33 Zoonotic infections in soil of recreational areas Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 Neotropical Helminthology Neotropical Helminthology, 2024, vol. 18 (1), 33-46 ORIGINAL ARTICLE / ARTÍCULO ORIGINAL PREVALENCE OF ANISAKIDS IN SARDINELLA AURITA AND MUGIL CUREMA FROM SUCRE STATE, VENEZUELAPREVALENCIA DE ANISÁKIDOS EN SARDINELLA AURITA Y MUGIL CUREMA DEL ESTADO SUCRE, VENEZUELA Erika Gomez-Martinez 1,3 , Del Valle Guilarte 2 , Zulay Simoni-Gonzalez 1 , Marielcip Bolivar 1 , Eurimar Rodriguez 1 ,Abdul Abner Lugo-Jimenez 4 , Grafe Oliveira- Pontes 6 *, Zeca Manuel Salimo 3,5 & Victor Irungu-Mwangi 3 ISSN Versión Impresa 2218-6425 ISSN Versión Electrónica 1995-1403 DOI: https://dx.doi.org/10.62429/rnh20241811721 Este artículo es publicado por la revista Neotropical Helminthology de la Facultad de Ciencias Naturales y Matemática, Universidad Nacional Federico Villarreal, Lima, Perú auspiciado por la Asociación Peruana de Helmintología e Invertebrados Af nes (APHIA). Este es un artículo de acceso abierto, distribuido bajo los términos de la licencia Creative Commons Atribución 4.0 Internacional (CC BY 4.0) [https:// creativecommons.org/licenses/by/4.0/deed.es] que permite el uso, distribución y reproducción en cualquier medio, siempre que la obra original sea debidamente citada de su fuente original. ABSTRACT Nematodes of the Anisakidae family were studied in Sardinella aurita Valenciennes, 1847 and Mugil curema Valenciennes, 1836, caught in Venezuelan coastal waters and sold in Cumaná’s main market in Sucre state, Venezuela. Fifty f sh of each species were analyzed, examined, and dissected and their internal cavities, cephalic organs, abdominal trunk, and caudal regions were inspected. T ey were also debrided, and muscles dissected in search of nematodes in their third larval stage (L 3 ) through simple visual examination. Detected nematode parasites were mechanically extracted from the organs 1 Instituto de Investigaciones em Biomedicina y Ciencias Aplicadas Dra. Susan Tai, Universidad de Oriente IIBCAUDO. Av. Universidad, CEP: 6101 Cumaná-Sucre, Venezuela. 2 Universidad de Oriente, Departamento de Bioanalisis Escuelas de Ciencia Núcleo de Sucre. Av. Universidad, CEP: 6101 Cumaná-Sucre, Venezuela. 3 Programa de Pós-Graduação em Medicina Tropical, Universidade do Estado do Amazonas, Av. Pedro Teixeira, s/n, Dom Pedro, 69040-000 Manaus-AM, Brasil. 4 Instituto Especializado de Estudios Superiores Loyola, San Cristobal, CEP: 91000 República Dominicana. 5 Faculdade de Ciências de Saúde, Universidade Lúrio, Nampula 364, CEP: 3100 Moçambique. 6 Fundação de Medicina Tropical Doutor. Heitor Vieira Dourado, Av. Pedro Teixeira, s/n, Dom Pedro, 69040-000, Manaus--AM, Brasil.* Corresponding author: grafeoliveira@gmail.comErika Gomez-Martinez: https://orcid.org/0000-0002-2467-8831Del Valle Guilarte: https://orcid.org/0009-0006-3000-2463Zulay Simoni-Gonzalez: https://orcid.org/0000-0002-5237-8238Marielcip Bolivar: https://orcid.org/0009-0005-4153-9285Eurimar Rodriguez: https://orcid.org/0009-0006-2763-3053Abdul Abner Lugo-Jimenez: https://orcid.org/0000-0002-7667-1260Grafe Oliveira-Pontes: https://orcid.org/0000-0003-0755-1086Zeca Manuel Salimo: https://orcid.org/0000-0002-0834-8728 Victor Irungu-Mwangi: https://orcid.org/0000-0002-2618-3816
34 Gomez-Martinez et al. Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 and tissues for identifcation and classifcation at the genus level. M. curema was the only fsh found to be infected with parasites. Distinct parasitic species were observed in 48/50 (96%) of the M. curema specimens analyzed. Te prevalence of helminths in these specimens included nematodes of the Anisakidae family, Acanthocephala, and other unidentifed parasites. Of the total specimens examined, the parasites were distributed as follows: Anisakidae family (66%) (33/50), Pseudoterranova sp. (44%) (22/50), Anisakis sp. (32%) (16/50) and Contracaecum sp. (12%) (6/50). Tese parasites were mainly located encysted in the caudal kidney (55.09%), the liver (28.81%), and the digestive viscera (inside the gut) (16.10%). Tis shows the existing human risk of acquiring anisakidosis larvae parasitism, depending mainly on the cultural habits of consuming raw or undercooked fsh. Keywords: Acanthocephalia – Anisakidae nematodes – Endoparasite – Saltwater fsh – Public health – Venezuela RESUMEN Se estudiaron nematodos de la familia Anisakidae en Sardinella aurita Valenciennes, 1847 y Mugil curema Valenciennes, 1836, capturados en aguas costeras de Venezuela y vendidos en el mercado principal de Cumaná en el estado Sucre, Venezuela. Se analizaron cincuenta peces de cada especie, se examinaron, diseccionaron y se inspeccionaron sus cavidades internas, órganos cefálicos, tronco abdominal y regiones caudales. También se desollaron y se diseccionaron los músculos en busca de nematodos en su tercer estadio larvario (L 3 ) mediante simple examen visual. Los parásitos nematodos detectados se extrajeron mecánicamente de los órganos y tejidos para su identifcación y clasifcación a nivel de género. M. curema fue el único pez encontrado infectado con parásitos. Se observaron distintas especies parasitarias en 48/50 (96%) de los especímenes de M. curema analizados. La prevalencia de helmintos en estos especímenes incluyó nematodos de la familia Anisakidae, Acanthocephala y otros parásitos no identifcados. Del total de especímenes examinados, los parásitos se distribuyeron de la siguiente manera: la familia Anisakidae (66%) (33/50), Pseudoterranova sp. (44%) (22/50), Anisakis sp. (32%) (16/50), y Contracaecum sp. (12%) (6/50). Estos parásitos se localizaron principalmente enquistados en el riñón caudal (55,09%), el hígado (28,81%) y las vísceras digestivas (dentro del intestino) (16,10%). Esto muestra el riesgo humano existente de adquirir larvas de anisakidosis, dependiendo principalmente de los hábitos culturales de consumir pescado crudo o poco cocido. Palabras clave: Acanthocephalia – Anisakidae nematodos – Endopárasitos – Peces de agua salada – Salud pública – Venezuela
35 Anisakids in Sardinella aurita and Mugil curema Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 INTRODUCTION In some countries, fsh is prominent source of animal protein in the diet. Specialists recommend it for its high nutritional value, easily digestible proteins, and rich content of minerals, vitamins, and polyunsaturated fatty acids that beneft the body, However, despite its many benefts, fsh carries a high risk of parasite transmission and cany (Gea, 2015; Rodríguez-Monje, 2019).Certain parasites can infect fsh and fshery products, afecting their muscles, skin, fns, and internal organs. However, only a few of these parasites pose health risks to humans. Sociocultural and behavioral factors, such as consuming raw or undercooked fsh, contribute to human infection. Tis can lead to ichthyozoonosis, a disease transmitted to humans by bacteria, viruses, and parasites through the ingestion of contaminated fsh and fshery products (Quijada et al ., 2005; Tejada & López, 2012). Te growing popularity of eating raw or uncooked seafood in western countries has resulted in the proliferation of certain parasitic infections in humans. One of the most severe fsh-transmitted infections is anisakiasis, which occurs when third-stage larvae belonging to the family Anisakidae are accidentally ingested (Rosales et al ., 1999).In Latin America, the high consumption of fsh and fshery products contributes to reported outbreaks of foodborne diseases (FBD), depending on the type of food involved (Balbachán et al ., 2000). In Venezuela, during the period from 1996 to 2004, the ingestion of fsh second and third leading cause of FBD (Maniscalchi et al ., 2015). While most ichthyoparasitoses do not cause pathologies in humans, some species can lead to serious disorders when infected fsh are consumed, posing a signifcant health risk that should not be underestimated (Puccio et al ., 2008; Tejada & López, 2012; Castellanos-Garzón et al ., 2019).In 1974, Anderson et al . described anisakids as belonging to the phylum Nemathelmintes, class Nematoda. Among the diferent genera are Anisakis , Pseudoterranova , Contracaecum and Hysterothylacium are notable (Anderson et al., 1974). Te species that stands out as the main infecting nematode due to its frequency and pathogenic action is Anisakis simplex , the etiological agent of the disease known as anisakiasis (Berruezo, 2000; Rello et al ., 2004; Akbar & Ghosh, 2005; Tuemmers et al ., 2014; Gea, 2015). Clinical signs and symptoms of anisakidosis include allergies and gastrointestinal symptoms. Gastric manifestations usually appear 12-48 hours after consuming fsh infected with the L 3 larval stage, which can be mistaken for gastritis. Intestinal manifestations are characterized by intense pain in the lower abdomen, accompanied by nausea, vomiting and occult blood in the feces. Allergic manifestations, on the other hand, occur a few hours after consuming infected fsh and present as pruritus, urticaria, asthma, and in some cases, anaphylaxis (Field-Cortazares & Calderón-Campos, 2009; Farias et al ., 2021).Human anisakidosis was frst reported in Japan in 1965 and later in Spain (Arenal et al ., 1991). Te number of reported cases of this parasitosis has increased since then. However, it is important to note that it remains an under-diagnosed condition, as it is often associated with exotic countries and cuisines (Rello et al ., 2004).In Latin America, cases of this parasitosis have been documented in Chile, Peru and Brazil. In Peru, eight cases were described, of which fve were confrmed to be caused by Anisakidae. In Chile, four cases were diagnosed, with two of them caused by the species Pseudoterranova dicipiens . Furher analysis revealed that the same species was responsible for seven cases in Chile. In Brazil, 85 fsh were randomly sampled from Colares Island and Vigia, Pará, using net fshing. It was found that 80% and 76% of the sampled fsh were were parasitized, particlularly the silver croacker ( Plagioscion squamosissimus Heckel, 1840), kumakuma ( Brachyplatystoma flamentosum Lichtenstein, 1819), and gilded catfsh ( Brachyplatystoma rousseauxii Castelnau, 1855) being the most parasitized species (Quijada et al ., 2005; Rodrigues et al ., 2015; da Silva et al ., 2021). In Venezuela, it has been demonstrated that anisakids infest marine mammals such as manatees, otters and dolphins, as well as numerous piscivorous birds in the diferent marshes, inlets, lakes, lagoons and deltas along the Venezuelan coast (Windevohxel, 2003). Anisakid infections have also been confrmed in fsh intended for mass consumption, in Caracas Capital District (Bandes et al ., 2005), Nueva Esparta state (Puccio et al ., 2008), the Médano Blanco coastal strip, Falcón state (Bracho-Espinoza et al ., 2013), and the north-eastern and insular coastal region of Venezuela (Maniscalchi et al ., 2015). Tese infections were found in fsh species of the Mugilidae family. Furthermore, in the Gulf of Paria, on the eastern coast of Venezuela, has reported the presence of Anisakis spp L 3 larvae in diferent varieties of croaker (corvine) fsh, specifcally Cynoscion virescens Cuvier, 1830 and Cynoscion microlepidotus Cuvier, 1830 species (Botto, 1981). Te State of Sucre is a fshing region where fsh such as sardines and mullet are consumed daily. In recent years,
36 Gomez-Martinez et al. Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 there has beena shift in the region’s consumption habits, with an increased consumption of preparations such as anchovies, sushi, ceviche and raw or undercooked canned f sh. Additionally, the sale of preferably uneviscerated f sh is another factor to consider regarding sanitary risks that can contribute to the presence of parasites in f shery products. Anisakidosis could become a public health problem in this area, as the local economy and food industry revolve around f shing. T is study aims to explore, document and analyze the presence of Anisakidae larvae and Acanthocephala in Sardinella aurita Valenciennes, 1847 and Mugil curema Valenciennes 1836, which are commonly consumed by the local population in Cumaná, Sucre State, Venezuela. T e study also highlights the potential risk of human anisakidosis associated with the consumption of these species. MATERIALS AND METHODS T e present study was carried out in the city of Cumaná, Sucre municipality, Sucre state, Venezuela. It involved the dif erent f sh outlets of the city’s main market, where var-ious varieties of marine species from artisanal shore and draft trawling are sold in the coastal area of the Gulf of Cariaco as shown in (Fig. 1). T e Gulf of Cariaco is lo-cated south of the Araya Peninsula, in the state of Sucre. Figure 1. Geographic location of the Gulf of Cariaco, Sucre municipality, Sucre state, Venezuela. Source: 10,51316° N, 63,97916° O, Google Maps. In the period of June-September 2019, a total of 100 specimens of f sh were processed: 50 of the S. aurita (pilchard) and 50 of the M. curema (mullet), which were acquired at dif erent f sh outlets in the main Cumaná market. T e samples were selected considering the following parameters: clearness of the eyes, presence of gills, presence of viscera, weight, size and f shing time. T ey were preserved in ice and then transferred to the laboratory at the Instituto de Investigaciones em Biomedicina y Ciencias Aplicadas Dra. Susan Tai, Universidad de Oriente (IIBCAUDO) to be evaluated using a previously established criterion (Cervigón, 1993; Osanz, 2001). For the parasitological analysis of the dif erent f sh species, the methods described by the United States Food and Drug Administration (USFDA, 2022), the Bacteriological Analytical Manual (BAM), the Center for Food Safety and Applied Nutrition (CFSAN) and the Canadian Health Canada OPFLP-02 laboratory procedures were used (Bier et al ., 2001).T e f sh were dissected and a simple visual examination of the organs was done to check for parasites. T ey were then placed into petri dishes with a 0.85% NaCl solution to be further examined under a stereoscopic microscope.T e collected nematodes underwent several processing steps including washing with a 0.85% NaCl solution, then f xation, and clarif cation followed by preservation in 70% glycerinated ethanol. T e Acanthocephalans were f xed, clarif ed, stained then preserved. T e taxonom-ic identif cation was done in accordance with the usual techniques in helminthology. Morphologic analysis was conducted using a Zeiss Axioscope light microscope and
37 Anisakids in Sardinella aurita and Mugil curema Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 mounted samples were photographed using the Zeiss Ax-ioscope light microscope. To calculate the parasitic indices of prevalence, mean intensity in each type of helminth, mean abundance of each genus of the family Anisakidae and Acanthoceph-ala mean intensity and mean abundance were done as described by Bush et al . (1997), and Morales & Pino (1995). T e number of parasites found and the degree of parasitic infestation was determined as described by Verján et al . (2001). Ethic aspects: T e authors point out that the study mate-rial - Sardinella aurita and Mugil curema - were acquired in a municipal market (Cumaná market), Venezuela for direct sale to the local population, not alive and without any type of interference by the researchers regarding to the way of f shing, interference in the behavior or in vivo manipulation of the specimens. We highlight that the anonymity of the vendors and f sh stalls in the Cumaná market was preserved. RESULTS Only the species M. curema were found to be parasit-ized with nematode larvae of the Anisakidae family and Acanthocephala, showing an overall prevalence of 88% and 58% respectively. T e hosts were each parasitized by at least one species of parasite. A total of 118 Ani-sakidae L 3 larvae were collected, including 37 Anisakis sp., 74 Pseudoterranova sp. and 7 of Contracaecum sp. Additionally, 33 Acanthocephala specimens were col-lected. In total, 5915 third-stage nematode larvae (L 3 ) were collected, with 5,666 being Anisakidae larvae and 249 being Acanthocephala larvae in the processed M. curema specimens. It is important to note that the simple visualization meth-od does not for the observation of newly formed larvae that may be encysted within the muscles, so the data de-scribed here may be underestimated.In this study, the Anisakidae family was found to para-sitize the digestive viscera ( Anisakis sp. 29.72%, Pseudo-terranova sp. 8.10% and Contracaecum sp. 28.57%), kid-neys ( Anisakis sp. 32.43%, Pseudoterranova sp. 68.91% and Contracaecum sp. 28.57%) and liver. ( Anisakis sp. 37.83%, Pseudoterranova sp. 22.97% Contracaecum sp. 42.86%). Acanthocephalans were found to parasitize the intestine (Fig. 2). Morphological characteristics of both Anisakidae and Acanthocephala were identif ed (Fig. 3). T ey were observed as immature larvae, whitish or pink-ish in color (pearly), with rounded unsegmented bodies measuring approximately 2.5 mm in diameter and be-tween 1 and 5 cm in length. T ese larvae were arranged in a spiral shape, and can be found in digestive viscera. T is is the f rst study on the presence of Anisakidae in Sucre state, Sucre municipality, which is of great zoonotic importance. In the Anisakidae family, the main species of public health importance are Anisakis simplex , Pseudoter-ranova decipiens and Contracaecum osculatum . Figure 2. Specimens of Anisakidae L 3 larvae in caudal kidney (a, b), intestine (c) and Acanthocephalans in intestine (d) collected from Mugil curema .
38 Gomez-Martinez et al. Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 T e Anisakidae family nematodes found in the M. cure-ma species were identif ed at the genus level. T ese nem-atodes were examined a light microscope to determine their morphological characteristics such as size, color, presence or absence of a perforating tooth and mucron, location of the excretory pore opening, and characteristics of the digestive system as previously outlined. Among the Anisakidae nematodes, Pseudoterranova sp. showed the Figure 3. Helminths of the family Anisakidae mechanically extracted from digestive viscera, kidneys and liver and Acantho-cephala extracted from digestive viscera, present in Mugil curema . (a) L 3 larvae of the family Anisakidae, (b) and (c) L 3 larvae of the family Anisakidae with cyst membrane, (d) and (e) Petri dish with L 3 larvae of the family Anisakidae and Acanthoceph-alans. Parasites from the Aniakidae family highlighted with red arrows. highest parasitic indices in terms of prevalence, mean in-tensity and mean abundance (Fig. 4). T is was followed by Anisakis sp. (Fig. 5), while Contracaecum sp. exhibited the lowest parasitic indices (Fig. 6). Overall, this study determined a prevalence of Anisakis sp. in 32% (16/50), Pseudoterranova sp. in 44% (22/50), and Contracaecum sp. in 12% (6/50) of the f sh examined (Table 1). Table 1. Parasitic indices of prevalence, mean intensity (MI) and mean abundance (MA) of Anisakidae, Acanthocephala and unidentif ed helminths collected from Mugil curema , from the Gulf of Cariaco, Vanezuela. GenusPrevalence (%) (MI)(MA)Anisakidae66 Anisakis 322.310.74 Pseudoterranova 443.361.48 Contracaecum 121.160.14Acanthocephala58--Unidentif ed helminths 22--
39 Anisakids in Sardinella aurita and Mugil curema Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 ct ep dl nr nr ic es es v ic a m Figure 4 . Pseudoterranova sp. L 3 larvae mechanically extracted from the digestive viscera, kidneys and liver, isolated from Mugil curema (a) ephalic end (ct: cuticular tooth, ep: excretory pore, dl: dorsal lip, nr: nerve ring) (b) ephalic end (nr: nerve ring, ic: intestinal cecum, es: esophagus) (c) Anterior end (es: esophagus, v: ventricle, ic: intestinal cecum) (d) Posterior end (a: anus, m: mucron). a es es m ct ep nr dl m Figure 5 . Anisakis sp. L3 larvae extracted mechanically from the Mugil curema (a) ephalic end (ct: cuticular tooth, ep: excretory pore, dl: dorsal lip, nr: nerve ring) (b, c) Anterior end (es: esophagus) (d) Posterior end (a: anus, m: mucron).
40 Gomez-Martinez et al. Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 Adult Acanthocephala specimens were identif ed as hav-ing a coloration that varied between cream, yellow and brown. T ey have rounded bodies with a proboscis or a i es m ct ep dl es ci ci ci nr es v va ga Figure 6 . Contracaecum sp. L 3 larvae extracted mechanically from the collected from Mugil curema (a, b) ephalic end (ct: cutic-ular tooth, ep: excretory pore, dl: dorsal lip, nr: nerve ring, es: esophagus, ic: intestinal cecum) (c) Anterior end (es: esophagus, v: ventriculus, ic: intestinal cecum, va: ventricular appendage, i: intestine) (d) Posterior end (a: anus, ga: ganglia, m: mucron). Figure 7. Acanthocephala specimens collected from Mugil curema (a-c) cephalic end showing proboscis and neck, the arrows show characteristics spines of the proboscis (d-e) Reproductive apparatus showing (f-g) Posterior end (h) Acanthocephalan eggs, arrows. invaginable tube at the cephalic end that is covered in characteristic spines. T e size of these specimens ranged from 2mm to 1-meter-long (Fig. 7).
41 Anisakids in Sardinella aurita and Mugil curema Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 Te tissue tropism of parasites in M. curema was found to be similar in all the specimens analyzed. Te parasites were primarily located in the kidneys (caudal kidney), with a higher parasite load of 55.09%. Tey were also present in the liver (28.81%) and other digestive viscera, such as the mesentery, stomach, pancreas, gallbladder and spleen (16.10%). It is interesting to note that despite the assumption that larvae in the digestive viscera provide greater development and migration, the parasite load in the kidneys was higher.Te parasitic coinfections between nematodes of the Ani-sakidae family, demonstrated the variety of parasites that can simultaneously infect fsh. We observed that 69.70% of the fsh analyzed had a monoparasitic infection ( Pseudoterranova sp.). Rates of biparasitism ( Anisakis sp/ Contracaecom sp) and triparasitism ( Pseudoterranova sp. / Anisakis sp./ Contracaecom sp.) were 27.27% and 3.03% respectively. Additionally, the parasitic coinfections be-tween Anisakidae and Acanthocephala were 100%. DISCUSSION Only the species M. curema was found to be parasitized with nematode larvae from the Anisakidae family, with an overall prevalence of 96%. Tis result is of great zoo-notic importance, as there have been several cases of Ani-sakiasis reported worldwide (Adams et al ., 1997). In Eu-rope, herring ( Clupea harengus Linnaeus, 1758), ancho-vy ( Engraulis encrasicolus Linnaeus, 1758) and sardines ( Sardina pilchardus Walbaum, 1792) are the fsh species most commonly associated with cases of anisakiosis (Au-dicana et al ., 2002; Audicana & Kennedy, 2008; Klimpel & Palm, 2011). Likewise, adult Acanthocephala worms were identifed and although they are not clinically signif-icant for humans, they are of great veterinary importance in various fsh species, and can also infect turtles, reptiles, and pigs (Drago, 2017).Tese results are consistent with those obtained previous studies conducted in markets alongs the northeastern coastal and insular regions of Venezuela (Maniscalchi et al ., 2015), the Médano Blanco coastal strip in Falcón State (Bracho-Espinoza et al ., 2013), Nueva Esparta State (Puccio et al ., 2008), and Caracas, the capital of Venezu-ela (Bandes et al ., 2005). Tese studies analyzed diferent species of fsh popular for consumption and found a pre-dominance of Anisakidae in the Mugilidae family (genus Mugil ). Unlike previous studies that showed a high in-fection rate of Anisakidae nematodes in M. curema spe-cies in the Gulf of Paria, in far eastern Venezuela. Botto (1981) also demonstrated the presence of Anisakis spp L 3 larvae in diferent varieties of corvine, specifcally in the Cynoscion virescens Cuvier 1830, and Cynoscion microlepi-dotus Cuvier 1830 species, further highlighting the risk of this parasitic infection in other commonly consumed fsh species. Mugil curema is abundant, along all coasts of Venezuela and is capable of adapting and subsisting on a wide va-riety of foods by adjusting its feeding habits according to envir on mental conditions (Cervigón, 1993; Franco & Bashirullah, 1992; Novoa, 2000; Guerra & Marín, 2002 ). It is worth noting that the M . curema species was also found to be parasitized by Acanthocephala helminths, with an infection rate of 58% in the fsh studied. Another study conducted in Lima, Peru, focusing on parasitesin marine fsh intended for human consumption, observed that the Mugil cephalus had an 86.70% infection rate with Ac-anthocephala, but did not fnd Anisakidae larvae in the specimens analyzed (Serrano et al ., 2017).Te processed M. curema species showed a parasitism rate of 66%. Tese results are similar to those reported by Maniscalchi et al . (2015) in a study conducted in the northeastern coastal and insular region of Venezue-la, where 90 out of 143 (62.94%) specimens of the M. curema species tested positive for nematodes from the Anisakidae family. Botto (1981) demonstrated in the Gulf of Paria that the genus Anisakis was prevalent in corvine species. Likewise, studies carried out along the Venezuelan coasts (Bandes et al ., 2005), in Caracas (Puccio et al ., 2008), in Nueva Es-parta state (Maniscalchi et al ., 2015), in the northeastern and insular Coastal Region of Venezuela, and the present study show a high prevalence of Anisakidae larvae in fsh species of the Mugilidae family, with prevalence rates of 76%, 100%, 62, 94% and 66% respectively (Franco & Bashirullah, 1992; Novoa, 2000). Live larvae has been found mainly in the viscera or encysted in the muscula-ture of a number of commercially important fsh species.Te infectious larval stages of Anisakis simplex and Pseudoterranova decipiens have been implicated in most cases of human anisakiasis. Several other species, such as Contracaecum osculatum Rudolphi, 1802 and Phocasca-ris spp . , also pose a public health concern (Adams et al ., 1997). Similar fndings were made in this study, where Anisakis sp. and Pseudoterranova sp. were the most prev-alent genera of nematodes. Consequently, they represent a possible source of infection of Anisakidae nematodes, constituting a public health risk to consumers. Tis risk can be preventable if the appropriate sanitary measures
42 Gomez-Martinez et al. Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 are applied. Moreover, it is important to mention that humans are paratenic hosts, as the larvae are unable to reach sexual maturity (López-Serrano et al ., 2000). Puc-cio et al . (2008) and, Maniscalchi et al . (2015) on the coasts of the northeastern and insular region of Vene-zuela obtained similar results. In these studies, larvae of nematodes of the Anisakidae family were found in fsh of the genus Mugil , with a higher prevalence for the genus Anisakis at 52% and 47.16%, respectively, followed by Pseudoterranova at 32% and 40.10%, and Contracaecum with 16% and 12.74%. Similarly, fsh of the genus Mugil from the Venezuelan coast have been observed to be in-fected with Anisakidae family with Pseudoterranova spp having the highest prevalence (56%), followed by Contra-caecum (48%) and Anisakis (16%) genera (Bandes et al ., 2005). Furthermore, a study carried out in Fálcon state revealed a high prevalence of anisakids, particularly of the genus Contracaecum (97%) with Pseudoterranova spp account-ing for only 3%, but no Anisakis (Bracho-Espinoza et al ., 2013). Shih & Jeng (2002) reported that in Golfete de Coro, specifcally in the Médano Blanco sector, hosted numerous migratory birds and reptiles that are defnitive hosts, of Contracaecum spp. parasite. Tis highlights that this area was conducive for the survival of nematodes from the Anisakidae family. Our results allow us to in-fer that if Contracaecum sp larvae are present in the fsh evaluated, adult parasites are certainly present in species of birds or reptiles from the place where the study was carried out.Globalization, cultural and gastronomy exchange have led to an increase in reported cases of anisakidosis world-wide, particularly in countries like Spain, Japan and Mex-ico. Tis poseses a potential risk to human health due to the rise in zoonotic parasitosis, which has become a signifcant public health concern in these countries (Chai et al ., 2005; Hochberg et al ., 2010). Venezuela is not im-mune to globalization, and the custom of consuming raw or seasoned fsh such as ceviches has made the fndings from our study a signifcant public health risk for the country.Our fndings on Pseudoterranova sp. mean intensity of infection and mean abundance were similar to those obtained by Bandes et al . (2005) and Maniscalchi et al . (2015), who reported 13.1 ± 5.2 and 2.26 ± 1.42 parasites per unit, respectively. Tese values are considered high when compared to the parameters set by the USFDA (2022). Tis highlights the risk of anisakid infection in humans in Sucre State, Venezuela if raw or undercooked fsh is consumed.Te high mean intensity and prevalence of parasites demonstrate the colonization ability of these three gen-era of Anisakidae in the population of M. curema. Tis categorizes mullet as a potential intermediate host in the biological cycle of Anisakidae, particularly Pseudoterrano-va sp. which has the highest prevalence and mean abun-dance, followed by Anisakis sp. and Contracaecum sp. On the contrary, studies conducted in Colombia found that Contracaecum spp. was the most prevalent, occurring in 100% of the fsh species analyzed, such as Salminus afn-is Steindachner, 1880, Hoplias malabaricus Bloch, 1794, and Pseudoplatystoma corruscans Spix & Agassiz, 1829 (Pardo et al ., 2007, 2008, 2009).Anatomically, the kidneys in the Mugilidae family fsh are attached to and run along vertebral column, covered by a conjunctival membrane. Tis suggests that the lar-vae have greater access to the vertebral column from the muscles adjacent to it, posing a higher risk of infection for humans due to possible larval migration towards the fsh musculature. Alternatively, it is hypothesized that the larvae’s tropism towards the kidney serves as a protective mechanism, preventing migration to the muscles. Tis may explain why no Anisakidae species were found in the musculature of the fsh under study, which difers from other studies that found only one specimen of Anisakidae in the musculature of M. curema (Bandes et al ., 2005; Maniscalchi et al ., 2015). Larval migrations in the mullet and blue grenadier fsh species are not well documented, so understanding this is essential for assessing the potential risk of consuming marine fsh and the need to quickly eviscerate the fsh to prevent larvae from migrating to the muscle tissues (Bandes et al ., 2005). Our fndings confrm the presence Anisakidae species, their ability to migrate, and their tro-pism for fsh muscle, as evidenced by the parasites found in close proximity to the muscles just a few hours after the fsh was purchased at the local municipal market. Further studies could evaluate the the long-term prefer-ence or tropism of these parasites. Parasites are consumers and therefore compete for re-sources. When multiple species of nematodes infect a host; there can be competition for nutrients, which is vital for parasite growth and reproduction. Tis is the case with Acanthocephala (intestinal helminths) where evidence suggests these species displace other worms (both adult and larval) from the best sites in the intestine, where nutrients are abundant (Silva-Melo et al ., 2021). In this study, all Acanthocephala isolated from M. curema were found in the intestines.
43 Anisakids in Sardinella aurita and Mugil curema Neotropical Helminthology, Vol. 18, Nº1, jan - jun 2024 Tis present describes the co-infection of nematodes from Anisakidae family, demonstrating the variety of parasites that simultaneously infect fsh. Trough our literature review, we found no published studies presenting simi-lar fndings on mixed parasitism. Tus, this study is the frst to reveal the presence of multiple infections by these three nematodes within the same fsh specimen, collected from the Gulf of Cariaco coast in Venezuela. In the coastal areas of Venezuela’s Sucre states, the Ani-sakidae L 3 larvae are present in M. curema fsh, indicat-ing a risk of infestation for residents who consume these fsh. It is also possible thst other fsh species inhabiting the Gulf of Cariaco may be similarly infested with these nematodes. We recommend conducting further research focusing on economically important fsh and other fsh consumed by humans to determine the extent of infesta-tion by these helminths.It is very important to note that identifying anisakid spe-cies solely on morphology is not reliable or efcient. Mo-lecular techniques are currently used to clarify the genetic and ecological diferences among anisakids, despite their morphological similarities (Mattiucci & Nascetti, 2008). Future studies should use molecular methods to identify and estimate the diferent Anisakid species in fsh from the coastal region of Venezuela.Te state of Sucre is rich in fsh, which is a primary pro-tein source for the area and a major economic pillar for the municipal market in Cumaná and surrounding areas. Te lack of sanitary control, combined with our fndings, is a signifcant public health concern. Additionally, the customs in the region could contribute to the spread of anisakidosis in coastal settlements and markets in Sucre, Venezuela. Tis study provides data on a food-borne par-asitic disease and ofers insights to prevent human infec-tions. Author contributions : CRediT (Contributor Roles Taxonomy)EGM = Erika Gomez-Martinez DVG = Del Valle Guilarte ZSG = Zulay Simoni-Gonzalez MB = Marielcip Bolivar ER = Eurimar Rodriguez AALJ = Abdul Abner Lugo-Jimenez GOP = Grafe Oliveira-Pontes ZMS = Zeca Manuel Salimo VIM = Victor Irungu-Mwangi Conceptualization : EGM, GOP, ZSG, DVG; Data curation : GOP, AAJL, ZMS, VIM; Formal Analysis : EGM, DVG, ZSG, GOP; Funding acquisition : EGM, AALJ, DVG; Investigation : EGM, DVG, ZSG, MB, ER; Methodology : EGM, DVG, ZSG, MB, ER; Project administration : EGM, DVG, ZSG; Resources : EGM, AALJ, DVG, ZSG; Software : EGM, DVG, ZSG, MB, ER; Supervision : EGM, ZMS, AALJ, GOP, VIM; Validation : EGM, GOP, ZSG; Visualization : EGM, GOP, VIM, ZMS; Writing – original draft : EGM, AALJ, GOP, VIM Writing – review & editing : EGM, GOP, VIM, AALJ, ZMS ACKNOWLEDGMENTS Tis work was fnanced by the Universidad de Oriente and the Consejo de Investigación, through the approval of the research work called: Evaluation of parasites of the Anisakidae family, producers of allergic symptoms in hu-mans, present in fsh for mass consumption, in diferent municipalities of the Sucre state, Venezuela, codigo CI-02-040400-1965-17. Tis work conclused em february, 2022. BIBLIOGRAPHIC REFERENCES Adams, A.M., Murrell, K.D., & Cross, J.H. (1997). Parasites of fsh and risks to public health. Revue Scientifque et Technique (International Ofce of Epizootics), 16, 652-660. Akbar, A., & Ghosh, S. (2005). Anisakiasis—a neglected diagnosis in the West. Digestive and Liver Disease, 37 , 7-9.
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