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Observed Parasites in the Gills and Gut of Anabas testudineus (Puyo) from
Kabacan, Cotabato, Philippines

Preprint · October 2021

DOI: 10.13140/RG.2.2.35314.02242

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A Laboratory Report on the Observed Parasites in the Gills and Gut of
Anabas testudineus (Puyo) from Kabacan, Cotabato, Philippines

In Partial Fulfillment for the Subject

BIO 621

Advanced Parasitology

Under Prof. Elma G. Sepelagio, RMT, MAppSci.

Submitted by:

John Paul A. Catipay, LPT

INTRODUCTION

According to Aypa (1995) aquaculture is deemed as the most promising

source of food in the years to come. Aquaculture systems depend on the use of

natural waters and natural food chains. As such, they are part of the environment

and two-way interactions are numerous (Encyclopedia of Life Support Systems,

2013). In Kabacan, which is mainly located in the landlocked province of

Cotabato, freshwater ecosystems are being explored and exploited. In 2018, the

country recorded a total of 826,059 tonnes of aquaculture production (FAO,

2018), where 39.05% of this encompass freshwater aquaculture production.

Aquaculture truly is one of the rising industries in the food production sector

feeding more than a hundred million Filipinos in a daily basis.

Although an established staple in Filipino households and gourmet, we

cannot overlook the fact that these aquatic organisms serve as hosts for

endoparasites which consumers may harbor via ingestion. Fish parasitism inflicts

considerable dilemmas to the aquaculture industry and human health, which

emerged from industrialization, and both economic and ecological problems.

Anabas testudineus (pupuyo) are known species in aquaculture and

delicatessens around the world, which may harbor parasitic nematodes (Mandal,

et al., 2016). In a previous study, it was found out that, there were six species of

nematodes found in 309 samples of A. testudineus out of 547 fish hosts (Das and

Goswami ,2014). In this laboratory report, endoparasites were observed to have

been infecting the freshwater and slightly brackish water fish Anabas testudineus,

locally known as puyo or pupuyo, from Kabacan, Cotabato, Philippines.

MATERIALS AND METHODS

Ten individuals of A. testudineus was examined in this laboratory activity.

The fish were obtained from the Kabacan wet market. Further questioning of the

vendor revealed that the fish were caught in the Brgy. Cuyapon side of the

Liguasan Marsh. Prior to dissection, the fish were weighed and its body size

measured with a ruler. Dissection was done using proper tools.

First, the operculum was examined for any attached parasites. This was done

through visual inspection, and parasites that were present were manually

extracted using forceps and blades.

Samples of the gills were cut and placed in glass slides for microscopic

examination. The fish were then cut open to obtain the gastrointestinal tract.

Dissection of the gut was done in clean petri dishes and was examined through

the aid of a stereomicroscope.

Extracted trematodes from the operculum of the fish were placed in glass

slides and flattened. These were stored in alcohol prior to staining.

Extracted nematodes from the gut of the fish were also placed in glass slides

and were examined morphologically with the aid of photomicroscopy.

RESULTS AND DISCUSSION

Six (8) out of the 10 fish specimen were positive for parasites. This makes up

80% of the total samples examined.

Through a thorough examination of the gut of A. testudineus, one nematode

species was obtained. Camallanus sp. was extracted from the small intestine of

the fish. A total of 3 out of the 6 positive fish samples were found to harbor

Camallanus sp. Specimens were placed on clean glass slides and were observed

using photomicroscopy (Figure 1). A total of 5 Camallanus sp. Individuals were

isolated from the three (3) positive specimen.

Figure 1. A screen grabbed photograph of the photomicroscopy of Camallanus sp.

obtained from A. testudineus
 Aside from Camallanus sp., 5 trematodes were also obtained from the

operculum of the fish. However, due to unforeseen circumstances, these

trematodes were not stained and properly identified. Microscopy was done last

May 2020, one year after the specimens were preserved. The trematode

specimens were dried out.

Species of the genus Camallanus is quite prevalent on fresh water fishes,

especially Camallanus cotti (Kim, Hayward, & Heo, 2002). Camallanus sp. was

also reported on littoral fish of Lake Taal (Capuchino et al., 2017).

Implications of Fish Parasite Infection

Parasites are significant group of pathogens that cause infection and

diseases for both freshwater and marine environments. Aquaculture parasitic

infection has drawn concerns with their effect on fish health management and

aquatic crop production worldwide. Hence, proper attention must be given to this

concern to maintain a sustainable aquaculture production (Chandra, 2006).

There is a need to have more researches on the economic impact of aquatic

animal diseases. Current researches show that socioeconomic impact includes

decrease in production, employment, income, market share, consumer

confidence, investment and even closure of the business. These concerns

contributed to the significance of fish diseases, especially in disease research

and control investments along with health management programs (Idowu,

Adedeji & Sogbesan, 2007).

 Parasites are found everywhere. They occur among fishes. Fishes harbor

ectoparasites and endoparasites which can cause different pathological diseases

in them. Parasites thrive in them because they form a close symbiotic relationship

with fishes by obtaining nutrition at the expense of the host's growth while

allowing the fish to live. Common parasites include the unicellular microparasites

such as viruses, bacteria, protozoans, and fungi. Multicellular parasites are listed

as fish parasites such as helminths and arthropods (Iyaji & Eyo, 2008).

Protozoans are a large paraphyletic group of organisms that commonly

parasitizes fishes. They include the flagellates, amoebae or rhizopodes,

alveolates, cercozoans and ciliates. They have simple life cycles and their

transmission is between and within hosts through contact. Others use vectors or

intermediate hosts. They can mass infect fishes causing weight loss, debilitation,

and mortality. However, majority are not of zoonotic concern. They are identified

based on their morphology of their different life stages (Austin & Newaj-Fyzul,

2017; Iyaji & Eyo, 2008).

Myxozoans are metazoans and classified as highly specialized cnidarian

parasites. Their life cycles alternate between vertebrate and invertebrate hosts in

aquatic environments. They are classified into two classes the myxosporea and

malacosporea. They are found as hosts in tropical, temperate, and polar regions.

They are not normally a human health concern but there were studies showing

myxozoans in stool samples. There were also cases where they cause food

poisoning through the consumption of raw olive flounder and allergic reactions

due to consumption of a fish infected with Kudoa sp. They are identified through
spore morphology, tissue tropism, clinical signs and host specify. (Austin &

Newaj-Fyzul, 2017; Patra et al., 2018).

Some fishes are infected with mesomycetozoea, fungi and fungal-like

organisms. These organisms belong to phyla Microsporidia, Ascomycota,

Oomycota and Chytridiomycota. They show host specificity but it varies

according to species. They were reported to infect all major animal groups but

those infecting aquacultured species are not detrimental human health. They are

classified based on morphological features. Further, molecular tools and

biochemical methods can be used on cell wall structure and presence of

pigments (Austin & Newaj-Fyzul, 2017; Gozlan et al., 2014).

Monogeans are mostly parasites of fish, frogs, and other aquatic animals in

both freshwater and marine habitats. They are significant ectoparasites of fish

which show relatively high host specificity. They are found on the skin and gills or

buccal cavity of the fish. They have a single definitive host in their life cycles.

There is no recorded monogean species that poses a threat on human health.

They are morphologically identified based on their opisthaptor and copulatory

apparatus (Austin & Newaj-Fyzul, 2017; Öztürk & Özer, 2014).

Digenea is considered as the largest endoparasitic metazoan group which

includes more than 18,000 nominal species. They are endoparasitic flatworms

infecting the alimentary canal and their associated organs. They have complex

life cycles which can include fish as the intermediate or definitive host. There are

more than 200 digenean species that are harmful to human health. They include
Schistosoma spp., Clonorchis sinensis, Paragonimus spp. and Opisthorchis

viverrini. They are identified based on their size, suckers and other body organs

(Orélis-Ribeiro, Arias, Halanych, Cribb & Bullard, 2014).

Cestodes or tapeworms are endoparasitic organisms that have a flattened

body which has an adhesive scolex, segmented neck and strobilum with a

number of hermaphroditic proglottids. They all occur in the digestive tracts except

Nesolecithus africanus which infects in the coelomatic cavity of the fish,

Gymnachus niloticus and Polyonchobothrium clarias in the gallbladder of Clarias

mossambicus. They have a heteroxenous life cycle. They pose a risk to human

health. Human infections are brought by the plerocercoid larvae through the

consumption of raw fish products. The most common freshwater cestode

parasites are Diphyllobothrium latum and D. dendriticum. They are

morphologically identified through the scolex, proglottids, and egg morphometry

(Austin & Newaj-Fyzul, 2017; Iyaji & Eyo, 2008).

Nematodes are widespread parasites both infecting wild and farmed fishes.

Adult nematodes can live within the intestinal lumen of the host fish while others

reside within the gonads. They damage the host by feeding on the host blood,

sera and tissue. Moreover, they cause mechanical damage by fixing to host

tissues, developing or migrating in them. They have a complex life cycle which

involves a fish either as an intermediate or definitive host. Some nematodes are

of human health concern. Common nematode parasites are the marine anisakid

nematodes belonging to Contracaecum, Pseudoterranova and Anisakis. Humans

are infected through the consumption of raw fish products. They can be identified
through their morphological characters such as the cephalic structures, papillae,

excretory pores, digestive system and other organs (Austin & Newaj-Fyzul, 2017;

Iyaji & Eyo, 2008; Kundu, Bandyopadhyay, Mandal & Gürelli, 2016).

Acanthocephalans are endoparasitic helminths commonly infecting both

marine and freshwater fishes worldwide. Their life cycles involve arthropods as

intermediate hosts and vertebrates as their definitive or paratenic hosts. They are

easily recognized by the presence of their evaginable proboscis crowned with

hooks. They are rarely reported as causing infection in human and such cases

are because of accidental infection. They are morphologically identified based on

their spines on their trunk, hooks on their proboscis and other structures.

Molecular methods are also used (Austin & Newaj-Fyzul, 2017; Iyaji & Eyo, 2008;

Sanil, Asokan, Lijo & Vijayan, 2011).

Arthropoda is a species-rich phylum with a complex taxonomy. Parasitic

stages of copepods are found on the external surfaces of their hosts. They can

cause damage to the host by their activities such as burrowing, attaching and

feeding on their host. In freshwater habitats, the most significant pathogens are

Salmincola salmoneus, Lernaea cyprinacea, Ergasilus sieboldi and Argulus

foliaceus. They are identified based on the morphology of both the adult male and

female arthropods (Austin & Newaj-Fyzul, 2017).

Management and Control of Fish Parasite Infections

Fish development programs depend mainly on fish parasite researches.

Parasites mainly helminths take fishes as their hosts. Fishes serve as carriers of
larval parasitic forms that can mature and cause diseases among vertebrates

specially man aside from their role as hosts of different parasites. Parasitic

infections among fishes contribute to weight loss, decreased growth rate,

emaciation, spread of diseases, inhibition of fish sexual maturity and mortality

(Chandra, 2006).

Mechanical damage includes the fusion of gill lamellae. Many parasites infect

the gills of fish causing mild or massive discoloration, numerous white spots and

mucus secretions. They can also occupy majority of the total area of an organ.

Physiological damage is observed in cell proliferation where parasites are

associated with bile duct proliferation in the liver. Parasites might act as agents in

fish carcinogenesis although this needs more evidence. Some parasites use the

host immune system in attaching to the fish host. In addition, they can also alter

their host's behavior for example trematode Euhaplorchis californiensis can lead

the California killifish (Fundulus parvipinnis) to swim slowly towards the water

surface to be eaten by birds which are their definitive hosts (Iwanowicz, 2011).

Early and fast diagnosis of fish parasites is important in controlling parasitic

diseases. Traditional diagnosis depends mainly on naked eye remaking or wet

mount examination using a light microscope. However, the biggest disadvantage

of this method depends mainly on the experience of the expert who is not always

accurate. As a result, new methods are being introduced for fish parasite

diagnosis. These are more accurate, safer and faster to manage parasitic

diseases in wild and cultured fishes. These include light microscope to examine

histopathological slides stained with histochemical stains. Tiny details of specific

parasites are observed using electron microscopes. Antigen-antibody reactions

are tested through immunological tests. Other new trends in diagnosis include

agar gel precipitation test, agglutination test, Enzyme Linked Immunosorbent

Assay (ELISA), latex agglutination test and fluorescent antibody test. In addition,

molecular techniques are also being employed which depend on DNA

sequencing of the parasite. Molecular diagnosis include Polymerase Chain

Reaction (PCR), Multiplex PCR, restriction enzyme digestion, DNA microarrays,

Loop Mediated Isothermal Amplification (LAMP) and Nested PCR. However,

conventional methods remain the standard way for the identification of the

parasites although non-conventional methods are slowly being applied in recent

studies (Noor El Deen, Zaki & Fawzi, 2018).

Fish health management are designated to prevent the propagation of fish

diseases. The goals are to prevent the introduction of diseases to healthy fishes,

inhibit the spread of existing disease agents, intensify the natural fish resistance

and yield high quality fishes. Most reported fish diseases are associated with

stress, hence, prevention practices should focus on good water quality, sanitation

and nutrition. These practices are useless if there is no farmer-oriented health

management programs. These include support on capacity building of farmers,

farm attendants, aqua culturist, fisheries officers, laboratory staff and

veterinarians on diagnostic methods and effective remedial action and public

awareness. Establishment of diagnostic centers is also highly recommended

(Idowu, Adedeji & Sogbesan, 2007).

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