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Veterinary Research Forum - Volume:9 Issue: 4, Autumn 2018

Veterinary Research Forum
Volume:9 Issue: 4, Autumn 2018

  • تاریخ انتشار: 1397/09/24
  • تعداد عناوین: 14
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  • Jaime Sard Arambur Junior email *, Tiago Luis Eilers Treichel, Saulo Tadeu Lemos Pinto Filho, Sergio Alexandre Gerhke, Alencar Kolinski Machado, Francine Carla Cadon, Ivana Beatrice Mânica da Cruz, Ney Luis Pippi Pages 293-299
    The aim of this study was to evaluate the potential use of a DNA comet assay, DNA fragmentation fluorimetric assay and reactive oxygen species levels as potential biomarkers of genome conditions of dental pulp stem cells (DPSCs) isolated from dog canine teeth. Mesenchymal stem cells were isolated from the dental pulp collected from dog teeth. The results obtained suggest the ideal moment for clinical application of cellular therapy for this type of cell. The cell culture was maintained with Dulbecco’s modified Eagle’s medium supplemented with 10.00% fetal bovine serum for eight passages. During each passage, cell proliferation, oxidative stress and level of DNA fragmentation were assessed by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay, testing 2,7 dichlorodihydro-fluorescein-diacetate and PicoGreen®, respectively. There were important differences among the first three DPSC passages compared to passages 4–8 and a large number of nuclei with some levels of DNA damage (30.00 to 40.00% in initial DPSC passages and > 50.00% in late passages), indicating in vitro DPSC genomic fragility. Within the limitations of this study, the results suggest these relatively simple and inexpensive approaches - comet and DNA fragmentation assays - could help sort stem cells with less DNA damage for use in research or therapies.
    Keywords: Dental pulp, DNA damage, Genotoxicity, Mesenchymal stem cell, Oxidative stress
  • Mobin Koohestani, Mehran Moradi *, Hossein Tajik, Armen Badali Pages 301-306
    This study was carried out to investigate the stability, antibacterial properties and biofilm removal potential of cell-free supernatant (CFS) of Lactobacillus acidophilus LA5 and Lactobacillus casei 431 against Staphylococcus aureus ATCC 25923. Antibacterial activity of both Lactobacillus strains was measured according to the agar spot method. The CFS was prepared by centrifugation of bacterial suspension at 4000 g for 10 min and the antimicrobial activity was measured using agar-well diffusion. The stability of CFSs during storage at 4.00 ± 2.00 °C and 25.00 ± 2.00 °C for a period of 4 weeks was measured based on the method of broth micro-dilution assay. Moreover, biofilm removal potential of CFS on 2-days-old biofilm of S. aureus developed on polystyrene and glass surfaces was also determined. The efficacy of CFS on bacterial biofilm established on the glass surface was also observed using fluorescence microscope. Results showed that inhibition zones of L. acidophilus (50.26 mm) were greater than L. casei (37.06 mm). The minimum inhibitory concentration of both CFSs remained stable (40 mg mL-1) during the storage for 28 days at 4.00 and 25.00 °C and storage temperature did not affect the antibacterial effectiveness of CFS. The addition of both CFSs significantly removed biofilm developed on both tested surfaces in a concentration-dependent manner. Biofilm removal property of L. acidophilus CFS was generally better than L. casei CFS which was confirmed by fluorescence microscope. The application of CFS of probiotic strains (i.e. Lactobacillus) as antibacterial and biofilm removal compounds could be very suitable to control the growth of food-borne pathogens.
    Keywords: Antimicrobial, Biofilm removal, Lactic acid bacteria, Probiotics
  • Homayoun Naderain, Neda Khanlarkhani, Iraj Ragerdi Kashani, Amirabbas Atlasi, Mohammad Ali Atlasi * Pages 307-313
    Steroids promote the myelination and regeneration in the peripheral nervous system. Whereas, little is known about the inducing effects by which the hormones exert their effects on Schwann cells differentiation. This could be revealed by the expression of Schwann cell markers in adipose-derived stem cells (ADSCs). The purpose of this study was to present the effects of progesterone and 17 β-estradiol on the Schwann cell markers in rat ADSCs. The mesenchymal stem cell markers (CD73, and CD90) were assayed by flow cytometry. Rat ADSCs were sequentially treated with β-mercaptoethanol, and all-trans-retinoic acid, followed by a mixture of basic fibrobroblast growth factor, platelet-derived growth factor, forskolin and heregulin. In experimental groups, forskolin and heregulin were substituted by progesterone and 17 β-estradiol. After induction, the expression of Schwann cell markers P0, and S-100 and the cellular immunocytochemical staining positive rate of anti-S100 and anti-glial fibrillary acidic protein (GFAP) antibodies were compared in the experimental and control groups. Progesterone and 17 β-estradiol triggered P0 and S-100 genes expression and induced a cellular immunocytochemical staining positive rate of S-100 and GFAP in rats ADSCs. Progesterone induced these changes stronger than 17 β-estradiol. Thus, progesterone may induce rat ADSCs toward Schwann-like cells by expression of Schwann cell markers and is more potent than 17 β-estradiol in the expression of these markers.
    Keywords: 17 ?–estradiol, Adipose tissue, Mesenchymal stem cells, Progesterone, Schwann cells
  • Mohammad Abbaszadeh Hasiri, Effat Baghaei Moghaddam *, Mohammad Reza Khalili, Amin Hossein Amini, Masoomeh Eghtedari, Mohammad Azizzadeh, Hooman Razmi Pages 315-321
    Uveitis is a major cause of vision loss. Methotrexate (MTX) has been widely used in uveitis due to its relatively safe profile. The purpose of this study was to evaluate the effects of two different dosages of MTX via intra-vitreal administration for treatment of endotoxin induced uveitis (EIU) in an experimental model. Thirty-five healthy rabbits were randomly divided into four groups and all animals were tolerated intra-vitreal injections. The first group received normal saline (NS), the second group received normal saline plus Salmonella typhimurium lipopolysaccharide endotoxin (LPS), (NS+LPS), the third group received 400 μg MTX plus LPS (LPS+MTX 400) and the fourth group received 800 μg MTX plus LPS (LPS+MTX 800). Intra-ocular inflammation was evaluated by clinical examination scoring during 7 post-injection days and histopathological examination at the end of study. Kruskal-Wallis and Mann-Whitney U tests were used to compare the histopathological and clinical scores. According to the clinical examinations, all groups demonstrated higher uveitis score than group 1 on first post-injection day. Also, groups 2 and 3 showed greater uveitis score than group 4. On the third, fifth and seventh post-injection days, clinical uveitis score in groups 2, 3 and 4 was significantly higher than group 1. The mean histopathological inflammation intensity scores in groups 2, 3 and 4 were significantly higher than group 1. Single intra-vitreal injection of 400 μg and 800 μg of MTX did not show significant anti-inflammatory effects on EIU in rabbits.
    Keywords: Inflammation, Intra-vitreal injection, Methotrexate, Uveitis
  • Sara khalili, Mehdi Mohebali *, Elaheh Ebrahimzadeh, , Parviz Shayan, Samira Mohammadi, Yeganeh, Mehrdad Moosazadeh Moghaddam , Samira Elikaee, Behnaz Akhoundi, Mohammad Kazem Sharifi, Yazdi Pages 323-328
    Zoonotic cutaneous leishmaniasis caused by Leishmania major is a most common type of vector-borne disease in Iran. The pentavalent antimonial drugs have been used in the treatment of cutaneous leishmaniasis for a long time, but drug resistance and some of serious side effects have been reported. Thus, discovery and development of new therapeutic candidates are needed. The CM11 peptide is one of these peptides that its anti-bacterial activity has been proven. This peptide is a short cecropin–melittin hybrid peptide obtained through a sequence combination approach. The aim of this study was to evaluate in vitro anti-leishmanial activity of CM11 peptide against amastigote forms of Leishmania major. In this study, amastigote forms of Iranian strain of L. major (MRHO/IR/75/ER) were cultured in the presence of different concentrations of meglumine antimoniate (Glucantime®) to find the most appropriate in vitro concentration of Glucantime® against L. major amastigotes. Then, the anti-leishmanial activities of various concentrations of CM11 peptide (8, 16, 32 and 64 µM) were evaluated for 24, 48 and 72 hr by DAPI staining. In addition, MTT assay was used to determine the cytotoxic effects of CM11 peptide on murine fibroblast cell line. The results showed that CM11 peptide has antimicrobial activity against Iranian isolate of L. major in the laboratory conditions. It seems that the CM11 peptide has significant potential to be used as a new anti-leishmanial agent.
    Keywords: Amastigote, CM11 Peptide, Leishmania major
  • Esmaeal Tamaddonfard *, Sina Tamaddonfard, Siamak Cheraghiyan Pages 329-335
    Vitamin B12 modulates pain at the local and peripheral levels. This study has investigated the effects of intracerebroventricular (ICV) injection of vitamin B12 on themuscle pain. We used diclofenac (cyclooxygenase inhibitor) and naloxone (opioid receptors antagonist) to clarify the possible mechanisms. For ICV injections, a guide cannula was implanted in the left lateral ventricle of the brain. Muscle pain was induced by intramuscular injection of formalin (2.50%; 50 µl) in the right gastrocnemius muscle and the number of paw flinching was recorded at 5-min blocks for 60 min. Locomotor activity was performed using an open-field test. Formalin induced a biphasic pain. Vitamin B12 (1.25, 2.50, 5.00 and 10.00 µg per rat) and diclofenac (12.50 and 25.00 µg per rat) significantly reduced both phases pain intensity. Significant antinociceptive effects were observed after combined treatments of diclofenac (6.25 and 12.50 µg per rat) with vitamin B12 (0.63 and 2.50 µg per rat), respectively. Prior ICV injection of naloxone (10.00 µg per rat) prevented vitamin B12 (10.00 µg per rat) and diclofenac (25.00 µg per rat) induced antinociceptive effects. All the above-mentioned chemicals did not alter locomotor behavior in an open-field test. The present results showed that the cyclooxygenase pathway and opioid receptors may be involved in the central antinociceptive effect of vitamin B12. In addition, opioid receptors might be involved in diclofenac-induced antinociception.
    Keywords: Diclofenac, Formalin-induced muscle pain, Opioid receptors, Rats, Vitamin B12
  • Salah Nadimi *, Mohammad Molazem, Seyedhosein Jarolmasjed, Mohammad Reza Esmaili Nejad Pages 337-341
    The objective of this study was to evaluate the pituitary gland dimensions due to age and weight using computed tomography (CT) in dogs and cats. The CT images of pituitary gland were assessed in 11 client-owned dogs (six males and five females; age range, 1 to 9 years) and 15 client-owned cats (eight males and seven females; age range, 1 to 14 years) with no evidence of pituitary diseases. The length, height, width and volume of the pituitary gland were measured in sagittal and transverse planes. Mean pituitary length, width, height and volume (± standard deviation: SD) were respectively 4.96 (± 0.69 mm), 3.62 (± 0.64 mm), 2.62 (± 0.05 mm) and 26.19 (± 7.99 mm3) in cats and were 7.00 (± 2.14 mm), 4.80 (± 1.20 mm), 3.80 (± 0.70 mm) and 77.53 (± 51.64 mm3) in dogs, respectively. Mean pituitary height-to-brain ratio (P:B ratio), (± SD) in cats and dogs was 0.28 (± 0.05) and 0.21 (± 0.03), respectively and mean percent of pituitary volume to brain volume (± SD) in cats and dogs was 0.10 (± 0.05) and 0.10 (± 0.07), respectively. There was no significant correlation between the size of pituitary gland and age, weight and body condition score (BCS) in dogs, however in cats, significant difference was found between height of pituitary gland and weight and BCS, pituitary width and weight and P:B ratio and BCS. These findings could be useful to identify abnormal pituitary gland enlargement in CT images. To be more accurate in the assessments, further studies are required.
    Keywords: Cat, Computed tomography, Dog, Pituitary gland
  • Reza Narenji Sani *, Bahram Hajigolikhani, Mahmood Ahmadi, Hamedani, Khatereh Kafshdouzan Pages 343-348
    The aim of the present study was to investigate the diagnostic evaluation of milk lactate dehydrogenase (mLDH) and alkaline phosphatase (mALP) activities by receiver operating characteristic (ROC) analysis curve in early lactation of ewes with subclinical mastitis (SCM) and determine the correlation between number of somatic cell count (SCC) and mLDH and mALP activities. A total of 196 udder half milk samples were collected within the first 6 weeks of lambing. The SCM was determined by positive milk bacterial culture and positive California mastitis test (CMT); SCC was determined by fossomatic method and enzyme activities were determined spectrophotometrically. The mLDH and mALP of SCM cases were positively correlated with SCC values. Values of mLDH, mALP and SCC were significantly higher in SCM than non-SCM udder halves. The optimum cut-off points of mLDH and mALP activities for SCM diagnosis were determined at 203.61 (U L-1) and 329.84 (U L-1), respectively. In conclusion, SCC has positive correlation with mALP and mLDH activities in SCM ewes and mLDH and mALP activities could be considered as reliable indicators for intra-mammary inflammation diagnosis.
    Keywords: Alkaline phosphatase, Early lactation, Ewe, Lactate dehydrogenase, Subclinical mastitis
  • Abdolrasoul Namjou *, Esfandiar Heidarian, Mahmoud Rafieian, kopaei Pages 349-355
    In this study, the effects of Urtica dioica hydro-alcoholic extract were investigated on the blood glucose and lipid profiles of female ovariectomized and non-ovariectomized rats. In total, 32 adult female rats were divided into four groups (eight each) including control and ovariectomy groups as well as non-ovariectomy and ovariectomy groups treated with 200 mg kg-1 of Urtica dioica extract orally in the last five weeks of the study starting from the week 56th. The duration of the study was 60 weeks. Glucose, serum lipid profiles and pancreatic pathological alterations were determined in these groups at the end of experiment. Serum glucose, triglyceride (TG), very-low-density lipoprotein (VLDL), and TG/high-density lipoprotein (HDL) ratio indicated a significant increase in the healthy female rats under treatment with Urtica dioica extract compared to others. The TG, cholesterol, HDL, low-density lipoprotein (LDL) and VLDL showed a significant increase in menopaused rats compared to others. The interaction of consuming Urtica dioica extract and ovariectomy caused significant decreases in glucose, TG, VLDL, HDL/LDL ratio and TG/HDL ratio. Consumption of Urtica dioica extract by non-menopaused rats damaged the beta cells in Langerhans islets. Results of the present study revealed that the consumption of Urtica dioica extract is not beneficial and has diabetogenic effects in female non-ovariectomized rats compared to ovariectomized ones.
    Keywords: Glucose, Lipid profiles, Ovariectomy, Rat, Urtica dioica
  • Javad Alikhani, Godratollah Mohammadi *, Gholamreza Shariati Pages 357-360
    The domestic water buffalo (Bubalus bubalis) is an economically important livestock species in many Asian and Mediterranean countries. Two types of water buffalos are river and swamp buffaloes. Genetically, the swamp buffalo has 48 chromosomes (2n = 48) and the river buffalo has 50 chromosomes (2n = 50). Khuzestan province is one of the important centers of buffalo farming in Iran. Since there was no cytogenetic information about Khuzestani buffalo breed, the present study was performed to provide karyotype and cytogenetic study in Khuzestan buffaloes. In this study, blood samples of 40 buffaloes were collected randomly from jugular vein via heparinized syringes. For metaphase smear, blood samples were cultured in RPMI 1640 medium containing fetal bovine serum and phitohemaglutination and then transferred to water bath for 69 hr. After stopping cell division by colcemide and using fixed solution, from each sample six slides were stained with Giemsa staining. Based on the results, number of chromosomes of the Khuzestani buffaloes was 50 (2n = 50). Among the 25 pairs of chromosomes, the first 5 pairs of autosomal chromosomes were sub-metacentric, 19 pairs of autosomal ones were acrocentric and the sex chromosomes (X and Y) were acrocentric. Furthermore, X and Y chromosomes were the largest and smallest acrocentric ones, respectively. Therefore, the buffaloes of Khuzestan province are considered as a river type.
    Keywords: Cytogenetic, Karyotyping, Khuzestan, River buffalo
  • Javad Aliakbarlu *, Sindokht Ghiasi, Behnaz Bazargani, Gilani Pages 361-365
    Barberry fruit is consumed in different forms including dried fruit, juice, jam and marmalade in Iran. This fruit is also used as a food additive (flavoring and colorant) in soup and rice dishes. In present study, antioxidant activities of acetone, ethanol and water (infusion and decoction) extracts of barberry (Berberis vulgaris) fruit were investigated using 2,2-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power methods. Total phenolic contents of the extracts were also estimated using Folin-Ciocalteu assay. In ABTS assay, acetone and ethanol extracts showed the highest radical scavenging activity, while in DPPH and reducing power methods, acetone extract and decoction exhibited the strongest antioxidant activity. Meanwhile, the antioxidant potential of water extracts increased with increasing heating time (antioxidant activity of decoction was higher than that of infusion). The highest total phenolic content was found in the acetone extract (92.75 mg GAE per g). It was concluded that the acetone extract and decoction of barberry fruit can be used as an effective natural antioxidant in food industry.
    Keywords: Antioxidant Activity, Barberry, Extract, Fruit
  • Eshrat Beigom Kia, Mitra Sharbatkhori *, Zahra Heidari, Farideh Tohidi, Bahareh Kamran Rashani, Farzaneh Zahabiun Pages 367-371
    A polycephalic larva of Taeniidae family isolated from abdominal cavity of a great gerbil (Rhombomys opimus) from Golestan province, northern Iran, was subjected to molecular analysis. Genomic DNA from the larva was obtained using a DNA extraction tissue kit. Polymerase chain reaction was performed for amplification of the partial 12S rRNA, cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes. BLAST analysis of DNA sequencing indicated 99.00% homology in 12S rRNA and cox1 genes and 98.00% homology in nad1 gene with Hydatigera krepkogorski (accession No. AB731762). The sequences of current isolate were deposited in GenBank by accession Nos. MF281971, MF281972 and MF281973 for 12 SrRNA, cox1 and nad1 genes, respectively. This study was the first report of molecular characterization of H. krepkogorski from Iran. Isolation and characterization of the adult stage from definitive host will help to better clarify incomplete life cycle and morphology data of this species in the world.
    Keywords: Hydatigera krepkogorski, Iran, Rhombomys opimus
  • Saeed Seifi *, Hojjatollah Shokri, Mahvash Karimi Madab Pages 373-378
    Fungal infections cause significant economic losses in the poultry industry either due to their direct infectious nature or due to mycotoxins production. Hatchery contamination with fungi can threaten chicken health. In this regard, geographical and seasonal distributions of airborne fungal contamination of 25 hatcheries in Mazandaran province, northern Iran, were investigated using an open plate method. The results of this study showed that hatcheries have various fungal contaminations, among which the most common were respectively Cladosporium (31.07%), Penicillium (24.00%), Aspergillus (20.63%), sterile hyphae (14.70%) and Alternaria (6.20%) from different regions. The results revealed that the highest level of fungal isolation was in spring and autumn. This study also showed that the concentration of fungal air spora in forest and seaside locations was significantly greater than mountainous ones. In spite of the regular disinfection in commercial hatcheries, fungal contamination was found in different parts.
    Keywords: Contamination, Fungus, Hatchery, Iran
  • Enrico Pierluigi Spugnini *, Andrea Lanza, Silvia Sebasti, Alfonso Baldi Pages 379-381
    A five-year-old female African hedgehog (Atelerix albiventris) was referred for a one month growing oral mass. The hedgehog was quiet, alert and responsive, with a 1.00 × 1.50 cm ulcerated lesion on the mandible. The patient was staged with total body radiographs, hematological and biochemical analyses and the mass was biopsied under anesthesia. The excised tissue was reported to be a squamous cell carcinoma. At this time, due to the extension of the mass, a surgical excision was not a therapeutic option and the tumor was palliated with electrochemotherapy (ECT). Monthly sessions of ECT were performed using intra-lesional bleomycin injection followed by trains of biphasic electric pulses. The treatment was well tolerated and the patient remained in partial remission for five months until tumor progression. At that time the pet was euthanized. The ECT resulted in improved local control and should be considered as a possible adjuvant treatment in exotic pets with advanced tumors.
    Keywords: African hedgehog, Bleomycin, Electrochemotherapy, Squamous cell carcinoma