Effect of prophylactic vitamin C administration on the efficiency of florfenicol or sulfadiazine-trimethoprim antimicrobial therapy in chickens with staphylococcal arthritis.

Septic arthritis (SA) in chickens shows improper response to antibacterial therapy. This study evaluates the effect of prophylactic vitamin C administration on the efficiency of sulfadiazine-trimethoprim (SDT) or florfenicol (FF) in broilers with experimental SA. Broilers (210) were randomly allocated into 7 equal groups: (I) negative control (NC) (normal birds); (II) positive control (PC) arthritic birds by injection of Staphylococcus aureus in tibiotarsal joint at the age of 35 days; (III) vehicle control (injected with sterile medium); (IV) arthritic FF-treated (20 mg/kg/day); (V) arthritic vitamin C + FF-treated (as above + vitamin C at 15 g/100L of D.W. from day 25 of age); (VI) arthritic SDT-treated (35 mg/kg/day); (VII) arthritic vitamin C + SDT-treated. Antibacterial therapy started at day 39 of age and lasted for 5 days. Samplings were performed at the age of 44 and 54 days. A long lasting SA with severe fibrinoheterophilic synovitis and reduced body weights developed in PC broilers as compared to NC group (p < 0.05). Oxidative stress was present at sampling 1. Arthritis was not reflected in IL-6 levels of synovial fluid of PC group. None of the antibacterials resulted in completely successful treatment. Vitamin C did not appreciably improve lameness and arthritis scores, although it decreased lipid peroxidation and improved weights of FF treated-arthritic birds. For SDT-treated birds, vitamin C only ameliorated histopathological changes. In conclusion, except for improving body weight in FF-treated birds, prophylactic administration of vitamin C is not associated with improvements in clinical outcome of antimicrobial therapy of broilers with SA, although it ameliorates oxidative stress and some histopathological changes.

Prevalence of Mycobacterium avium subspecies paratuberculosis IS 900 DNA in biopsy tissues from patients with Crohn's disease: histopathological and molecular comparison with Johne's disease in Fars province of Iran.

BACKGROUND: Crohn's disease is a chronic enteritis of humans that affects the gastrointestinal tract, especially the terminal ileum, cecum and colon. The etiology of this disease is still unknown but seems to be multifactorial. There are reports about the potential link between Crohn's disease in humans and the causative agent of Johne's disease in ruminants. Because of the prevalence of Johne's disease in the Fars Province of Iran, the aim of this study was to investigate the prevalence of MAP in the biopsy tissues of patients affected by Crohn's disease in this area. METHODS: The study was performed from April 2015 to June 2017 at Namazi Hospital, Shiraz University of Medical Sciences, and School of Veterinary Medicine, Shiraz University, Shiraz, Iran. Intestinal biopsies of 30 patients (12 male and 18 female; mean age, 34 years; range 4-77 years) with the confirmed diagnosis of Crohn's disease and 30 patients diagnosed as non-inflammatory bowel disease (19 male and 11 female; mean age, 38 years; range 13-68 years) were studied by molecular, histopathological and histochemical methods. Also, similar numbers of adult goats affected by Johne's disease were studied, comparatively. DNA extractions of tissue specimens were subjected to PCR to amplify a 413-bp sequence of the IS900 gene. RESULTS: Using IS900-PCR, the overall prevalence of MAP in patients affected by Crohn's disease and non-inflammatory bowel disease were 47 and 13%, respectively. In addition, the prevalence of MAP in goats affected by Johne's disease was 70%. Using acid-fast histochemical staining, only 7% of Crohn's disease patients were weakly positive as paucibacillary and 43% of Johne's disease cases were moderate to strongly positive as multibacillary. Histopathologically, granulomatous enteritis (83 and 90%), lymphoplasmacytic enteritis (17 and 14%), edema and lymphangiectasia (67 and 96%), and vasculitis (20 and 73%) were common findings in Crohn's and Johne's diseases, respectively. CONCLUSION: Our findings demonstrate a remarkable association between MAP and CD in this population, and support an etiologic relationship between MAP infection in humans and the development of CD. MAP infection in human tissue may display species-specific pathologic findings, as occurs with other zoonotic pathogens.

Molecular characterization of Peste des Petits ruminants virus isolated from four outbreaks occurred in southern Iran.

BACKGROUND: Peste des petits ruminants (PPR) is a severe infectious disease in both domestic and wild small ruminants. Due to its heavy economic burden and hence social and health impacts on human populations, Food and Agriculture Organization of the United Nations (FAO) and The World Organization for Animal Health (OIE) have targeted PPR for eradication by 2030. In order to plan and implement a successful eradication program, factual status assessments prior to devising disease control strategies is a vital criterion. The aim of this study was to isolate and characterize PPR virus from a rising wave of outbreaks in southern Iran. RESULTS: Twenty-one clinical samples, including blood as well as oral, nasal and ocular swabs were collected from ten sick animals in 4 various herds and were examined with ELISA and RT-PCR for the presence of PPR virus antigen and genome, respectively. The virus was successfully isolated in primary lamb kidney cell culture and identified by RT-PCR. Phylogenetic analysis of the sequenced N genes revealed that, while the earliest reports of Iran's outbreaks were grouped into clusters with Saudi Arabia, Turkey and Africa, in this study reported sequences were grouped with samples from Pakistan, Tajikistan and China in particular. This observation suggests a shift in PPRV flow from the western borders of the country to the eastern neighboring countries. CONCLUSIONS: Lineage IV of PPR virus is presently circulating in Iran, with certain levels of genetic diversity. Present study along with previous reports demonstrates the dispersal patterns and movements of PPR virus, which highlights the reversal pattern of virus flow in recent years. Such information is necessary to understand PPRV molecular epidemiology and to develop more proper control strategies to eradicate the disease in the planned time.

Comparison of the regenerative effect of adipose-derived stem cells, fibrin glue scaffold, and autologous bone graft in experimental mandibular defect in rabbit.

BACKGROUND/AIMS: One of the main concerns for maxillofacial and orthopedic surgeons is finding a method to improve regeneration of large craniofacial bone defects. The aim of this study was to investigate the healing and regenerative effects of fibrin glue associated with adipose-derived stem cells (ADSCs) and fibrin glue scaffold alone with autologous bone grafts in experimental mandibular defects of the rabbit. METHODS: Bilateral uni-cortical osteotomies were performed in the mandible of 20 male Dutch rabbits. The animals were randomly divided into 2 equal groups. In one group, the defect on the right side was treated by fibrin glue associated with ADSCs and the defect on the other side remained as the control. In another group, the defect on the right side was treated with fibrin glue and on the left side with autologous bone graft. After 28 and 56 days, five rabbits from each group were evaluated by computed tomography (CT) and histopathological examinations. RESULTS: Coronal CT showed a remarkable reconstruction of cortical bone in the fibrin glue associated with ADSCs group at 28 and 56 days post-surgery. Histopathologically, new cortical bony bridge formation was seen increasingly in the fibrin glue, fibrin glue associated with ADSCs, and autologous bone graft groups after 28 days. Statistical analysis of the thickness of new cortical bone in the treatment versus control groups showed a significant difference between fibrin glue alone and fibrin glue associated with ADSCs groups (P = 0.02). No significant difference was found between the fibrin glue associated with ADSCs and the autologous bone graft groups (P > 0.05). CONCLUSIONS: The healing process had a significant increase in the thickness of new cortical bone when fibrin glue scaffold associated with ADSCs was used.

The beneficial effects of spraying of probiotic Bacillus and Lactobacillus bacteria on broiler chickens experimentally infected with avian influenza virus H9N2.

This study investigated the clinical, antiviral, and immunological effects of spraying Bacillus spp. and Lactobacillus spp. as a single or mixture probiotic compound on experimentally infected broiler chickens with AIV H9N2. Two hundred and forty 1-day-old broilers were randomly assigned to 6 groups as follows: Ctrl- (no challenge AIV; no spray probiotic), Ctrl+ (AIV challenged; no spray probiotic), AI+B (AIV challenged; daily spraying of probiotic Bacillus spp.), AI+L group (AIV challenged; daily spraying of probiotic Lactobacillus spp.), AIV+BL (AIV challenged; daily spraying of probiotic Bacillus spp. and Lactobacillus spp.), and G-DW (daily spraying of normal saline; no AIV challenged). The birds were reared for 35 d. On the 22nd day of age, broiler chickens were challenged by AIV H9N2. The probiotics were sprayed at 9×109 CFU/m2 daily for 35 d. Growth performance, clinical signs, virus shedding, macroscopic and microscopic lesions were evaluated at various days in all groups. Spraying with probiotics improved the body weight gain and food conversion ratio in the AI+B, AI+L, and AI+BL groups compared to the Ctrl+. The severity of clinical signs, gross lesions, pathological lesions and viral shedding in the probiotic treatment groups was lower than in the Ctrl+ group. The findings of this study suggest the daily spraying of Lactobacillus and Bacillus probiotics alone or in combination during the rearing period reduce clinical and nonclinical aspects of H9N2 virus infection; so, it can be effective as a preventive protocol for controlling the severity of AIV H9N2 infection in broilers.

Prevalence and pathology of dermatophilosis in camels (Camelus dromedaries) in Iran.

Gross and histopathologic lesions of the integumentary system were studied in 103 camels (Camelus dromedarius) slaughtered in Yazd Province of Iran. Dermatophilosis was diagnosed in 14 (13.6%) of the studied camels. Grossly, patchy thickening of the skin with dark brown scabs or crusts were observed on the rump, flanks, abdomen, shoulders, and neck. A heavy infestation with brown hard ticks was present on the lesions in almost all of the affected camels. On parasitology examination, all the ticks were identified as Hyalomma spp. Histopathologic examination of the affected skins revealed prominent superficial thickening of the epidermis as orthokeratotic and parakeratotic hyperkeratosis, degenerating neutrophils, serous fluid, and bacterial filaments. A lot of keratinaceous debris associated with an exudate rich in neutrophils were seen as remarkable features of the lesions on the superficial epidermis. Dermal inflammation was mild with infiltration of mononuclear inflammatory cells, particularly lymphocytes, around superficial vessels. A Gram-positive filamentous organism which was divided into parallel rows of cocci was observed in the crusts by Brown and Brenn Gram stain. This is the first report of the occurrence of dermatophilosis in the camels of Iran. With regard to infestation of the lesions by Hyalomma spp., it seems that this tick may have an important role in the pathogenesis of dermatophilosis in the camels of this region.

Effect of prophylactic administration of vitamin C in chickens with staphylococcal septic arthritis.

BACKGROUND: Septic arthritis (SA) due to Staphylococcus aureus is a major cause of lameness in poultry with improper response to antimicrobial therapy. OBJECTIVES: The study evaluates the effect of prophylactic administration of vitamin C on SA induced by methicillin resistant S. aureus in chickens. METHODS: One hundred and twenty chickens were randomly assigned into four groups: I. Negative control (NC), II. Positive control (PC) with SA induced at the age of 35 days by intra articular injection of S. aureus. III. Vehicle control (VC) and IV. Arthritic vitamin C-treated (VitC) group (15 g/100 L of drinking water from day 25 to the end of the experiment). Samplings were performed on day 44 (sampling 1) and day 54 (sampling 2) of age. RESULTS: Arthritic birds showed an obvious decrease in body weight with severe clinical arthritis and lameness which were not significantly affected by vitamin C administration at both samplings. Moreover, marked increase in serum malondialdehyde (MDA) concentration of the PC group was observed in sampling 1. Administration of vitamin C successfully reduced MDA concentration at both samplings. In sampling 2, birds in the VitC group showed significantly higher total antioxidant capacity (TAC) than NC birds (p < 0.05). Interleukin-6 concentration in synovial fluid of chickens remained statistically similar among groups in both samplings, while histopathological changes were ameliorated in the VitC group in sampling 2. CONCLUSIONS: Prophylactic administration of vitamin C especially for relatively longer period can ameliorate oxidative stress and histopathological changes due to staphylococcal arthritis in chickens, although it is not associated with a significant effect on clinical manifestations of the disease.

Changing patterns of acute phase proteins and inflammatory mediators in experimental caprine coccidiosis.

This experiment was conducted to assess the changing patterns and relative values of acute phase proteins and inflammatory cytokines in experimental caprine coccidiosis. Eighteen newborn kids were allocated to 3 equal groups. Two groups, A and B, were inoculated with a single dose of 1×10(3) and1×10(5) sporulated oocysts of Eimeria arloingi, respectively. The third group, C, received distilled water as the control. Blood samples were collected from the jugular vein of each kid in both groups before inoculation and at days 7, 14, 21, 28, 35, and 42 post-inoculation (PI), and the levels of haptoglobin (Hp), serum amyloid A (SAA), TNF-α, and IFN-γ were measured. For histopathological examinations, 2 kids were selected from each group, euthanized, and necropsied on day 42 PI. Mean Hp concentrations in groups A and B (0.34 and 0.68 g/L) at day 7 PI were 3.2 and 6.3 times higher than the levels before inoculation. The mean SAA concentrations in groups A and B (25.6 and 83.5 µg/ml) at day 7 PI were 4.2 and 13.7 times higher than the levels before inoculation. The magnitude and duration of the Hp and SAA responses correlated well with the inoculation doses and the severity of the clinical signs and diarrhea in kids. These results were consistent with the histopathological features, which showed advanced widespread lesions in group B. In both groups, significant correlations were observed for TNF-α and IFN-γ with SAA and Hp, respectively. In conclusion, Hp and SAA can be useful non-specific diagnostic indicators in caprine coccidiosis.

Improvement of Rat Spinal Cord Injury Following Lentiviral Vector-Transduced Neural Stem/Progenitor Cells Derived from Human Epileptic Brain Tissue Transplantation with a Self-assembling Peptide Scaffold.

Spinal cord injury (SCI) is a disabling neurological disorder that causes neural circuit dysfunction. Although various therapies have been applied to improve the neurological outcomes of SCI, little clinical progress has been achieved. Stem cell-based therapy aimed at restoring the lost cells and supporting micromilieu at the site of the injury has become a conceptually attractive option for tissue repair following SCI. Adult human neural stem/progenitor cells (hNS/PCs) were obtained from the epileptic human brain specimens. Induction of SCI was followed by the application of lentiviral vector-mediated green fluorescent protein-labeled hNS/PCs seeded in PuraMatrix peptide hydrogel (PM). The co-application of hNS/PCs and PM at the SCI injury site significantly enhanced cell survival and differentiation, reduced the lesion volume, and improved neurological functions compared to the control groups. Besides, the transplanted hNS/PCs seeded in PM revealed significantly higher migration abilities into the lesion site and the healthy host tissue as well as a greater differentiation into astrocytes and neurons in the vicinity of the lesion as well as in the host tissue. Our data suggest that the transplantation of hNS/PCs seeded in PM could be a promising approach to restore the damaged tissues and improve neurological functions after SCI.

Lumpy skin disease, an emerging transboundary viral disease: A review.

Lumpy skin disease is an emerging bovine viral disease, which is endemic in most African countries and some Middle East ones, and the elevated risk of the spread of disease into the rest of Asia and Europe should be considered. The recent rapid spread of disease in currently disease-free countries indicates the importance of understanding the limitations and routes of distribution. The causative agent, Capripoxvirus, can also induce sheeppox and goatpox. The economic significance of these diseases is of great concern, given that they threaten international trade and could be used as economic bioterrorism agents. The distribution of capripoxviruses seems to be expanding due to limited access to effective vaccines and poverty within farming communities. This is largely due to the economic effects of the Covid-19 pandemic and the imposition of crippling sanctions in endemic regions, as well as an increase in the legal and illegal trade of live animals and animal products, and also global climate change. The present review is designed to provide existing information on the various aspects of the disease such as its clinicopathology, transmission, epidemiology, diagnosis, prevention and control measures, and the potential role of wildlife in the further spread of disease.

The evaluation of attenuated Neospora caninum by long-term passages on murine macrophage cell line in prevention of vertical transmission in mice.

To date, there is no effective vaccine to prevent abortion or vertical transmission associated with neosporosis in cattle. In the present study, the efficacy of a live experimental vaccine of Neospora caninum attenuated (NCa) by long-term serial passages on a murine macrophage cell line was evaluated in the prevention of vertical transmission and abortion in the mouse model. Forty non-pregnant mice were randomly divided into four equal groups including non-immunized/challenged (injected with PBS); positive control (inoculated with un-attenuated NC-1 tachyzoites); immunized/challenged (inoculated with NCa attenuated strain) and immunized/non-challenged or vaccinated (inoculated with NCa) groups. Following pregnancy synchronization, both the immunized and control mice were challenged with virulent live NC-1 tachyzoites (2.5 × 106) in the mid-pregnancy stage. The number of abortions and post-natal pup mortalities was recorded. Serological, molecular, and histopathologic examinations were employed to evaluate the efficacy of the vaccine and the vertical transmission rates. Results indicated that the live attenuated N. caninum strain (NCa) could significantly reduce the risk of abnormal parturitions and fetal mortality in the vaccinated group (20 %) compared to the non-immunized/challenged group (80 %). Also, the NCa strain reduced the lesion score in the brain of the offspring (0.3 vs 1.9) compared to the non-immunized/challenged group (P < 0.05). The molecular assay showed a decrease in the parasite DNA detection rates from 83 % and 77 % in the non-immunized/challenged group to 27 % and 0 % in the vaccine group in the brain and liver tissues, respectively. While in the immunized/non-challenged group no parasite DNA was detected in the brain tissue samples of the pups. Serological analyses showed that NCa strain was able to stimulate the humoral immunity and create effective protection against neosporosis with a moderate systemic IFN-γ response. In conclusion, the NCa strain could significantly (P < 0.05) reduce the risk of vertical transmission and proved to be a safe vaccine while conferring significant levels of protection in the laboratory mice.

Lentiviral vector-mediated transduction of adult neural stem/progenitor cells isolated from the temporal tissues of epileptic patients.

OBJECTIVES: Neural stem/progenitor cells (NS/PCs) hold a great potential for delivery of therapeutic agents into the injured regions of the brain. Efficient gene delivery using NS/PCs may correct a genetic defect, produce therapeutic proteins or neurotransmitters, and modulate enzyme activation. Here, we investigated the efficiency of a recombinant lentivirus vector expressing green fluorescent protein (GFP) for genetic engineering of human NS/PCs obtained during brain surgery on patients with medically intractable epilepsy. MATERIALS AND METHODS: NS/PCs were isolated from human epileptic neocortical tissues. Three plasmids (pCDH, psPAX2, pMD2.G) were used to make the virus. To produce the recombinant viruses, vectors were transmitted simultaneously into HEk-293T cells. The lentiviral particles were then used to transduce human NS/PCs. RESULTS: Our in vitro study revealed that lentivirus vector expressing GFP efficiently transduced about 80% of human NS/PCs. The expression of GFP was assessed as early as 3 days following exposure and remained persistent for at least 4 weeks. CONCLUSION: Lentiviral vectors can mediate stable, long-term expression of GFP in human NS/PCs obtained from epileptic neocortical tissues. This suggests lentiviral vectors as a potential useful tool in human NS/PCs-based gene therapy for neurological disorders, such as epilepsy.

Tissue Engineering in Maxillary Bone Defects.

BACKGROUND: Restoration of craniofacial bone defects has been a concern for oral and maxillofacial surgeons. In this study, the healing effect of fibrin glue scaffold was compared with autologous bone graft in mandibular defects of rabbit. METHODS: Bilateral unicortical osteotomy was performed in the diastema region of 10 male Dutch rabbits. The subjects were randomly divided into 2 equal groups. The mandibular defect on the right side was treated with fibrin glue scaffold and the defect on the left side with autologous bone graft provided from iliac crest. After 4 and 8 weeks, five rabbits from each group were sacrificed and the defects were evaluated morphologically, by coronal computed tomography scanning (CT-scan) and by histological examinations. RESULTS: The healing effect of fibrin glue scaffold and autologous bone graft was similar with appropriate osteogenesis in comparison to the control group. CONCLUSION: Using fibrin glue can be a non-invasive treatment of choice in mandibular defects and maxillofacial surgeries when compared with autologous bone graft.

Cerebral astrocytoma in a sheep.

Astrocytoma as one of the most common central nervous system (CNS) tumors is rarely reported in veterinary literature. A 7-year-old Persian Lori-Bakhtiari ewe was presented to the clinic with a two months history of progressive blindness, nystagmus to the right, bilaterally decreased pupillary reflexes, head pressing and paddling. At necropsy, a whitish well-circumscribed mass with dimensions of 3.50×2.50×1.50 cm was observed in the dorsal parietal lobe of the left cerebral hemisphere. Microscopically, the mass was well-circumscribed and highly cellular, consisted of round to elongated cells with scant and vacuolated cytoplasm with few, flaccid processes. The nuclei were round to oval with densely stippled chromatin and indistinct nucleoli. Immunohistochemical analyses showed positive staining for vimentin, S100 and glial fibrillary acidic protein. Definitive diagnosis of cerebral protoplasmic astrocytoma was made on the basis of the histopathological and immunohistochemical findings. This type of neoplasm should be included in the differential diagnosis of CNS lesions in the sheep.

An overview on autologous fibrin glue in bone tissue engineering of maxillofacial surgery.

The purpose of this review is to have an overview on the applications on the autologous fibrin glue as a bone graft substitute in maxillofacial injuries and defects. A search was conducted using the databases such as Medline or PubMed and Google Scholar for articles from 1985 to 2016. The criteria were "Autograft," "Fibrin tissue adhesive," "Tissue engineering," "Maxillofacial injury," and "Regenerative medicine." Bone tissue engineering is a new promising approach for bone defect reconstruction. In this technique, cells are combined with three-dimensional scaffolds to provide a tissue-like structure to replace lost parts of the tissue. Fibrin as a natural scaffold, because of its biocompatibility and biodegradability, and the initial stability of the grafted stem cells is introduced as an excellent scaffold for tissue engineering. It promotes cell migration, proliferation, and matrix making through acceleration in angiogenesis. Growth factors in fibrin glue can stimulate and promote tissue repair. Autologous fibrin scaffolds are excellent candidates for tissue engineering so that they can be produced faster, cheaper, and in larger quantities. In addition, they are easy to use and the probability of viral or prion transmission may be decreased. Therefore, autologous fibrin glue appears to be promising scaffold in regenerative maxillofacial surgery.

A retrospective study of sporadic bovine abortions, stillbirths, and neonatal abnormalities in Atlantic Canada, from 1990 to 2001.

In a retrospective study on 265 cases of sporadic bovine abortions, stillbirths, and neonatal deaths in Atlantic Canada (1990 to 2001), an etiological diagnosis was made in 117 cases (44.2%). The cases were divided into 2 groups: 234 abortions, and 31 stillbirths and neonatal deaths. Identified causes of abortion were bacteria (24.4%), fungi (6.8%), viruses (6.0%), protozoa (Neospora spp.) (2.1%), congenital anomalies (0.4%), and miscellaneous conditions (1.3%). In addition, placentitis without demonstrable infectious agents was observed in 17 (7.3%). Of the 31 cases of stillbirth and neonatal death, identified causes were dystocia (22.5%), congenital anomalies (22.5%), meconium aspiration syndrome (16.1%), and miscellaneous conditions (6.5%). No etiological diagnosis was made in 59% of abortions and 32.4% of stillbirths and neonatal deaths. The 3 most common identifiable causes of abortion in this study were bacterial, fungal, and viral infections.

Hematological and serum biochemical analyses in experimental caprine coccidiosis.

In order to provide information on hematological and biochemical changes in Eimeria arloingi infection, eighteen newborn kids were allocated to three equal groups. Two groups, A and B were inoculated with a single dose of 1 × 10(3) and 1 × 10(5) sporulated oocysts of E. arloingi, respectively. The third group C remained uninfected as control. Blood samples were collected from jugular vein before inoculation as control and at 7, 14, 21, 28, 35 and 42 days post inoculation (DPI). Total erythrocyte and total leukocyte counts, hemoglobin (Hb), packed cell volume (PCV) and differential leukocyte counts were determined. Serum biochemical parameters including total protein (TP), albumin, chloride (Cl(-)), sodium (Na(+)), potassium (K(+)), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma glutamyltransferase (GGT) were measured. All infected animals showed symptoms of clinical coccidiosis including diarrhea and oocyst excretion after the prepatent period that varied from 16 to 18 days after inoculation. The oocysts output did not stop until the end of the experiment. The diarrhea was associated with a reduction in ALP activity, increases in PCV and Hb and decreases in Na(+), Cl(-) and K(+). No significant differences were found in AST, ALT, GGT, albumin and TP during 42 DPI. There was no hepatic damage.

Comparison of use of Vero cell line and suspension culture of murine macrophage to attenuation of virulence of Neospora caninum.

In this study the tachyzoite yields of Neospora caninum were compared in two cell lines: Vero (African Green Monkey Kidney) and suspension culture of murine macrophage (J774) cell lines. Then, N. caninum were continuously passaged in these cell lines for 3 months and the effect of host cells on virulence of tachyzoites was assessed by broiler chicken embryonated eggs. Inoculation was performed in the chorioallantoic (CA) liquid of the embryonated eggs with different dilutions (0.5 × 10(4), 1.0 × 10(4), 1.5 × 10(4)) of tachtzoites isolated from these cell cultures. The mortality pattern and pathological changes of the dead embryos and hatched chickens were noted. Tissue samples of brain, liver and heart were examined by histopathological and detection of DNA of parasite by polymerase chain reaction (PCR). Also, consecutive sections of the tissues examined histologically were used for immunohistochemical (IHC) examination. Embryos inoculated with tachyzoites derived from Vero cell line (group V) showed a higher mortality rate (100%) than the embryos that received tachyzoites derived from J774 cell line (group J) (10% mortality rate). The results of this study indicated that the culture of N. caninum in J774 cell led to a marked increase in the number of tachyzoite yields and rapid attenuation in comparison to Vero, so the results were confirmed by IHC and PCR. This study is the first report of the significant effect of host cell on the attenuation of virulence of N. caninum tachyzoites. These findings could potentially provide a practical approach in the mass production of N. caninum tachyzoites, and also in producing live attenuated vaccine.

Experimental caprine coccidiosis caused by Eimeria arloingi: morphopathologic and electron microscopic studies.

The progressive morphohistopathologic changes, distribution pattern of lesions and ultrastructural characteristics in Eimeria arloingi infection were investigated in experimentally infected kids. The 18 newborn animals allocated to 3 equal groups. Two of groups, A, B were inoculated with a single dose of 1 × 10(3) and1 × 10(5) sporulated oocysts of E. arloingi, respectively. At 7, 14, 21, 28, 35, and 42 days postinoculation (DPI), 1 kid from each group was necropsied for pathologic and ultrastructural studies. Progressive lesions were present at 21, 28, 35 and 42 DPI in the jejunum, ileum, cecum with fewer in the duodenum and proximal colon. The oocysts shedding begin between 16 to 18 DPI. Grossly, minimal changes were observed at 21 DPI as few whitish plaques or nodules and advanced lesions at 42 DPI as pseudoadenomatous pattern in the mucosa and a cerebriform pattern on the serosal surface of jejunum and ileum. Early histopathologic lesions due to schizogony phase were including presence of intracytoplasmic developmental stages of the parasite such as trophozoites, immature to mature schizonts and mild infiltration of inflammatory cells. In late lesions due to various stages of gametogony, the histological pattern was mainly remarkable hyperplasia of the villi and crypts epithelial cells, eventually developed into papillary projections of reactive epithelium. The mesenteric lymph nodes showed a few numbers of large schizonts in the cortical lacteals. This study showed E. arloingi as a highly pathogenic species for kids, the incubation period was 16-18 days and the main target organ was jejunum with characteristic morphohistopathologic lesions.

Immunohistochemical and polymerase chain reaction studies in Neospora caninum experimentally infected broiler chicken embryonated eggs.

The diagnostic characteristics of immunohistochemistry (IHC) and polymerase chain reaction (PCR) methods were studied in the tissues of broiler chicken embryos experimentally infected by Neospora caninum. An infection with N. caninum NC-1 isolate was conducted in 70 broiler chicken embryonated eggs randomly divided into seven equal groups. After 8 days of incubation, six groups were inoculated with 10, 10(2), 10(3), 10(4), 10(5), and 10(6) doses of tachyzoites/embryonated egg. The 7th group was considered as control. The mortality rate and pathological changes of the dead embryos and hatched chickens up to 60 days old were noticed. Consecutive sections to those used for histopathological examination including the liver, heart, brain, and chorioalantoic (CA) membrane were subjected to IHC. The intensity and distribution of the immunostaining was graded as highly to mildly positive. For PCR procedure, DNA was extracted from 50mg of the tissues and primer pair Np21/Np6 was used for amplification of the Nc-5 gene. The results of the immunosignaling ranged from variable degrees of mild to moderate staining as dark-brown to brown and coarsely to finely granular, mostly within the cytoplasm of infected cells such as the endothelial cells of blood vessels. The parasite aggregation was more predominant in the heart than other tissues. Immunoreactivity for N. caninum antigen was multifocally moderate positive in the heart, liver and CA of the 10(3) dose, and also heart, liver, brain and CA of the 10(4) dose. IHC showed mildly positive in the liver and heart of the chicken embryos infected with 10 and 10(2) tachyzoites, as well. The results of the PCR confirmed the existence of the parasite in all of the examined tissues from the 10(3) and 10(4) doses. In conclusion, the results indicate a good agreement between IHC and PCR in diagnosis of neospora antigen in the infected tissues.