Detection of Neoascarophis mariae (Nematoda: Cystidicolidae) parasitic in Parupeneus forsskali and Mulloidichthys flavolineatus (Perciformes: Mullidae) from the Red Sea in Egypt.

The nematode parasite Neoascarophis mariae was collected from the intestine of Parupeneus forsskali and Mulloidichthys flavolineatus from the Red Sea in Egypt and was described using both light and scanning electron microscopy. It is mainly characterized by the presence of four submedian labia surrounded the oral aperture in the outer side, two sublabia and two pseudolabia in the inner side, rounded cephalic end, and short vestibule in both sexes. The females vulva is located near the posterior end of the body, their oval uterine eggs have a smooth surface devoid of filaments. Males have bifurcated deirids, a large left spicule (85-425 µm) with a tapered tip, and a short right spicule (50-181 µm) with a broad and rounded tip.

Epidemiological, Morphological, and Morphometric Study on Haemonchus spp. Recovered From Goats in Egypt.

Goats can be infected by multiple groups of external and internal parasites. Haemonchus spp. are among abomasal parasites that can result in higher mortality and several considerable economic losses in goats. Early detection of parasites and better understanding of the major risk factors associated with infection are among the main strategies for controlling the infection. Considering this, information on hemonchosis in goats from Egypt, and the contribution of goats in the maintenance of the epidemiological foci of the disease is limited. This study investigated the prevalence of Haemonchus species among 240 abomasum samples collected during postmortem examination of goat carcasses from Assiut Governorate, Egypt. Moreover, the association of the major risk factors to describe the epidemiological pattern of the disease was explored. This study demonstrated that 16.66% of abomasa samples harbored Haemonchus species. Additionally, age, sex, and sampling season were the most significant risk factors associated with infection. Following the variable factors under study, goats aged 1 year or older were at higher risk, with an infection rate of 22.14% (31 of 140), than those younger than 1 year (9%) [p = 0.008; odds ratio (OR) = 2.87; 95% confidence interval (CI), 1.30-6.35]. The infection rate was 25% (19 of 76) in males and 12.8% (21 of 164) in females [p = 0.024; odds ratio (OR) = 2.26; 95% confidence interval (CI), 1.13-4.53]. Moreover, the exposure to infection was higher in summer (22.22%) than in winter (8.33%) (p = 0.007; odds ratio (OR) = 0.318; 95% confidence interval (CI), 0.139-0.725). More importantly, three species of the parasite-Haemonchus contortus, Haemonchus placei, and Haemonchus longistipes-were identified for the first time, and the confirmation of the identification and morphological characterization of the worms was performed using light microscopy and SEM. Collectively, this study reveals interesting epidemiological, morphological, and morphometric findings associated with the occurrence of hemonchosis among goats in Egypt. This study suggests further research for exploring the major circulating species of the parasite in Egypt, which is mandatory for controlling the disease.

Epidemiological, Morphometric, and Molecular Investigation of Cystic Echinococcosis in Camel and Cattle From Upper Egypt: Current Status and Zoonotic Implications.

Cystic echinococcosis has been considered one of the major parasitic zoonoses which is associated with severe economic losses. The present study was undertaken to investigate the occurrence, organ distribution, cyst fertility, and viability of cystic echinococcosis in slaughtered camels and cattle from various abattoirs in Assiut Governorate, Egypt. The work also involved morphological, morphometric, and molecular identification of the parasite. The occurrence of hydatid cysts was investigated in total number of 100 lungs of camels and 574 liver and lungs of cattle admitted to three slaughterhouses at Assiut Governorate, Egypt. Moreover, several individual variable factors, including organ involvement, age, sex, and hydatid cyst characteristics, were studied to identify their possible association with the occurrence of the disease. Genomic DNA was extracted from the hydatid cysts, followed by molecular identification of the parasite through amplification of ribosomal DNA internal transcribed spacer (ITS) regions. Hydatid cysts were found in 6 camels (6%) out of 100 inspected camels, while 5 hydatid cysts (0.87%) were detected in a total number of 574 cattle examined. The parasite was detected exclusively in lungs of camels, while lungs were the main organ infected by the parasite in cattle and one hydatid cyst was found in the liver (0.17%). In camel, 66.7, 16.65, and 16.65%of detected cysts were fertile, sterile, and calcified, respectively, while in cattle, these percentages were 60, 20, and 20%, respectively. None of the studied variable factors were significantly associated with the occurrence of the disease in camels, with the exception that all cysts were found in the lung. Conversely, we found a significant association (P < 0.05) between the age and sex of the slaughtered cattle and the occurrence of hydatid cysts. In this respect, the rate of infection was higher in female cattle and those cattle more than 5 years (P < 0.05). The morphological, morphometric, and molecular studies confirmed the presence of the parasite. Taken together, our results concluded that camels and cattle play a potential role in maintaining the transmission cycle of this zoonotic parasite.

Unusual opecoelids from Red Sea triggerfishes with special reference to characteristic concepts of the Opistholebetinae Fukui, 1929 (Digenea: Opecoelidae).

Three digeneans belonging to the Opecoelidae are reported and described from triggerfishes (Tetraodontiformes: Balistidae) collected in the northern Red Sea off Egypt. Both Macvicaria longicirrata (Manter, 1963) Aken-Ova, Cribb Bray, 2008 and Neopycnadena tendu (Bray Justine, 2007) n. comb. were recovered from the intestine of the titan triggerfish, Balistoides viridescens (Bloch Schneider)-each represents a new host record-and Gaevskajatrema balistes n. sp. was found parasitizing the lower intestine of the Picasso triggerfish, Rhinecanthus assasi (Forsskål). We continue to support synonymy of Gaevskajatrema ponticum (Koval, 1966) Machkevsky, 1990 with Gaevskajatrema perezi (Mathias, 1926) Gibson Bray, 1982, not as a differentiated species. We adopt the restricted posterior extension of the ceca and vitellarium to the testicular zone, without extension of either into the post-testicular space, as diagnostic in distinguishing Gaevskajatrema. Gaevskajatrema balistes n. sp. differs from G. perezi based on its substantially smaller body size with fewer eggs, a longer cirrus-pouch reaching ovarian level and it parasitizes a distinct host group from a structurally and ecologically different ecosystem. Neopycnadena n. gen. is erected for Pseudopycnadena tendu Bray Justine 2007 based on its possessing a large broadly oval cirrus-pouch with a massive field of prostatic cells occupying the entire volume of the cirrus-pouch, a wide, cup-shaped and thick-walled ejaculatory duct, distinct dorsal position of the excretory pore, the bifurcal dextral position of the genital pore, its report from a distinct host group and distant locality and its phylogenetic uniqueness compared with Pseudopycnadena fischthali Saad-Fares Maillard 1986. Neopycnadena n. gen. is ecologically similar to opistholebetines in their life-cycles and morphology; however, phylogenetically separate from opistholebetines as well as from the Polypipapiliotrematinae Martin, Cutmore Cribb in Martin, Sasal, Cutmore, Ward, Aeby Cribb, 2018 and members of Clade [C] of Martin and colleagues, thus we conclude that Neopycnadena n. gen. is unique. Neopycnadeninae n. subfam. is proposed to accommodate Neopycnadena n. gen. We consider that the probable characterization of tetraodontiform specialist taxa (as indicated by the presence of a muscular post-oral ring) and the specificity of the Opistholebetinae Fukui, 1929 sensu stricto with a tetraodontiform host are no longer reliable characters differentiating Gaevskajatrema and Macvicaria Gibson Bray, 1982. The nature of the post-oral structure is discussed and it is adopted to be a diagnostic feature at the generic level among taxa of the Opistholebetinae sensu latu. It is concluded that the expanded concept of the Opistholebetinae is more supported than the restricted one, Birendralebes Srivastava Ghosh, 1972 remains incertae sedis within the Opecoelidae Ozaki, 1925 rather than in the Opistholebetinae, and we provide a generic key to the Opistholebetinae.

Reliability of heterophyid antigens in heterologous protection against human schistosomiasis.

Schistosomiasis is a neglected tropical parasitic disease which has been controlled by praziquantel for many decades; however, reemergence of praziquantel resistant strains has been a continuous threat. Therefore, the development of reliable antischistosomal vaccine is significantly demanded for optimal control. In the present study, comparison among Schistosoma haematobium, Schsitosoma mansoni and Pygidiopsis genata crude antigens was carried out by Sodium dodecyl sulphate polyacrylamide gel electrophoresis. Hyperimmunization of rabbits with tested parasites' crude antigens was done to obtain hyperimmune sera. Western blotting was applied to show cross reactivity between parasites' crude antigens and either homologous or heterologous sera. Although there was no cross reaction between P. genata crude antigens and sera of both Schistosoma species and vice versa; it is supposed that the immunogenic band at 79 kDa might develop cross reactivity with Schistosma spp. SEA fractionation if used in future studies.

Re-examination of the phylogenetic relationships within the Gyliauchenidae Fukui, 1929 (Digenea) based on morphological and molecular evidence with a proposal for Paragyliaucheninae n. subfam. and a description of Flagellotrema convolutum Ozaki, 1936.

Flagellotrema convolutum Ozaki, 1936 was found parasitising the intestine of two new host fish species, the Indian sail-fin surgeonfish, Zebrasoma desjardinii (Bennett) (Acanthuridae), and the Picasso triggerfish, Rhinecanthus assasi (Forsskål) (Balistidae), from the northern Red Sea off Egypt. Another description of this species is provided with detailed morphological observations made of the genital systems. Using newly acquired molecular data from the D1-D3 regions of 28S rDNA, the phylogenetic relationships of subfamilies and genera within the Gyliauchenidae Fukui, 1929 are elucidated with morphological support. The Petalocotylinae Ozaki, 1934 and the Robphildollfusiinae Paggi & Orecchia, 1963 are recognized as valid subfamilies within the Gyliauchenidae. The Apharyngogyliaucheninae Yamaguti, 1942 and the Ichthyotreminae Caballero & Bravo-Hollis, 1952 remain junior synonyms of the Gyliaucheninae Fukui, 1929. Based on its unique position relative to all gyliauchenid subfamilies and its distinct separation from all other gyliauchenine genera, the Paragyliaucheninae n. subfam. is erected to contain Paragyliauchen Yamaguti, 1934. Paragyliauchen differs from all other gyliauchenine genera by having a pharynx differentiated into two, well-developed muscular regions: an anterior region composed of a ring with indented projections anteriorly and a posterior region that is ellipsoidal or barrel-shaped. Modified and/or new keys to the four subfamilies we recognize within the Gyliauchenidae as well as the genera within each subfamily are presented, and we discuss the evolutionary development and etymology of the unique anatomy of the anterior of gyliauchenids.

HISTOPATHOLOGICAL ANALYSIS OF SCHISTOSOMIASIS IN THE GASTROINTESTINAL TRACT WITH FIRST RECORD OF SCHISTOSOMAL APPENDICITIS FROM SOHAG, UPPER EGYPT.

Schistosomiasis is a chronic granulomatous inflammation that affects many systems in the body including the gastrointestinal tract. Appendiceal schistosomiasis is also described and can be a precursor lesion of schistosomal appendicitis. The present study was done to make a retrospective analysis of histopathological changes in the gastrointestinal tract affected by Sckistosoma mansoni among patients attending Sohag University Hospital, Sohag Governorate between June 2013 and June 2016. A total of 150 colon and 30 appendix specimens were collected through out the period from male infected pa- tients aged between 35-50 years and suffering from abdominal pain and dysentery. Histopathological examination of the tissue biopsies was performed. 5p tissue sections were prepared and examined microscopically. Ten specimens were documented to have intestinal schistosomiasis, nine of the colon 9/150 (6%) were diagnosed as chronic schistosomal colitis and one of the appendix 1/30 (3.3%) as chronic schistosomal appendicitis. Microphotographs of the tissue sections were prepared for histopathological observations. Histopathological examination of all specimens revealed degenerated pinkish and calcified bluish bilharzial eggs in the submucosa and even musculosa with surrounding granulomatous reaction. Bilharzial polyps of the colon were detected in two specimens (20%) and bilharzial worms within venules of the muscle layer in two specimens (20%). During the present study, S. mansoni was documented as a not uncommon cause of chronic colitis and for the first time from Sohag as a cause of chronic appendicitis. All specimens did not show any malignant or premalignant cells.

Pelopscreadium aegyptense n. gen., n. sp. and Pelopscreadium spongiosum (Bray & Cribb, 1998) n. comb., (Digenea: Lepocreadiidae), each from disjunct populations of the Yellow boxfish, Ostracion cubicus Linnaeus (Ostraciidae).

Bianium spongiosum Bray & Cribb, 1998 (Lepocreadiidae), described from the yellow boxfish, Ostracion cubicus Linnaeus (Ostraciidae), off Lizard Island, Queensland, Australia, possesses a combination of the following three morphological features which distinguishes it from all the other species currently assigned to the genus: (1) large internal patches of large cells forming sponge-like pads we have termed "pelops"("pelop" sing.) laterally in the forebody extending from near the anterior extremity to about the level of the intestinal bifurcation rather than possessing a scoop; (2) ceca that reach to near the posterior extremity where they end blindly without ani; and (3) a vitellarium which is present laterally but not dorsal to the ceca. Based on this we propose the erection of Pelopscreadium n. gen. (Lepocreadiidae) with the assignment of B. spongiosum to this new genus as the type-species, Pelopscreadium spongiosum (Bray & Cribb, 1998) n. comb. Pelopscreadium aegyptense n. sp., also from the yellow boxfish but from the Red Sea off Sharm El-Naga, Egypt, is described as the second member of the new genus because it shares these three characteristics with P. spongiosum.

First case of intestinal acariasis from Egypt.

We are hereby reporting a case where the eggs and adults of the mold mites; Tyrophagus putrescentiae (Shrank) and the trophozoites of Blastocystis sp. were found in stool of three years old child from Minia City, Egypt. Intestinal mite infection was diagnosed after repeated identification of mite' stages from six consecutive stool samples to exclude the possibilities of contamination and spurious infection. The patient was suffering from severe colicky abdominal pain and burning sensation around the anus one month ago. All other members of his family were having the same acarine in their feces, but were all symptomless. The patient was treated with ivermectin 200 µg/kg body weight once every 10 days for three doses. His cure indicated that he was having asymptomatic blastocystosis.

Present status of protozoan pathogens causing water-borne disease in northern part of El-Minia Governorate, Egypt.

Little is known about the role of different water supplies in the diversity and public health significance of pathogenic protozoan parasites. Most of these organisms have been ubiquitous in waters worldwide. The numbers of waterborne infections indicate a significant risk for their transmission even by drinking water. Hence, a total of 336 water samples were collected during 2009-2013 from different water sources from different areas of northern part of El-Minia Govemnorate, Egypt and were investigated for pathogenic protozoa. They were examined by direct microscopy followed by Modified Ziehl- Neelsen and Giemsa stains. 140 samples (41.7 %) were positive (statistically significant; P value P<0.0003). Prevalence rates were in Summer (66.7%/), Spring (51.1%), Fall (26.2 %) and Winter (22.6%). These data were statistically significant (P<0.0001). The commonest protozoa detected as a single infection was Cryptosporidium sp. (53.17%) found in all water supplies, followed by Blastocystis hominis (15.87%), Cyclospora caytenensis (11.9%), Entamoeba hiseolytica/dispar (8.73%) Giardia lamblia (6.35%() and Naegleria sp., (3.97%). Moreover, there were 14 samples with mixed parsitic infection: they were Cryptosporidium sp. and B. hominis in six samples, Cryptosporidum sp. and C. caytenenisin five samples and Cryptosporidium sp. and E histolytica/dispar in three samples. The most common contamninated water source was ponds where 32 samples (66.7%) were-positive followed by canal water 30 samples (62.5%). The results were discussed and the recommendations were offered.

Comparative study of the sensilla on antenna and maxillary palpsof five culicine mosquitoes in Sohag Governorate, Egypt.

Mosquitoes are unquestionably the most medically important arthropod vectors of disease. The maintenance and transmission of the pathogens that cause malaria, lymphatic filariasis, and numerous viral infections are absolutely dependent on the availability of competent mosquito vectors. Although the medical community has known for over a century the role played by mosquitoes in the transmission of malaria and lymphatic filariasis, these diseases continue to have a devastating influence on less privileged populations throughout the tropical and subtropical regions of the world. The present study was done to identify the different culicine mosquitoes in four different districts in Sohag Governorate and to describe and compare the various types of sensilla located on their antenna and maxillary palps using scanning electron microscopy.

Genetically confirmed Fasciola hepatogigantica n.sp.

Identification of liver fluke species cannot be achieved by clinical, pathological, coprological or immunological methods. However, the differential diagnosis between F. hepatica and F. gigantica infection is very important because of their different pathological manifestations. Moreover, in countries where the two species co-exist, morphologically intermediate forms were reported. The present study aimed to identify these forms by the use of molecular characterization of DNA sequence. Based on morphometric criteria, adults of Fasciola hepatica, F. gigantica and intermediate forms were collected from naturally infected sheep and cattle from various regions of Sohag Governorate. A simple and rapid new method (QIAamp DNA Mini Kit) was used to isolate DNA from the worms and their RELP patterns were obtained after digestion of the PCR products with AvalI restriction enzymes. The result of a regular PCR experiment for the amplification of the selected 28S rDNA fragment with the designed primer set yielded identical 618- bp-long PCR products for the three types of Fasciola where the RFLP profile obtained from F. hepatica revealed four fragments of 628, 575, 165 and 95 bp, and F. gigantica generated three fragments corresponding to 628, 358 and 300 bp fragments whereas the intermediate forms revealed four fragments of 628, 541, 358 and 300 bp, which were similar to those ofF. gigantica but with a distinctive fragment of 541. These results confirmed that three species are present in our locality: F. hepatica, F. gigantica and an intermediate form which was named F. hepatogigantica n.sp. on basis of having few morphometric characters from F. hepatica (length and pattern of uterine coils) but genetically they were more related to F. gigantica.

Histological and PCR xenomonitoring of culicine mosquitoes for filarial infestation in Sohag Governorate, upper Egypt.

The nematodes Wuchereria bancrofti and Brugia malayi; the principal etiologic agents of lymphatic filariasis are mosquito dependant in the biological transmission. Dirofilariasis is essentially a disease of canines which can also be transmitted to humans by culicine mosquitoes. The present work studied the histological and PCR xenomonitoring filarial infestation in culicine mosquitoes in Sohag Governorate. One hundred female mosquitoes of the 5 culicine spesies present in the selected localities, were dissected and histological sections of thoracic muscles were examined for filarial larvae. Also 50 female Culex pipiens molestus were collected from the same areas and tested for 3 filarial worms using PCR. Cx. pipiens molestus was the only culcine mosquito harbouring larvae of Wuchereria bancrofti and Dirofilaria spp. Results were compared and it was found that PCR proved easier to do and it gave better data as it was able to differentiate dirofilarial species. The results indicated a clear success of health authorities in anti-filariasis control measures and pointed out to avoid the hazard of possible occurrence of future cases of human dirofilariasis in Sohag Governorate.

Histochemical and ultrastructural studies on the calcareous corpuscles and eggs of Taenia taeniaeformis and Dipylidium caninum.

Calcareous corpuscles were noticed by several previous workers to be present in larval and adult cestodes without knowing their function. However, nothing was mentioned in the available literature about distribution of these corpuscles and their density, structure and composition in different parts of the body of different cestodes. Hence, in the present work, a comparative study of their distribution, density, histochemical and ultrastructural characters in different parts of the body was performed in Taenia taeniaeformis and Dipylidium caninum. Due to the presence of the eggs in their gravid segments, their histochemical and ultrastructural characteristics were also studied. It was found that the size, location and density of the calcareous bodies were different in different body parts of the same and the other cestode. Histochemically, the main component of these corpuscles was calcium; while other constituents as polysaccharides, lipids, protrins and mucopolysaccharides were found in their outer rim. Ultrastructurally, they were quite similar in the two studied cestodes and different stages of their development were exhibited. Histochemically, the eggs of both cestodes were similar in their contents. However, some ultrastructural differences have been demonstrated particularly in relation to the size and shape of the rods in the embryophore and the structures in between the embryophore and onchosphere.

Development and hatching mechanism of Fasciola eggs, light and scanning electron microscopic studies.

Both light microscopy and scanning electron microscopy were used in the description. There were variable measurements of eggs from the same fluke and there was no relationship between the size of the fluke and size of eggs. Light microscopy revealed that the operculum has different shapes in Fasciola gigantica and F. hepatica. Under normal laboratory conditions of temperature (26 ± 1 °C), miracidia of F. gigantica developed within 12-16 days period, but those of F. hepatica developed within a period of 13-15 days. The miracidia of F. gigantica measured 110 × 70 µm and that of F. hepatica measured 136 × 74 µm. The life span of the miracidium in F. gigantica ranged between 9 and 12 h, while in F. hepatica the life span did not exceed 10 h. Miracidial epidermal plates in miracidium of F. gigantica were found to be 20 plates arranged in four tiers of six, four, six and four. Hatching process was recorded using scanning electron microscope, and it indicated partially opened operculum.

Comparative morphological studies on three Sarcocystis species in Sohag, Egypt.

Samples from the tongue, heart, oesophageal and skeletal muscles were collected from 100 cow and 100 buffalo from Sohag slaughterhouses. Macroscopic examination for sarcocysts was followed by microscopic ones on impression smears and compressed muscles. Histological sections and transmission electron microscope (TEM) studies were done on positive cases. Sarcocystis infection rate of cows were 84%. Two species of Sarcocystis were detected: 1- S. cruzi with a membrane provided with hair-like villar protrusions folded over the surface of the cyst. These protrusions were rod-like, round, oval or irregular in shape and were approximately parallel to the cyst surface. 2-S. hominis characterized by a cyst wall consisting of cylindrical finger-like villar protrusions and having microfilaments. The protrusions were perpendicular on the cyst surface with broad tips and contain microfilaments. Buffaloes' muscle samples revealed an infection rate of 28%. The macroscopic fusiform-shaped species only was detected (Sarcocystis fusiformis). The role of cyst wall ultrastructure and thickness were discussed regarding identification and pathogenicity.

The pattern of bilharzial infection with hepatitis B virus associated with delta agent.

Delta hepatitis is a relatively common in Egypt especially in patients with chronic liver disease. Delta virus infection is associated with a high frequency of hepatic decompensation and it seems to impose additional burden on liver function. It does not seem to be more prevalent in particular type of cirrhosis but the CAH pathology was found to be mostly associated with virus infection. HBV replication seems to be inhibited by the presence of delta virus as evidenced by the low frequency of HBe Ag in serum and HBs Ag in liver among patients with HDV infection.

Studies on certain serum enzymatic activities in hepatosplenic bilharziasis.

Determination of serum enzymatic activities in hepatosplenic bilharziasis (H.S.B.) were conducted in 100 bilharzial patients, in different stages and 30 controls SGot, SGPT, ALK pH, ALD and SLDH with its both fractions heat stable and labile.). Early elevation of serum enzymatic activities of SGOT, SGPT and SALK. pH, may be considered as a sensitive parameter for functional changes in H.S.B. rather than other conventional liver function tests. The elevated enzymic activity of total LDH in this study was associated with elevation in its both fractions. In particular, the changes in the total activity was in parallel with that of its heat labile fraction. The latter may be considered as a confirmatory test for marked deterioration of liver functions in H.S.B. The changes in the heat stable fraction was inconstant and may be attributed to extrahepatic bilharzial dissimilation. Significantly high serum enzymatic activity of SALD was found in cases of H.S.B. particularly those showing striking muscle wasting.