Recent insights into the morphology, molecular characterization and tissue localization of the caprine Sarcocystis species infecting domestic goats (Capra hiricus): Sarcocystis moulei, Sarcocystis capracanis, and Sarcocystis hircicanis.

Ninety-seven (64.67%) out of 150 domestic goats (Capra hiricus) carcasses were found to be infected by Sarcocystis moulei, Sarcocystis capracanis, and Sarcocystis hircicanis sarcocysts. Sarcocystis moulei macrosarcocysts were detected in the cardiac, esophageal, skeletal, lingual, and diaphragmatic muscles of seven goats (4.67%) out of the 150 examined animals, whereas the microscopic Sarcocystis species were found in (90/150 = 60%). Two morphotypes of S. moulei were observed. Morphotype (I) macrosarcocysts were large-sized oval, ovoid, spherical, and measured 2-7 mm in length x 2-6 mm in width. Sarcocystis moulei morphotype (II) macrosarcocysts were spindle-shaped, spheroid, sometimes elongated, and measured 1.8-6 x 0.5-2 mm. By TEM, all S. moulei morphotypes were ultrastructurally the same and had a sarcocyst wall that was characterized by highly branched or cauliflower-like villar protrusions (VP) with dumbbell-like structures. The VP interior was packed with well-developed microtubules in longitudinal and cross arrangements. Sarcocystis moulei cyst wall was 3-6 µm thick. Sarcocystis capracanis microsarcocysts detected herein had a cyst wall that ranged from 4-8 µm in thickness. The VP was upright finger-like or cylindrical. The PVM had electron-dense corrugations in the region of the VP. Few amounts of microfilaments were detected inside the cores of VP. Sarcocystis hircicanis had a thinner cyst wall (~1-3 µm) with hairy long VP that ranged from 1 to 7.5 µm in length. Microtubules were missing inside the cores of the VP. The three caprine Sarcocystis species were molecularly characterized on the level of the 18S rRNA, 28S rRNA, and Cox1 genes.

Morphologic and molecular identification of three macroscopic Sarcocystis species infecting domestic sheep (Ovis aries) and cattle (Bos taurus) in Egypt.

In a survey study on the macroscopic species of Sarcocystis infecting domestic sheep (Ovis aries) and cattle (Bos taurus) in Egypt, the macrosarcocysts of Sarcocystis gigantea and Sarcocystis medusiformis were detected in the carcasses of 33 domestic sheep out of a total of 250 (13.20%), whereas Sarcocystis hirsuta macrosarcocysts were found in 17 out of 150 cattle (11.33%) slaughtered at the municipal abattoirs of two different provinces in Egypt. The sarcocysts of each species were thoroughly described morphologically through gross inspection, histopathologic and transmission electron microscopic (TEM) examination. By TEM, S. gigantea primary cyst wall was 6-7.5 µm thick and had irregular highly branched cauliflower-like villar protrusions (VP).The VP contained microtubules (mt) and multiple electron dense granules (edg) that were dispersed inside the cores of the branched VP. Besides, the parasitophorous vacuolar membrane (PVM) had minute blister-like invaginations all over the entire surface of the sarcocyst. S. medusiformis cyst had a thin sarcocyst wall (~2 µm thick) as compared to that of S. gigantea. The cyst wall had trapezoidal or nearly pyramidal VP that were surrounded by thick PVM in addition to a ground substance GS that contained electron-dense fine particles. S. hirsuta sarcocyst wall was 7-9 µm thick and possessed rhomboid-shaped VP that contained microtubules (mt) and electron-dense granules (edg) of variable sizes. The edg were arranged in rows and running parallel to the longitudinal axis of the protrusions. The VP had characteristic narrow neck-like constrictions at their bases, dilated middle portions, and tapered distal ends. The detected macrosarcocysts were eventually confirmed by molecular characterization on the levels of 18S rRNA, 28S rRNA, and Cox1 sequences. Phylogenetic analyses based on the sequences of the 18S rRNA and Cox1 genetic markers gave rise to robust associations of the currently identified isolates of S. gigantea, S. medusiformis, and S. hirsuta within a major clade of Sarcocystis species with felines as presumed or known definitive hosts.

S-Methylcysteine (SMC) Ameliorates Intestinal, Hepatic, and Splenic Damage Induced by Cryptosporidium parvum Infection Via Targeting Inflammatory Modulators and Oxidative Stress in Swiss Albino Mice.

Cryptosporidiosis has been proposed to be one of the major causes of diarrhoeal disease in humans worldwide that possesses zoonotic concern. Thereby, this study investigated the potential effects of s-Methylcysteine (SMC) on the parasite in vivo followed by the measurement of cytokines, oxidative stress parameters, and an investigation of the major histopathological changes. Sixty male Swiss albino mice weighing 20-25 g were allocated equally into five groups and orally administered saline only (control), SMC only (SMC50) (50 mg/kg b.w.), and 104Cryptosporidium parvum oocysts per mouse via an esophageal tube (C + ve untreated). The fourth and fifth groups (C + SMC25, C + SMC50) administrated 104C. parvum oocysts combined with SMC25 (low dose) and 50 (high dose) mg/kg b.w., respectively. At days 7 and 14 post-infection (PI), the feces was collected from each group in order to count C. parvum oocysts. After two weeks of treatment, the animals were euthanized and the serum was collected for biochemical analysis. Next, the intestinal, spleen, and liver sections were dissected for histopathological examination. The results revealed lower oocyst numbers in the C + SMC25 and C + SMC50 groups compared to the infected untreated group. Moreover, higher doses of SMC treatment significantly reduced the enteritis induced by C. parvum in a dose-dependent manner. The hepatic lesions were also mitigated as demonstrated in C + SMC25 and C + SMC50 groups unlike the infected group via lowering the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) enzymes and increasing albumin and globulin serum levels. SMC administration also reduced cytokines production (SAP, TNF-α, IL-6, and IFN-γ) mediated by Cryptosporidium infection in contrast to the infected untreated group. There were marked lymphoid depletion and amyloidosis observed in the infected untreated group, while the treated groups showed obvious increase in the lymphoid elements. Moreover, the scoring of intestinal parasites, hepatic, and splenic lesions in the SMC-treated groups exhibited significantly lower pathological lesions in different organs in a dose-dependent manner, compared to the infected untreated group. Our results also revealed a significant change in the malondialdehyde content with an elevation of glutathione and superoxide dismutase in the intestines collected from C + SMC25 and C + SMC50 mice relative to the untreated group. Taken together, our results indicated that SMC could be a promising effective compound for treating and declining C. parvum infestation via restoring structural alterations in different tissues, enhancing antioxidant enzymes, and suppressing the cytokines liberation.

The anticoccidial activity of the fluoroquinolone lomefloxacin against experimental Eimeria tenella infection in broiler chickens.

Coccidiosis is a crucial parasitic disease of the poultry industry. As a result of the enormous global economic losses and the increased resistance to the conventional anticoccidial agents, there is a continuous need to find new anticoccidials. Here, the anticoccidial effect of the fluoroquinolone lomefloxacin versus diclazuril in experimentally infected broilers was tested for the treatment of Eimeria tenella infection. Ninety 14-day-old Cobb strain broiler chickens were allocated into five groups, each with 18 chicks. Group 1 (G1) was separated as an uninfected negative control and received no treatment; group 2 (G2), infected untreated (positive control); group 3 (G3), infected and treated with lomefloxacin at a dose rate of 100 ppm in drinking water; group 4 (G4), infected and treated with diclazuril at a dose rate of 2.5 ppm in drinking water; group 5 (G5), infected and treated with lomefloxacin at a dose rate of 100 ppm plus diclazuril at dose rate of 2.5 ppm in drinking water. Clinical signs, mortality rates, number of oocysts per gram of faeces (OPG), growth performance parameters (weight gain: WG and feed conversion ratio: FCR), lesion scoring, haematological and serum biochemical analyses, antioxidant biomarkers and histopathologic inspection of the caeca were used as evaluation criteria for the anticoccidial efficacy of both lomefloxacin and diclazuril. The findings herein showed that administration of lomefloxacin and/or diclazuril improved growth performance parameters (WG, FCR) and significantly (P ≤ 0.05) reduced OPG, and diminished the severity of bloody diarrhoea and mortalities. Additionally, haematological indices and serum biochemical parameters such as ALT, AST, ALP, creatinine, uric acid, total proteins, albumin and globulin were improved. Finally, a significant elevation in the levels of the antioxidant biomarkers was observed in the chicks of G3, G4 and G5 as compared with those of G2.

Ultrastructural and Molecular Identification of the sarcocysts of Sarcocystis tenella and Sarcocystis arieticanis Infecting Domestic Sheep (Ovis aries) from Egypt.

BACKGROUND: In spite of the global economic significance of sheep production, little is known about the prevalence of various Sarcocystis spp. infecting the domestic sheep (Ovis aries) in Egypt. MATERIALS AND METHODS: Muscle samples were collected from 175 sheep (> 2 years) slaughtered at El-Mahalla El-Kubra abattoir, Gharbia governorate, Egypt. Samples were initially examined by naked eye for the existence of macrosarcocysts. The microscopic sarcocysts were detected and identified using the light microscopy and the Transmission electron microscopy (TEM). Different microscopic species of ovine Sarcocystis were molecularly confirmed by PCR, sequence analyses and phylogeny. RESULTS: Preliminary light microscopic inspection of the muscle specimens revealed the existence of only the microscopic sarcocysts of Sarcocystis tenella and Sarcocystis arieticanis in 152 (86.8%) out of the175 examined animals. Sarcoysts of S.tenella had striated thick cyst wall that amounted from 3.5-5.5 µm in thickness whereas, S.arieticanis sarcocysts had a thin cyst wall that ranged from 1-3 µm in thickness. S.tenella sarcocysts were detected in 115 sheep (65.7%), and were more prevalent than those of S.arieticanis, observed only in 68 sheep (38.8%). No macroscopic sarcocysts were observed in any of the examined carcasses. Transmission electron microscopy (TEM) of the cyst wall of S.tenella revealed the existence of the short stubby villar protrusions (VP) with the characteristic disk-like structures at the tips of the (VP). While, TEM of S.arieticanis showed that the cyst wall had elongated tubular protrusions that measured approximately 5-7 µm in length. Each (VP) consisted of a dome-shaped base (0.3-0.9 µm in diameter), a relatively thick middle portion (0.1-0.3 µm) in width, and a thin hair-like distal portion that measured about (0.03 x 1-4.5 µm). CONCLUSION: Comparative analyses of the sequences of the four genetic markers (18S rRNA, 28S rRNA, mitochondrial cox1 and ITS-1) for S.tenella and S.arieticanis isolates detected herein, revealed genetic variations of 95% and 95- 96% among the different isolates on the level of the 18S rRNA and 28S rRNA, respectively. Whereas, the cox1 and ITS-1 shared sequence identities of 76-78% and 70-73%, respectively. S.tenella was strongly related to S.capracanis infecting goats (Capra hircus). Sequence identity of 98% on the level of 18S rRNA, 28S rRNA genes was observed between the currently identified isolates of S.tenella and the formerly GenBank deposited isolates of S.capracanis. While, cox1 sequences shared identities of 92-93%. Furthermore, S.arieticanis isolates identified here were closely related to the formerly published sequences of S.hircicanis. The 18S rRNA and 28S rRNA sequences of S.arieticanis shared 98% and 94-95% identities with those of S.hircicanis, respectively. However, 87-88% homologies were observed between the cox1 sequences of S.arieticanis and S.hircicanis. Consequently, cox1 and ITS-1 gene sequences act as better genetic markers than 18S rRNA and 28S rRNA sequences for the characterization of ovine Sarcocystis spp. Maximum parsimony analyses based on the sequences of three genetic markers, (18S rRNA, 28S rRNA and mitochondrial cox1), yielded the same placement of the currently identified isolates of the two taxa (S.tenella and S.arieticanis) within a clade of Sarcocystis species with carnivorous animals as known, or assumed, final hosts.

Poxvirus infection in a Steller's sea eagle (Haliaeetus pelagicus).

A severely emaciated adult Steller's sea eagle (Haliaeetus pelagicus) was found dead with electrocution-induced severe wing laceration, and with multiple cutaneous pock nodules at the periocular regions of both sides nearby the medial canthi and rhamphotheca. Histopathological examination of the nodules revealed hyperplasia of the epidermis with vacuolar degeneration and intracytoplasmic inclusion bodies (Bollinger bodies). The proventriculus was severely affected by nematodes and was ulcerated. Nucleotide sequencing of a PCR-amplified product of Avipoxvirus 4b core gene revealed 100% identity to the sequence of Avipoxvirus derived from other eagle species. This report describes the first detection of Avipoxvirus clade A from a Steller's sea eagle.

Morphologic and molecular characteristics of Sarcocystis atraii n. sp. (Apicomplexa: Sarcocystidae) infecting the common coot (Fulica atra) from Egypt.

A single morphologic type of Sarcocystis cysts found in two out of 43 examined common coots, Fulica atra, is considered to represent a new species for which the name Sarcocystis atraii n. sp. is proposed and its description is provided. Coots were hunted from the vicinity of Brolos Lake located at KafrElsheikh province, Egypt. The structural morphology of the revealed sarcocysts was described using light and transmission electron microscopy. Sarcocysts were found in the leg and thigh muscles. The cysts were microscopic and measured 165-850 µm in length × 50-85 µm in width. Histologically; the sarcocyst wall was wavy and had minute undulations. Ultrastructurally, it measured 1-3 µm in thickness and possessed many mushroom-like villar protrusions sometimes originating from other mushroom-like villar protrusions that measured approximately 0.5-2 µm in length and up to 2 µm in width, with the presence of electron dense ground substance of 300 nm to 1 µm thick. The bradyzoites were elongated, banana-shaped and measured 7.5-14 × 1.5-2.5 µm, with centrally or terminally located nuclei. The ultrastructural features of the cyst wall belonged to type 24. On the basis of sequencing and phylogenic analyses for 18S rRNA , 28S rRNA genes and ITS-1 region; S. atraii n. sp. is considered a genetically distinct species, being most closely related to avian Sarcocystis spp. whose definitive hosts are predatory mammals.

Morphologic identification of a new Sarcocystis sp. in the common moorhen (Gallinula chloropus ) (Aves: Gruiformes: Rallidae) from Brolos Lake, Egypt.

Sarcocystis species are among the most common and widespread protozoan parasites of mammals and birds. The current study provides the first record of infection with Sarcocystis species in the common moorhens from Brolos Lake, KafrElsheikh province, Egypt. Morphology of the parasite cysts was described using light and transmission electron microscopy. Out of 25 examined birds, sarcocysts were found in neck, thigh, and legs muscles of two birds. The cysts were microscopic and measured 150-650 µm in length×45-185 µm in width. Histologically, the sarcocyst wall appeared striated and characterized by the presence of radial spines. Ultrastructurally, it measured 2-4.5 µm in thickness and had irregularly shaped crowded finger-like villar protrusions that measured 1.5-4 µm in length and up to 0.4-2 µm in width with the presence of dense electron ground substance of 200-750 nm thick. Several septa derived from the ground substance were present and divided the cyst into compartments containing both bradyzoites and metrocytes. The bradyzoites were banana-shaped and measured 6-12 × 1-2 µm with centrally or posteriorly located nuclei. The ultrastructural features of the cyst wall belonged to type 10 cyst wall according the classification of Dubey et al. (1989) and Dubey and Odening (2001).

Prevalence and diversity of Hepatozoon canis in naturally infected dogs in Japanese islands and peninsulas.

Canine hepatozoonosis is a worldwide protozoal disease caused by Hepatozoon canis and Hepatozoon americanum and is transmitted by ixodid ticks, Rhipicephalus and Amblyomma spp., respectively. H. canis infection is widespread in Africa, Europe, South America, and Asia, including Japan. The objective of this study was to study the distribution pattern and diversity of H. canis in naturally infected dogs in nine Japanese islands and peninsulas. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011 and the ages and sexes were identified. Direct microscopy using Giemsa-stained blood smears revealed H. canis gametocytes in the peripheral blood of 45 (23.6%) dogs. Polymerase chain reaction (PCR) was performed on EDTA-anticoagulated blood, initially with the common primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, and then with the specific primer set (HepF and HepR) amplifying about 660 bp fragments of the same gene. Based on PCR, 84 (42.9%) dogs were positive using the common primer and 81 (41.3%) were positive using the specific primer. The current investigation indicated that all screened areas, except for Sado Island and Atsumi Peninsula, were infected. Yaku Island had the highest infection rate (84.6% in males and 100.0% in females), while Ishigaki Island showed the lowest infection rates (8.3% in males and 17.7% in females). Both sexes were infected with no significant difference. However, diversity of infection among the surveyed islands and peninsulas was significantly different (P < 0.05). Although H. canis has previously been reported in dogs in Japan, the higher infection rate described in the current study and the diversity of infection in a wide range of islands strongly encourage prospective studies dealing with the prevention and treatment of the infection in dogs, as well as control of ticks.

Coenurus cerebralis cysts in the left lateral cerebral ventricle of a ewe.

A three-and-a-half year-old female Rahmani ewe was presented suffering from nervous symptoms. Grossly, a large cyst measuring 7 × 4 cm and weighing 145 g occupied the dilated left lateral ventricle. The overlying cerebral tissue was thin, atrophied and congested. It tore easily, and the cyst was evacuated spontaneously. Microscopically, liquefactive necrosis surrounded by aggregations of macrophages, eosinophils, lymphocytes, fibroblasts and giant cells was predominantly observed. Hyperplasia and severe necrosis of the ependymal cell lining of the lateral ventricle were observed. Extensive subependymal inflammatory cell infiltrations, accompanied by neovascularization and fibroblastic proliferation, were seen. Based on the gross and histopathological lesions and cyst morphology and location, the cyst was diagnosed as Coenurus cerebralis. This report describes a rare case of coenurus cyst in the left lateral cerebral ventricle of a ewe and the associated lesion.