Identification of Eimeria acervulina (Apicomplexa: Eimeriidae) infecting the broiler chicken Gallus gallus domesticus through morphology and molecular analyses.

Coccidiosis is an intestinal protozoan disease that affects the poultry industry worldwide. The severity of this disease varies depending on the identity of the infectious agents. Therefore, this study was carried out to identify the Eimeria species that affect broiler chickens, Gallus gallus domesticus, through morphological and molecular phylogenetic analyses. Twenty-five faecal samples were collected from the broiler chickens in a commercial poultry farm in Riyadh (Saudi Arabia). Using the floatation technique, faeces were examined microscopically for the Eimeria occurrence. Identification of Eimeria species was performed based on morphological criteria and molecular tools (DNA amplification for the partial small subunit ribosomal RNA (18S rRNA), internal transcribed spacer (ITS)-1, and mitochondrial cytochrome c oxidase I (COI) genes. In this study, 32% (8 out of 25) of collected samples were found to be positive for coccidiosis. After sporulation in potassium dichromate (K2Cr2O7), the sporulated oocysts were observed as ovoid and measured 18.37-23.19 µm (19.87) long and 15.07-18.67 µm (16.46) wide, with the anterior location of a polar granule and absence of micropyle. These Eimeria oocysts were assumed to size and shape characteristics of Eimeria acervulina. Molecular analysis was conducted on the sequences of the polymerase chain reaction products from the three genes studied (18S rRNA, ITS-1, and COI). At the three genes, results showed that the resultant sequences clustered with E. acervulina from different regions confirming morphological description. This study highlighted the importance of molecular techniques to detect avian Eimeria species more than the traditional morphology-based tool to optimise the appropriate anticoccidial strategies for long-term control in the studied area.

Identification of sheep eimerian parasites, Eimeria crandallis and Eimeria faurei, employing microscopic and molecular tools.

Coccidiosis, caused by apicomplexan Eimeria species, is a protozoan disease that affects various species of wild and domestic animals. However, data available on Eimeria diversity in ruminants in Saudi Arabia is meagre. Therefore, this study was designed to investigate some eimerian parasites infecting sheep (Sawakni and Harrie breeds) using microscopy and molecular methods for the first time in Saudi Arabia. Twenty-four fecal samples were collected from sheep farms. Based on the floatation technique, eimerian oocysts were observed in 8 of the 24 (33.33%) fecal samples. The coccidian-positive samples were subjected to fecal culture in a shallow layer of 2.5% potassium dichromate (K2 Cr2 O7 ). Detected eimerian oocysts were described micromorphometrically as the basis for traditional oocyst identification. Morphologically, the sporulated oocysts were similar to those of sheep eimerian parasies; Eimeria faurei and Eimeria crandallis. PCR products from the two eimerian species detected from Sawakni and Harrie breeds were sequenced and were found to be distinct from each other with mutations at five positions. One of them clustered with E. crandallis with 99.8%-100% identity with sequences available in GenBank. E. crandallis was obtained from two Sawakni sheep and two Harrie sheep. The other sequences grouped with E. faurei with 99.8% identity with the only sequences available in GenBank. E. crandallis was detected from both Sawakni and Harrie breeds whereas E. faurei was detected only from Sawakni sheep. The findings of this study have implications for the importance of morphometric identification with advanced molecular tools to confirm the identities of sheep Eimeria species and to address the taxonomic study of this eimeriid parasite at the species level.

Morphological and mitochondrial approaches of Hatschekia sargi (Copepoda: Hatschekiidae) as a parasite of Epinephelus chlorostigma.

Little information is available until now about the copepods infecting different fish species. Therefore, this study aimed to provide light on siphonostomatoids infecting Epinephelus chlorostigma. Twenty fish specimens were taken from the Red Sea coast (Jeddah, Saudi Arabia), and ectoparasitic copepods were investigated. Light microscopy and molecular tools were used to examine copepods isolated from fish. Parasitological indexes were calculated and showed that 60% of the examined fish were infected with a mean intensity of 12 parasite/fish. Morphological examination revealed that this copepod species is characterized by all unique features of the genus Hatschekia with special reference to Hatschekia sargi. The taxonomic position of the recovered species in the Hatschekiidae family within Siphonostomatoida was confirmed using phylogenetic analysis based on partial mitochondrial cytochrome c oxidase subunit I (mt COI) gene sequences. The mt COI gene query revealed that the recovered Hatschekia species is closely related to Hatschekia maculatus (gb| JQ664005.1). This study discovers a new host for Hatschekia species isolated from Saudi Arabia and conducts the first genomic investigation of the mt COI gene.

Persea americana extract protects intestinal tissue from Eimeria papillata-induced murine Infection.

Coccidiosis is the most prevalent disease-causing widespread economic loss among farm and domestic animals. Currently, several drugs are available for the control of this disease but resistance has been confirmed for all of them. There is an urgent need, therefore, for the identification of new sources as alternative treatments to control coccidiosis. The present work aimed to study the effect of the Persea americana extract (PAE) as an anti-coccidial, anti-oxidant, and anti-apoptotic modulator during murine intestinal Eimeria papillata infection. A total of 25 male mice were divided into five groups, as follows: Group1: Non-infected-non-treated (negative control), Group2: Non-infected-treated group with PAE (500 mg/kg b.w). Group3: Infected-non-treated (positive control), Group4: Infected-treated group with PAE (500 mg/kg b.w.), and Group5: Infected-treated group with Amprolium (120 mg/kg b.w.). Groups (3-5) were orally inoculated with 1 × 103 sporulated E. papillata oocysts. After 60 min of infection, groups (4 and 5) were treated for 5 consecutive days with the recommended doses of PAE and amprolium. The fact that PAE has an anti-coccidial efficacy against intestinal E. papillata infection in mice has been clarified by the reduction of fecal oocyst output on the 5th day post-infection by about 85.41%. Moreover, there is a significant reduction in the size of each parasite stage in the jejunal tissues of the infected-treated group with PAE. PAE counteracted the E. papillata-induced loss of glutathione peroxidase (GPx), superoxide dismutase (SOD), and total antioxidant capacity (TCA). E. papillata infection also induced an increase in the apoptotic cells expressed by caspase-3 which modulated after PAE treatment. Moreover, the mRNA expression of the goblet cell response gene, mucin (MUC2), was upregulated from 0.50 to 1.20-fold after treatment with PAE. Based on our results, PAE is a promising medicinal plant with anti-coccidial, anti-oxidant, and anti-apoptotic activities and could be used as a food additive.

Morphological, molecular, and histopathological studies on Hadjelia truncata from Columba livia domestica and its role as an environmental biological indicator.

Pigeons are a cosmopolitan group of birds with abundant and large populations associated with human activities. This study focused on determining parasitic infections within domestic pigeons (Columba livia domestica). Forty-eight pigeons were examined for infections, of which 29.16% were infected with a nematode parasite, identified as Hadjelia truncata (Habronematidae), under the koilin layer of their gizzards. The population of nematodes in infected gizzards did not exceed 20 adult worms. DNA from the gizzard worms was extracted and subjected to PCR using primers that amplify the partial 18S rDNA and cytochrome C oxidase subunit I (COX I) regions. Identification of this parasite based on microscopic study revealed the presence of trilobed lips with cephalic papillae and amphidial pores, as well as other characteristic features. In males, spicules were unequal with the presence of six pedunculated pairs of caudal papillae (4 pre- and 2 post-anal) and a tail surrounded with caudal ala. In females, the vulva was a rounded aperture located in front of the posterior end of the esophagus and uteri, which was filled with numerous embryonated eggs. DNA Sequences from partial 18S rDNA were homologous to sequences obtained from H. truncata in GenBank with a high percentage of identity. DNA sequences from mitochondrial gene COX I, however, were unique, and they were the first sequenced for H. truncata, since no sequences for this taxon were previously available in GenBank. Histopathological examination revealed enlargement of infected gizzards in comparison to non-infected ones, with the presence of necrosis and interstitial infiltration in the koilin layer. Concentrations of heavy metals (Fe, Cu, Zn, Cd, Cr, and Co) were measured using inductivity-coupled plasma in tissues (liver, muscles, and gizzards) from infected and non-infected pigeons as well as their parasites. Results showed different affinities of metals to tissues. Recovered parasites can minimize element concentration from their pigeon tissues. In Saudi Arabia, this study was considered the first report identifying pigeon nematodes and evaluating of the effects of their pathogenicity on the animals' welfare, as well as their application as a useful tool for monitoring environmental pollution.

Blood Biomarkers of Neonatal Sepsis with Special Emphasis on the Monocyte Distribution Width Value as an Early Sepsis Index.

Background and Objectives: Early detection of neonatal sepsis is critical because it is potentially fatal. Therefore, sepsis biomarkers of sufficient sensitivity and specificity are needed. This study aimed to evaluate the utility of peripheral blood parameters as neonatal sepsis biomarkers and the diagnostic performance of the monocyte distribution width (MDW) in sepsis in a neonatal intensive care unit. Materials and Methods: A cross-sectional study was conducted from September 2019 to August 2020 at the King Saud University Medical City in Riyadh, Saudi Arabia. Samples were collected and organised as follows: 77 study cases were subdivided into two subgroups (other health complication (49) and sepsis (28)), and there were 70 controls. The causative microorganisms of neonatal sepsis were isolated. Peripheral blood samples were collected from each neonate in an ethylenediaminetetraacetic acid tube for a complete blood count and a leukocyte differential count. Moreover, the receiver operating characteristic (ROC) curve analysis was used to measure the diagnostic performance of the MDW. Results: The haematological parameters and neonatal sepsis cases had a considerable correlation. The MDW was the most significant haematological parameter. The ROC analysis of the MDW demonstrated that the area under the curve was 0.89 (95% confidence interval: 0.867 to 0.998) with a sensitivity of 89.3%, a specificity of 88.2%, and a negative predictive value of 97.2% at the cut-off point of 23. Conclusions: The use of haematological parameters is feasible and can be performed rapidly. Neonatal sepsis showed a strong correlation with leukopenia, anaemia, thrombocytopenia, and an elevated MDW value. Moreover, the ROC curve analysis confirmed the high diagnostic ability of the MDW in neonatal sepsis prediction.

Molecular identification of the rodent-borne pathogen Rodentolepis nana using the genetic markers of ITS-1, 18 S, and 28 S rDNA.

BACKGROUND: Rodentolepis nana (syn. Hymenolepis nana), the most common cyclophyllid tapeworm infecting rodents, is a well-studied gastrointestinal parasite in mice and belongs to the family Hymenolepididae. METHODS: The present study focuses on the molecular analysis for the nuclear genes (ITS-1, 18 S, and 28 S rDNA) used for the accurate recognition of the recovered Rodentolepis species. RESULTS: The annotated partial ITS-1, 18 S, and 28 S rDNA gene regions were deposited in GenBank (gbǀ MW310394.1, gbǀ MW327585.1, and gbǀ MW324479.1, respectively) and further used in the maximum likelihood method (ML) to clarify their genetic relationships at the species level. The interrogation sequence of R. nana was aligned and belonged to the family Hymenolepididae, in the same group as all Hymenolepis species, which were distinct from Cyclophyllidea cestodes, especially species belonging to Anoplocephalidae and Taeniidae. Sequence data support the paraphyly of Hymenolepis species. CONCLUSIONS: The phylogeny supports the availability of the ITS-1, 18 S, and 28 S rDNA genes as reliable genetic markers for evolutionary relationships.

Evaluation of Nephrotoxicity in Oreochromis niloticus After Exposure to Aluminum Oxide Nanoparticles: Exposure and Recovery Study.

Nephrotoxic impacts of Al2O3 nanoparticles (NPs) were studied in Oreochromis niloticus after seven days of exposure and fifteen days of recovery periods. Fish were classified as group I (dechlorinated water); group II (2 mg/L Al2O3NPs); group III (4 mg/L Al2O3NPs); group IV (8 mg/L Al2O3NPs). Blood creatinine and uric acid levels showed marked increases in groups III and IV. A dose-dependent disturbance in renal antioxidant components was recorded as indicated by elevated catalase, superoxide dismutase, thiobarbituric acid reactive substances levels, and decreased glutathione reduced concentration. Renal histopathology was recorded with the highest % of appearance in group IV. A reduction in renal Al content, kidney function biomarkers (excepting group IV), and enhanced antioxidant status were observed after applying a recovery strategy. Several structural damages were identified following the recovery period, but the alteration frequencies indicated regressive histopathological changes. The Al2O3NPs-induced nephrotoxicity can be reduced after applying a suitable recovery period.

Silver Nanoparticles Induce Time- and Tissue-Specific Genotoxicity in Oreochromis niloticus: Utilizing the Adsorptive Capacities of Fruit Peels to Minimize Genotoxicity.

Fish were exposed to a sub-lethal concentration of silver nanoparticles with and without orange and banana peels water treatment for 24, 48, and 96 h. The adsorption of AgNPs on both peels was recognized by scanning electron microscopic, energy dispersive X-ray spectroscopy, and laser ablation imaging. The % of DNA damage in liver and muscle tissues (comet assay) showed significant elevations in all studied groups with the maximum level in liver tissues after 24 h. DNA damage was markedly decreased after 48, and 96 h signifying the presence of an effective repairing mechanism. Micronucleus and nine nuclear anomalies were recorded in the peripheral blood cells. All anomalies were observed in all studied groups with a maximum induction rate after 96 h. of exposure. Based on the % of DNA damage and the frequencies of nuclear anomalies, water treatment with orange and banana peel succeeded to reduce AgNPs-induced genotoxic damage.

Long-Term Exposure to the Water of Wadi El-Rayan Lakes Induced Testicular Damage and Endocrine Disruption in Mugil cephalus.

This study aimed to investigate the testicular function of Mugil cephalus that inhabit Wadi El-Rayan lakes. Testes of fish inhabiting the upper lake (site 2) and the lower lake (site 3) of Wadi El-Rayan showed significant decreases in gonadosomatic index, high accumulation levels of six metals, and eight organochlorine pesticide residues. Compared to reference fish, high percentages of histological alterations as testicular degeneration, germ cell reduction, testicular inflammation, vacuolization, and loss of tubular arrangement were observed in sites 2 and 3. Moreover, endocrine disruption signs were recorded based on the percentage of ovotestis appearance and the ovotestis severity index values. The maximum defective testicular antioxidant mechanisms were recorded in site 3 as indicated by sharp decreases in catalase, superoxide dismutase, glutathione reduced levels, and high thiobarbituric acid reactive substances. Finally, long-term exposure to Wadi El-Rayan water may impair the reproductive health of fish via testicular oxidative damage and endocrine disruption.

The phylogenetic position of Arhaphe deviatica within Hemipteran insects: A potential model species for eco-devo studies of symbiosis.

Insecta is known to be the most diverse group of species, exhibiting numerous forms of endosymbiotic associations. Molecular techniques have provided significant indicators for insect-microbe interactions. The present study aimed to register one of the true bugs of pentatomomorpha and clarify its taxonomic position through phylogenetic analysis of the partial 16S rRNA gene region. A maximum likelihood analysis retrieved a generally well-supported phylogeny based on Tamura 3-parameter model. Based on the partial mitochondrial 16S rRNA gene sequences, a phylogenetic study of suborder Heteroptera relationships within Hemipteras' order was constructed. Sequences of 221 bases of the 3' end of the gene from 28 species within 16 families were analyzed. This analysis and bootstrap confidence revealed two major clades comprising four suborders within Hemiptera, with a close relationship between Heteroptera + (Sternorrhyncha + (Auchenorrhycha + Coleorrhyncha)). Infraorder Pentatomomorpha is forming a sister group with a substantial bootstrap value to Cimicomorpha. Pyrrhocoroidea forms a sister relationship with Lygaeoidea + Coreoidea. There is a close relationship between Largidae and Pyrrhocoridae within Pyrrhocoroidea. The results show that the present species is firmly embedded in the genus Arhaphe with 94.35% sequence resemblance to its congeners. Besides, the recovered hemipteran species considered a potential model group for studying different symbionts. We propose both phylogenetic and ecological evolutionary developmental biology viewpoints for a more synthetic understanding of insect populations' molecular evolution.

Heteromicrocotyla polyorchis Unnithan, 1961 (Monogenea: Heteromicrocotylidae), a gill parasite of the yellow-spotted trevally, Carangoides fulvoguttatus (Carangidae) from Saudi Arabia: Morphology and phylogeny.

Little information for parasitic infections of Carangoides fulvoguttatus was recorded. The present study was intended to investigate the gill parasite Heteromicrocotyla polyorchis of this fish and to provide a full morphological description and clarify its taxonomic status through phylogenetic analysis of the 28S rRNA gene region. A total of sixty fish specimens have been collected from the studied area (the Red Sea in Jeddah Province, Saudi Arabia) and gills were isolated and examined for identification of parasites. Using light electron microscopy, the recovered monogenean parasite's morphology was exhaustively characterized and described. Microscope examinations found that this parasite species represent Heteromicrocotyla polyorchis, and it could be distinguished from congeners of the same genus by armed genital atrium and cirrus sac, follicular post-ovarian testes, unique shape and number of clamps on both haptor sides, and the dorsally curved tip of the male copulatory organ. Morphological features were combined with molecular analysis of the 28S rRNA gene region. The selected gene for the isolated Heteromicrocotyla species was analyzed using appropriate primers to assist in phylogeny with those in the GenBank database. The present monogenean species was characterized by unique genetic sequences that were analyzed and deposited in the GenBank for the first time under the accession number MW406473. Phylogenetic analyses reported that the maximum identity between the current Heteromicrocotyla species and taxa of Heteromicrocotylidae was between 91.42 and 92.09% and confirmed its taxonomic status in this family with a well-distinct clade. The present study supported the second report of H. polyorchis as carangid fish ectoparasites and investigates the first appearance in C. fulvoguttatus inhabiting Saudi Arabia.

Cucullanus bulbosus (Lane, 1916) Barreto, 1918 (Nematoda, Cucullanidae) from the common ponyfish Leiognathus equulus (Leiognathidae): Morphology and molecular study.

The common ponyfish Leiognathus equulus is a marine fish species with very high commercial value. Little information is available about its parasitic infections. Based on light and scanning electron microscopy, as well as sequencing and analysis of the partial regions of the ITS-1, 18S rRNA, COX1 genes, were employed for the systematic evaluation of a nematode parasite, which it first isolated from L. equulus in Jeddah Province, Red Sea, Saudi Arabia. Results revealed that this nematode parasite closely resembles the previously described Cucullanus bulbosus. Microscopic examination showed that it distinguished from congeners by the unique structure of hemispherical elevation at pseudobuccal capsule level, the ratio of esophagus/body length, spicules size, presence of pre-cloacal sucker, rod-shaped gubernaculum, and the arrangement of caudal papillae in males. Phylogenetic analyses based on ITS-1, 18S rRNA, and COX1 gene regions were constructed to investigate phylogenetic relationships between this parasite species and other related taxa. Results supported that Cucullanus bulbosus resembles a sister of Cucullanus genypteri, Cucullanus pulcherrimus, Cucullanus bourdini, Cucullanus extraneus, and Cucullanus hainanensis by using different genetic markers. This study provides more information about combining morphological and molecular data to identify Cucullanus species with the first natural occurrence in the common ponyfish inhabited in Saudi Arabia.

Anti-proliferative and apoptosis induction activities of curcumin on Leishmania major.

Leishmaniasis is a major vector-borne disease triggered by an obligate intracellular protozoan parasite of the genus Leishmania and transmitted by the bite of phlebotomine female sand flies. This parasite causes a wide range of human diseases, from localized self-healing cutaneous lesions to fatal visceral infections. The aim of this study was to investigate the cytotoxic, antiproliferative, and apoptotic effects of curcumin on Leishmania major promastigotes (MHOM/SA/84/JISH) and to assess these effects on the cell cycle of promastigotes. The MTT colorimetric assay was used to evaluate the cytotoxicity and proliferation of promastigotes. Additionally, flow cytometry was used to analyze the cell cycle. The Annexin V/propidium iodide staining technique followed by flow cytometry was used to study the cell death induced by curcumin. In this study curcumin showed a potent antileishmanial effect, exhibiting cytotoxicity against L. major promastigotes. At 80µM, the survival in curcumin treated promastigotes reached 22%; however, the median lethal concentration of curcumin (LC50) was 35µM. The drug exerted its cytotoxic effect by inducing apoptosis. Curcumin-induced cell death in promastigotes reached 82.5% at 80µM concentration. In addition, curcumin delayed the cell cycle in the S-phase inhibiting cell proliferation. Thus, curcumin was shown to be effective against L. major promastigotes. Therefore, curcumin merits further research studies to demonstrate its efficacy in treating cutaneous leishmaniasis.

Morphology and molecular taxonomy of the tongue worm, genus Raillietiella (Pentastomida) from the lungs of berber skinks Eumeces schneideri (Scincidae): First report.

In the present study, pentastomids belonging to the order Cephalobaenida were isolated from the lungs of Berber skinks Eumeces schneideri (Famiy: Scincidae), which were morphologically described by light and scanning electron microscopy and taxonomically justified by 18s rDNA molecular analyses of the parasites. Seventeen host specimens were collected from well-vegetated wadis at high altitudes, Jizan, Saudi Arabia as new type locality; twelve specimens (70.59%) were infected. All of the recovered parasites were adults, possessed small broadly triangular cephalothorax flattened on the ventral surface and merged smoothly with a uniformly thick and squat abdomen and terminated in a pair of divergent lobes. The results obtained indicated that the parasites belong to the sharp-tipped posterior-hook Raillietiella spp. distinguished from other raillietiedids of the same group some important characteristic features including annulus number, shape and dimensions of the buccal cadre, copulatory spicules, and anterior and posterior hooks. The anterior hook of the female specimens (n=5) had a blade length (AB) of 135±5 (110-146) µm and shank length (BC) 158±5 (150-169) µm while the posterior hook was much larger with AB measuring 221±5 (200-236) µm and BC 286±6 (280-289) µm. For the male specimens (n=5), the anterior hook had an AB of 73±3 (72-75) µm and a BC 102±5 (100-103) µm. The posterior hook was much larger with AB 190.6±5 (190-191) µm and BC 221±5 (280-289) µm. The morphological characterization of the recovered parasites was closely similar to R. aegypti previously isolated from the same host. Sequence alignment by the maximum likelihood analysis for the data obtained from the 18S rDNA analysis of the parasites exhibits identities ranging between 88-95% with pentastomid genera recovered from the GenBank. The phylogenetic tree supported the inclusion of the parasites within the monophyletic Pentastomida clade with maximum identity to the raillietiellid species. The recovered sequences from the present study were deposited in GenBank under Accession number MK970649.1. The present molecular analysis was the first to confirm the taxonomic position of R. aegypti isolated from the host examined.

First record of third-stage Terranova larval type II (Nematoda, Anisakidae) in the common ponyfish Leiognathus equulus Forsskål.

The current study was carried out to investigate the natural occurrence of nematode parasites that infect the common ponyfish Leiognathus equulus from Jeddah, Saudi Arabia. Third-stage nematode larvae were found to be encysted in the peritoneum of the fish studied, with the prevalence of infection being 25%. Light microscopy revealed that this parasite belongs to the Anisakidae family within the genus Terranova by having all the generic characteristic features. Based on the intestinal caecum ratio to the length of the ventriculus being 2:1, the excretory pore with ventral location below the boring tooth, the body ended with a conical tail; the larvae found in the present study were identified as Terranova larval type. To validate its taxonomic position within Anisakidae, this Terranova species' morphological features were combined with the ITS-1 gene's molecular analysis. It demonstrated sequence similarities 94.38-76.57% with taxa of Anisakidae. A preliminary genetic comparison between the present parasite and other ascaridoids placed it as a putative sister taxon to the previously described Terranova species. The first record of the current anisakid larvae in the common ponyfish with a unique genetic sequence for the partial sequence of the ITS-1 gene was observed in this study. Its taxonomic position was confirmed in Anisakidae.

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, Molecular, and Pathological Appraisal of Hymenolepis nana (Hymenolepididae) Infecting Laboratory Mice (Mus musculus).

Hymenolepis nana, typically a parasite found in conventionally established mouse colonies, has zoonotic potential characterized by autoinfection and direct life cycle. The objective of this study was to determine the rate of parasite infection in laboratory mice. The hymenolepidide cestode infected 40% of the 50 mice sampled. The rate of infection in males (52%) was higher than in females (28%). Morphological studies on the cestode parasite showed that worms had a globular scolex with four suckers, a retractable rostellum with 20-30 hooks, and a short unsegmented neck. In addition, the remaining strobila consisted of immature, mature, and gravid proglottids, irregularly alternating genital pores, lobulated ovaries, postovarian vitelline glands, and uteri with up to 200 eggs in their gravid proglottids. The parasite taxonomy was confirmed by using molecular characterization based on the sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (mtCOX1) gene. The parasite recovered was up to 80% identical to other species in GenBank. High blast scores and low divergence were noted between the isolated parasite and previously described H. nana (gb| AP017666.1). The phylogenetic analysis using the COX1 sequence places this hymenolepidid species of the order Cyclophyllidea.

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.

Light microscopic study of four plagiorchiid trematodes infecting marine fish in the south-eastern Mediterranean Sea, Alexandria City, with descriptions of two new species.

During the present investigation, a total of 220 fish specimens belonging to three different species, namely, little tunny Euthynnus alletteratus, African snook Lates niloticus, and striped red mullet Mullus surmuletus, were collected from January-November 2016 from the coasts off Abu Qir landing site, Alexandria City, south-eastern Mediterranean Sea, Egypt. The collected fish samples were dissected and examined for the presence of helminth parasites. Twenty-three out of 220 (10.45%) fish specimens were found to be naturally infected with four species of trematode parasites belonging to three different families of the order Plagiorchiida. The recovered parasite species were collected and identified by applying light microscopic examinations. The present study recorded two new parasite species, namely, Stephanostomum alletterani sp. nov. and Bathycreadium mulli sp. nov., belonging to the families Acanthocolpidae and Opecoelidae and infecting E. alletteratus and M. surmuletus, respectively and re-descriptions of the two remaining species, namely, Acanthostomum spiniceps and Aponurus mulli of the families Acanthostomatidae and Opecoelidae, respectively, to clarify the measurements of some body parts. Morphological and morphometric characterizations revealed some differences between the present species and other related species detected previously. Future studies are recommended to include advanced molecular characteristics for these species.