of Antioquia, Medellín, Colombia under ID occurrence (178-PA-FCA-UdeA). The nucleotide sequences isolated from Contracaecum reported in this study are available in the GenBank database under accession numbers PP213326, PX127524, PX127523 and PX102573, respectively.

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