Anthelmintic and therapeutic effects of the biogenic zinc oxide nanoparticles against acute kidney injury induced by Parascaris equorum Infection in rats.

Complications of parasite infections, especially kidney disease, have been linked to poorer outcomes. Acute kidney damage, glomerulonephritis, and tubular dysfunction are the most prevalent renal consequences of Parascaris equorum infection. The purpose of this study was to determine the pharmacological effects of green-produced zinc oxide nanoparticles (ZnO NPs) on P. equorum infection in male Wistar rats. Thirty-six male rats were divided into two groups of 18 each: infected and non-infected. Both groups were separated into three subgroups, each of which received distilled water, 30 mg/kg ZnO NPs, and 60 mg/kg ZnO NPs. After 10 days of ZnO NPs administration, four larvae per gram of kidney tissue were present in the untreated infected group. While, no larvae were present in ZnO NPs (30 mg/kg) treated group, and one larva/g.tissue was present in ZnO NPs (60 mg/kg) treated group compared to untreated infected animals. P. equorum infected rats had increased kidney biomarkers (creatinine, urea, uric acid), malondialdehyde, and nitric oxide, with a significant decrease in their antioxidant systems. On the other hand, infected treated rats with green-produced zinc oxide nanoparticles had a substantial drop in creatinine, urea, uric acid, malondialdehyde, and nitric oxide, as well as a significant rise in their antioxidant systems. P. equorum infection in rats caused severe degenerative and necrotic renal tissues. On the other hand, there were no detectable histopathological alterations in rats treated with ZnO NPs (30, 60 mg/kg) as compared to the infected untreated animals. When compared to infected untreated mice, immunohistochemical examination of nuclear factor-kappa B showed a significant decrease during treatment with ZnO NPs (30, 60 mg/kg). Green-produced zinc oxide nanoparticles are a viable therapeutic strategy for Parascaris equorum infection due to their potent anthelmintic activity, including a significant decrease in larval burden in infected treated rats.

Correction: Anthelmintic and therapeutic effects of the biogenic zinc oxide nanoparticles against acute kidney injury induced by Parascaris equorum Infection in rats.

[This corrects the article DOI: 10.1007/s12639-023-01637-z.].

Anthelmintic and Hepatoprotective Activities of the Green-Synthesized Zinc Oxide Nanoparticles Against Parascaris equorum Infection in Rats.

MAIN CONCLUSIONS: Green-synthesized zinc oxide nanoparticle is a promising treatment modality against parasitic infection through its powerful anthelmintic, antioxidant, healing promotion, and anti-inflammation effects. BACKGROUND: Nanoparticles have many properties, depending on their size, shape, and morphology, allowing them to interact with microorganisms, plants, and animals. OBJECTIVES: Investigation of the therapeutic effects of green-synthesized zinc oxide nanoparticles (ZnO NPs) on Parascaris equorum infection in rats. METHODS: Thirty-six rats were divided into two divisions: the first division is noninfected groups were allocated into three groups. Group 1: Control, group 2: ZnO NPs (30 mg/kg), and group 3: ZnO NPs (60 mg/kg). The second division is infected groups were allocated into three groups. Group 1: vehicle, group 2: ZnO NPs (30 mg/kg), and group 3: ZnO NPs (60 mg/kg). FINDINGS: Ten days post-infection, two larvae per gram of liver tissue were present in the vehicle group compared to the control group. No larvae were recovered from ZnO NPs (30 mg/kg), and one larva/g.tissue from ZnO NPs (60 mg/kg)-treated groups compared to untreated infected animals. Green-synthesized ZnO NPs caused a significant decrease in liver functions, low-density lipoprotein (LDL), cholesterol, triglycerides, malondialdehyde (MDA), and nitric oxide (NO). While it caused a significant increase in hemoglobin (HB), high-density lipoprotein (HDL), butyrylcholinesterase (BCHE), glutathione (GSH), catalase (CAT), and glutathione S-transferase (GST) in infected treated rats. The histological inflammation and fibroplasia scores showed a significant enhancement during the treatment with ZnO NPs (30, 60 mg/kg) compared to the infected untreated animals that scored the highest pathological destruction score. Immunohistochemical markers of NF-κB showed a significant decrease during the treatment with ZnO NPs (30, 60 mg/kg) compared to the infected untreated animals.

Morphological and molecular identification of third-stage larvae of Anisakis typica (Nematoda: Anisakidae) from Red Sea coral trout, Plectropomus areolatus.

Anisakidosis is a foodborne zoonotic infection induced by members of the family Anisakidae via the consumption of raw or undercooked fish such as sushi and sashimi. Identifying anisakid larval species is critical for the epidemiology and diagnosis of diseases caused by them. This study aimed at identifying Anisakis larvae collected from marine fish in Egyptian waters based on morphological characteristics and molecular analysis. Thirty marine fish coral trout, Plectropomus areolatus, were collected from Hurghada, Red Sea, Egypt, to investigate larval nematodes of the genus Anisakis. The larvae were detected encapsulated in the peritoneal cavity and muscle of the fish host. This examination revealed that anisakid larvae naturally infected 19 fish specimens with a prevalence of 63.33% and a mean intensity of 4.1 ± 0.40. Most of them (68 larvae: 71.57%) were found in the musculature. Morphological and morphometric analyses using light and scanning electron microscopy revealed a head region with a prominent boring tooth, inconspicuous lips, and a characteristic protruded cylindrical mucron. All larvae in this study possessed the same morphology as Anisakis Larval type I. Molecular analysis based on ITS region using maximum likelihood and Bayesian phylogenetic methods confirmed them as Anisakis typica. This is the first study to identify A. typica larvae from the commercial fish coral trout P. areolatus in Egyptian waters using morphological and molecular methods.

Pharyngodon mamillatus (Nematoda: Pharyngodonidae) in the ocellated skink Chalcides ocellatus (Squamata: Scincidae) from Egypt: Light and scanning electron microscopic studies.

Twenty-one Pharyngodon mamillatus were recovered from the large intestine of 4 out of 12 ocellated skink Chalcides ocellatus collected from the South Sinai desert, Egypt. The prevalence of infection was 33.33% (intensity 2-4). The recovered pharyngodonid was re-described based on light and scanning electron microscopy. According to the morphology, it was characterized by regular transverse annulations extending from the posterior margin of the lips to the level of the anus, triangular mouth opening was surrounded by three large bilobed lips possessing three papillae, oval-shaped excretory pore, slit-like vulva opening, and slit-like anal ventral opening at the posterior end which terminates with a tail filament. Here, the morphometric measurements of adult female P. mamillatus were given in this study. It has no intraspecific morphological variations from the previously described P. mamillatus, but it appeared with small range on biometric features. These variations may be accounted by host/environment attributes since they exhibit wide host and geographical spectra. The present study was applied for detailed morphological knowledge of the recovered P. mamillatus for strengthening their taxonomic validity. HIGHLIGHTS: The Pharyngodon mamillatus naturally parasite of the intestinal region of Chalcides ocellatus. The present study clarifies the morphological key for the description of P. mamillatus.

Immunotoxical, neurotoxical, histopathological and immunohistopathological alterations of Nerium oleander and Tecoma stans methanolic extract on Biomphalaria alexandrina snails.

Schistosomiasis is a severe illness that caused socioeconomic problems. The present study aimed to investigate the molluscicidal activities of the methanolic extract of Nerium oleander and Tecoma stans on B. alexandrina snails. The present results showed that N. oleander had the higher molluscicidal effect (LC50: 138.6 mg/l) than T. stans methanolic extract (LC50: 256.0 mg/l). These concentrations had no mortality effects on Daphnia magna during the first 12 h of the exposure, while, they had a cercaricidal activity. Exposure of B. alexandrina snails to the sub lethal concentrations (LC10 and LC25) of the methanolic extract of either N. oleander or T. stans caused a concentration- dependent significant decrease in their mean total number of hemocyte and hyalinocytes percent, while, both the round small and the granulocytes were increased than the control group. Exposure of B. alexandrina snails to LC25 of the methanolic extract of N. oleander or T. stans, caused morphological alterations in the hemocytes that were studied by both light and electron microscopy. The sub lethal concentration (LC25) significantly decreased the acetyl cholinesterase activities, acid and alkaline phosphatase levels and the protein content. Histopathological changes occurred in the digestive and the hermaphrodite glands of exposed B. alexandrina snails to LC25 of the methanolic extracts. These alterations were confirmed by Immunohistochemistry for PCNA and Cyclin D1 expressions. Conclusively, these plants could be used to decrease the spread of schistosomiasis as they are cheap and environmentally safe to replace the synthetic molluscicides for snail control.

Prevalence and Molecular Epidemiology of Cryptosporidium Infection in Clarias gariepinus Fish in Egypt.

PURPOSE: This study investigated the prevalence and molecular detection of Cryptosporidium spp. in catfish (Clarias gariepinus). METHODS: A total of 300 Carias gariepinus fish were collected from two freshwater sources: the Nile River (180) and drainage canals (120). The stomach and intestine epithelium of each individual fish sample were screened by modified Ziehl-Neelsen (mZN) staining technique for the detection of Cryptosporidium oocysts followed by the serological survey for detection of Cryptosporidium antibodies using Enzyme-Linked Immunosorbent Assay (ELISA) and molecular characterization using complemented DNA polymerase chain reaction (cPCR). RESULTS: ELISA showed higher prevalence of 69.3% than that prevalence obtained by mZN, 64% for the total examined Clarias gariepinus fish. Also, higher prevalence of Cryptosporidium infection 65.5% and 75.8% obtained by ELISA than 61.1% and 68.3% by mZN, in both fish groups from Nile River and Drainage canal, respectively. PCR analysis revealed the expected positive bands at 1056 bp. DNA sequencing and phylogenetic analysis proved that the positive-PCR Cryptosporidium isolate identified in the present study was Cryptosporidium molnari. CONCLUSION: Freshwater fishes (Clarias gariepinus) are subjected to a high infection rate with Cryptosporidium spp.; the drainage canals obtained fishes showed higher prevalence than that collected from Nile River which indicates an important public health problem and a potential risk of drainage canals in Egypt. ELISA showed higher prevalence of cryptosporidiosis than mZN, for the total examined Clarias gariepinus fish and phylogenetic analyses confirmed this protozoal organism to be a novel species of Cryptosporidium molnari.

Histological technique to detect the physiological age of the malaria vector mosquito Anopheles pharoensis (Diptera: Culicidae).

The risk of malaria recurrence increases due to the main vector, Anopheles pharoensis. The physiological age of the mosquito population is needed to expect malaria vector dynamics. The number of completed gonotrophic cycles is of great importance in determining the physiological age of females. A technique has been described that focuses on the number of dilatations remaining in the ovarioles after each oviposition to determine how many blood meals have been taken. At each gonotrophic cycle, the chances of infection of the vectors are repeated. The histological changes that occur immediately in the ovarioles and ovulation itself were studied. Under the influence of the contractions of the ovarian muscles, the eggs begin to move over the distal end of the ovariole into the inner oviduct. The terminal pedicle is markedly dilated near the diameter of the eggs. After the expulsion of the mature eggs, ovariole dilations were found at the point of their development in the terminal pedicle due to the accumulation of nurse cell remnants and follicular epithelium. The results were used to develop epidemiological localization and to evaluate the effectiveness of antimalaria interventions. The ovarian inspection often provides a technique to distinguish nulliparous from parous female anophelines. In addition, this study can provide basic entomological knowledge on the physiological age of mosquitoes by considering the histological changes in the ovaries, which allow the evaluation of vector management strategies in the field.

Haemogregarines and Criteria for Identification.

Apicomplexa is a phylum that includes all parasitic protozoa sharing unique ultrastructural features. Haemogregarines are sophisticated apicomplexan blood parasites with an obligatory heteroxenous life cycle and haplohomophasic alternation of generations. Haemogregarines are common blood parasites of fish, amphibians, lizards, snakes, turtles, tortoises, crocodilians, birds, and mammals. Haemogregarine ultrastructure has been so far examined only for stages from the vertebrate host. PCR-based assays and the sequencing of the 18S rRNA gene are helpful methods to further characterize this parasite group. The proper classification for the haemogregarine complex is available with the criteria of generic and unique diagnosis of these parasites.

Morphological description and phylogenetic assessment of 28S rRNA for Thelandros chalcidiae sp. nov. from Chalcides ocellatus.

Thelandros is a genus of oxyurid nematodes which parasitize both omnivorous and herbivorous hosts. Thelandros chalcidiae sp. nov. is a new taxa described from the large intestine of the ocellated skink, Chalcides ocellatus, from the South Sinai Desert in Egypt. The recovered parasite species was examined at both morphological and molecular levels in order to determine the exact taxonomic position within Pharyngodonidae family. The current pharyngodonid species is characterized by a mouth opening bounded by three bilobed lips; male worms are characterized by the presence of two pairs of cloacal papillae (pre- and adcloacal) and one single caudal papillae (postcloacal) and caudal alae in an auricular form and females with post-equatorial vulva, amphidelphic ovary in which anterior ovary extended to level of excretory pore and posterior ovary extended posteriorly up anal opening. The recovered nematodes were compared with other known species from different hosts and it was found to be morphologically different from them. Molecular characterization based on the partial 28S rRNA nuclear ribosomal gene sequence showed sequence identities ≥ 83.15% with taxa under family Pharyngodonidae, 74.84-87.37% with Oxyuridae, 80.54% with Heteroxynematidae, and 75.98-77.72% with Thelastomatidea. Phylogenetic analysis showed that parasite sequence in conjunction with existing data facilitate placement of this species within Oxyurida. The present species is deeply embedded in genus Thelandros with close relationships to previously described Thelandros sp. and T. galloti in same taxon. This study highlighted importance of combining morphological and genetic data with taxonomy in pharyngodonid species.

Morphological redescription and phylogenetic assessment of Spauligodon aspiculus (Oxyuroidea: Pharyngodonidae) infecting the white-spotted gecko, Tarentola annularis (Squamata: Gekkonidae).

Thirty white-spotted geckos, Tarentola annularis, from the South Sinai desert in Egypt, were examined for helminth parasites. Spauligodon aspiculus was observed to infect 19 geckos with 63.33% as a prevalence of parasitic infection. The present nematode species is separated from congeners by morphological and metrical characteristics such as lateral alae, aspinose filamentous tail, and no spicule, and three pairs of caudal papillae with posterior pair excluded from envelopment by the caudal alae in the male worms, and knobbed eggs, and postbulbar vulva in females. It compared morphometrically with other Spauligodon species described previously and showed few differences in measurements. Molecular characterization based on the partial 28S rRNA nuclear ribosomal gene sequence showed that there was a close identity, up to 72%, with other sequences retrieved from GenBank. Phylogenetic analysis showed that the parasite sequence in conjunction with existing data facilitates the investigation of the placement of this pharyngodonid species within Oxyuridae. The present species is deeply embedded in the genus Spauligodon with close relationships to previously described Spauligodon nicolauensis (gb| JN619349.1, and JF829243.1) as more related sister taxa. This study highlights the importance of combining genetic and morphological data with taxonomy in pharyngodonid species.

Ultrastructure and phylogenetic characterization of the microsporidian parasite Heterosporis lessepsianus n. sp. (Microsporidia: Glugeidae) infecting the lizardfish Saurida lessepsianus (Pisces: Synodontidae) inhabiting the Red Sea.

Heterosporosis is an increasingly important microsporidian disease distributed worldwide, impacting wild and farmed-raised fish in both fresh and marine water environments. Twenty three out of 130 (17.69%) of the lizardfish Saurida lessepsianus were found to be naturally infected with microsporidian parasites. The rate of parasitic infection was increased during winter reaching 29.23% (19/65) and fall to 6.15% (4/65) in summer. The infection was recorded as numerous macroscopic whitish cysts reached 3.8-6.5 mm in diameter embedded in the abdominal cavity, skeletal muscles and mesenteric tissues of the infected fish inducing an enormous hypertrophy of infected tissues. Light microscopic examination revealed that parasitic foci were encapsulated by a host-derived fibrous membrane containing different developmental stages of the parasite. Spores were oval to pyriform in shape. Transmission electron microscopic study showed the presence of smooth membranes of the sarcoplasmic reticulum forming a thick, amorphous coat surrounding the various developmental stages of the examined parasite (meronts, sporont, sporoblasts, and spores). Mature spores were electron dense and uninucleate. The anchoring disk was found in a central position at the anterior end of the spore and a large vacuole was located at the posterior end. There was a definite number (7-8) of the polar filament turns. Molecular analysis based on the 16 small subunit (SSU) rDNA gene was performed to determine the phylogenetic position of the present parasite species. A 615 bp region of the 16SSU rDNA gene of the studied parasite was sequenced and deposited in GenBank under the accession number MF769371. Multiple sequence alignment demonstrated a high degree of similarity (>82%) with other twenty microsporidian species isolated from different aquatic hosts. The most closely related sequence was provided by the GenBank entry JF745533 for Heterosporis saurida isolated from the marine fish Saurida undosquamis with the highest percentage of identity (98%) and lowest divergence value (0.9). The ultrastructural characteristics and phylogenetic analysis support the recognition of a new species, herein named Heterosporis lessepsianus sp. n.

Altered renal immune complexes deposition in female BWF1 lupus mice following Plasmodium chabaudi infection.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease that has a mysterious relationship with malaria infection. The current study was designated to compare between the effect of the live and the gamma irradiated Plasmodium chabaudi infection on BWF1 lupus murine model. A total of 30 female BWF1 mice were randomly divided into three groups (10 mice/group) as follows: group (I) lupus group (lupus non infected); group (II) live malaria infected group (lupus + live malaria infection); and group (III) irradiated malaria-infected group (lupus + gamma irradiated malaria infection). Live P. chabaudi infection was accompanied with a decrease in survival rate and food consumption in comparison to the control group of mice while gamma irradiated P. chabaudi -infection was unable to do this effect. Additionally, live P. chabaudi infection was accompanied with an increased level of proteinuria and increased rate of immune complexes deposition in kidney. Moreover, infection with live, but not gamma-irradiated P. chabaudi was accompanied with an increase in nitric oxide (NO), hydrogen peroxide (H2O2), and malondialdehyde (MDA) levels in plasma of lupus mice. The levels of both total cholesterol and triglycerides in plasma of lupus mice after live P. chabaudi infection were obviously decreased in comparison to the control group. On the other hand, gamma-irradiated P. chabaudi infection resembled the control group. Our data revealed that infection of lupus mice with live but not gamma-irradiated P. chabaudi has several histological and biochemical effects.

Morphological Re-Description and 18 S rDNA Sequence Confirmation of the Pinworm Aspiculuris tetraptera (Nematoda, Heteroxynematidae) Infecting the Laboratory Mice Mus musculus.

Aspiculuris tetraptera is a heteroxynematid nematoda infecting most of the laboratory animals, occasionally mice which represent the mostly used animal for biological, medical, and pharmacological studies. The present study aimed to investigate the prevalence of nematode parasites infection in the laboratory mice Mus musculus in Egypt. Morphologically, this oxyurid possessed four distinct cephalic papillae on a cephalic plate, with three small rudimental lips carrying two sessile poorly developed labial papillae and one pair of amphidial pores. Esophagus divided into cylindrical corpus and globular bulb. Distinct cervical alae interrupted at the level of esophago-intestinal junction forming an acute angle. At the caudal end, twelve caudal papillae in male worms while an ovijector apparatus opening and a vulva surrounded by protruded lips in females were observed. The general morphological criteria include this nematode with other Aspiculuris species which were compared in the present study. Molecular characterization based on 18SSU rDNA sequencing performed to confirm the taxonomic position of this species and to documents the morphological data. Sequence alignment detects a percent of identity up to 88.0% with other Heteroxynematidae species. Phylogenetic analysis showed that the present recorded is a putative sister taxon to A. tetraptera recorded in a previous study. The SSU rDNA sequence has been deposited in the GenBank under the accession no. MG019400.

Light microscopy and surface topography of Urotrema scabridum and Renschetrema indicum (Digenea) from Rhinopoma hardwickii (Chiroptera): first report in Egypt.

This report introduced the description of two different species of digenean parasites isolated from the intestine of Rhinopoma hardwickii with new host and locality records in Egypt. The recovered helminthes were studied morphologically and morphometrically by light microscopy and the surface topography of the two species was elucidated by scanning electron microscopy (SEM). Urotrema scabridum had an elongated body, testes were tandem, ovaries were pretesticular, and vitelline follicles were observed in 2 lateral fields. SEM showed that the anterior half was covered with random and backwardly directed tegument spines. The lumen of the oral sucker was as a longitudinal slit encircled with type I dome-shaped papillae. The ventral sucker was wrinkled and covered by tongue-shaped tegument spines and several scattered papillae. Renschetrema indicum had a fusiform body with minute spines densely distributed in the anterior part of the body; testes sub-triangular, ovary fusiform; vitellaria were randomly distributed around the ceca and genital organs. SEM showed that the fore-body was ventrally concave and surrounded by cytoplasmic ridges equipped with numerous closely packed claw-shaped spines. The oral sucker was externally surrounded by two circles of papillae while the lip of the ventral sucker was rounded and surrounded by three papillae located in its upper end and anterolaterally.

Light microscopy and surface topography of Urotrema scabridum and Renschetrema indicum (Digenea) from Rhinopoma hardwickii (Chiroptera): first report in Egypt / Microscopia de luz e topografia superficial de Urotrema scabridum e Renschetrema indicum (Digenea) de Rhinopoma hardwickii (Chiroptera): primeiro relato no Egito

Abstract This report introduced the description of two different species of digenean parasites isolated from the intestine of Rhinopoma hardwickii with new host and locality records in Egypt. The recovered helminthes were studied morphologically and morphometrically by light microscopy and the surface topography of the two species was elucidated by scanning electron microscopy (SEM). Urotrema scabridum had an elongated body, testes were tandem, ovaries were pretesticular, and vitelline follicles were observed in 2 lateral fields. SEM showed that the anterior half was covered with random and backwardly directed tegument spines. The lumen of the oral sucker was as a longitudinal slit encircled with type I dome-shaped papillae. The ventral sucker was wrinkled and covered by tongue-shaped tegument spines and several scattered papillae. Renschetrema indicum had a fusiform body with minute spines densely distributed in the anterior part of the body; testes sub-triangular, ovary fusiform; vitellaria were randomly distributed around the ceca and genital organs. SEM showed that the fore-body was ventrally concave and surrounded by cytoplasmic ridges equipped with numerous closely packed claw-shaped spines. The oral sucker was externally surrounded by two circles of papillae while the lip of the ventral sucker was rounded and surrounded by three papillae located in its upper end and anterolaterally.

Resumo O trabalho descreve duas espécies diferentes de parasitas digêneos isolados do intestino de Rhinopoma hardwickii, com novos registros de hospedeiros e localidade no Egito. Os helmintos recuperados foram estudados morfologicamente e morfometricamente por microscopia óptica, e a topografia superficial das duas espécies foi verificada por microscopia eletrônica de varredura (MEV). Urotrema scabridum apresenta um corpo alongado, testículos em tandem, ovários localizados pré-testiculares e os folículos vitelinos foram observados em 2 campos laterais. A microscopia eletrônica (ME) mostrou que a metade anterior do tegumento estava coberta com espinhos aleatoriamente e voltados para trás. O lúmen da ventosa oral apresentava-se como uma fenda longitudinal cercada de papilas em forma de cúpula tipo I. A ventosa ventral estava enrrugada e coberta por espinhos no tegumento em forma de língua e com várias papilas dispersas. Renschetrema indicum tinha um corpo fusiforme com espinhos diminutos densamente distribuídos na parte anterior do corpo; testículos sub-triangulares, ovários fusiformes; as glândulas vitelínicas estavam distribuídas aleatoriamente em torno do ceco e dos órgãos genitais. À ME a região anterior mostrou-se ventralmente côncava e cercada por cristas citoplasmáticas equipadas com numerosos espinhos em forma de unhas muito próximos. A ventosa oral apresentou-se externamente cercada por duas fileiras de papilas, enquanto a borda da ventosa ventral mostrou-se arredondada e cercada por três papilas localizadas na sua extremidade superior e antero-lateralmente.

Hematological, physiological and genotoxicological effects of Match 5% EC insecticide on Biomphalaria alexandrina snails.

Freshwater snails are used as brilliant biomarkers of aquatic ecosystem pollution by chemical compounds. The objective of this study is to highlight the ecotoxicological impacts of the insecticide Match 5%EC (its active ingredient is lufenuron 5% EC) on Biomphalaria alexandrina snails the intermediate host of Schistosoma mansoni in Egypt. The present investigation recorded a remarkable molluscicidal effect of lufenuron 5% EC on these snails and there was a decrease in total number of their hemocytes after exposure. Three morphologically distinct populations of circulating hemocytes were identified (round small cells, granulocytes and hyalinocytes) and results showed that some hyalinocytes had a shrunk nucleus and some were degenerated. Significant increase of transaminases (ALT and AST), while, a decrease of the total protein and albumin content in hemolymph was recorded. The results of alkaline comet assay in the present study demonstrated that lufenuron 5% EC has a genotoxic effect especially when its concentration increases. It can be concluded that Biomphalaria alexandrina snails can be used as bio monitor to screen the deleterious effects of lufenuron 5% EC insecticide as a cause of the environmental pollution, and this insecticide can be used in controlling schistosomiasis because of its molluscicidal effects on B. alexandrina snails.

Rhabdias bufonis (Rhabdiasidae) from the lung of the African common toad, Amietophrynus regularis (Bufonidae) in Egypt: new data on the basis of light and scanning electron microscopic study.

BACKGROUND AND AIMS: Rhabdias sp. (Rhabdiasidae) is a nematode parasite of family Rhabdiasidae infecting the lung of amphibians. The present study provides new morphological details for Rhabdias bufonis isolated from the lungs of the African common toad, Amietophrynus regularis based on observations of light and scanning electron microscopy (SEM). METHODS: Forty specimens were collected from its natural habitat: the damp, moist fields and gardens at Giza governorate, Egypt. Worms were isolated from the lungs, fixed and then preserved. They were examined using light and SEM with further line drawings. RESULTS: Fourteen specimens (35%) were found to harbor Rhabdias with an intensity of three to five worms per host. Bodies of the gravid females were elongated, slender, measured 3.22-9.86 (5.64 ± 0.03) long and 0.09-0.48 (0.23 ± 0.02) wide at mid-length. Buccal capsule was with cylindrical lumen and sclerotized walls. Ovaries were almost straight. The uteri were located anterior and posterior to the vulva. Uterus were filled with a large number of eggs (17-42). Eggs were oval in shape and some of them were with fully developed larvae inside. The tail was comparatively short, gradually tapered. SEM showed that worms possessed an oval anterior end with a simple, slit like oral opening. The lipless edges of the mouth opening were bordered with tiny cuticular elevations, radiated outwards. Two pairs of submedian cephalic papillae were observed around the oral opening as well as two amphids. CONCLUSION: The current study presents new morphological details for R. bufonis isolated from the African common toad. Also, the morphology of the slit-like mouth opening, the two pairs of cephalic papillae, two amphids and the three pairs of cuticular elevations supporting the area around mouth opening were investigated.

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.