EC CLINICAL AND EXPERIMENTAL ANATOMY Research Article Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study

Abstract

The aim of the current study was to estimate the metacercarial infection in Ruby-Red-Fin Shark (Rainbow Shark), Epalzeorhynchos frenatum (Teleostei: Cyprinidae). Samples of fish were commercially purchased from an ornamental shop in Assuit city, Egypt and brought to the laboratory. Fish samples were used for light histological analysis. All fish measured and deeply anesthetized with benzocaine (4 mg/L). Fish were ranging from 10 to 12 cm in standard body length. Fish were deeply anesthetized with benzocaine (4 mg/L). Using Histological examination, we identified a heterophyid parasite metacercaria; Ascocotyle (Ascocotyle sp.). The metacercarial cyst was located inside the cartilaginous support of the gill arches which were surrounded by Thick fibrous perichondrium. Cartilage enlargement occurs as a result of invaginations of the perichondrium into the cartilage mass and kl. proliferation of the chondrocytes progeny. As well as hyperplasia, hypertrophy, and fusion were present in the affected gill filaments. Erosion of the cartilage was reported with formation of tunnels in the cartilage. Increase rodlet cells in the branchial epithelium and in the underlying connective tissue. The disappearance of the gill lamellae could affect respiratory efficiency. Lymphatic spaces were rich in numerous Lymphatic cells. Poor controls of diseases permit the transport of parasites between different countries and increase the prevalence

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OPEN ACCESS EC CLINICAL AND EXPERIMENTAL ANATOMY
Research Article
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of
Ruby-Red-Fin Shark (Rainbow Shark) Epalzeorhynchos frenatum (Teleostei:
Cyprinidae): Light Microscopic Study
Sara Abdelhakiem1, Soha A Soliman2, Hanan H Abd-Elhafeez3*, Enas Abdel- Hafez4 and Rania S Zaki5
1Lecturer, Department of Zoology, Faculty of Science, Assuit University, Egypt
2Asistant Professor, Department of Histology, Faculty of Veterinary Medicine, South Valley, Qena, Egypt
3Assistant Professor, Department of Anatomy, Embryology and Histology, Faculty of Veterinary Medicine, Assuit University, Egypt
4Professor, Department of Anatomy, Embryology and Histology, Faculty of Veterinary Medicine, Assuit University, Egypt
5Lecturer, Department of Meat Hygiene, Faculty of Veterinary Medicine, New Valley University, Egypt
Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
*Corresponding Author: Hanan H Abd-Elhafeez, Assistant Professor, Department of Anatomy, Embryology and Histology, Faculty of
Veterinary Medicine, Assuit University, Egypt.
Received: May 07, 2019; Published: August 17, 2019
Introduction
Trematodes of the family Heterophyidae are widely distributed in different piscivorous [1]. Ascocotylinae have been described as a
subfamily of heterophyid species which include three subgenera, Ascocotyle, Phagicola and Leighia [2]. The metacercaria of the genus As-
cocotyle   
  
-
Abstract
The aim of the current study was to estimate the metacercarial infection in Ruby-Red-Fin Shark (Rainbow Shark), Epalzeorhyn-
chos frenatum

benzocaine (4 mg/L). Fish were ranging from 10 to 12 cm in standard body length. Fish were deeply anesthetized with benzocaine
Ascocotyle (Ascocotyle sp.). The meta-

Cartilage enlargement occurs as a result of invaginations of the perichondrium into the cartilage mass and kl. proliferation of the
-
tilage was reported with formation of tunnels in the cartilage. Increase rodlet cells in the branchial epithelium and in the underlying

Lymphatic cells. Poor controls of diseases permit the transport of parasites between different countries and increase the prevalence
of infections.
Keywords: Ascocotyle; Ruby-Red Fin Shark; Chondrocytes; Proliferation

Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
297
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
lid and hydrobiid snails in which eggs are developed to cercaria that leaves and swims out of the snail. The second intermediate hosts

The cercaria of Ascocotyle-
-

gills to infection is considerable due to their external location contacted to water medium.
-

Epalzeorhynchos frenatum is commonly known

Materials and Methods
 
from each shop) in Assiut City, Egypt and brought to the laboratory. The standard length body for each sample was measured from the top
of snout to caudal peduncle. Fish were deeply anesthetized with benzocaine (4 mg/L) for histological analysis.
Histological investigations

      
Hoffman [11] keys. The histological sections were stained with Haematoxylin and eosin [12]. Combined Alcian blue Ph 2.5 and PAS stain
was used to differentiate between neutral and acidic polysaccharides. All staining was cited in Bancroft’s theory and practice of histologi-
cal [13]. Stained sections were examined by Leitz Dialux 20 Microscope. Photos were taken using a Canon digital camera (Canon Power-
shot A95).
Preparations of resin embedding samples

processed according to description in paper of Abd-Elhafeez and Soliman [9].
Results
    Ascocotyle (Ascocotyle
sp.) Price (1935). Metacercarial cysts were located inside the cartilaginous support of the gill arches which were surrounded by thick

The perichondrium of the cartilage was enlargement as a result of the proliferation of the chondrocytes progeny (Figure 1C). Cysts

Erosion of the cartilage was reported with formation of tunnels in the cartilage (Figure 1D and 1E). As well as hyperplasia (Figures 2A

   
were observed in the connective tissues and in the lymph space between the epithelial cells (Figures 2C, 2H and 2I).

Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.

Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
We demonstrated a Large number of granular stages of rodlet cells using the combined Alcian blue pH 2.5 and periodic acid sheiff



Figure 1: Metacercarial cyst in gill arch.



Cartilage enlargement occurs as a result of invaginations (arrow) of the perichondrium into the cartilage mass and proliferation of
the chondrocytes progeny (arrowheads). D: Cysts pyriform shaped (pyriform body, arrowhead) surrounded by thin-walled (thin wall,

anterior margin of ventral sucker (vs) and excretory bladder (bladder, arrowhead) Y –shaped with short arms. E and F: invaginations of


Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
299
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
Figure 2: Changes of tissue responding to Metacercarial cyst.

of gill arches showing hyperplasia (double arrowheads), Increase number of granular stages of rodlet cells (blue arrows) and a low

Hypertrophy of pavement cells (pv, arrowheads). F: a Large number of transitional stage of rodlet cells (arrowheads) and extravasation



massive invasion of lymphocytes (arrowheads). Note: In I the congestion of blood vessels (double arrowheads).

Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
300
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
Figures 3:



Note, Increased granular stages of rodlet cells (blue arrows, a mix of blue and magenta stained) and mucous cells blue
stained (Acid mucous) double arrowheads, (C) Example of infected gill and (D) healthy gill demonstrating the difference in
numbers of granular stages of rodlet cells (blue arrows). Note the mucous cells with positive staining to alcian blue pH2.5



Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
301
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
Discussion

and algae [15]. The occurrence of Ascocotyle (Ascocotyle Sp.) metacercaria encysted in the musculature of the gill arches is remarkable.
The present study revealed that the occurrence of heterophyid species of Ascocotyle (Ascocotyle sp.) in the gills of ruby shark, Epalzeo-
rhynchos frenatum. These results agreed with Salgado-Maldonado., et al. [16] who found metacercariae of Ascocotyle (Ascocotyle sp.) in

The distribution of Ascocotyle         
Ascocotyle
rates from one study to another is related to local conditions which endure the parasite life cycle. Salem., et al. [17] reported that Asco-
cotyle
-

Ascocotyle
among Egyptian dogs (Asmaa., et al. 2016), poor sanitary conditions and disposal of waste in water bodies permit the transmission of
     


Ascocotyle species [5], Despite these parasites did
not cause acute disease, but in long-term, they appear economical loss due to damaging of gills with metacercariae. Lesions of the gills as
a result of the presence of parasitic infection were varied with the agent, host, and density of infection (Robert 2001). The structure of the
normal gill was demonstrated by many authors [24,25].
In the current study, gills respond to the parasitic infestation by attracting and activation of several types of immune cells including
lymphocytes and rodlet cells. A considerable number of the lymphocytes were observed in the lymph space between the epithelial cells
and in the underlying connective tissue. Granular stages of rodlet cells have been observed as demonstrated by Abd-Elhafeez and Soliman
[9]. The predominance of the acidic polysaccharides stained by alcian blue and diminishing of the neutral polysaccharides is remarkable
features of infestation by the metacercarial cyst. This may indicate that the acidic components affect the stability of the metacercarial
 Fried and Johnson [26] who described that breaching of the metacercarial cyst occurs in the acid-pepsin
medium. The infection of the gills in the present study with trematode metacercariae caused hyperplasia, hypertrophy, and a fusion of gill

-

Ascocotyle (Ascocotyle
as heart, gill arches and mesentery [29]. Ascocotyle metacercariae were also reported encystment in different organs as gills, heart, kidney
and muscle [30].
-
    


Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
302
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
Conclusion
       

Authors’ Contributions
Abdel-Hakeem and Abdel-Hafez designed and coordinated the study, shared in sampling and wrote the manuscript. Soliman assisted
in work, histological analysis, manuscript writing and data analysis, Zaki participated in samples collection, data analysis and revising the


Competing Interests
The authors declare that they have no competing interests.
Availability of Data and Materials
All the data regarding this study are demonstrated in the manuscript.
Consent for Publication
Not applicable.
Ethics Approval
The method of consent and the animal work were approved by the ethics committee of Assiut University, Egypt.
Funding
This research did not receive any research fund.
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Citation: Hanan H Abd-Elhafeez., et al. “Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark
(Rainbow Shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study”. EC Clinical and Experimental Anatomy 2.7
(2019): 296-304.
303
Occurrence of Metacercarial Cyst of Ascocotyle (Ascocotyle Sp.) in the Gills of Ruby-Red-Fin Shark (Rainbow Shark)
Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
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Epalzeorhynchos frenatum (Teleostei: Cyprinidae): Light Microscopic Study
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