Abortion is considered an important disease problem of small ruminants in Borana pastoral area. A cross-sectional study was conducted to estimate the prevalence and risk factors of chlamydiosis, coxiellosis (Q-fever) and brucellosis in small ruminants in selected districts of Borana zone. A total of 506 sheep and goats were tested using serological tests. Fifty (9.88%; 95% CI: 7.42, 12.82), 144 (28.46%; 95% CI: 24.56, 32.61) and none (0.00%; 95% CI: 0.00, 0.59) of them were positive for chlamydiosis, coxiellosis and brucellosis, respectively. History of abortion was recorded in 136 (32.00%; CI: 27.59, 36.67) of sheep and goats in the study area. The logistic regression analysis, however, showed that statistically significant difference ccurred among districts and between the species of small ruminants. The prevalence odd of antibodies against C. abortus was significantly lower in Miyo, Dire and Teltelle districts compared to Dillo. The odd of infection with this bacterium was lower in sheep than goats. Similarly the odd of infection with C. burnettii was significantly higher in Dillo district than the rest of the districts studied, higher in goats than sheep and higher in adult animals than young ones. High prevalence of abortion is observed in sheep and goats in the study area. High seropositivity of C. burnetii and C. abortus in both sheep and goats tested implies risks of human infection by both diseases. Thus, attention needs to be paid to further study of both diseases in animals and humans in the area.

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Seroprevalence and associated risk factors for chlamydiosis, coxiellosis and brucellosis in sheep and goats in Borana pastoral area, southern Ethiopia.

Parasites that infect multiple species cause major health burdens globally, but for many, the full suite of susceptible hosts is unknown. Proactive disease surveillance involves gathering host-parasite association data, predicting missing links, and targeting efforts towards the most likely undocumented interactions. Using the largest global network of mammal host-parasite interactions amalgamated to date (>29,000 interactions), we predict undocumented links and conduct targeted literature searches. We find evidence for many of the top "missing" links, including parasites of humans, domesticated animals, and endangered wildlife, and identify regions such as tropical and central America as likely hotspots of undocumented associations. This approach of iterated prediction and targeted surveillance can efficiently guide the collection of host-parasite interaction data critical for developing broad-scale theories in disease ecology and evolution, help to identify previously undocumented hosts, and inform predictions of future host-parasite interactions.

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Predicting missing links in global host-parasite networks

Despite previous detection of Chlamydia pecorum in sporadic ovine abortions, published descriptions of naturally occurring infections with fetoplacental lesions are lacking. This report provides the first descriptions of severe necrosuppurative chorionitis with vasculitis, and fetal pyelonephritis and enteritis in late-term abortions of maiden ewes. Chlamydial infection was detected using a Chlamydia genus-specific qPCR (quantitative polymerase chain reaction) on tissue extracts from 3 fetuses. C. pecorum was identified using a targeted qPCR assay, which also determined infectious load within fetal tissues. The presence of viable C. pecorum in fetal samples was confirmed by cell culture. Multilocus sequence typing (MLST) data indicated that the C. pecorum strains from each fetus were identical and of sequence type (ST) 23. Chlamydia sp. immunohistochemistry showed strong positive immunolabeling of fetoplacental lesions. Other infectious abortigenic agents were excluded with specific testing. This report confirms C. pecorum as a likely cause of ovine abortion and provides the first descriptions of associated fetoplacental lesions in naturally infected sheep.

Chlamydia pecorum-Associated Sporadic Ovine Abortion.

参照GenBank中登录的牛肠道病毒（ BEV ）全基因组序列，针对其3 D基因设计合成了1对特异性引物，提取病毒RNA，逆转录为cDNA，进行PCR扩增，经条件优化，建立了BEV的RT-PCR检测方法，并对该方法的特异性、敏感性进行验证。结果显示，该方法从分离的牛肠道病毒中扩增出了732 bp的特异性目的片段；且对牛传染性鼻气管炎病毒（ IBRV）、牛病毒性腹泻病毒（ BVDV）、牛轮状病毒（ BRV）、牛冠状病毒（ BCoV）等相关病毒均无交叉反应；其检出敏感度达10-1 TCID50。应用该方法对山东地区84份临床疑似发病牛样品进行检测，21份为阳性，阳性检出率为25％。

牛肠道病毒 rt -pcr 检测方法的建立及初步应用

Escherichia coli is one of the most important bacterial pathogens of human and poultry. Avian pathogenic Escherichia coli (APEC) and uropathogenic Escherichia coli (UPEC) isolates cause diseases outside intestine environment. These strains share some common traits. It has been suggested that transfer of these commonalities between different hosts is of importance in the epidemiology of disease in poultry and human. In the present study, presence of 4 virulence associated genes (sitA, iutA, traT, and tsh) were investigated among 26 APEC and 25 UPEC isolates by polymerase chain reaction (PCR) assay. Statistical analysis was also carried out using minitab 15. The frequencies of the presence of these selected genes in APEC and UPEC isolates were 96.2% and 64% for traT, 88.5% and 76% for sitA, 84.6% and 68% for iutA, and 61.5% and 16% for tsh, respectively. traT and sitA were the most prevalent genes among APEC and UPEC isolates, respectively. Diverse combinations of virulence associated genes were also detected among these strains. Combinations of virulence genes of iutA-sitA-tsh-traT and iutA-sitA-traT were the most frequent in APEC and UPEC isolates, respectively. It supports possibility of virulence genes transfer between poultry and human. Further studies are needed to ascertain the importance of such possibilities in the epidemiology of the infection in human and poultry.

تشخیص ملکولی ویروس هپاتیت دلتا در اهداکنندگان خون با روش rt-pcr

Background and Objectives: Enzootic abortion of ewes (EAE) is caused by infection of sheep and goats by Chlamydia abortus bacterium. Chlamydial abortion in bovine could occur by Chlamydia abortus, Chlamydia psittaci and Chlamydia pecorum. C. psittaci is the causative agent of psittacosis or ornithosis disease in humans and birds. It also causes acute pneumonia in cattle and sheep. The present study aimed at surveying the role of chlamydial agents in ruminants abortion. Materials and Methods: A total of 117 aborted material samples (Cotyledon, liver, spleen, and abomasal contents of fetus) from 9 cattle and 100 sheep in Shahr-e-Kord and 8 sheep from Bagh-e-Malek were collected from different herds with abortion history during the lambing periods from 2014 to 2016. After DNA extraction, the samples were tested by species-specific PCR to detect C. abortus, C. pecorum and C. psittaci. Results: Out of 117 clinical sample (108 sheep and 9 cattle), chlamydial infection was detected in 66 (56.41%) samples by Chlamydiales order-specific primers. A total of 24 (36.36%) and 24 (36.36%) samples indicated positive forms of C. abortus and C. psittasi infections, respectively. Only 1 (1.5%) C. pecorum was identified from cattle using nested PCR during this study. Among 66 Chlamydiales - positive samples, 20 (30.30%) samples with coinfection of C. abortus and C. psittaci were detected, however, infection of 3 species was not detected in the samples. Conclusion: Because of the high percentage of chlamydial infection in these regions and probability of coinfection, conducting epidemiological studies on the role of different animals is highly recommended.

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The role of zoonotic chlamydial agents in ruminants abortion.

Background: One disadvantage of expressing heterologous proteins in Escherichia coli is that the proteins are frequently expressed as insoluble inclusion bodies. To avoid this problem, heterologous proteins are typically expressed as a fusion protein. Maltose binding protein (MBP) is one of the widely used partners for production of recombinant fusion proteins in E. coli. MBP is among the most effective solubility enhancers. In addition, MBP can be used as an affinity tag for purification of recombinant proteins on a column of amylose resin. Objectives: The purpose of this study was to investigate the use of rice flour, a natural source of amylose, for purification of MBP fusion proteins. Materials and Methods: MBP and a fusion protein of MBP and avian influenza virus nucleoprotein (MBP-NP) were expressed in E. coli and subjected to purification by rice flour and a commercial amylose resin. The purified proteins were analyzed by SDS-PAGE. Results: The results indicated that MBP and MBP-NP, both were successfully purified by rice flour. Conclusions: Rice flour can be used for purification of MBP fusion proteins. Although the efficiency of purification by rice flour is less than amylose resin, however, the yield is sufficient to obtain a quantity of protein required for research purposes.

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Using rice flour for purification of maltose binding fusion proteins expressed in escherichia coli

The outer membrane protein of Aeromonas hydrophila is a potential candidate for vaccine development. In this study, after cloning and expression of ompTS, 270 common carp, weighing 44 ± 5.7 g divided into five groups, were injected intraperitoneally twice with 3‐week intervals. Groups included the following: PBS, PBS plus Freund's adjuvant, recombinant protein, recombinant protein plus Freund's adjuvant and 20 fish as negative control. Two weeks after the second injection, 30 fish of each group were challenged with a dose of 2 × LD50 of Aeromonas hydrophila and RPS was measured. The antibody level was measured using ELISA test. The protection of recombinant protein in the immunized fish with and without adjuvant, respectively, was about 82.61% and 78.26% (the protection of recombinant protein electroeluted from an SDS–PAGE with and without adjuvant, respectively, was about 78.62% and 69.57%). The average of antibody level in recombinant protein with and without adjuvant was significantly higher than the PBS group (p < .05). The ability of recombinant ompTS to increase the antibody level and to protect the fish from challenge by A. hydrophila demonstrated that recombinant ompTS protein injection can be used to immunize common carp against A. hydrophila infection.

Investigation on protective effect of recombinant protein (OmpTS) of Aeromonas hydrophila in Common carp (Cyprinus carpio)

Background: Avian influenza viruses (AIVs) including the subtype H9N2 cause considerable financial losses to poultry industries. Rapid and accurate diagnosis of avian influenza (AI) infection is important in control and eradication programs. OBJECTIVES: The aim of this study was to produce monoclonal antibodies (MAbs) specific for the nucleocapsid protein )NP (of AIV H9N2 subtype to improve diagnostic assays. METHODS: Recombinant NP protein was expressed in Escherichia coli and purified using amylose resin chromatography column and used as an antigen for mice immunization. Spleen cells of the immunized mice were fused with SP2/0 myeloma cells. Next, culture supernatants of primary hybridoma clones were screened by indirect ELISA. After three rounds of sub cloning, the reactivity of the MAbs with recombinant and natural antigens was assessed by Western blotting. RESULTS: Six MAbs showed specific binding to recombinant and natural NP from AIV H9N2 in Western blot analysis, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay. Cross-reactivity with genetically non-related including Newcastle viruse (Paramyxoviridae family) was not detected. CONCLUSIONS: Based on the results, the MAbs generated in this study could be used for the development of rapid diagnostic assays for recognition of AIV.

https://ijvm.ut.ac.ir/article_66061_9b4e073e03236567e85cc5ff79d3d03a.pdf

Production of monoclonal antibodies against recombinant nucleoprotein of avian influenza virus, serotype H9N2.

The Purpose of this study was to evaluate the prevalence of Canine Coronavirus (CCV) in diarrheic dogs of Ahvaz district. Fecal samples were collected from 58 diarrheic dogs between 2005 and 2006. Dogs were grouped by age (less than and above 6 months), sex, breed, geographical location (urban and rural) and quality of diarrhea (hemorrhagic and non-hemorrhagic) to determine whether these factors were associated with coronavirus infection, using Fischer's exact test. Prevalence to CCV antigens in these dogs was 3.45% (2 of 58) by means of immunochromatography assay (IC) indicating that this virus is present in the environment. Though the infection had more prevalence in dogs less than 6 months, rural dogs and German shepherds, but there were no significant differences between different sexes, age groups, breeds, location and quality of diarrhea (p>0.05). CBC in 2 dogs that were affected with coronavirus infection was normal. This study that is the first report on prevalence of CCV in Iran showed that CCV infection in the Ahvaz dogs may be attributed as a cause of viral diarrhea in dogs. A vaccine is recommended, especially for open population and kennel dogs that are at a high risk for contacting the virus. Key words: Canine Coronavirus, Immunochromatography assay, dog, diarrhea.

https://journals.ut.ac.ir/article_27370_a3fc95ea5b313c072df87d8e6ea2b2e8.pdf

Masoud Ghorbanpoor Najafabadi

Papers

The role of zoonotic chlamydial agents in ruminants abortion.

Using rice flour for purification of maltose binding fusion proteins expressed in escherichia coli

Investigation on protective effect of recombinant protein (OmpTS) of Aeromonas hydrophila in Common carp (Cyprinus carpio)

Production of monoclonal antibodies against recombinant nucleoprotein of avian influenza virus, serotype H9N2.

Antigenic detection of Canine Coronavirus in diarrheic dogs in Ahvaz