Rewaida Abdel-Gaber

Morphological re-description and molecular characterization of Kudoa pagrusi (Myxosporea: Multivalvulida) infecting the heart muscles of the common sea bream fish Pagrus pagrus (Perciformes: Sparidae) from the Red Sea, Egypt

In the present study, 100 samples of different sizes of the common sea bream fish Pagrus pagrus were collected from the Egyptian water along the Gulf of Suez, Red Sea and examined for the prevalence of myxosporidian parasites in general and Kudoa spp. in particular. Fish samples were thoroughly externally examined. After dissection, all the internal organs were removed and examined. A total of 60 out of 100 fish specimens were found to be infected with Kudoa stages. Parasitic infection was restricted to the heart muscles of the examined fish. None of the other organs was found to be infected. Macroscopic cysts (plasmodia) heavily infested the different parts of the heart muscles. Each plasmodium measured 1.2–2.5 (1.53 ± 0.2) mm × 0.63–0.80 (0.65 ± 0.2) mm. Mature spores are quadratic in shape in the apical view showing four equal valves and four symmetrical polar capsules. Fresh spores were 5.0–7.1 (5.7 ± 0.2) μm long × 5.4–8.5 (6.1 ± 0.3) μm wide. On the basis of spore morphology, the present species was identified as Kudoa pagrusi. Morphometric characterization revealed that the relatively small size of this Kudoa species was the distinctive feature that separates it from all previously described species. Molecular analysis based on small subunit ribosomal DNA (SSU rDNA) sequences revealed that the highest percentage of identity was observed with K. scomberomori and followed by K. shiomitsui, K. hypoepicarclialis, K. amamiensis, and K. kenti. The kudoid spores showed morphometric variations to some extents but had essentially identical nucleotide sequences of the SSU rDNA gene sequences closest to those of K. scomberomori and K. shiomitsui recorded from elasmobranchs in the Indo-Pacific Ocean. The present findings support the identification of an ancestral marine origin of the present Kudoa species.

Morphological re-description and phylogenetic relationship of five myxosporean species of the family Myxobolidae infecting Nile tilapia

Freshwater fish have a major economic and nutritional importance worldwide. Myxosporeans are highly dangerous parasites that infect different fish species, causing severe damage to a large number of economically important species, especially in aquaculture. We conducted a survey of myxosporean parasites infecting Nile tilapia Oreochromis niloticus (Perciformes: Cichlidae) collected from different localities along the River Nile passing through Giza province, Egypt. Out of 100 fish specimens collected, 45 were found to be naturally infected with these parasites in the region of the trunk kidney. Light microscopic examination revealed the presence of 5 distinct myxosporean species belonging to 2 different genera, viz. Myxobolus and Triangula, belonging to the family Myxobolidae; all 5 species have been previously described. Morphological characteristics, host specificity and geographical distribution, tissue tropism, and molecular analysis of the partial sequence of small subunit ribosomal DNA gene revealed that the recovered myxosporean species described herein were genetically distinct from other myxozoan species but had 95% sequence similarity to M. cerebralis. Also, phylogenetic analysis placed the present myxosporean species in the freshwater Myxobolus clade, which is a sister group of freshwater Myxobolus/Henneguya species.

Morphological re-description of Electrotaenia malapteruri (Cestoda: Proteocephalidae) and Dujardinnascaris malapteruri (Nematoda: Heterocheilidae) infecting the Electric catfish Malapterurus electricus and heavy metal accumulation in host and parasites in relation to water and sediment analysis in Lake Manzala, North Delta, Egypt

Parasites are one of the most serious limiting factors in aquaculture. The Electric catfish Malapterurus electricus was subjected to study the prevalence and mean intensity of parasitic infections throughout the whole year of 2015. Heavy metals accumulation in host fish and parasites were determined in relation to water quality and sediments of two different sites of Lake Manzala (Manzala and Bahr El-Baqar), Egypt. A total of 100 specimens of Electric catfish were collected and examined for the presence of helminth parasites. Two parasite species were recovered and morphologically identified. These were cestoda Electrotaenia malapteruri and nematode Dujardinnascaris malapteruri. Heavy metal analysis in water and sediments showed that measured heavy metals in Bahr El-Baqar were found in risky levels higher than permissible limits and Manzala site. Sediments were found to contain a higher level of metals than water samples. Heavy metals accumulation in recovered parasites and their host were also determined and showed significantly higher concentrations in parasites compared to their host tissues. According to bioconcentration factors, E. malapteruri showed that highest accumulation rate for all recorded elements up to 302. Essential elements like Cu and Fe were found in significantly higher concentrations in D. malapteruri, whereas E. malapteruri accumulated elements Cd, Pb, Ni, Mn, Zn and Ca to a significantly higher degree. Accordingly, the ratios (C[D. malapteruri]/C[E. malapteruri]) for most essential elements were higher than 0.5. Therefore, fish cestodes can be regarded as useful bio-indicators more than nematodes when evaluating the environmental pollution of aquatic ecosystems by heavy metals.

Morphological, molecular and pathological appraisal of Callitetrarhynchus gracilis plerocerci (Lacistorhynchidae) infecting Atlantic little tunny (Euthynnus alletteratus) in Southeastern Mediterranean

The Atlantic little tunny, Euthynnus alletteratus, is widely distributed in temperate and tropical waters of the Atlantic Ocean, Black and Mediterranean Seas. In this study, wild-caught little tunny from Egypt, were found to be naturally infected with trypanorhyncha metacestodes, and the overall prevalence rate of infection was 38.7%. The blastocysts were either loosely attached to the mesentery of infected fish, or firmly attached and deeply embedded within the hepatic parenchyma. These encysted plerocerci are identified as Callitetrarhynchus gracilis (Trypanorhyncha, Lacistorhynchidae) based on its morphological and molecular characterization. The morphological characteristics of C. gracilis including scolex shape; the bothridia groove; the presence of frontal glands; the length of post-larval (appendix); metabasal armature; the existence of ‘Chainette’ and satellite hooks of different size were studied and described by Light and Scanning electron microscope. The phylogenetic analysis of lsrDNA gene of plerocerci confirmed the identification of the species to be deeply embedded in genus Callitetrarhynchus. The histopathological examination revealed severe pathological changes in the affected organs, including necrosis, inflammatory reactions, fibrosis and migratory tracts of the parasitic larvae together with marked visceral organs adhesions. To the best of our knowledge, this is the first report describing the detection of C. gracilis in little tunny collected from the Abu Qir landing site in Alexandria, Egypt.

Characterization of 28S rRNA sequences of cestoda parasite Electrotaenia malapteruri Fritsch, 1886 from the Electric catfish Malapterurus electricus (Siluriformes: Malapteruridae)

Proteocephalids are cestoda parasites that mostly infect freshwater fish. The present study was carried out to investigate the presence of proteocephalids infecting the electric catfish Malapterurus electricus from Lake Manzala, Kafr El-Sheikh Governorate, Egypt. Morphological characterization revealed the present parasite is a cestoda belonging to the genus Electrotaenia. Morphologically, the recovered worms were characterized by an elongated body measuring 100-127 (120 ± 2) mm long and 0.92-2.11 (2.76 ± 0.1) mm wide. The anterior part of the worm was obvious terminated at a spherical scolex measured 1.12-1.91 (1.72 ± 0.01) mm long and 1.12-1.65 (1.42 ± 0.01) mm wide with a rostellum-like apical organ equipped by 5-6 irregular rows of minute hooklets, as well as four uniloculate suckers with a diameter of 0.13-0.15 (0.14 ± 0.01) mm and covered with microtriches. A long unsegmented neck was observed followed by acraspedote and anapolytic strobila consisted of 85-120 proglottids divided into 50-58 immature, 12-19 mature, and up to 49 gravid proglottids. Molecular characterization based on 28S rRNA sequences was done to confirm the taxonomy of this parasite based on its morphology. It was observed that there was a close identity up to 72.0% with other protocephalid species obtained for comparison from the GenBank. Also, the data obtained revealed that there was high blast scores and low divergence between the present parasite and previously described Electrotaenia malapteruri (acc. no. JX477434). Phylogenetic analysis showed that the parasite sequence in conjunction with existing data investigates the placement of this protocephalid species within Proteocephalidea. It was shown that the present species is deeply embedded in the genus Electrotaenia with close relationships to other Electrotaenia malapteruri as a putative sister taxon.

First molecular characterization and morphological aspects of Sarcocystis fusiformis infecting water buffalo Bubalus bubalis in Egypt

Fresh muscle samples from water buffalo (Bubalus bubalis) aged 2–15, from Giza Province, Egypt; were examined for Sarcocystis infection. Macroscopic ovoid sarcocysts embedded in the muscle tissues of the examined buffaloes were detected; they measured 152–230 (210 ± 7) μm in length and 37–119 (95 ± 3) μm in width. The esophagus was the most infected organ followed by the diaphragm, and tongue, while the heart muscles were the least infected. The cyst cavity was compartmentalized by septa derived from the ground substance located under the primary cyst wall. Using transmission electron microscopy, the primary cyst wall bordered sarcocysts were determined to be 0.08–0.22 μm in thickness, raised from the parasitophorous vacuolar membrane, and surrounded by a secondary cyst wall of host origin. The primary cyst wall had irregular wall folds with numerous cauliflower-like projections of variable sizes and shapes accompanied by knob-like electron-dense elevations. 18S rRNA gene expression studies confirmed that the present parasite isolates belonged to the genus Sarcocystis. The sequence data showed significant identities (>90%) with archived gene sequences from many Eimeriidae organisms, and a dendogram showing the phylogenetic relationship was constructed. The most closely related species was Sarcocystis fusiformis KR186117, with an identity percentage of 98%. The recovered sequences were deposited in the GenBank under the accession number MG572125. The present study, to our knowledge, is the first collective ultrastructural and molecular study that confirmed the taxonomy of sarcocysts isolated from water buffaloes in Egypt as Sarcocystis fusiformis.

Morphological description of Cosmocerca sp. (Nematoda: Cosmocerdidae) from the Mascarene grass frog Ptychadena cf. mascareniensis (Amphibia: Ptychadenidae). A light and scanning electron microscopic studies

The Mascarene grass frog Ptychadena cf. mascareniensis is a species of frog with a vast area of distribution in Africa. A total of 300 frog specimens were collected from different localities at El-Giza province, Egypt; then dissected and examined for the presence of parasitic infection. Only eighty six (28.66%) specimens were found to be naturally infected with nematode parasite. Seasonally, the prevalence of infection was reached its maximum value of 74.66% during summer and minimum values of 26.66% (20/75), 13.33% (10/75) during spring and autumn, respectively; while no records were observed during winter season. The morphology of the recovered parasite was studied by using light and scanning electron microscopy. The adult worm characterized by anterior extremity with small mouth opening being surrounded by three lips provided with four sub-median cephalic papillae and one pair of lateral amphids. Body measurements showed that male worms were smaller than females measuring 1.22–2.43 (2.21 ± 0.1) mm in length and 0.21–0.34 (0.29 ± 0.01) mm in width. Females measured 1.9–3.7 (2.8 ± 0.1) mm in length and 0.24–0.42 (0.38 ± 0.01) mm in width. Comparing the present parasite with other species of the same genus described previously, several similarities were observed. However, peculiar new characteristics such as the arrangement of plectanes and somatic papillae, the presence of gubernaculum, the position of nerve ring, excretory pore, and vulval opening make it reasonable belongs to the family Cosmocercidae and identified as Cosmocerca sp. In addition, the present study was the first report for occurrence of cosmocercid species from the Mascarene grass frog in Egypt.

Morphological re-description and new geographical records for three digenean parasites infecting African sharptooth catfish Clarias gariepinus (Pisces: Clariidae) in Egypt

Sixty specimens of African sharptooth catfish Clarias gariepinus (Clariidae) were collected during the period of March 2014–February 2015 from boat landing sites and fishermen of the River Nile at Beni Suef Governorate, Egypt, and examined for the presence of digenean parasites. The morphology of these parasites was studied by using light microscopy to describe morphological and morphometrically measurements for different body parts. Three digenean species belonging to three different genera were collected from various organs within the examined fish species. A certain degree of site specificity was also observed, with Thaparotrema botswanensis being found only in the gall bladder of C. gariepinus, while both Pseudoholorchis clarii and Glossidium pedatum were found only in the intestine and thus seem to occupy a certain niche within their host. This study represent as the first record of T. botswanensis and G. pedatum from C. gariepinus, as well as the first report of the genera from the River Nile in Egypt. In addition, re-description of P. clarii clarifies measurements for some body parts.