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Detection of Coxeilla brunetii in bulk tank milk samples from dairy bovine farms using nested-PCR in Qom, Iran, 2011
2013
Ghalyanchi Langeroudi, Arash | Babkhani, Narges | Zolfaghari, Mohammad Reza | Majidzadeh Arbadili, Keivan | Morovvati, Abbas | Soleimani, Mohammad
BACKGROUND: Q fever is a zoonotic disease caused byCoxiella burnetii, a species of bacteria that is distributedworldwide. In cattle, Coxiella burnetii infections are generallyasymptomatic but can also be associated with reproductivedisorders. OBJECTIVES: The aim of this study was to achievemolecular detection of Coxiella burnetii in dairy bovine milkfarms using Nested PCR in Qom province, Iran. METHODS:From January to February 2011 (winter) and July to September2011(summer) a total of 100 bovine bulk milk samples wereequally collected from five areas of Qom. The nested PCR assayused to screen for C. burnetii was designed from the nucleotidesequence of the com1 gene encodin a 27-kD outer membraneprotein (OMP). RESULTS: In this study, 14% (14 of 100) of bulkmilk were positive. CONCLUSIONS: These results support thehypothesis of high prevalence and endemic pattern of Q fever inQom province of Iran.
Показать больше [+] Меньше [-]Blood and tissue levels of diazinon in rabbit following a subacute dermal exposure to incremental doses
2013
Arab, Hoseinali | Goudarzi, Masoud | Koohi, Mohammadkazem | Shams, Gholamreza
BACKGROUND:Uncontrolled application of diazinon (DZN)can cause environmental contamination and adverse healtheffects on humans or animals. OBJECTIVES:This study aimed toinvestigate the toxic effects and the level of DZN in serum andtissues of rabbits following a sub acute dermal exposure totoxicant. METHODS:Different doses of DZN were applied dailyto New Zealand rabbits through the ear skin in incremental dosesfor 4 weeks. Blood samples were collected at the beginning andthe end of each dose-week period. Tissue samples were collectedfrom brain, muscle, kidney and liver on day 28, after euthanizingthe rabbits. DZN contents of the blood and tissue samples weremeasured using a reversed phase HPLC system. RESULTS:Clinical observations indicated signs of toxicity in the animalsexposed to DZN as shown by diarrhea and body weight loss fromday twenty. The level of DZN in the blood elevated withenhancing exposure time and reached the highest level at the endof the fourth week (0.620±0.26ppm). The highest level of DZNwas found in the brain tissue (0341±0.015 ppm). CONCLUSIONS:The results of this study revealed the tissue accumulation andsubsequent toxic effects of DZN following the subacute dermalexposure to the toxicant. It suggests that the determination of thetoxicant level in the serum or tissue can be a monitoring methodfor the detection of the contamination rate.
Показать больше [+] Меньше [-]The mRNAexpression study on small amount of Theileria annulata lymph node biopsy sample using SMART-cDNA technology
2013
Sadr Shirazi, Nastaran | Shayan, Parviz | Eckert, Berigiteh | Ebrahimzadeh, Elaheh | Jafari, Sedigheh
BACKGROUND: A major issue in many gene expressionstudies utilizing small amount of biological materials is thelimited quantity of RNApurified from clinical samples, which isoften used for RT-PCR or standard Northern blot analysis.OBJECTIVES: The SMART cDNA synthesis method and subsequentSMART-cDNA-PCR technique was used to analyse 3genes in macroschizonts of Theileria annulata in small lymphnode biopsy material. METHODS: The SMART-cDNA of TaSpgene was cloned in pTZ57R/T-vector and sequenced. We focusedon genes encoding surface proteins TaSp, TaD and HSP70.RESULTS: Our results showed that SMART cDNA dependablyreproduces the expression profile found in messenger RNA. TheRT-SMART-PCR showed the amplification of the processedmRNAs. The sequencing analysis showed that the amplifiedcDNA was coded for TaSp protein in Theileria annulata.CONCLUSIONS: It was concluded that the SMART PCRtechnique is practical for amplification of complete sequence ofmRNAs in the form of cDNAs, and therefore for gene expressionstudies if only small amounts of starting material are available.
Показать больше [+] Меньше [-]Molecular characterization of non-structural gene of H9N2 subtype of avian influenza viruses isolated from broiler chickens in Iran
2013
Emadi Chashmi, Hesam-aldin | Vasfi Marandi, Mahdi | Bozorgmehrifard, Mohammad Hasan | Bashashati, Mohsen | Barin, Abbas
BACKGROUND: The H9N2 subtype of avian influenzaviruses (AIVs) have spread in Asia and Middle East countriesand have become a serious threat to poultry industry in Iran.OBJECTIVES:Characterization of genes of H9N2 subtype involvingin pathogenicity and diagnosis are crucial in control of avianinfluenza outbreaks. The Nonstructural (NS) gene and its proteinproducts (NS1 & NS2) are important as diagnostic marker, lifecycle and pathogenicity of AIVs. METHODS:The NS gene of fivestrains, isolated from 1998 to 2010, were completely sequencedand analyzed. RESULTS:All of the examined strains were composedof 890 nucleotides with 230 amino acids. In this regard, onlytwo Iranian strains from GeneBank had 217 amino acids in NS1protein. All Iranian H9N2 strains subdivided into two distinctsublineages including I and II. Comparative analysis of NS genesof Iranian strains showed that since 2003, they might haveoriginated from Pakistan H7N3 strains; whereas from 2008 theycould be originated from Pakistan H9N2 strains. CONCLUSIONS:Although the low pathogenic H9N2 subtype has been permanentlycirculating from 1998 to the present in Iran, phylogeneticanalysis of NS genes revealed that sublineage II has circulatedmore in poultry industry of Iran. These epidemio-logicallyvariations could be related to vaccination pressure due tomassive vaccination or NS gene reassortment in rural andbackyard chickens.
Показать больше [+] Меньше [-]An outbreak of abortion in Afshari sheep with probable involvement of Campylobacter fetus
2013
Saleh, Masoumeh | Harkinezhad, Mohammad Taher | Marefat, Alireza | Salmani, Vahid
BACKGROUND:Abortion is one of the most important factorsreducing lambing rate and consequently profitability of sheepfarms. In addition to financial losses, it is also important from azoonotic point of view. OBJECTIVES: The aim of this study wasto investigate bacterial abortifacient agents in an outbreak ofabortion occurring in Afshari sheep in the northwest of Zanjanprovince. METHODS:Vaginal swab samples were collected from217 Afshari ewes (129 samples were taken from aborted ewes, 3samples from ewes with crippled and deformed lambs, and 85samples from animals that had given birth to healthy lambs) fromreported flocks involved in outbreak. Swabs were examined byPCR assay to detect DNAfrom Coxiella burnetii, Chlamydophilaabortus, Salmonella enterica, Yersinia enterocolitica, Campylobacterfetus, Brucella ovis and Leptospira interrogans. RESULTS:Based on the results, only DNA of Campylobacter was detectedin the samples. A 266 bp fragment specific for Campylobacterwas amplified from 51.52% and 34.12% samples belonging toaborted and non-aborted ewes, respectively. CONCLUSIONS:Significant presence of the bacterium in aborted ewes (p<0.001)compared to the non-aborted groups with odd ratio of 3,emphasizes that Campylobacter could be involved in theoutbreak of the abortion. Considering the importance of thedisease, prophylactic measures are needed to reduce the disease.However, further investigations are required to determine theimpact of this bacterium in prevalence of abortion in sheep inother areas.
Показать больше [+] Меньше [-]Histological study of cartilaginous cells in the body of penis in one-humped camel (camelus dromedarius)
2013
Yousefi, Mohammad Hasan | Gilanpour, Hasan | Salimi-Bejestani, Mohammad Reza
BACKGROUND:Several investigations showed cartilaginouscells in fibrous tissue of the free part of the penis in one humpedcamel. OBJECTIVES: The aim of this study was accurate assessmentof existence of cartilaginous cells in penis shaft of onehumpedcamel. METHODS: Six camel penises from maturedcamels more than 3 years-old were collected from an abattoir.Different specimens were prepared from each penis and kept in10% formalin container for fixation. After passing differentstages of histotechnique methods, several slides were preparedfrom each specimen, stained with Haematoxylin Eosin andstudied. RESULTS: Results showed that the majority of cartilaginouscells were inside the collagen fibers of tunica albuginea andaround corpus cavernosum and corpus spongiosum of penis andtheir distributions were dissimilar in different parts of the penisshaft. This survey further showed that in penis shaft length, themajority of cartilaginous cells were inside tunica albuginea,which is surrounded by corpus spongiosum and particularly, theventral surface of urethra. CONCLUSIONS: The number ofcartilaginous cells decreased gradually from distal extremitytowards the proximal extremity of the body of the penis andincreased gradually from external layer of tunica albugineatowards the internal layer of tunica albuginea and centre ofcorpus cavernosum penis. Existence of cartilaginous cells insidethe leaf tissue of the penis was seen with aging and puberty.
Показать больше [+] Меньше [-]Isolation of bovine spermatogonial cells and co-culture with prepubertal sertoli cells in the presence of colony stimulating factor-1
2013
Shafiei, Shiva | Tajik, Parviz | Ghasemzadeh-nava, Hamid | Movahedin, Mansoureh | Talebkhan Garoussi, Massoud | Qasemi-Panahi, Babak | Rahimi Feyli, Peyman
BACKGROUND: Spermatogonial stem cells (SSCs) are infrequentself-renewing cells among the type A spermatogoniawithin the seminiferous tubules and are the basis of spermatogenesisin mammalian testis. An adequate number of SSCs is aprimary requirement for the study of their behavior, regulation, andfurther biomanipulation. OBJECTIVES: In this paper, we studiedthe development of the primary co-cultures of type A spermatogoniaand prepubertal bovine sertoli cells in the presence of ColonyStimulating Factor 1 (CSF1), a potential contributor in the SSCniche. METHODS: The effect of different concentrations of CSF1(0, 10, 50 and 100 ng/mL) on the colonization activity of spermatogonialcells was assessed 4, 7 and 11 days after the beginning of theculture by counting the total number of colonies and measuring theirarea in each group of the present experiment. Immunofluorescentstaining against OCT4 and vimentin led to the confirmation of thenature of both the SSCs and sertoli cells. RESULTS: Results showedthat the total number of colonies from day 4 to 11 increasedsignificantly in all groups, independent of CSF1 concentration. Inaddition, the total number and total area of colonies were higher (notsignificant) in 10 and 50 ng/mL CSF1 treatments than the controland 100 ng/mL CSF1 groups in all the three evaluations during theexperiment. However, this difference was only significant (p<0.05)between the total area of colonies in the control and 10 ng/mLCSF1groups at day 4 of co-culture. CONCLUSIONS:It was concluded thatCSF1 can be a suitable growth factor for improving SSCs colonizationin vitro, particularly during the first days of culture whereaccompanying sertoli cells still have not proliferated sufficiently tosupport the propagating spermatogonial cells.
Показать больше [+] Меньше [-]The submandibular salivary gland microscopic morphology of the adult African giant pouched rat (Cricetomys gambianus, waterhouse-1840)
2013
Ikpegbu, E. | Nlebedum, U.C. | Nnadozie, O. | Agbakwuru, I.O.
BACKGROUND: In the present study, the submandibularsalivary gland microscopic morphology of the adult Africangiant pouched rat was investigated. This study was carried out toprovide the basic histology of this organ in the giant pouched rat,to accompany the dearth of information of its microscopicarchitecture in the available literature. This becomes of evenhigher importance when considering the possible use of thisspecies of rodent as a future laboratory animal to replace theWinster rat, because of its bigger size and the possibility ofdomesticating the giant pouched rat as a ready source of animalprotein. In addition, the need to understand the digestive biologyto help animal nutritionists in feeding formulation may also beachieved. The histology revealed the presence of both serous andmucus secretory acini. Some mucus cells showed serousdemilumes. The myoeithelial cells were seen around thesecretory cells and the intercalated ducts. The serous glandregion with more relatively profuse intralobular ducts was largerin size than the mucus gland region. The intralobular ducts ofintercalated and striated ducts were lined by simple cuboidal andsimple columnar cells, respectively. The excretory duct waslined by the stratified cuboidal cells. The large serous glandularregion reflects need for more enzymic action in the oral cavity,while the mucus glands will help produce mucin that willlubricate the digestive tract. This study, for the first timedocuments the normal histology of submandibular salivarygland in this species, hence filling the knowledge gap that willhelp further research especially on the role of myoepihelial cellsin the secretory glands tumours.
Показать больше [+] Меньше [-]Isolation, characterization and transduction of canine bone marrow-derived mesenchymal stem cells (cBM-MSCs)
2013
Rezaei, Mahdieh | Jamshidi, Shahram | Saffarpour, Anna | Ashouri, Mahdi | Sharifi, Davood | Zamankhan Malayeri, Hamed | Tamimi, Naqa | Rokn, Amir Reza
BACKGROUND: Stem cell therapy in small animal medicineis still in its infancy and few in vitro and in vivo research projectsregarding animal Mesenchymal Stem Cells (MSCs) have beencarried out. On the other hand, Cell tracking is the first step of thecell-based therapies and is essential to recognize cell fate posttransplantation. OBJECTIVES: The aim of this study was toisolate, characterize, and transduce cBM-MSCs. METHODS:Canine Bone Marrow-derived Mesenchymal Stem Cells (cBMMSCs)were isolated from bone marrow of dogs andcharacterized based on morphology, differentiation capacities,and surface marker expressions. For the first time, we labeledcBM-MSCs by GFP-encoding lentiviral vector to track them.RESULTS: cBM-MSCs were successfully isolated and proliferated.Morphologically, these cells were similar to otherMSCs from other sources and species and were able todifferentiate into osteocytes and adipocytes. cBM-MSCsexpressed surface marker CD44 but were not able to expressCD34. Approximately, 70% of cells efficaciously expressedGFPafter labeling; CONCLUSIONS:We found that GFP labelingis an easy and effective technique to track transplanted cBMMSCs.Our results also provide fundamental information aboutcanine BM-MSCs in order to use in veterinary medicine.
Показать больше [+] Меньше [-]Antibiotic residues and aflatoxin M1 contamination in milk powder used in Tehran dairy factories, Iran
2013
Noori, Negin | Karim, Guity | Raeesian, Mahyar | Khaneghahi Abyaneh, Hamid | Bahonar, Alireza | Akhondzadeh Basti, Afshin | Ghadami, Freshteh
BACKGROUND: The presence of aflatoxin M1 (AFM1) andantibiotic residues in milk and milk products is a public healthconcern. Milk and milk powder have the potential forintroducing AFM1 and antibiotic into human diet. In recentyears, milk powder has been used on a large scale in dairyfactories. Consequently, antibiotic residues and aflatoxincontamination control in these products has gained importance.OBJECTIVES: The aim of this survey was to determine the levelof β-lactam and tetracycline antibiotic residues and also AFM1contamination of milk powder used in Tehran dairy factories.METHODS: During 12 months (September 2011 to September2012), 240 samples of milk powder were collected from tenTehran dairy factories. All samples were analyzed for thepresence of AFM1 using ELISA technique. In addition,antibiotic residues were determined by BetaStar Combo test, arapid assay for both β-lactam and tetracycline antibiotics.RESULTS: The samples depicted positive results i.e. 30% and17.5% for β-lactam and tetracycline antibiotics, respectively.Also, AFM1 was found in 155 cases (64.6%) with an averageconcentration of 29.85 ± 18.99 ng/ L. CONCLUSIONS:The resultsshowed the milk powder used by dairy factories is safe in respectof AFM1 contamination and antibiotic residues in Tehran.
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