Mohie A. M. Haridy

Pathological and immunohistochemical study of chickens with co-infection of Marek's disease virus and chicken anaemia virus

Chicken anaemia virus (CAV) is the most important confounding pathogen in Mareks disease virus (MDV) infection. The effect of CAV co-infection at 4 weeks of age after inoculation of virulent MDV (vMDV, KS strain) or very virulent MDV (vvMDV, Md/5 strain) in 1-day-old chicks was investigated by pathological and immunohistochemical studies. CAV increased the mortality rates induced by vMDV or vvMDV. The packed cell volume was reduced significantly in vMDV–CAV infection; however, no reduction or non-significant reduction was observed in vMDV infection. Bone marrow hypoplasia was related to CAV co-infection and none of the birds inoculated with vMDV or vvMDV had hypoplasia. Severe atrophy of the thymus and bursa of Fabricius was observed in the vvMDV–CAV and vvMDV groups. Complete regeneration of the thymus cortex and bursa of Fabricius in the vMDV group was noted and was in contrast to sequential lymphoid depletion after CAV inoculation in the vMDV–CAV group. The spleen was either regenerated, lymphoid depleted or had lymphoproliferative lesions. Lymphoid depletion in the spleen was not detected in the vMDV group; however, it was prominent in the vMDV–CAV and vvMDV–CAV groups during the first 2 weeks after CAV inoculation. CAV inclusions and antigens were detected in the thymus cortex and spleen of vMDV–CAV and vvMDV–CAV groups during the experiment. Severe depletion of CD8+ T cells was observed in depleted spleen and thymus. The neoplastic foci appeared around splenic arterioles and venules, and stained mainly by CD4 antibody; however, CD8+ T cells were singly dispersed or were present in clusters. It could be concluded that CAV was responsible for bone marrow hypoplasia, severe anaemia and hindrance of lymphoid organ regeneration in MDV–CAV co-infection.

Fullerene C 60 nanoparticles ameliorated cyclophosphamide-induced acute hepatotoxicity in rats

Cyclophosphamide (CP), a chemotherapeutic agent, induces hepatotoxicity as one of its side effects. Therefore, the aim of the present study is to investigate the potential hepatoprotective effects of fullerene C60 nanoparticles (C60) against the high toxic dose of CP. Twenty five albino rats were randomly assigned to 5 groups (n=5 per group). Group 1 served as a control. Group 2 received 200mg/kg of CP once intraperitoneally, while group 3 treated with the same CP dose plus C60 (4mgkg, orally) daily for 10days. Group 4 exposed CP and ZnCl2 (4mgkg, orally) daily for 10days. Group 5 exposed to CP and co-treated with C60 and ZnCl2. One day after last treatment, blood and livers were collected for hematological, biochemical and histopathological investigations. C60 normalized significantly RBCs, HB, PCV, WBCs and platelets numbers compared to CP-exposed rats. Moreover, liver enzymes namely ALT, AST and ALP revealed that CP elevated their levels and C60 significantly (p<0.05) reduced them to basal levels. The level of oxidative stress marker namely, MDA was elevated upon CP exposure and normalized by C60 treatment. In addition, antioxidant systems e.g. GSH, CAT and SOD were depleted from liver tissue due to CP toxicity these were recovered by C60 administration. The hepatoprotective effects of C60 on tested parameters were comparable with ZnCl2 and neither additive nor synergistic effect was observed. Histopathogically, severe liver degeneration was recorded after CP treatment, however, only mild changes were observed after C60 administration. Our data suggest that C60 improves both blood and hepatic parameters altered by cyclophosphamide-induced toxicities. The current study is of clinical relevance particularly, application of C60 as a monotherapy or in combination to ameliorate the CP side effects in cancer-treated patients.

Hepato- and nephroprotective effects of bradykinin potentiating factor from scorpion (Buthus occitanus) venom on mercuric chloride-treated rats

Bioactive peptides such as bradykinin potentiating factor (BPF), have, anti-oxidative, anti-inflammatory, immunomodulatory and ameliorative effects in chronic diseases and play a potential role in cancer prevention. It is known that the liver and kidney accumulate inorganic mercury upon exposure, which often leads to mercury intoxication in these organs. In this study, we investigated the effect of bradykinin potentiating factor (BPF), a scorpion venom peptide, on mercuric chloride-induced hepatic and renal toxicity in rats. We used 20 adult male Albino rats divided into four equal groups: the first group was injected with saline (control); the second group was administered daily with mercuric chloride (HgCl2) for 2 weeks; the third group was administered with BPF twice weekly for 2 successive weeks, while the fourth group was exposed to BPF followed by HgCl2. We observed that HgCl2 treated rats had a significant increase in serum ALT, AST, ALP, creatinine and urea levels compared to control. Furthermore, HgCl2 treated rats showed a marked decrease in total proteins, albumin and uric acids compared to control. The previously studied parameters were not significantly changed in BPF pretreated rats compared to control. Moreover, a significant decrease in the activities of glutathione perioxidase (GSH), superoxide dismutase (SOD), and catalase (CAT), in addition to a significant increase in the level of malondialdehyde (MDA) were observed in hepatic and renal tissues of rats after HgCl2 treatment. In contrast, the HgCl2/BPF treated rats showed a significant elevation in the activity of GSH, SOD, and CAT accompanied with a significant regression in the level of MDA compared to the HgCl2 exposed rats. We conclude that treatment with BPF is a promising prophylactic approach for the management of mercuric chloride-induced hepato- and nephro-toxicities.

Impacts of fullerene C60 and virgin olive oil on cadmium-induced genotoxicity in rats

Currently, cadmium is considered to be one of the major environmental pollutants. Environmentally, cadmium is released in various forms e.g. oxide, chloride and sulphide. The aim of the present study was to examine the genotoxic impact of fullerene nanoparticles C60 (C60) and virgin olive oil (VOO) on cadmium chloride (CdCl2)-induced genotoxicity in rats. To evaluate these effects on DNA damage and chromosomal frequency, 25 albino rats were randomly assigned to 5 groups (n=5 per group): Group 1 served as a control; Group 2 received a single intraperitoneal dose of CdCl2 (3.5mg/kg); Group 3 animals were treated with C60 (4mg/kg, orally) every other day for 20days; Group 4 received a single intraperitoneal dose of CdCl2 (3.5mg/kg) and an oral dose of C60 (4mg/kg); and Group 5 received a single intraperitoneal dose of CdCl2 (3.5mg/kg) and oral doses of VOO every other day for 20 consecutive days. Genotoxic and anti-genotoxic effects of C60 and VOO were evaluated in the liver, kidney and bone marrow using molecular and cytogenetic assays. As expected, CdCl2 and C60 administration was associated with band number alterations in both liver and kidney; however, C60 pretreatment recovered to approximately basal number. Surprisingly, C60 and VOO significantly attenuated the genotoxic effects caused by CdCl2 in livers and kidneys. In bone marrow, in addition to a reduction in the chromosomal number, several chromosomal aberrations were caused by CdCl2. These chromosomal alterations were also reversed by C60 and VOO. In conclusion, molecular and cytogenetic studies showed that C60 and VOO exhibit anti-genotoxic agents against CdCl2-induced genotoxicity in rats. Further studies are needed to investigate the optimal conditions for potential biomedical applications of these anti-genotoxic agents.

Effect of a bradykinin-potentiating factor isolated from scorpion venom (Leiurus quinquestriatus ) on some blood indices and lipid profile in irradiated rats

Bradykinin appears to be an important regulator of cardiovascular function. It is also being increasingly noted as a participant in actions of drugs that affect the liver, kidney, and circulation. In our previous studies, bradykinin-potentiating factor (BPF) isolated from scorpion venom (Leiurus quinquestriatus) has been shown to be protective against hepato- and nephrotoxicity as well as healing skin burns by reducing oxidative stress in hyperglycemic conditions. Therefore, we aim to evaluate the ability of BPF in treating irradiated rats. A group of rats was exposed to γ-irradiation and subsequently treated with BPF injections aiming to elucidate the possibility of BPF to rescue γ-irradiation harmful effects. As controls, we used γ-irradiation exposed, BPF-injected, and untreated rats. The data obtained showed that the irradiated animals suffered from marked changes of many important blood parameters including red blood cells, leukocytes, platelets, hemoglobin, packed cell volume, high-density cholesterol, total cholesterol, triglycerides, and low-density cholesterol. Interestingly, BPF was able to rescue the deleterious effects of irradiation in rats and normalized their blood parameters to the basal levels. We conclude that BPF could ameliorate irradiation damaging effects.

Coenurus cerebralis cyst in the orbit of a ewe.

A 4-year-old Rahmani breed ewe was presented for surgery to the Veterinary Teaching Hospital, South Valley University, Egypt with enlargement and protrusion of the eye ball, blepharitis and congestion of the conjunctiva. On examination, a cyst 2.5 cm x 3.5 cm in diameter containing sandy fluid was detected in the perioptic nerve fat. Histopathological examination revealed that the epithelial lining of the conjunctiva was necrotic and severely infiltrated by neutrophils. The underlying connective tissue was oedematous, hyperaemic and severely infiltrated by neutrophils. Desquamation of the corneal epithelium was seen, together with oedema of the stroma. The tissue surrounding the cyst was compressed and the lacrimal glands revealed pressure atrophy. The muscular tissue was atrophied and infiltrated by fat cells. The cyst wall was lined with white scolices protruding from the inner wall. Based on the gross and histopathological characteristics of the cyst observed, the cyst was diagnosed as Coenurus cerebralis. This is the first report of orbital coenurosis in a sheep.A 4-year-old Rahmani breed ewe was presented for surgery to the Veterinary Teaching Hospital, South Valley University, Egypt with enlargement and protrusion of the eye ball, blepharitis and congestion of the conjunctiva. On examination, a cyst 2.5 cm x 3.5 cm in diameter containing sandy fluid was detected in the perioptic nerve fat. Histopathological examination revealed that the epithelial lining of the conjunctiva was necrotic and severely infiltrated by neutrophils. The underlying connective tissue was oedematous, hyperaemic and severely infiltrated by neutrophils. Desquamation of the corneal epithelium was seen, together with oedema of the stroma. The tissue surrounding the cyst was compressed and the lacrimal glands revealed pressure atrophy. The muscular tissue was atrophied and infiltrated by fat cells. The cyst wall was lined with white scolices protruding from the inner wall. Based on the gross and histopathological characteristics of the cyst observed, the cyst was diagnosed as Coenurus cerebralis. This is the first report of orbital coenurosis in a sheep.

Efficacy of Caltropis procera and Ficus sycomorus extracts in treating MRSA (methicillin-resistant Staphylococcus aureus)-keratitis in rabbit.

MRSA-induced keratitis in rabbit was used to evaluate the therapeutic effect of F. sycomorus leaves and C. procera latex extracts. Within the 6 rabbit groups tested, group 1 received sterilized saline, while other groups (2 to 6) received 100 μl of intrastromal injections of 1.5×103 colony forming unit (cfu) ml-1 of methicillin-resistant Staphylococcus aureus (MRSA). After 12 hours, groups 3 to 6 also received chloramphenicol, aqueous extract of C. procera latex, aqueous and alcoholic extracts of F. sycomorus leaves, respectively 3 times daily for 12 successive days. The tested extracts inhibited MRSA growth in vitro (i.e. on culture medium). Colony counts in cornea discs from groups 3 to 6 were significantly reduced (P ≤ 0.001) compared to group 2 (untreated). Clinical signs of keratitis were observed on group 2 until the end of experiment. In groups 3 to 6, gradual recovery was observed and signs disappeared by the 12th DPI (days post inoculation). Only mild symptoms persisted in group 5 (aqueous extract of leaves). In group 3 and 5, cornea, iris, ciliary body and conjunctiva showed mild leukocytic infiltration and depigmentation of melanin cells while recovery of cornea and iris was observed in groups 4 and 6. In conclusion, the used extracts have potential therapeutic effects on MRSA-induced keratitis in rabbit.

Pathological Findings of Tenacibaculum maritimus Infection in Black Damselfish, Neoglyphieodon melas and Picasso Triggerfish, Rhinecanthus assasi in Red Sea, Egypt

An outbreak of a disease with external body lesions in Picasso triggerfish (Rhinecanthus assasi) and black damselfish (Neoglyphieodon melas) has been observed in the indoor aquarium of NIOF, Hurghada, Egypt. Thirty Picasso triggerfish and thirty-five black damselfish were clinically affected with gross skin lesions. The affected fishes were euthanized for gross, histological and bacteriological examinations. The disease onset started after exposing the fish to catching and indoor rearing stress. Anorexia, erratic swimming and hemorrhagic skin and mouth ulcers and fin rot were the main symptoms and lesions among Picasso triggerfish and black damselfish. Single species of bacteria was isolated from skin ulcer, liver and spleen and was diagnosed as Tenacibaculum maritimus. The gross lesions were irregular shallow erosions and deep ulcers on the dorsal and lateral skin, sometimes on the head, mouth and fins, and occasionally, on the cornea. Histological findings, the hepatic tissue revealed fatty degeneration and focal liquefactive necrosis. Congestion, hemorrhage, hemosidrosis and lymphoid depletion was observed in spleen. The deep skin ulcers characterized by epidermal and muscular necrosis. Picasso triggerfish and black damselfish had lesions similar to those of natural infection and mortality rates of 50 and 60%, respectively in experimental reproduction of the disease. The natural T. maritimus infection in Picasso triggerfish and black damselfish in Red Sea is not only a superficial skin lesion but also a systemic infection.

Protective Effects of Fullerene C60 Nanoparticles and Virgin Olive Oil against Genotoxicity Induced by Cyclophosphamide in Rats

The potential effects of the fullerene C60 nanoparticle (C60) as well as virgin olive oil (VOO) against the cyclophosphamide- (CP-) induced cytotoxic and mutagenic effects were evaluated by two main methods: molecular intersimple sequence repeat (ISSR) assay and cytogenetic biomarkers. Thirty adult male rats were divided to five groups (control, CP, C60, CP + C60, and CP + VOO). CP was i.p. injected with a single dose of 200 mg/kg; C60 and VOO were given orally (4 mg/kg dissolved in VOO and 1 ml, resp.) in alternative days for 20 days. The ISSR analysis revealed an increased in the DNA fragmentation level for liver and heart tissues represented by 21.2% and 32.6%, respectively, in the CP group. The DNA polymorphism levels were modulated and improved in CP + C60 (8.9% and 12%) and CP + VOO (9.8% and 12.7%) for hepatic and cardiac tissues, respectively. The bone marrow cytogenetic analysis revealed that C60 and VOO had significantly decreased the frequency of CP-induced chromosomal aberrations (chromosomal ring, deletion, dicentric chromosome, fragmentation, and polyploidy). Fullerene C60 and VOO have ability to reduce DNA damage and decrease chromosomal aberrations. In conclusion, fullerene C60 and VOO have protective effects against the CP-induced mutagenicity and genotoxicity. Fullerene C60 and VOO open an interesting field concerning their potential antigenotoxic agents against deleterious side effects of chemotherapeutics.

Candida parapsilosis and Candida tropicalis infections in an Okhotsk snailfish (Liparis ochotensis)

An Okhotsk snailfish (Liparis ochotensis) kept at Nagoya aquarium exhibited sudden death. Microscopically, the fish showed multiple granulomatous foci in the gills, liver and kidney. Multiple yeast-like organisms as well as pseudohyphal elements were observed within granulomatous lesions. Immunohistochemically, the hyphae were negative for both Asperigullus and Mucor spp., and a weak positive for Candida sp. The seminated-PCR product was consistent with Candida parapsilosis and C. tropicalis. This is the first record of disseminated mycotic granulomatous lesion due to C. parapsilosis and C. tropicalis infection in fish.