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Cloning and expression of Eimeria necatrix microneme5 gene in Escherichia coli
2016
Mayahi, Mansour | Jolodar, Abbas | Masaeli, Sharouz | Hamidinejat, Hosein | Seyfi Abad Shapouri, Masoud | Moori Bakhtiari, Naghme
Background: Coccidiosis caused by Eimeria necatrix has the most economic impact onpoultry production. Micronemal proteins in Eimeria necatrix are thoughtto be critical ligands determining host cell specificity at the time ofinvasion. OBJECTIVES: Isolation and purification of Eimeria necatrix oocysts from Khuzestan province of Iran was performed. AcDNA encoding microneme 5 (EnMIC5) protein was cloned and expressed asrecombinant protein before the evaluation of its immunogenicity by Westernblotting. METHODS: A primer pair was designed based on the publishednucleotide sequence of Eimeria necatrix LZ strain micronem5 gene. APartial cDNA sequence fragment of 758 bp coding for microneme 5 protein(EnMIC5) was amplified by semi- Nested RT-PCR. PCR products were cloned andexpressed in a Maltose Binding protein (MBP) containing expression vector(pMAL-c2x) in Escherichia coli. The cDNA which is encoded for 252 aminoacids shows high degree of conservation. It contains the adhesive plasmapre-kallikrein and seven hydrophilic motifs. RESULTS: The results of SDS-PAGErevealed that the recombinant protein with a molecular weight of 70 kDa wasover-expressed after induction with IPTG. Western blotting results revealedthat the expressed recombinant protein was reacted with sera of the chicksinfected with Eimeria necatrix. It was suggested that this proteinshould have a good immunogenicity and can be used for further studies. CONCLUSIONS: In conclusion, the high degree of sequence homology indicates that thisprotein is immunogenic and might be aninteresting vaccine target, and deserves further investigation
Mostrar más [+] Menos [-]Identification of enterototxin harboring gene among Clostridium perfringens isolates with different toxin types in Iran
2016
Jabbari, Ahmad-Reza | Esmaelizad, Majid | Samimi, fariba
Background: Clostridium perfringens is known as the most widely distributed pathogenic microorganism in nature. It is an extremely important pathogen of human and domestic animals. In a commonly used classification scheme, C. perfringens is divided into five toxinotypes (A to E) based on the production of four major toxins (alpha, beta, epsilon, and iota). Enterotoxin is not usually used for C. perfringens typing but it is a fatal toxin with necrotic activity. Based on our knowledge there is no published scientific report regarding identification of enterotoxin positive C. perfringens isolates from animals in Iran. OBJECTIVES: To study the presence and frequency of enterotoxin gene among C. perfringence isolates with different types. METHODS: A specific single PCR assay was developed and used for detection of cpe gene to identify the entrerotoxin harboring isolates among different types of C. perfringens isolated from animal enteric diseases in Iran. RESULTS: It was found that cpe gene presents among C.perfringens isolates types A, B, C and D with 63.6% (7/11), 25% ( 5/20), 21.4% (3/14), 53.3% (8/15), respectively. Totally 23 of 60 ( 38.3%) isolates screened by PCR were cpe-positive. CONCLUSIONS: This is the first report of cpe- positive isolates of C.perfringens causing enterototoxemia in animals in Iran. Further studies to investigate the synergistic effect of CPE toxin in pathogenesis of enteric diseases in animals is suggested.
Mostrar más [+] Menos [-]Dopamine- induced hypophagia is mediated via NMDA and mGlu1 receptors in chicken
2016
Taherian, Mohammadreza | Baghbanzadeh, Ali | zendehdel, morteza
Background: Feeding behavior is regulated by a complex network which interacts via diverse signals from central and peripheral tissues. It is known dopaminergic and glutamatergic systems have crucial role on food intake regulation but scarce reports exist on their interaction in appetite regulation in broilers. OBJECTIVES: The present study was designed to examine the role of glutamatergic system on dopamine-induced hypophagia in neonatal meat-type chicken. METHODS: In experiment 1, chicks received ICV injection of control solution, dopamine (40 nmol), MK-801 (NMDA glutamate receptors antagonist, 15 nmol) and co-injection of dopamine + MK-801. In experiment 2, birds were ICV injected with saline, dopamine (40 nmol), CNQX (AMPA glutamate receptors antagonist, 390 nmol) and co-injection of dopamine + CNQX. In experiment 3, chicks received ICV injection of control solution, dopamine (40 nmol), AIDA (mGLUR1 glutamate receptors antagonist, 2 nmol), dopamine + AIDA. Experiments 4 and 5 were similar to experiment 3, except birds were injected with LY341495 (mGLUR2 glutamate receptors antagonist, 150 nmol) and UBP1112 (mGLUR3 glutamate receptors antagonist, 2 nmol) instead of AIDA. Then the cumulative food intake was measured until 120 min post injection. RESULTS: According to the results, ICV injection of dopamine significantly decreased food intake (p<0.001). Co-injection of dopamine and MK-801 decreased dopamine induced hypophagia (p<0.001). Moreover, the food intake of chicks was significantly increased by co-injection of AIDA and dopamine (p<0.001). CONCLUSIONS: These results suggest dopamine-induced hypophagia is mediated via NMDA and mGlu1 receptors in chicken.
Mostrar más [+] Menos [-]Mechanical behavior measurement of the sheep small intestine using experimental tests
2016
Rassoli, Aisa | Fatouraee, Nasser
Background: There is no consistent data on the mechanical properties of sheep intestine. OBJECTIVES: We performed a series of biaxial strain measurement experiments and extracted the constitutive model to describe the mechanical characteristics of the sheep intestinal tissue. METHODS: Eleven specimens were obtained freshly from sacrificed sheep and the planar biaxial tests were performed on the tissue specimens by applying simultaneous loads along the circumfer-ential and longitudinal directions. Then the measured data were fitted into the anisotropic four-parameter Fung-type model and also to the modified Mooney-Rivlin model. RESULTS: The specimens showed some degree of anisotropy; the stiffer direction is not gener-ally predictable. Some of the specimens were stiffer in the circumferential direction, and the others in the longitudinal direction. However, the average results state the circumferential di-rection as the stiffer orientation. CONCLUSIONS: It can be concluded that sheep intestine be-haves normally as a nonlinear anisotropic tissue which is well-characterized by the modified Mooney-Rivlin model.
Mostrar más [+] Menos [-]Anatomical study of the Iranian brown bear’s skull (Ursus arctos): A case report
2016
Yousefi, Mohammad Hasan
\The Brown bear (Ursusarctos) is a species at risk of extinction. It is considered the largestcarnivore and lives in northern Iran. Several studies on the structure of skullhave been accomplished in differentanimals. The aim of this study was the inscription of gross anatomicalcharacteristics of skulls of three Iranian adult male brown bears that weretransferred to the Anatomical Departmentof the Faculty of Veterinary Medicine of Semnan University. After processing,that included cleaning, degreasing and bleaching, skulls were studied from thedorsal or frontal, ventral, lateral, rostral, caudal and medial views. Thefacial part of the brown bear’s skull from dorsal view was small and thecranium was seen quadrilateral and larger than the dogs. The facial part oflacrimal bone and also the optic groove of presphenoid were absent. There wasnot articulation between maxillary and nasal bones. The interincisive canal waspresent and situated inter palatine processes of incisive bones. The orbitalcavity was small in brown bear. Lacrimal canal was formed by lacrimal andmaxillary bones. The interparietal bone and external sagittal crest in brownbear were seen as being shorter than the dogs. Tympanic bulla was very smalland jugular foramen rounded. The external acoustic meatus was formed bysquamous and tympanic part of temporal bone. In conclusion, the brown bear’sskull has different important macroscopic characteristics compared to othercarnivores
Mostrar más [+] Menos [-]Molecular evaluation of M2 protein of Iranian avian influenza viruses of H9N2 subtype in order to find mutations of adamantane drug resistance
2016
Malekan, Mohammad | VasfiMarandi, Mehdi | Barin, abas | Mokhtari azad, Talat | Ranjbar, Mohammad Mehdi | Bashashati, Mohsen
Background: The H9N2 subtype of influenza A viruses is considered to be widespread in poultry industry. Adamantane is a group of antiviral agents which is effective both in prevention and treatment of influenza A virus infections. These drugs inhibit M2 protein ion channel which has role on viral replication. OBJECTIVES: The main objective of this study is to evaluate M gene of avian influenza viruses (AIVs) of H9N2 subtype in order to find adamantane drug resistance mutations. METHODS: Over 100 suspected samples were collected from different geographical regions of Iran during 2012-2013. Samples were injected via allantoic sac of 9-11 day-old chicken embryos. A total of 11 out of 100 were AIV. The H9N2 subtype was confirmed by specific RT-PCR. The RT-PCR was conducted for full length M gene. PCR amplified products were purified and then conducted for commercial direct sequencing. Finally, sequences were checked for possible sites of adamantane resistance mutations. RESULTS: Overall, 8 out of 11 viruses harbored the adamantane resistance-associated mutations. Of which, four viruses were isolated in 2013 and four viruses in 2012. Two different resistance-associated mutations were observed during different years. CONCLUSIONS: The present study provided clear evidence concerning resistance AIVs of H9N2 subtype that were circulating in Iranian poultry sector. This concern is always present as M segment might be introduced into human influenza viruses by reassortment phenomenon.
Mostrar más [+] Menos [-]Preparation and evaluation of a thermosensitive liposomal hydrogel for sustained delivery of danofloxacin using mesoporous silica nanoparticles
2016
Kiani, Katayoun | Rassouli, Ali | Hosseinzadeh Ardakani, Yalda | Akbari Javar, Hamid | Khanamani Falahatipour, Sakineh | Khosraviyan, Pegah | Zahraee Salehi, Taghi
Background: Sustained release delivery system can reduce the dosage frequency and maintain the therapeutic level of drugs for a longer time. Biodegradable, biocompatible and thermosensitive chitosan-beta-glycerophosphate (C-GP) solutions can solidify at body temperature and maintain their physical integrity for a longer duration. OBJECTIVES: To develop a novel delivery system based on the integration of liposomes in hydrogel using mesoporous silica nanoparticles (MSNs) for sustained release of danofloxacin in farm animals. METHODS: The MSNs were prepared using N-cetyltrimethylammonium bromide and tetraethylortho silica. The liposomes were prepared by thin film hydration method. C-GP solution containing danofloxacin-loaded MSN liposomes underwent different in-vitro tests, including evaluation of the entrapment efficiency, gelation time, morphology, drug release pattern as well as antimicrobial activities against S. aureus and E. coli. RESULTS: The mean pore size of MSNs was 2.8 nm and the mean MSN entrapment efficiency was 45%. Kinetics of danofloxacin release from liposomal hydrogel followed the Higuchi’s model. This formulation was capable of sustaining the danofloxacin release for more than 96 h. The FTIR studies showed that there were no interactions between danofloxacin and hydrogel excipients. Scanning electron microscopy (SEM) showed that the formed gel had a continuous texture, while the swelled gel in the phosphate buffer had a porous structure. Microbiological tests revealed a high antibacterial activity for lipomosal hydrogel of danofloxacin-loaded MSN comparable with danofloxacin solution. CONCLUSIONS: The liposomal hydrogel solidified at body temperature, effectively sustained the release of danofloxacin and showed in vitro antibacterial effects.
Mostrar más [+] Menos [-]Effect of Lipia Citridora leaves powder on growth performance, carcass traits, blood metabolites and meat quality of broilers
2016
Mehrparvar, Marziyeh | Mazhari, Mozhgan | Esmaeilipour, Omidali | Sami, Masoud
Background: Since the prohibition of antibiotic growth promoter in poultry ration by the European Union, poultry producers have been trying to replace them by other growth promoters like medicinal herbs. Lipia citridora is a medicinal herb enriched by several powerful phenolic compounds and with antioxidant and antimicrobial properties which lead to enhancing appetite and growth performance of broiler. OBJECTIVES: The aim of this study was to investigate the effect of Lipia citridora leaves powder (LCLP) on growth performance, carcass traits, blood metabolites, and meat quality of broilers. METHODS: The experiment was carried out with 160 one-day-old male Ross 308 broiler chickens in a completely randomized design. The chicks were allocated to four diets including basal diet and treatment diets which were supplemented with 3 levels of LCLP (0.25, 0.5 and 1.0% of diet) in 16-floor pens with 10 chickens each. At the end of the experiment, 2 chicks/pen were selected and the assigned parameters were evaluated. RESULTS: All levels of LCLP improved (p<0.01) feed intake, body weight gain and feed conversion ratio in all periods except for starter phase. Also, 1% LCLP improved FI (1.49%), BWG (4.20%) and FCR (2.59%) compared to control group during the whole period. Birds fed herbal additive had a significantly higher relative weight of carcass, breast, bursa of fabricius and spleen in 1% LCLP group compared with the control group. Addition of 1% of LCLP decreased (p<0.05) LDL (15.85%), cholesterol (8.73%), triglyceride (8.82%), and increased (p<0.01) white blood cells (8.04). Meat quality enhanced via an intense reduction (p<0.01) in thiobarbituric acid (50.15%) and cooking loss (14.16%). CONCLUSIONS: Our finding showed that the highest level of LCLP in this study (1%) performed the best as improved growth performance, ameliorated serum lipids, increased white blood cells and enhanced the meat quality.
Mostrar más [+] Menos [-]Acute toxicity evaluation of five herbicides: paraquat, 2,4-dichlorophenoxy acetic acid (2,4-D), trifluralin, glyphosite and atrazine in Luciobarbus esocinus fingerlings
2016
alishahi, mojtaba | Tulaby Dezfuly, Zahra | Mohammadian, Takavar
Background: Evaluation of herbicide pollution in aquatic environments needs the great concern and the most important echo-pollutant effects of herbicides are related to their effects on non target aquatic organisms. Native fish can serve as a proper bio-indicator for evaluation of pollution on aquatic ecosystems. OBJECTIVES: To find environmentally friendly herbicides, in this study the acute toxicity of five widely used herbicides in Iran as aquatic ecosystems pollutants on Luciobarbus esocinus were investigated. METHODS: Acute toxicity (96 h LC50) of five herbicides (Paraquat, 2,4-dichlorophenoxy acetic acid, Trifluralin, Glyphosite and Atrazine) were determined via OECD standard method. L.esocinus exposed to Serial concentrations (more than 6 in triplicates) of each herbicide. Mortalities at 24, 48, 72 and 96 hours after exposure were recorded and the LC50 were calculated using Probit software. RESULTS: Results showed that acute toxicity of these herbicides are significantly different in L. esocinus. The 96 h LC50 of Paraquat, 2,4-D, Trifluralin, Glyphosite and Atarzine in L.esocinus were 54.66, 138.8, 1.09, 716.83 and 44.30 mg/l respectively. Glyphosite showed lowest toxicity in Luciobarbus esocinus among the five herbicids. The highest toxicity of herbicides in L. esocinus belongs to Trifluralin. The mortality rate of exposed fish to herbicides enhanced either by increasing herbicides concentration or duration of exposure. Mortality patterns during 96 hours of toxicity evaluation were similar in all five herbicides. CONCLUSIONS: Regarding the high application and similar efficacy of herbicides in most of the cane farms of Khouzestan province, and based on different toxicities of these five herbicides for fish as a non targeting organism, Glyphosite is highly recommendable as a proper alternative to Trifluralin, Atrazine, Paraquat and 2,4-dichlorophenoxy acetic acid.
Mostrar más [+] Menos [-]Metastatic cutaneous neuro- myofibroblastic sarcoma induced by avian leukosis virus subgroup J in a rooster (Gallus gallus domesticus)
2016
Norouzian, Hassan | Dezfoulian, Omid | Hosseini, Hossein
An adult native cock (Gallus gallus domesticus) referred to the aviary clinic with multiple different sizes of round dermal nodules. The bird died few days later, and was then submitted for further evaluation. Macroscopic and microscopic examinations as well as a PCR test were done to identify type and cause of the tumor. In histopathological assessment of biopsy specimen, it consisted of interlacing bundles of fibroblasts that orientated in different directions with plump or elongated spindle shaped nuclei and fairly abundant cytoplasm. At necropsy several large white nodules were implanted in lung and liver. Microscopically the proliferated fibroblastic cells were invaded to both organs, and were similar to those described for skin lesion. The tumor cells had immunoreaction for alpha smooth muscle actin, vimentin and S100 protein, whereas they were negative for desmin and pancytokeratin, suggesting a diagnosis of metastatic neuro-myofibroblastic sarcoma. A PCR test specific for avian leukosis virus subgroup J (ALV-J) confirmed the presence of that virus in tumor specimens. Sequencing and phylogenetic analysis showed a relatively low similarity in the LTR segment (90%) of the studied virus with other ALV-J strains. It might be the first report of cutaneous neuro-myofibroblastic sarcoma, potentiated to metastasis to other organs induced by ALV-J.
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