ARCHIVES OF RAZI INSTITUTE
Year:2013 | Volume:68 | Issue:2|Papers Count:12

Scorpionism is a known significant problem of medical and social importance in many tropical and subtropical regions including the Middle East. In Iran, highest prevalence of scorpion sting about 60% of all the stings has been reported from Khuzestan province. Among the 21, 000 cases of reported scorpion stung patients, 12% were caused by H. lepturus, but contributed to 95% of all moralities in scorpion stung patients. The sting of H. lepturus does not produce an immediate pain as does the sting of other scorpions, rather cause delayed swelling that may diffuse and is often accompanied by late necrosis at the sting site suggestive of less significant role of the nervous system stimulation. Since the venom from H. lepturus is cytotoxic in nature and the renal response and blood toxicity are normally simultaneously manifested, it is suggested that the toxin binds to kidney tissue and potentially induce acute renal failure in stung patients. Pharmacokinetic analysis revealed that Intramuscular (i.m) injection of antivenoms is ineffective in neutralizing the action of venoms. Although some reports mention the slow distribution rate of H. lepturus venom following sting, but since the cytotoxic effect of venom from this scorpion is irreversible by antivenom once it occurs, it is recommended to use antivenom through intravascular (i.v) route. Antivenoms of F (ab) 2 fraction are the best choice of treatment for their fast extravasation, their ample distribution into the extracellular space, and their prolonged retention time.

Acute bee paralysis virus (ABPV) is a small single stranded RNA virus recently classified within the family Dicistroviridae, genus Cripavirus. Here, we describe the first study of ABPV in unhealthy bee colonies, which has been an unusual loss in adult bee population and significant honey bee mortality during the year. The aim of this study was evaluation of ABPV infection in honey bee colonies in apiaries with loss of population in different geographical provinces in Iran. Adult bee samples were collected between July - September 2011 and 2012 and were originated from 23 provinces with different geographic areas of Iran. Following the RT-PCR reaction with the specific primers on the isolated RNA, an approximately 618 bp product was detected. We demonstrated the presence of ABPV RNA in 9 (5.62 %) out of 160 samples collected from Iranian apiaries.

In diagnostic and research bacteriology settings with budget and staff restrictions, fast and cost-effective genome extraction methods are desirable. If not inactivated properly, cellular and/or environmental DNA nucleases will degrade genomic material during the extraction stage, and therefore might give rise to incorrect results in PCR experiments. When crude cell extracts, proteinase K-treated templates and purified DNAs prepared by phenol-chloroform-isoamylalcohol method as well as a commercial extraction kit were subjected to the Salmonella enterica Enteritidis specific STM2 PCR, with exception of crude cell extract, PCR products from all other three methods saved their integrity for 28 days post-generation. This work aimed to find out whether improvement to boiling method can guaranty stability of PCR products. As results showed, treatment of crude cell extracts from S. Enteritidis with proteinase K offers an inexpensive, fast and effective DNA extraction method suitable for high-throughput laboratories.

Brucellosis is a debilitative disease that imposes costs on both economy and society. It is shown that although the vaccine can prevent abortion, it does not provide complete protection against infection. In Iran, Brucella melitensis is a common causative agent for brucellosis and BP26 protein of this bacterium having a good antigenesity and an important vaccine candidate. In this study B. melitensis bp26 gene was cloned first in to PTZ57R/T vector and accessed on the PET28a vector and sequenced. Recombinant vector transformed and expressed in to E. coli BL21 (DE3) and then recombinant protein was purified with Ni-NTA column of chromatography against His tag. Obtained rOmp28 could be used as a research experimental tool to find its potential as a detection kit and vaccine candidate.

Mycoplasma synoviae expressed variable lipoprotein haemagglutinin (VlhA) is believed to play a major role in pathogenesis of the disease by mediating adherence and immune evasion. The aim of this study was sequencing Iranian M. synoviae isolates for the detection of nucleotide variation in the M. synoviae vlhA gene. Using oligonucleotide primers complementary to the single-copy conserved 5’ end of vlhA gene, amplicons of ~400 bp were generated from 10 M. synoviae isolated from commercial broiler chicken farms in Iran, afterward the conserved domain of the vlhA gene of M. synoviae was sequenced and analyzed for Iranian isolates. The results showed that, there was a complete concordance between all Iranian isolates nucleotide sequence (1-386 nt). In comparison with vaccine MS-H strain sequence, all Iranian isolates; entire vlhA sequence downstream of nucleotide 386 was different. It was also observed in all Iranian M. synoviae isolates, point mutations and frame-shift mutation. This study was demonstrated a difference between Iranian isolates and live commercial vaccine MS-H strain. Furthermore, these data indicated that changes in the vlhA gene sequence could introduce into the expressed vlhA gene amino acid codons and effective in pathogenesis rate in flocks.

Filariasis in dogs is caused by several species of filariids. Because of importance of this infection in veterinary medicine and public health, it is necessary to carry out an epidemiological and cross sectional studies in various geographical areas and use of well-adapted diagnosis methods. In this study 205 capillary and whole blood samples were collected from doges in various counties of East-Azerbaijan province. Samples after preparation were examined by Knott’s test and light microscope for presence of microfiler. In molecular identification, Pan-filarial and species-specific PCR perimers was used to differentiate among Dirofilaria immitis, Dirofilaria repens, Acanthocheilonema reconditum, Acanthocheilonema dracunculoides, Brugia malayi and Brugia pahangi. Descriptive statistics were used for data analysis. Total infection prevalence with the microscopic evaluation was 77 (37.5 %) and in PCR test was 94 (45.8 %). The most common species of canine filarial parasite identified in this study was D. immitis 54 (57.4 %) followed by Acanthocheilonema species 40 (42.6 %). The molecular evidence on the sequence of the ITS-2 region provided strong evidence that the canine microfilariae discovered in this study belong to a novel species of Acanthocheilonema. Information about infection prevalence helps us to improve disease management practices in the studied area, apply new hygiene policy and reduce extra costs of therapeutic agents and PCR is a quick and accurate molecular genetics method for detection of filarial species.

In the last decades researchers had focused on developing a vaccine against tick based on protective antigen. Recombinant vaccines based on concealed antigen from Boophilus microplus have been developed in Australia and Cuba by the name of TICKGARD and GAVAC (De La Fuente and Kocan, 2006). Further studies on this antigen have shown some extent of protection against other species (De Vos et al., 2001). In Iran most important species is Hyalomma anatolicum and limited information about its control are available. This paper reports structural and polymorphic analysis of HA03 as an Iranian candidate concealed antigen of H. a. anatolicum deposited in Gen-Bank. (Aghaeipour et al. GQ228820). The comparison between this antigen and other mid gut concealed antigen that their characteristics are available in GenBank showed there are high rate of similarity between them. The HA03 amino acid sequence had a homology of around 89%, 64%, 56% with HA98, BM86, BM95 respectively. Potential of MHC class I and II binding region indicated a considerable variation between BM86 antigen and its efficiency against Iranian H. a. anatolicum. In addition, predicted major of hydrophobisity and similarity in N-glycosylation besides large amount of cystein and seven EGF like regions presented in protein structure revealed that value of HA03 as a new protective antigen and the necessity of the development, BM86 homolog of H. a. anatolicum HA03 based recombinant vaccine.

In recent years, encapsulation of drugs and antigens in hydrogels, specifically in calcium alginate particles, is an interesting and practical technique that was developed widespread. It is well known that alginate solution, under proper conditions, can form suitable nanoparticles as a promising carrier system, for vaccine delivery. The aim of this study was to synthesis alginate nanoparticles as protein carrier and to evaluate the influence of various factors on nanoparticles properties. Alginate nanoparticles were prepared by ionic gelation method. Briefly, various concentrations of CaCl2 were added to different concentrations of sodium alginate dropwisly by homogenizing magnetically at 1300 rpm. The effects of homogenization time and (-) rate were investigated on nanoparticle feature. Nanoparticles were characterized for their morphology and size distribution. Evaluation of loading capacity and loading efficiency of nanoparticles were performed by using various concentration of BSA. The concentration of 0.3%w/v sodium alginate and 0.1%w/v CaCl2 solution, homogenization time 45 min and homogenization rate 1300 rpm were observed as suitable condition - to prepare optimized nanoparticles. It can be concluded that the properties of nanoparticles are strongly dependent on the physicochemical conditions. The optimum concentrations of alginate and CaCl2 and appropriate condition led to forming desirable nanoparticles that can be used as carrier for drug and vaccine delivery.

Most of the systematic studies used in the development of human embryo culture media have been done first on mouse embryos. The general use of NMRI outbred mice is a model for toxicology, teratology and pharmacology. NMRI mouse embryo exhibit the two-cell block in vitro. The objective of this study was to evaluate and compare the effects of four kinds of culture media on the development of zygotes (NMRI) after embryo vitrification. One-cell mouse embryos were obtained from NMRI mice after superovulation and mating with adult male NMRI mice. And then randomly divided into 4 groups for culture in four different cultures media including: M16 (A), DMEM/Ham, F-12 (B), DMEM/Ham's F-12 co-culture with Vero cells (C) and DMEM/Ham's F-12 co-culture with MEF cells (D). Afterward all of the embryos were vitrified in EFS40 solution and collected. Results of our study revealed, more blastocysts significantly were developed with co-culture with MEF cells in DMEM/Ham's F-12 medium. More research needed to understand the effect of other components of culture medium, and co-culture on NMRI embryo development.

Leishmaniasis is caused by parasitic protozoa of the genus Leishmania. Cutaneous leishmaniasis (CL) is a complex disease with wide spectrum of clinical manifestations. In order to identify leishmania species causing CL in Isfahan by a definite molecular technique (PCR method), this study was undertaken over 2010- 2011.124 Patients with suspicious lesion of Leishmaniasis and positive direct smear from lesion were selected. Samples were cultured in NNN and RPMI 1640 media Negative and positive control and clinical samples was applied for PCR in the same condition. In the next step, standard PCR was carried out using classic protocol. From 124 patients, 111 (89.51%) cases were infected as L. major and 12 (9.67%) cases were infected by L. tropica, However only in one patient simultaneous infectious with both L. major and L. tropica was identified by PCR techniques which could not be possible in microscopy. L.major was the most prevalent species in the studied patients (p-value<0.001).

A molecular survey was performed for identification Theileria spp in sheep and ticks during from 2010-2011 in Fasa and Kazeroun areas, Fars province, Iran. A total of 100 sheep from different flocks were clinically examined and blood samples with ixodid ticks collected. The prepared blood smears from capillary vein of ear were stained with giemsa methods and examined by using light microscope. The collected ticks were separated into tick pools with five ticks according to their species and sex. Then, the salivary glands were dissected out in 0.85% saline solution under stereomicroscope. The boold and tick salivary glands samples were examined by using semi-nested PCR. The Theileria spp infection was observed in 46% of blood smears, while 76 % of blood samples were positive by using semi-nested PCR. T.ovis, T. lestoquardi and mixed infection were detected in 43 (43%), 3 (3%) and 30 (30%) of positive samples, respectively. Any significant difference was not observed between the frequency of Theileria spp infection in sheep of Kazeroun and Fasa areas. In the present study, the most prevalent ticks were R. turanicus 48.8% and followed by H. a. anatolicum 42.2% and H. marginatum 8.8%. The results were shown that one pool belong to salivary glands of H. turanicus were infected with T. ovis. Based on the obtained results, it is concluded that T. ovis have high prevalence with compared to T. lestoquardi and also, R. turanicus could be the vectors T. ovis in this area.

Leishmaniasis is an important and common zoonotic disease with a great impact on public health. In the present study, a total of 195 companion cats of different ages were examined for serum antibody detection against Leishmania infantum by immunochromatography assay. The cats were selected from those referring to Veterinary Hospital of Ahvaz University, south west of Iran from May 2009 to March 2012. Classification was made by age, sex, breed, region and season. The studied cats were divided based on age into three groups (<1 year, 1-3 years and>3 years) and based on area into five regions (north, east, west, south and central). The results were analyzed by using Chi-square analysis, Fischer's exact test and Z test. Eighteen of 195 serum samples (9.23%) had antibody against Leishmania infantum (%95 Confidence Interval: 5.1-13.3%). Prevalence was significantly higher in adult cats above 3 years (28.57%; 14 out of 49) compared with mean-age cats 1- 3 years (3.57%; 3 out of 84) (Odds Ratio: 10.8) and less than 1 year (1.61%; 1 out of 62) (OR: 24.4) (P<0.001). Prevalence was higher in male cats (9.82%; 11 out of 112) than females (8.43%; 7 out of 83), the spring season (13.04%; 6 out of 46) and north region (13.89%; 5 out of 36), but the difference was not significant between the prevalence of infection relative to host gender, season and region (P>0.05). In conclusion, it is necessary to control cat's population in these area particular adult cats to reduce risk of infection transmission to other animals and human.