Molecular identification and phylogenetic analysis of Lactobacillus and Bifidobacterium spp. isolated from gut of honeybees (Apis mellifera) from West Azerbaijan, Iran

Available online: 15 December 2016Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) and phylogenetic analysis were used for molecular identification of lactic acid bacteria (LABs) isolated from Apis mellifera. Eighteen honeybee workers were collected from three different apiaries in West Azerbaijan. LABs from the gut of honeybees were isolated and cultured using routine biochemical procedures. Genomic DNA was extracted from LABs and a fragment of 1540 bp in size of 16S rRNA gene was amplified. PCR products were digested using HinfI endonuclease and digested products with different RFLP patterns were subjected to nucleotide sequencing and phylogenetic analysis. The results revealed that Lactobacillus and Bifidobacteria spp. are were the most abundant LABs in honeybee gut. Phylogenetic analysis showed that both Lactobacillus and Bifidobacterium were closely clustered with high similarity percentage with the same bacteria isolated from honeybees’ gut elsewhere. It was concluded that LABs isolated from honeybees had low sequence divergence in comparison with LABs isolated from other sources such as dairy products.Mohammad Farouq Sharifpour, Karim Mardani, Abdulghaffar Ownag

Multimeric recombinant M2e protein-based ELISA: A significant improvement in differentiating avian influenza infected chickens from vaccinated ones

Killed avian influenza virus (AIV) vaccines have been used to control H5N1 infections in countries where the virus is endemic. Distinguishing vaccinated from naturally infected birds (DIVA) in such situations however, has become a major challenge. Recently, we introduced the recombinant ectodomain of the M2 protein (M2e) of H5N1 subtype as a novel tool for an ELISA based DIVA test. Despite being antigenic in natural infection the monomer form of the M2e used in ELISA had limited antigenicity and consequently poor diagnostic capability. To address this shortcoming, we evaluated the use of four tandem copies of M2e (tM2e) for increased efficiency of M2e antibody detection. The tM2e gene of H5N1 strain from Indonesia (A/Indonesia/CDC540/2006) was cloned into a pMAL- p4x expression vector and expressed in E.coli as a recombinant tM2e-MBP or M2e-MBP proteins. Both of these, M2e and tM2e antigens reacted with sera obtained from chickens following live H5N1 infection but not with sera from vaccinated birds. A significantly stronger M2e antibody reaction was observed with the tM2e compared to M2e antigen. Western blotting also supported the superiority of tM2e over M2e in detection of specific M2e antibodies against live H5N1 infection. Results from this study demonstrate that M2e tetramer is a better antigen than single M2e and could be more suitable for an ELISA based DIVA test.Farshid Hadifar, Jagoda Ignjatovic, Simson Tarigan, Risa Indriani, Esmaeil Ebrahimie, Noor Haliza Hasan, Andrea McWhorter, Sophie Putland, Abdulghaffar Ownagh, Farhid Hemmatzade

Slaughterhouse Wastewater Treatment by Combined Chemical Coagulation and Electrocoagulation Process

Slaughterhouse wastewater contains various and high amounts of organic matter (e.g., proteins, blood, fat and lard). In order to produce an effluent suitable for stream discharge, chemical coagulation and electrocoagulation techniques have been particularly explored at the laboratory pilot scale for organic compounds removal from slaughterhouse effluent. The purpose of this work was to investigate the feasibility of treating cattle-slaughterhouse wastewater by combined chemical coagulation and electrocoagulation process to achieve the required standards. The influence of the operating variables such as coagulant dose, electrical potential and reaction time on the removal efficiencies of major pollutants was determined. The rate of removal of pollutants linearly increased with increasing doses of PACl and applied voltage. COD and BOD5 removal of more than 99% was obtained by adding 100 mg/L PACl and applied voltage 40 V. The experiments demonstrated the effectiveness of chemical and electrochemical techniques for the treatment of slaughterhouse wastewaters. Consequently, combined processes are inferred to be superior to electrocoagulation alone for the removal of both organic and inorganic compounds from cattle-slaughterhouse wastewater

Treatment of Vaginal Candidiasis by Ethanolic Extract of Propolis in Rabbit

Background & aim: Propolis is a resinous substance collected by honeybees from buds and leaves of trees and plants, possessing various biological properties. This study aimed to assess the anti-fungal activities of Ethanolic extract of Propolis (EEP) in an experimental infection of vaginal candidiasis. Methods: In this study, twenty-four female rabbits were used for experimental infections. Animals were randomly divided into six equal groups: positive control group, negative control group, and four treatment groups. The four test groups were given doses of 125, 250, 500 and 1000 µg/ml of Propolis Ethanolic extract respectively while the positive control group received the standard Nystatin treatment and negative control group were given saline, locally, two times a day for 20 days. The data were analyzed, using descriptive statistics. Results: Among all groups, the results showed that dose of 1000 µg/ml of EEP were the most effective dose in treatment of experimental vaginal candidiasis. Clinical signs of vaginal candidiasis were healed in Nystatin group after 9 days. No effect was recorded after 20 days treatment of infection for others doses of EEP that were used in this study. Conclusion: It can be concluded that Ethanolic extract of Propolis in 1000 µg/ml concentration could treat experimental vaginal candidiasis in rabbits in a shorter time in comparison with standard nystatin treatment.

The effects of Ethanol Extract of Propolis (EEP) on the experimentally induced Candida keratitis in rabbits

"n 800x600 Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman","serif";} Background: Propolis (bee glue) is a resinous substance obtained from bee hives living on various plant sources. The purpose of this study was to evaluate the effects of ethanol extract of propolis (EEP) on the experimentally induced Candidial keratitis in rabbits."n"nMethods: The alcoholic extract of propolis was prepared by 80% ethyl alcohol. After suppressing the immune system of 24 male rabbits, experimental Candida albicans keratitis was induced in the animals under local anesthesia and sterile conditions. The animals were later divided into four groups including the control or glycerin group and a nystatin and two 500 and 1000µg/ml EEP groups. Treatment continued for 21 days and after sacrificing the animals by humane methods, histopathological samples of the rabbits' eyes were prepared."n"nResults: Keratitis was developed in the eyes of all rabbits a week after the yeast inoculation. In the control group in which animals received glycerin, keratitis persisted until day 21. Clinical signs of keratitis disappeared in the Nystatin and 1000µg/ml EEP groups after 14 and 21 days, respectively. The clinical signs of keratitis partially ameliorated in the animals receiving 500µg/ml EEP. Histopathological examination revealed no differences between groups receiving nystatin or 1000µg/ml EEP."n"nConclusion: It is concluded that, ethanol extract of propolis could completely treat Candida albicans keratitis in 1000µg/ml concentrations. This extract can be used as a safe antifungal agent against Candida albicans and it is a good substitute for synthetic antifungal agents like nystatin

Phylogenetic and molecular analysis based on genes 16S-rRNA, OMPA and POMP to identify Chlamydia abortus infection occurrence at the milk samples of goats and sheep in west Azerbaijan of Iran.

Background and Objectives: Enzootic abortion in sheep and goats, also called ovine enzootic abortion (OEA) or enzootic abortion of ewes (EAE), is caused by Chlamydia abortus. The disease has a major economic impact as it represents the most important cause of lamb loss in sheep in parts of Europe, North America and Africa. This serious and potentially life-threatening zoonosis can also affect pregnant women after contact with lambing ewes, leading to severe febrile illness in pregnancy and loss of the foetus. Materials and Methods: The present study was conducted to the Phylogenetic and Molecular Analysis based on Genes 16S-rRNA, OmpA and POMP of C. abortus in milk samples collected from sheep and goats in West Azerbaijan province, Iran. During 2018, a total number of 360 milk samples were collected from sheep (n = 180) and goats (n = 180) of different regions of the province. All milk samples were subjected to DNA extraction and examined by PCR. Results: Among 360 milk samples collected from sheep and goats, 31 (8.611%; 95% CI=6.13-11.96) were positive for Chlamydia spp. The helicase, 16S-rRNA and ompA genes were examined and resulted in 8, 31, 31 of positive samples respectively. The accession numbers have been deposited in GenBank (NCBI) (MT367602 and MT367603). Conclusion: Phylogenetic analysis based on the gene of helicase showed that most of the isolates shared similarity > 99.97%

The effect of cytosolic extract of Alternaria aternata fungus on Monocyte-derived dendritic cell maturation and T-lymphocyte polarization in the presence of myelin basic protein

Background: Multiple Sclerosis (MS) is an autoimmune disease with impairment in function of central nervous system. Macrophages and dendritic cells play important roles in alleviating or progression of the disease. These cells can cause inflammation and damage to the myelin of nerve cells by realizing of harmful substances when these cells get matured. We studied the effect of Alternaria alternata extract on maturation of monocyte- derived dendritic cell (modc) and T-cell responses in the presence of Myelin Basic Protein (MBP) as a laboratory model of multiple sclerosis (MS). The purpose of this study is suitable dendritic cells production for usage in MS immunotherapy.Methods: For this study plastic adherent monocytes were cultured with granulocyte/ macrophage- colony stimulating factor (GM-CSF) and interleukin -4 for converting these cells to modc and pulsed with MBP and matured in the presence of monocyte-conditioned medium (MCM) in control group and MCM + Alternaria alternata extract in treatment groups. Anti-CD14, anti-CD83, anti-human leukocyte antigen-DR (anti HLA-DR) monoclonal antibody were carried out for phenotyping. Autologos T cell responses and cytokine production were evaluated.Results: The results showed that the expression of CD14 decreased and CD83, HLA-DR increased in treatment groups in comparison with control groups. The production amount of IL-10 overcame IL-12 and in T cell the production of cytokines, IL-17 and Interferon-&gamma; (IFN-&gamma;) decreased and IL-4 was increased (P<0.05). These effects escalated with increasing of dosage from 50 to 100 (mg/ml) (P<0.001).Conclusion: Alternaria alternata extract can cause maturation of MBP-pulsed modc and skewing of T- lymphocyte toward Th2 and thereby can evolve into a new strategy in immunotherapy of MS

Molecular detection of Coxiella burnetii in horse sera in Iran.

Coxiella burnetii is a zoonotic bacterium that can infect a wide range of animals including horses. However, its circulation dynamics in and through horses are still unclear. The aim of this study was to evaluate prevalence of C. burnetii and its genomic characteristics in horse sera samples in the North of Iran (Golestan Province). The samples were collected in 2018 and the age, sex, and breed of each animal were recorded. Nested-PCR was used to detect C. burnetii based on the presence of the transposable gene IS1111. The results showed that 7.50 % (P 6 years old (p-value <0.05, 95 %, CI: 7 %-19.8 %). In previous studies, it was concluded that the horses' population in Golestan Province should be considered as an important factor in the epidemiology of Q fever and consequently in public health. Further studies should be implemented to evaluate if horses may be relevant indicators of zoonotic risk in urban and suburban endemic areas