Immunological evaluation of a novel multi-antigenic DNA vaccine encoding SAG1, SAG3, MIC4, GRA5, GRA7, AMA1and BAG1 against Toxoplasma gondii in BALB/c mice.

Many recent studies have been conducted to find new DNA vaccines based on Toxoplasma gondii antigens. DNA vaccines encoding complex of different antigens showed better immune responses compared to single antigen vaccine. In this study, we constructed a DNA vaccine encoding SAG1, SAG3, MIC4, GRA5, GRA7, AMA1 and BAG1 against T. gondii, and evaluated the immune response it induced in BALB/c mice. For this purposes, thirty BALB/c mice were randomly divided into three groups containing tenmice each. There were two negative control groups (PBSand pVAX1 vector) and one vaccination group (pVAX1-MAF, Multantigenic Fragment). On days 0, 14 and 28, the mice were immunized intramuscularly, and 5 weeks later they were challenged with T. gondii RH strain. The immune responses were evaluated using lymphocyte proliferation assay, T-cell subsets detection, and measurement of antibody and cytokine levels. The results showed that mice immunized with pVAX1-MAF developed high levels of IL-2, IL-12, IgG and IFN- γ as well as CD3+CD4+ T cells. In addition, the survival time of mice immunized by pVAX1-MAF was longer than that control mice. In conclusion, our results show that the multiple DNA vaccine encodingSAG1, SAG3, mic4, GRA5, GRA7, AMA and BAG1effectively enhanced humoral and cellular immune responses, and prolonged the survival time. Together this would suggest that further investigation may result in a promising candidate vaccine to treat toxoplasmosis.

Genomic Detection of Mycobacterium avium subspecies paratuberculosis in Blood Samples of Patients with Inflammatory Bowel Disease in Southern Iran.

BACKGROUND: Inflammatory bowel disease (IBD), of which Crohn's disease (CD) and ulcerative colitis (UC) are the two main clinicopathological subtypes, is a group of digestive system diseases of unknown etiology. Risk factors for IBD are environmental factors, genetics, and immune system agents. Mycobacterium avium subspecies paratuberculosis (MAP) is one of the most important infectious factors and a suspected cause of IBD. The present study aimed to determine the prevalence of MAP in both IBD patients and non-IBD people as well as to investigate the relationship between the presence of this bacterium and IBD. METHODS: A cross-sectional study was conducted during May-December 2017 among 146 IBD patients (32 with CD and 114 with UC) at the Motahari Clinic affiliated to Shiraz University of Medical Sciences, Shiraz, Iran. For comparison, the blood samples of 146 non-IBD volunteers (the control group) were tested for the presence of MAP using the polymerase chain reaction method (specific IS900 fragment). The data were analyzed using the SPSS software (version 19.0). The Kolmogorov-Smirnov test was used to evaluate the normal distribution of variables. The χ2 test was used to compare the qualitative variables between the groups. RESULTS: MAP was present in 104 (71.2%) IBD patients out of which 24 (75%) had CD and 80 (70.2%) had UC. In the control group, MAP was present in 63 (43.2%) non-IBD volunteers. There was a significant association between the presence of IBD and MAP (P<0.001). CONCLUSION: A high prevalence of MAP was observed in the South of Iran. MAP DNA was detected in the blood samples of CD and UC patients as well as non-IBD volunteers. The high prevalence of MAP indicated a possible role of MAP in stimulating IBD.

Application of high-resolution melting-curve analysis on pvpA gene for detection and classification of Mycoplasma gallisepticum strains.

Mycoplasma gallisepticum (MG) is an avian species pathogen which causes heavy economic losses in the poultry industry. The purpose of this study was to determine genomic diversity of 14 MG field strains from chicken, Chuker partridge and peacock collected during 2009-2012 in Iran by polymerase chain reaction and partial sequencing of the pvpA gene. A High-Resolution Melting (HRM) technique was also developed and applied to differentiate between field and vaccine strains. Sequencing of the pvpA gene revealed a 51 nucleotide deletion, within DR-1 and DR-2, among MG strains from chicken and partridge whilst 63 nucleotides were deleted in MG strain from peacock. One nucleotide substitution was also observed among chicken MG strains. Phylogenetic analysis of the sequences clustered all of the Iranian MG strains into two clades or phylogeny groups; the strains from chicken and partridge in one group (group 1) and the strain from peacock into another group (group 4). HRM analysis has also produced comparable outcome to those of sequencing; four distinct melting curves which correspond to the three MG strains from chicken, Chukar partridge and peacock and ts-11 vaccine strain. Overall, findings of this study point towards a single source of infection for the chicken and partridge MG strains and likelihood of the strains being native and endemic in Iran. Peacock considered as an exotic species in Iran, hence the genetic distance for the pvpA gene. MG can be transmitted easily among different avian species and this distinct peacock strain may pose a threat to poultry industry. Our findings also show that molecular variation among pvpA gene of MG strains could be revealed using the relatively rapid and affordable HRM technique.

Molecular detection of virulence genes in Pseudomonas aeruginosa isolated from children with Cystic Fibrosis and burn wounds in Iran.

Pseudomonas aeruginosa possesses various virulence factors which contribute to the bacterial invasion and toxicity. Moreover, children suffered from Cystic Fibrosis (CF) and burn wounds are at a high risk of various bacterial infections. The aim of this study was to determine the prevalence of virulent genes in P. aeruginosa isolated from children with CF and burn wounds and comparing their virulence genes to figure out the role of every virulent factor in the infections. P. aeruginosa were isolated from sputum, oropharyngeal swabs, and broncho-alveolar lavage (BAL) specimens from CF and burn wounds between June 2013 and June 2014 in Tehran's hospitals. Bacterial genomic DNAs were extracted and uniplex, duplex and multiplex PCR were performed for detection of toxA, algD and plcN, exoS, lasB, plcH genes, respectively. The prevalence rate of virulence genes in P. aeruginosa isolated from CF was; toxA (63.1%), algD (64.6%), plcH (87.7%), plcN (60%), lasB (95.4%) and exoS (70.8%) and virulence genes in P. aeruginosa from burn patients were: toxA (36.9%), algD (70.1%), plcH (79%), plcN (63.1%), lasB (82%) and exoS (21.1%). The prevalence of three virulent genes in P. aeruginosa was higher in CF comparing to burn wound infections. We found that the number of toxA, lasB and exoS were significantly higher in the bacteria which were isolated from children with CF. This finding shows that these virulence factors play an important role in CF infections by P. aeroginosa.

Detection and phylogenetic characterization of Columbid circoviruses in Chaharmahal va Bakhtiari province, Iran.

The virus genus Circovirus belongs to the family Circoviridae and contains virus species with circular single-stranded DNA genomes. The viruses are known to infect vertebrate species, including pigs, dogs, pigeons and ducks. In this study a nested polymerase chain reaction (PCR) was applied to investigate prevalence of circoviruses in pigeons in the Chaharmahal va Bakhtiari province, Iran. A total of 50 faecal samples were subjected to nucleic acid extraction, PCR amplification and DNA sequencing. Nested PCR primers were designed to amplify a 508 base pair segment of the pigeon circovirus (PiCV) capsid gene. Of the 50 faecal samples examined, 12 (24%) produced the expected DNA amplicons with identical DNA sequences. Phylogenetic analysis has grouped the viruses with those classified as group A circoviruses. The viruses were closely related to PiCVs found in Poland, Northern Ireland, the USA, Nigeria and Hungary. To the authors' knowledge, this is the first report of molecular detection and genomic characterization of PiCV from Iran.

The impact of integrating rabbit haemorrhagic disease virus (K5) release with pindone baiting on wild rabbit populations.

Several conventional and recently available tools are available for an integrated control of European rabbits in Australia. We quantified the impact of the release of rabbit haemorrhagic disease virus K5 (RHDV K5, hereafter K5) and pindone (2-pivalyl-1,3-indandione) baiting at 13 sites within Cudlee Creek fire scar in the Adelaide Hills, South Australia. K5 release was followed by pindone baiting between December 2021 and March 2022; the application of both control methods followed industry best practice. We counted rabbits using spotlights before and after the application of both control methods. Fly samples and livers from dead rabbits were collected to track K5 transmission within and between sites, and to detect the natural circulation of rabbit haemorrhagic disease virus 2 (RHDV2). K5 release had minimal impact on rabbit populations, with treated populations increasing by a mean of 65.5% at 14 days post-release and 27.9% at 77 days post-K5 release across all sites, comparable to the changes at control sites. K5 detection in flies up to 77 days post its release, and its detection in rabbit livers, demonstrates that it can survive and transmit in the environment for prolonged periods and that it can lethally infect some rabbits. This limited impact of K5 is consistent with previous studies and may be explained by pre-existing RHDV/RHDV2 immunity in the target populations or the presence of young rabbits with natural innate RHDV immunity. The detection of K5 in flies from control sites demonstrates that it was vectored beyond its release location. A reduction in rabbit counts post-pindone baiting was observed at most treatment sites, with a mean population reduction of 36.6% across all sites. Landholders need to carefully and strategically plan their integrated rabbit control programmes. Not all combinations of controls, even if theoretically logical, achieve meaningful outcomes for rabbit management.

Neospora caninum Suspects as One of the Most Important Causes of Abortion in Large Dairy Farms in Isfahan, Iran.

BACKGROUND: This study aimed to reveal the serological prevalence of Neospora caninum in large dairy farms in Isfahan Province, central Iran. METHODS: Serum samples were collected from 1500 cattle living in four large dairy farms in Isfahan Province, Iran during 2014-2015 and examined for anti N. caninum IgG antibodies. Overall, 113 serum samples were also collected from the dogs living in these areas; suspecting to be risk factors for this infection. All the serum samples were investigated to find IgG antibodies by using ELISA. Dogs' sera were also analyzed by indirect fluorescent antibody test. RESULTS: Totally, 395 out of 1500 bovine samples (26.33%) were positive for N. caninum: 34%, 21.61%, 23.03% and 29.01% in four investigated clusters (farms). Infection rate was significantly more in cows with the history of abortion. The infection rate in dogs was 17.69%: (20 out of 113). CONCLUSION: The results show a high seroprevalence of the infection and possibly the role of the dogs in horizontal transmission of the infection.

Genomic identification of human vaccinia virus keratoconjunctivitis and its importance as a laboratory-acquired infection.

CONTEXT: Vaccinia virus (VACV) is a member of orthopoxvirus genus of the family Poxviridae. VACVs are enveloped, double-stranded DNA viruses. Several species of this family, for example, molluscum contagiosum, smallpox, deerpox, horsepox, rabbitpox, and VACVs may cause conjunctivitis. AIMS: Given the high incidence of keratoconjunctivitis in Iran (approximately 3.6%-53.9%) and insufficient clinical diagnostic measures, laboratory tests for detection of its causes and determination of accurate keratoconjunctivitis/conjunctivitis prevalence due to different pathogens are essential. SETTINGS AND DESIGN: In this research, conjunctival samples collected from 100 patients with keratoconjunctivitis signs were referred to an eye hospital of Iran. SUBJECTS AND METHODS: After DNA extraction, polymerase chain reaction (PCR) was carried out for detection of VACV. PCR-positive products were further subjected to DNA sequencing. STATISTICAL ANALYSIS USED: The results were analyzed using Chi-square test. RESULTS: In this study, 28% of the samples were positive and a statistically significant relationship obtained between working in medical or research laboratories and VACV prevalence (P < 0.05). CONCLUSIONS: This study showed a high rate of VACV keratoconjunctivitis, and therefore, further studies for its prevention and control are necessary.

Prevalence of Human T-Lymphotropic Virus Types I and II in Patients With Hematological Disorders in Isfahan, Iran.

BACKGROUND: Human T-cell lymphotropic virus types Ι and ΙΙ (HTLV-Ι and HTLV-II) are deltaretroviruses which may cause leukemia, lymphoma and HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP). In addition, HTLV-1 may be related to thalassemia and hemophilia cases after blood transfusion. OBJECTIVES: The aim of this study was evaluation of the prevalence of HTLVs in patients with hematological disorders (leukemia, thalassemia, lymphoma and hemophilia). PATIENTS AND METHODS: This cross-sectional study was conducted during April to October 2012. A total of 101 serum samples were collected from patients and were stored at -20ºC. DNA was extracted from serum by an extraction kit. The extracted DNA was tested by polymerase chain reaction (PCR) for detection of HTLV-Ι and HTLV-II pol and tax gene sequences, respectively. Samples were collected from 67 (66.33%), 20 (19.80%), 4 (3.96%), and 10 (9.90%) patients with thalassemia, leukemia, lymphoma and hemophilia, respectively. RESULTS: One thalassemia sample was HTLV-Ι positive, but none of the samples contained the genome of HTLV-II. The prevalence of HTLV-Ι in this study in patients with hematological disorders was 0.99%. CONCLUSIONS: The prevalence of HTLV-Ι in hematological disorders was similar to that of other parts of Iran. The present study revealed that HTLV-Ι screening should be performed before blood transfusion to reduce the risk of virus transmission in patients with hematological disorders. More study should be performed to detect these viruses in blood donors.

Detection of Antiseptic-Resistance Genes in Pseudomonas and Acinetobacter spp. Isolated From Burn Patients.

BACKGROUND: Quaternary ammonium compounds (QAC), which contain benzalkonium chloride as the most widely used agent, are employed as wound and skin antiseptics, as well as disinfectants in hospitals. The resistance mechanism to disinfectants is usually determine by genes which are related to resistance to quaternary ammonium compounds, namely, qacE, qacΔE1, qacΔE1 that are found in Gram-negative bacteria. OBJECTIVES: The aim of this study was to determine the incidence of antiseptic resistance genes, qacE and qacΔE1, in clinical isolates of Pseudomonas aeruginosa and Acinetobacter bumanii. MATERIALS AND METHODS: In this study, 83 clinical isolates of Pseudomonas aeruginosa, and 5 isolates of Acinetobacter baumannii from burn hospitals in Tehran and Isfahan provinces in 2010-2011, were tested by the PCR method. RESULTS: Out of the 83 clinical isolates of Pseudomonas aeruginosa, 49 isolates (50%) had the qacE gene, and 76 isolates (91.5%) had the qacΔE1 gene. In addition, in 5 isolates of Acinetobacter bumanii, 2 isolates (40%) had the qacE gene, and 4 isolates (80%) had the qacΔE1 gene. CONCLUSIONS: This study shows that the genes which harbored resistance to quaternary ammonium compound antiseptics are widespread among Pseudomonas aeruginosa and Acinetobacter bumanii isolates in burn patients.

Relationship between Crimean-Congo hemorrhagic fever virus strains circulating in Iran and Turkey: possibilities for transborder transmission.

Crimean-Congo hemorrhagic fever (CCHF) is an important zoonotic viral disease that is asymptomatic in infected livestock, but poses a serious threat to humans. The high fatality rate may be due to phylogenetic variations in the virus, transmission routes, and a lack of an efficient surveillance system for the disease. The geographical features of the eastern and southeastern borders of Turkey may facilitate transmission of viruses between countries of the region. Therefore in this study we focused on the genetic relationship between Turkish and Iranian CCHF viruses based on their S-segment sequences. The research was performed on a total of 104 blood samples from small ruminants reared in southwest Iran. The results of phylogenetic analysis showed that Iranian CCHF virus isolates were closely related to human-originating Turkish Group II viruses from a European lineage reported previously.