Microbiology
Hassan Tizfahm Tikmehdash; Alireza Dehnad; Nader Mosavari; Behroz Naghili Hokmabadi; Sanaz Mahmazi
Articles in Press, Accepted Manuscript, Available Online from 13 March 2024
Abstract
Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and ...
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Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and fourth quarters of 2020, throughout 2021, and in the first and second quarters of 2022, the complement fixation test (CFT) was performed on 350 horses. The malleination was used to confirm the positive CFT cases. Blood samples were taken for culture and for preparing serums to perform the ELISA. Deep eye discharge, nostril, cutaneous ulcers, and lymph fluid swabs were cultured, and finally, PCR was carried out. Eleven (3.14%) horses were CFT-positive. Based on the malleination on these 11 horses, six (1.7%) were affected by glanders, five (1.42%) were not affected (false positive), and one horse (0.28%) was CFT-negative despite exhibiting clinical signs. It was positive by malleination, ELISA, and PCR. A total of seven positive cases of glanders were diagnosed. B. mallei could not be isolated, but the B. cepacia complex was isolated in one case. Except for three cases (14.28%), the results of the CFT, mallein, and ELISA tests were consistent (71.42%). The amount of confidence interval (CI) was 95%. It is suggested that ELISA be used as a complement to CFT in screening and, if positive results are observed in one of the tests, the entire herd be examined more accurately using the mallein and western blot confirmatory tests.
Hossam Farouk Ahmed; Yamen Mohammed Hegazy; Sameh Abdelmageed Ibrahem
Volume 12, Issue 4 , December 2021, , Pages 409-414
Abstract
A total number of 62 clinically healthy dairy animals of three farms located in Kafr El Sheikh governorate, Egypt, were subjected to field screening surveys of subclinical mastitis (SCM) using California mastitis test (CMT). The obtained results revealed that 38.80% of quarter milk samples were positive ...
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A total number of 62 clinically healthy dairy animals of three farms located in Kafr El Sheikh governorate, Egypt, were subjected to field screening surveys of subclinical mastitis (SCM) using California mastitis test (CMT). The obtained results revealed that 38.80% of quarter milk samples were positive to CMT. The most frequently major causative agents isolated from the positive CMT samples were Escherichia coli, Staphylococcus aureus, and environmental streptococcus spp. Acute-phase proteins (APPs), as immunological biomarkers for SCM, including milk serum amyloid A (mSAA) and haptoglobin (Hp) were measured using ELISA. A significant positive correlation was found between the severity of the mammary infection of cow's quarter milk samples represented in somatic cell count (SCC) and each of APPs and pH values. The correlation coefficient (R) between SCC and mSAA, Hp and pH were 0.54, 0.38 and 0.73, respectively. On the other hand, there was a significant negative correlation between casein percentage in milk of SCM cases, and each of APPs, pH and the presence of bacterial pathogens in the milk samples. The obtained results threw light on the inter-relationship between SCC, mSAA, pH value and casein percentage in milk of cows and buffalo suffered from SCM. The percentage of casein in milk is considered a significant accurate tool for diagnosis of SCM and this finding offers the farmers a cheap and fast selection for diagnosis of such disease. These results presented a specific structured view on the efficacy of different diagnostic tools of SCM in dairy herds.
Parasitology
Majid Khodaverdi; Gholamreza Razmi
Volume 10, Issue 2 , June 2019, , Pages 119-123
Abstract
The aim of the present study was to determine seroprevalence of Toxoplasma gondii infectionin stray cats and correlation with oocyst shedding and IFN-γ concentration. From April to August 2016, one hundred fifty-nine stray cats were captured from various localities in Mashhad area. The blood and ...
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The aim of the present study was to determine seroprevalence of Toxoplasma gondii infectionin stray cats and correlation with oocyst shedding and IFN-γ concentration. From April to August 2016, one hundred fifty-nine stray cats were captured from various localities in Mashhad area. The blood and fecal samples were collected from each cat. The serum samples were examined to detect antibodies against T. gondii infection by ELISA assay and the fecal samples were microscopically examined for T. gondii oocyst detection. The concentration changes of IFN-γ in serum samples of seropositive and seronegative cats were measured using ELISA kit. The results showed that59.12% (94/159) of cats had antibodies against T. gondii infection. The seroprevalence of T. gondii infection in the adult cats above three years olds was higher than other groups. Regarding gender, month and region factors, the difference of seroprevalence of T. gondii infection was not significant. In this study, the Toxoplasma/Hammondia like oocyst (THLO) were detected in 2.56% (4/156) in fecal samples of one seropositive and three seronegative cats. Results also showed that the mean value for IFN-γ concentration in the seropositive cats was significantly higher than that of the seronegative cats. Based on the results, the high percentages of stray cats were infected with T. gondii in this area. The IFN-γ concentration of seropositive cats was higher than that of the seronegative cats.
Parasitology
Hossein Rezvan; Amir Khaki; Mehdi Namavari; Roya Abedizadeh
Volume 10, Issue 1 , March 2019, , Pages 79-84
Abstract
Neospora caninum is an obligate intracellular parasite causing abortion and reproductive failure in ruminants. Here, the seroprevalence of Neospora DNA and anti-Neospora antibodies and the correlation between the DNA and the antibody using polymerase chain reaction (PCR) and a new developed whole cell-based ...
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Neospora caninum is an obligate intracellular parasite causing abortion and reproductive failure in ruminants. Here, the seroprevalence of Neospora DNA and anti-Neospora antibodies and the correlation between the DNA and the antibody using polymerase chain reaction (PCR) and a new developed whole cell-based enzyme-linked immunosorbent assay (ELISA) in water buffalo (Bubalus bubalis) were investigated. To determine the level of anti-Neospora antibody, 83 serum samples were collected from buffaloes in the northwest of Iran. Plates were coated with 2 × 106 whole Neospora tachyzoites and the anti-Neospora antibody level was determined by calculating the ratio of sample/positive control (S/P) optical densities (ODs) in the ELISA. All samples with the ration of 0.50 or above were accounted as positive. To confirm the presence of Neospora DNA, the serum samples were directly subjected to PCR and nested PCR for detection of Neospora NC5 gene without the DNA isolation process. A total number of 83 buffalo serum samples were examined for the presence of anti-N. caninum immunoglobulin G and Neospora DNA. All samples with the S/P ratio of 0.50 or above (16 samples, 19.27%) were also positive for Neospora DNA. All samples with OD less than 0.50 (34 samples, 40.96%) were negative for Neospora DNA. However, 33 samples with the S/P ratio of bellow 0.50 (39.75%) showed a significant level of antibody. A 100% correlation was observed between high levels of the anti-Neospora antibody and Neospora DNA in the serum of water buffalo, and the whole N. caninum tachyzoites have the potency to be used as antigens for detection of the parasite in ELISA.
Immunology
Masoud Reza Seyfi Abad Shapouri; Mohammad Rashno; Pezhman Mahmoodi; Mahshid Ariya
Volume 9, Issue 1 , March 2018, , Pages 67-72
Abstract
Monoclonal antibodies (MAbs) are invaluable molecules which have several advantages over polyclonal immunoglobulins (Igs) including consistency and higher specificity and hence can be used in biological researches, diagnosis and treatment of diseases. The present study was conducted to produce monoclonal ...
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Monoclonal antibodies (MAbs) are invaluable molecules which have several advantages over polyclonal immunoglobulins (Igs) including consistency and higher specificity and hence can be used in biological researches, diagnosis and treatment of diseases. The present study was conducted to produce monoclonal antibody against chicken IgG.TheIgG molecules were purified from chicken serum and used as antigens to immunize several mice. Thereafter, a well-immunized mouse was chosen and used for fusion process. After production of hybridoma cells, several rounds of cloning were carried out and produced MAbs were examined by various immunological assays including enzyme-linked immunosorbent assay (ELISA) and western and dot blotting. Assessment of grown hybridomas indicated that only one clone (5B8) has produced desired MAb against chicken IgG. Meanwhile, using an indirect ELISA, it was shown that this MAb successfully recognizes chicken IgG molecules attached to influenza virus nucleoprotein. Evaluation of cross reactivity of MAb 5B8 with several avian serum samples revealed that this molecule specifically identifies chicken antibody molecules. However, it also recognized turkey antibodies with less affinity. In addition to research applications like isolation and purification of chicken and turkey IgG molecules, such a MAb can be applied to design and development of various immunoassays (e.g. ELISA) in these avian species.
Immunology
Fereshteh Yazdani; Mehran Bakhshesh; Majid Esmaelizad; Zohre Azita Sadigh
Volume 8, Issue 3 , September 2017, , Pages 209-213
Abstract
Bovine ephemeral fever is an acute and arthropod-borne viral disease of cattle and water buffalo which occurs seasonally in most of the world tropical and subtropical regions. The epizootic feature of the disease has been reported in Iran with serious economic consequences. The surface glycoprotein G ...
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Bovine ephemeral fever is an acute and arthropod-borne viral disease of cattle and water buffalo which occurs seasonally in most of the world tropical and subtropical regions. The epizootic feature of the disease has been reported in Iran with serious economic consequences. The surface glycoprotein G of bovine ephemeral fever virus (BEFV) is composed of 4 antigenic sites (G1-G4) and plays the main role for eliciting neutralizing antibodies and protective immunity. The G1 – epitope is a linear antigenic site and conserved among BEFV strains. In order to develop an ELISA test based on G1-epitope as coating antigen, this study was carried out to express the recombinant G1-epitope of BEFV in prokaryotic system. Using PCR and specific primers, a length of 88 amino acid of the G glycoprotein of BEFV including G1- epitope was amplified and cloned into the expression vector pGEX-4T-1, with the GST moiety. The recombinant plasmid (pGEX-4T-1-G1) was then transformed into Escherichia coli BL21 and expression of fusion protein was induced by 0.10 mM IPTG. The maximum expression of the fusion protein was obtained at 16 hr post induction as verified by SDS-PAGE electrophoresis, and it was also confirmed that this protein bearing G1- epitope is sufficiently biologically active to bind to anti-BEFV serum in western blot experiment.
Epidemiology
Behnaz Norouzi; Alireza Taghavi Razavizadeh; Mohammad Azizzadeh; Ashraf Mayameei; Vahid Najar Nezhad Mashhadi
Volume 6, Issue 3 , September 2015, , Pages 245-249
Abstract
Maedi-Visna (MV) virus and caprine arthritis encephalitis (CAE) virus known as small ruminant lentiviruses (SRLVs) cause chronic diseases in susceptible animals. The main reservoirs of these viral agents are sheep and goat. In sheep, MV virus causes a disease as the same name of the virus. This is the ...
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Maedi-Visna (MV) virus and caprine arthritis encephalitis (CAE) virus known as small ruminant lentiviruses (SRLVs) cause chronic diseases in susceptible animals. The main reservoirs of these viral agents are sheep and goat. In sheep, MV virus causes a disease as the same name of the virus. This is the first seroprevalence survey of SRLVs in sheep population of Khorasan-e-Razavi province in Iran. Two hundred and twenty sheep from 30 flocks in 12 regions of the province were selected by random cluster sampling method. Serum samples were analyzed for the presence of antibodies against MV/CAE viruses. The seroprevalence in sheep was 34.5% (95.0% CI: 28.3 to 40.7%). Totally, the seroprevalence was in the range of 6.7 to 72.2 %. In 26 flocks of sheep (89.6%; 95.0%CI: 74.4 to 98.8%), at least one seropositive case was detected. The relationship between seropositivity and age, sex, flock size and breeds of sheep were statistically analyzed. In logistic regression model, only age was correlated with SRLV seroprevalence (p < 0.05). This study showed relatively high seroprevalence against SRLVs in sheep population in this area of the country. Due to difficulty in clinical diagnosis, chronic course of the disease, the absence of effective vaccine and treatment and huge economic loss, more epidemiological studies with regards to prevention and control of the disease are necessary.
Mohammad Sabaghan; Mahdi Pourmahdi Borujeni; Masoud Reza Seifi Abad Shapouri; Aria Rasooli; Milad Norouzi; Sayeh Samimi; Siavash Mansouri
Volume 5, Issue 4 , December 2014, , Pages 325-328
Abstract
Bluetongue (BT) is a viral disease of ruminants transmitted by Culicoides biting midges and has the ability to spread rapidly over large distances. The disease occurs almost worldwide between latitudes approximately 35˚ S and 50˚ N. Among the numerous diseases of ruminants, BT has gained considerable ...
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Bluetongue (BT) is a viral disease of ruminants transmitted by Culicoides biting midges and has the ability to spread rapidly over large distances. The disease occurs almost worldwide between latitudes approximately 35˚ S and 50˚ N. Among the numerous diseases of ruminants, BT has gained considerable importance in recent years as one of the best examples of the effects of climate change on disease spread. Sheep are major livestock species in Iran, but studies of BT have not gained the priority compared to other diseases. Thus, the objective of this study was to describe the distribution and seroprevalence of bluetongue virus (BTV) infections in sheep in Kohgiluyeh and Boyer-Ahmad province of Iran, and to identify factors associated with the exposure of these sheep to BTV infection. Sera from 262 apparently healthy sheep were collected during the year 2011. The collected sera of the animals were screened with competitive enzyme like immunosorbent assay (c-ELISA). Two hundred and three (77.48%) out of 262 sera tested were positive to BTV antibodies. Statistically significant differences were found in the seroprevalence BT, between sex and age of sheep (p < 0.001). No statistically significant differences were observed in BTV seroprevalence among different seasons, nor among recently aborted and normally delivered.
Aliasghar Bahari; Jamal Gharekhani; Masoumeh Zandieh; Ali Sadeghi-Nasab; Hesameddin Akbarein; Ahmad Karimi-Makhsous; Morteza Yavari
Volume 4, Issue 2 , June 2013, , Pages 111-114
Abstract
A cross-sectional study with a random cluster sampling design was carried out to estimate the seroprevalence of bovine herpesvirus type 1 (BHV-1) in non-vaccinated dairy herds in Hamedan province, west of Iran. Simple random sampling was used for selection of cattle in each herd. Informative data about ...
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A cross-sectional study with a random cluster sampling design was carried out to estimate the seroprevalence of bovine herpesvirus type 1 (BHV-1) in non-vaccinated dairy herds in Hamedan province, west of Iran. Simple random sampling was used for selection of cattle in each herd. Informative data about each herd and selected animals were recorded by the farm manager in a provided questionnaire. Blood samples were collected from 492 animals in 41 industrial herds. A commercial indirect ELISA test was used to determine the seropositivity against BHV-1. The individual and herd seroprevalence for BHV-1 were 58.74% and 82.93%, respectively. The intra-herd prevalences were ranged from 16.70% to 100%. Geographical characteristics of Hamedan province may explain the high sero-prevalence rates found in this study compared to those of others obtained from different parts of the country. The proportion of seropositive cows were increased with age (p <0.05). Animals from large and moderate sized herds had higher odds of seropositivity than those of small size herds. These findings could be related to the presence of a considerable number of BHV-1 carriers in this region. The high herd and animal prevalence found in the present study suggested necessity of implementing an intensive control program for reducing BHV-1 infection rates.