Objective: The present research was to identify the prevalence of Anisakis sp on Epinephelus sp in the waters of East Nusa Tenggara, East Indonesia.
Materials and methods: A total of 50 fish of Epinephelus sp were collected at Kupang Fish Market, East Nusa Tenggara. Identification of Anisakis sp with morphologically Observed at the shape of ventriculus, boring tooth, and mucron in a binocular microscope. The anisakids morphologically identified were counted to estimate the prevalence.
Results: Prevalence of Anisakis sp in Epinephelus sp is 22%. Organ that become predilection Anisakis sp are abdominal cavity, gonad, intestines, and muscles.
Conclusion: This study suggest that the prevalence of Anisakis sp in the Epinephelus sp can raise awareness of the potential for foodborne zoonoses due to Anisakis sp and gastro-allergic condition of this parasite.
Keywords: Anisakis sp, East Nusa Tenggara, Epinephelus sp
Anisakiasis is fish-borne parasitic zoonoses for the most part to populations living development Country (Ivanovic et al., 2015). Anisakiasis is an emerging zoonosis caused by Anisakis sp, one of the fish parasitic (Audiciana and Kennedy, 2008). The number of annual anisakiasis cases is estimated to be high (Bao et al., 2017). Species Anisakis pathogens are Anisakis simplex and Anisakis pegreffii (Mattiucci et al., 2013; Cipriani et al. 2016). For allergy, the only implicated parasite in fishery products is the nematode Anisakis simplex (Ivanovic et al., 2015). This disease is always associated with foodborne and very important in the globalization of the seafood industry (Šimat et al., 2015). The importance of disease can cause early symptoms, sensitization to allergic and chronic conditions in humans (Baird et al., 2014).
The main source of infection in humans because it consumes raw fish containing larvae Anisakis sp (Acha and Szyfres, 2003). Anisakis simplex can cause severe hypersensitivity reactions and true anaphylactic reactions can also occur exposure to allergens from dead worms by food-borne, airborne, or skin contact routes (Audicana and Kennedy, 2008). Anisakis can cause Gastrointestinal manifestations and abdominal pain (Vaughan et al., 2015) as well as adhesive intestinal obstruction and Asymptomatic colonic disorders (Takamizawa and Kobayashi, 2015; Tamai and Kobayashi, 2015). Several species of fish reported infected with Anisakis sp include in Auxis thazar, Rastrelliger kanagurta, Katsuwonus pelamis from East Indonesia Waters (Uga et al. 1996, Anshary et al. 2014; Hibur et al. 2016). One of the most numerous fish in Indonesian waters is Epinephelus sp (Jefri et al. 2015). The objective of our study was to identify the prevalence of Anisakis sp on Epinephelus sp in the waters of East Nusa Tenggara.
MATERIALS AND METHODS
A total of 50 fish of Epinephelus sp were collected at Kupang Fish Market, East Nusa Tenggara. The fish were placed in individual bags, labeled and stored in an icebox for transfer to the Parasitology Laboratory at the Faculty of Veterinary Medicine, University of Nusa Cendana and at the Fish Quarantine Station, Ministry Of Marine And Fisheries, Republic Of Indonesia, in Kupang (East Nusa Tenggara, Indonesia). The fish were examined externally to identify the presence of nematode parasites. They were then sectioned using scissors and tweezers and an internal organ revision was carried out under a stereoscopic microscope. The nematodes found were then extracted and stored in Eppendorf tubes with 70% alcohol until identification. The offal or parts of the organs in the examined fish are muscle, liver, spleen, and intestine was collected. The organ is placed in a petri dish and given a physiological solution and then examined the presence of parasitic infection Anisakis sp.
Identification of Anisakis sp with morphologically Observed at the shape of ventriculus, boring tooth, and mucron in a binocular microscope. According to Shiraki in 1974, that for the genus Anisakis, larval morphology classification is divided into Type I or Type II with specific characteristics (D’Amelio et al., 2010). To differentiate between the genera of the Anisakidae family, the following morphological characteristics were noted: excretory pore position, and presence, extent, and position of the intestinal cecum and the ventricular appendix (Fukuda et al., 1988). The anisakids morphologically identified were counted and the prevalence was calculated following the methodology of Castellanos et al., (2017).
RESULTS AND DISCUSSION
Morphological parasites Anisakis sp.
Based on the results of research on Epinephelus sp, Anisakis larva type I and type II larva larvae were found. Anisakis larvae type 1 has the characteristic that is boring tooth, ventriculus, posterior there is mucron located at the tail end (Figure 1). The larvae showed a posterior end with a conical termination with absence of mucron, characteristic of Anisakis Type II larvae (Figure 2).
Anisakis nematode parasite is divided into two types namely type I and type II. The most distinguishing morphological difference is that type 1 has a mucron while type 2 does not have a mucron (Castellanos et al., 2017). The boring tooth function is found on the anterior part to perforate the intestinal wall. Boring tooth also serves as a larval defense tool for holding onto the mucosa of the small intestine when there is intestinal contraction in digesting food (Murata et al., 2011). The third stage has boring tooth, ventriculus, posteriorly there is a mucron located on the posterior. Excretory pore serves as a discharge hole located between the ventrolateral lips, while the esophagus serves as a link between the mouth with the ventriculus. Ventriculus is a part that is directly related to the intestine and characterizes the Anisakis sp. And other nematodes (Fukuda et al., 1988).
The posterior part of Anisakis sp. Larvae has anus and mucron located at the posterior end. The results obtained in Epinephelus sp. Also found the existence of type II Anisakis larvae with a total of 5 larvae. Anisakis larvae type II has a characteristic that is boring tooth, ventriculus, and posterior part does not have mucron (Mattiucci and Nascetti, 2006). This result is supported by the statement of (Castellanos et al., 2017) which suggests that larva type Anisakis I have mucron while cirri Anisakis type 2 has no mucron at the posterior end and has a shorter and wider ventriculus than Anisakis type I.
Prevalence of Anisakis sp in Epinephelus sp
Larvae of anisakid nematodes were identified parasitizing Epinephallus sp as larvae of infective stage L3 of Anisakis on 11 of 50 Epinephallus sp. The results showed that The Prevalence of Anisakis larvae in Epinephelus sp was 22%. Of the 11 samples found 22 larvae L3 with details of 17 larval anisakis in the abdominal cavity, 1 anisakis larvae on Gonad, 6 larvae anisakis on the intestines, and 3 larvae anisakis on muscles (Figure 3).
Based on the results of the study there are several predilection organs that become the infection of Anisakis larvae in the abdominal, intestinal, gonad and muscle cavities. Anisakis spill predilection organ sp influenced by the convenience factor for Anisakis sp (Mladineo et al., 2016). The risk of zoonoses by parasites from fish usually enters the human body in the form of a third-stage larva that enters with the fish eaten. Humans can be infected if swallowing larvae in the third stage when consuming raw or undercooked fish (Acha and Szyfres, 2003).
On a life cycle, an adult nematode form of Anisakis sp can be found in the lumen of the marine mammal gut, which subsequently lays eggs and is excreted through faeces (Chai et al., 2005), the floating eggs become the larval stage of L1 and L2. The L2 larvae are then ingested by small crustaceans, in which the L3 stage is reached. Infected crustaceans are then ingested in fish, where L3 larvae are trapped in their digestive tract. The life cycle of this nematode is completed when the fish is infected swallowed by marine mammals (L4), and further into adult forms (Castellanos et al., 2017). In the human, body larvae will live and the larvae penetrate the intestinal mucosal tissue. Cases of infection are generally symptoms of Anisakis sp. Subclinical but larval when the larvae of life can come out with vomiting or in the feces. This study adds data on the incidence of zoonotic diseases in eastern Indonesia, particularly anisakis prevalence data as in some reports on parasite study in fish in Indonesia in several fish species (Hadidjaja et al., 1978; Anshary et al., 2014; Detha et al., 2015).
Prevalence of Anisakis sp in Epinephelus sp is 22%. Organ that become predilection Anisakis sp are abdominal cavity, gonad, intestines, and muscles.
Thanks to the Faculty of Veterinary Medicine, University of Nusa Cendana and Directorate General of Higher Education of the Republic of Indonesia who has provided financial support for this research.
CONFLICT OF INTEREST
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
AD, DAW, JA, YR, CM perpetrated the prevalence of Anisakis sp in Epinephilus sp. PRN also drafted the manuscript. YR and KM were involved in writing up of manuscript with PRN, DAW and JA as well as refined English of the draft. Finally, the manuscript was read and commissioned by all authors.