Nancy Belem Beltrán Maldonado, Rigoberto Hernández Castro, Erika Margarita Carrillo Casas, Alejandro BenÃtez Guzmán and Beatriz Arellano Reynoso
Canine brucellosis caused by Brucella canis is a worldwide distributed zoonosis. Infection often results in abortion, orchitis, epididymitis, and discospondylitis. The 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) is currently the gold standard diagnostic tool for B. canis. Although it has been a widely used test, it detects IgG and IgM antibodies and has low sensitivity and specificity. The antigen used in this diagnostic test commonly cross-reacts with other pathogens like Escherichia coli O157: H7, Francisella tularensis, Vibrio cholerae, Salmonella N group y Pseudomonas maltophilia, and its production require a level III biosafety laboratory. As a limiting factor, this test is not commercially available in our country. For this reason, it is necessary to seek additional antigen candidates for the diagnosis of canine brucellosis with a methodology of easy access for use in the veterinary clinic. Our group has demonstrated high levels of the GroEL protein in the serum of animals experimentally infected with B. canis, suggesting its role as a candidate protein for detection in diagnostic tests. For recombinant protein preparation, specific primers were designed to amplify the GroEL gene and later cloning into the pQE60 plasmid. Further protein expression requires the E. coli M15 strain that harbors the plasmid pREP4 (lacIq repressor protein). Protein semiquantification by Western Blot will compare recombinant GroEL with native protein. Purification by FPLC (fast protein liquid chromatography) and assessment as the capture antigen by indirect ELISA will be performed with serum from experimentally infected dogs.
Recent Publication
Thamer Mutlag Jasim, Qasim Mohammad Banja, Mohammad Al K-Araji and Qays Al-Khafaji
Recurrent pregnancy loss (RPL) is the most frustrating and challenging field in reproductive medicine because the aetiology is often unknown and there are few evidence based diagnoses and treatment. The cytomegalovirus (CMV) has a ubiquitous DNA herpes virus, as with other herpes viruses, it becomes latent after primary infection but can reactivate with renewed viral shedding. The aim of the present study is to estimate the role of transforming growth factor beta 1 (TGFB1) to CMV immunoglobulin. The study was done in Kamal Al- Smarrai hospital in Baghdad, Iraq, from the period of October 2016 to February 2017. This study was performed on 88 pregnant women attended, 24 with unsuccessful abortion (two or more abortion) and 27 had single abortion and compared with 37 women with normal pregnancy were control, no recurrent abortion). Serum levels of TGF, IgG, IgM and IgG avidity for anti - CMV virus were measured in the serum. They used ELISA reader and electrochemiluminescence for CMV IgG avidity. There were no significant differences between the studied groups in their age, family history of abortion. Serum anti- CMV IgG was significantly higher in RPL and single abortion group compared to IgG TGFB1 in the studied groups. There was no significant difference in the median of IgG and IgM among different groups. There was no significant difference among different groups in their IgG avidity. There is inverse weak correlation between IgM and anti CMV IgG with TGF B1 in control group. There was no correlation between IgG IgM and IgG avidity with TGF B in recurrent abortion group. The current study showed a high proportion of pregnant women with past CMV infection. The RPL, anti-CMV IgM and TGFB were correlated directly with RPL patient compared with healthy control.
Recent Publications
P Branco, S Arantes, A Candeias, A T Caldeira and M González-Pérez
Bio deterioration of cultural heritage (CH) materials (e.g., paper, marble, lime, mortar, parchment, metal, glass) was neglected for a long time since it was previously believed that it was only due to chemical and physical processes. However, over the last decades, it has been proven that the action of microorganisms is a critical factor in the deterioration process. Biodeteriogenic microorganisms cause serious aesthetical and structural damages in CH materials of inestimable value. Some of them can synthesize carotenoid compounds causing pink staining such as Rhodotorula sp., this yeast has been associated with the deteriorative effects observed in the Évora Cathedral, Portugal. To distinguish Rhodotorula sp., from other microorganisms that produce the same type of alterations on CH materials, proper identification methods must be applied. RNA-fluorescence and in situ hybridization (RNA-FISH) has the potential to specifically identify the target microorganism of interest in complex microbial communities (it is based on hybridization of fluorescently-labeled oligonucleotide probes targeting to specific regions of the ribosomal RNA). Thus, the aim of this study was to design a novel genus specific RNA-FISH probe against Rhodotorula sp., and to evaluate its specificity and performance both in silico and experimentally. This will contribute for facilitating Rhodotorula sp., identification in degraded CH materials by RNA-FISH. A novel probe for Rhodotorula sp., (L-S-Rh160-a-A-19-ATTO 647N, Rh160-ATTO 647N) was designed using decipher program. Its specificity was analysed in silico by nucleotide BLAST and its performance in terms of characteristics of the probe e.g., GC content and temperature of hairpin structures by oligonucleotide properties calculator oligocalc and hybridization efficiency by math FISH program. The experimental performance and specificity were evaluated by constructing the fluorescence-signal-response / formamide concentration curve for the target and non-target yeast (Cryptococcus adeliensis) and by testing the probe against several other non-target CH biodeteriogenic microorganisms. To do this, a previously described RNA-FISH procedure was applied and the results were analysed by flow cytometry (FC) and by epifluorescence microscopy (EM). In silico analyses of Rh160-ATTO 647N indicated that this probe has a high coverage and specificity for the target genus (796 matches of the target organism in 1000 sequences and only one match for organisms from the same ecosystem of the target organism, Cryptococcus sp.,); fulfills the criteria for being an RNA-FISH probe e.g., 52.60% of GC content and temperature of hairpin structures below hybridization temperature and shows a high maximal theoretical hybridization efficiency with Rhodotorula sp., (99.92% with 0% of formamide). The experimental results were in agreement with these in silico analyses revealing that Rh160-ATTO 647 N probe has a high specificity and performance without formamide. Maximal strong and intense fluorescence signals were detected by FC for the target and absence of signal for all the non-target microorganisms tested. Therefore, this study contributes to an easy and fast identification of Rhodotorula sp., yeast involved in the CH bio deterioration process by RNA-FISH. This will be advantageous for CH safeguard.
Charambira K, Mutunzi H, Zishiri C, Sandy C and Ncube R
Background: Zimbabwe conducted a second anti-tuberculosis drug resistance survey (TB-DRS) in 2015/16 using the Xpert MTB/RIF assay. This assay uses molecular beacons in five overlapping regions of the rpoB DNA region. The probes detect mutations in the codons 507 to 511 (Probe A), 511 to 518 (Probe B), 518 to 523 (Probe C), 523 to 529 (Probe D), and 529 to 533 (Probe E). We report the frequencies of mutations in rpoB gene of the Mycobacterium tuberculosis (MTB) among the TB-DRS samples with rifampicin resistance tuberculosis (RR-TB) detected.
Method: A retrospective review of data collected through the TB-DRS and using the GxAlert platform to check the actual probe details for those tests from samples that had RR-TB strains. Sputum smear positive samples had an Xpert MTB/RIF assay done followed by phenotypic culture and drug susceptibility testing (DST) in those that had RR-TB detected.
Results: A total of 60 specimens had RR-TB detected on Xpert MTB/RIF assay. Of these, 50 (83.3%) had Mycobacterium tuberculosis growth on culture with 48 (96.0%) confirmed RR-TB on phenotypic DST. Among those confirmed RR-TB on phenotypic DST, 23 (47.9%) had rifampicin mono-resistance (RMR) detected and 25 had additional isoniazid resistance (MDR-TB). Probe E mutations occurred in 46% (23/50), probe D 34% (17/50), probe B 10% (5/50), probe A 2% (1/50) and probe C 2% (1/50) of the specimens. Among the RMR, probe D mutation occurred in 54.5% (12/22), probe E 36.4% (8/22), probe A 4.5% (1/22) and probe B 4.5% (1/22).
Conclusion: There is increase in the RMR prevalence from zero percent to 48% between the 1994/5 and 2015/6 TB-DRS. Rifampicin (RMP) seems to be associated with mutations in codons 523 to 529 of the rpoB gene of MTB DNA. GxAlert makes it possible to conduct such surveillance remotely and there is need for further studies to cement this.
The S- The aureus isolates belonged to different types of sequences (STs) with the most common ST15 and ST152. All isolates carried the blaZ gene, and also observed low prevalence of tetK and dfrG genes. All the isolates were negative about mecA. The genes of the pvl were common and observed in distinct lines revealing diverse Sa2int phages. The genomics data at Korle-Bu Teaching Hospital revealed multiple transmission events involving S. Auroreus ST15 involves contamination of various surfaces in the emergency pediatric ward where the outbreak occurred.
Conclusion: Dissemination pattern of the ST15 clone in the Korle-Bu Teaching Hospital emergency ward illustrates a fundamental issue with the hospital's disinfection of environmental surfaces. Various phage populations rather than a single highly transmissible type of phage is likely to mediate the high prevalence of pvl genes among the genes S. aureusisolates.
Habibu U Abdu, Yusuf Y Deeni, Andrew J Spiers, Hapca S and Mukhtar M Dauda
Mosquito breeding sites were grouped into two different study zones (A and B) on the basis of human related activities taking place in and around the breeding sites. An. gambiae larvae collected from ecologically contrasting breeding sites were reared to adults in the laboratory. Adults from the F1 progeny were assayed for resistance against 4% DDT, 0.75% permethrin and 0.1% bendiocarb using the WHO adult insecticide susceptibility bioassay protocol. During mosquito sampling a survey was carried out in each site with the aim of documenting the most widely used insecticide. The levels of the physicochemical environmental factors were measured from the anopheline breeding sites. Results shows that pyrethroids (cypermethrin, lambda-cyhalothrin and cyfluthrin) and organophosphates (dichlovos, dimethoate and chloropyrifos) were most commonly used for crop protection in the agricultural sites, organochlorine (endosulfan and fipronil) and carbamates (carbofuran and carbaryl) were also used to a lesser extent. On the other hand, interview in the residential sites revealed indoor residual sprays (IRS), piya piya sprays (piya piya sprays are formulations produced locally as insecticeds sprays and without government approval) and coils containing pyrethroid insecticides with cypermethrin, lambda-cyhalothrin and cyfluthrin as common active ingredients were mainly used for personal protection. The results of measurement of physicochemical parameters showed little variation in the levels of the physical environmental factors (pH and temperature) across the sampling sites in the two zones studied. However, the levels of nitrates, nitrites, phosphates, sulphates and carbon content were higher in sites located in zone A than those in zone B. Overall, zone A is significantly different from zone B (p=0.000). There was evidence of high insecticides resistance among the mosquitoes tested from all the sampling sites. However, mosquitoes from agricultural sites (zone A) recorded higher insecticide resistance when compared to those from residential sites (zone B). These high levels of resistance are probably related to extensive pesticide usage in the zone. This is further supported by higher levels of the environmental chemicals recorded in zone A compared to zone B. These observations could have a significant impact on the environmental management and insecticide based approach to malaria vector control in Nigeria.
Observed in distinct lines revealing diverse Sa2int phages. The genomics data at Korle-Bu Teaching Hospital revealed multiple transmission events involving S. Auroreus ST15 involves contamination of various surfaces in the emergency pediatric ward where the outbreak occurred.
Conclusion: Dissemination pattern of the ST15 clone in the Korle-Bu Teaching Hospital emergency ward illustrates a fundamental issue with the hospital's disinfection of environmental surfaces. Various phage populations rather than a single highly transmissible type of phage is likely to mediate the high prevalence of pvl genes among the genes S. aureus isolates.
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