scholarly journals Genetic Resistance to Brucella abortus in the Water Buffalo (Bubalus bubalis)

2006 ◽  
Vol 74 (4) ◽  
pp. 2115-2120 ◽  
Author(s):  
Giorgia Borriello ◽  
Rosanna Capparelli ◽  
Michele Bianco ◽  
Domenico Fenizia ◽  
Flora Alfano ◽  
...  
Keyword(s):  
Brucella Abortus ◽  
Water Buffalo ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Genetic Resistance ◽  
Genetic Selection ◽  
Bubalus Bubalis ◽  
Prolonged Incubation ◽  
Gradient Gel Electrophoresis ◽  
Selection For

ABSTRACT Brucellosis is a costly disease of water buffaloes (Bubalus bubalis). Latent infections and prolonged incubation of the pathogen limit the efficacy of programs based on the eradication of infected animals. We exploited genetic selection for disease resistance as an approach to the control of water buffalo brucellosis. We tested 231 water buffalo cows for the presence of anti-Brucella abortus antibodies (by the agglutination and complement fixation tests) and the Nramp1 genotype (by PCR-denaturing gradient gel electrophoresis). When the 231 animals (58 cases and 173 controls) were divided into infected (seropositive) and noninfected (seronegative) groups and the Nramp1 genotypes were compared, the seropositive subjects were 52 out of 167 (31%) in the Nramp1A + (Nramp1AA or Nramp1AB) group and 6 out of 64 (9.4%) in the Nramp1A − (Nramp1BB) group (odds ratio, 4.37; 95% confidence limits, 1.87 to 10.19; χ2, 11.65 for 1 degree of freedom). Monocytes from Nramp1BB subjects displayed significantly (P < 0.01) higher levels of Nramp1 mRNA than Nramp1AA subjects and also a significantly (P < 0.01) higher ability in controlling the intracellular replication of several Brucella species in vitro. Thus, selection for the Nramp1BB genotype can become a valuable tool for the control of water buffalo brucellosis in the areas where the disease is endemic.

Post-warming hatching and birth of live calves following transfer of in vitro-derived vitrified water buffalo (Bubalus bubalis) embryos

2004 ◽  
Vol 61 (7-8) ◽  
pp. 1429-1439 ◽  
Author(s):  
Danilda Hufana-Duran ◽  
Prudencio B Pedro ◽  
Hernando V Venturina ◽  
Rogelio D Hufana ◽  
Apolinario L Salazar ◽  
...  
Keyword(s):  
Water Buffalo ◽  
Bubalus Bubalis

scholarly journals Evaluation of the effects of tropical tanniferous plants on rumen microbiota using qRT PCR and DGGE analysis

2013 ◽  
Vol 58 (No. 3) ◽  
pp. 106-116 ◽  
Author(s):  
C. Longo ◽  
A.L. Abdalla ◽  
J. Liebich ◽  
I. Janzik ◽  
J. Hummel ◽  
...  
Keyword(s):  
Denaturing Gradient Gel Electrophoresis ◽  
Bactericidal Effect ◽  
Methanogenic Bacteria ◽  
Gas Production ◽  
Microbial Populations ◽  
Methanogenic Community ◽  
In Vitro Gas Production ◽  
Gradient Gel Electrophoresis ◽  
Relative Gene

Tanniferous forages may have bacteriostatic and/or bactericidal effect on different rumen microbial populations. We investigated the influence of the tropical tanniferous plants Styzolobium aterrimum (STA), Styzolobium deeringianum (STD), Leucaena leucocephala (LEU), and Mimosa caesalpiniaefolia (MIC) containing 20, 64, 56, and 105 g condensed tannis (CT)/kg dry matter (DM) and Cynodon spp. cv. Tifton 85 (CYN) as control on Fibrobacter succinogenes and methanogenic microbes in rumen liquor from sheep using the in vitro gas production technique (Hohenheim gas test). The relative gene expression of F. succinogenes at t<sub>&frac12;</sub> (time point when 50% of the maximal gas production has been reached) analyzed by quantitative PCR was 0.20- and 0.28- fold lower than the control when LEU and STA was applied and 0.91- and 0.85-fold lower with MIC and STD. Methanogenic population was 0.29- and 0.58- fold reduced with STA and LEU compared to the control, but 5.50- and 1.43- fold higher with MIC and STD. At 24 h, F. succinogenes was reduced for all legumes as well as methanogenic bacteria, except for MIC which increased 4.15-fold. Denaturing gradient gel electrophoresis (DGGE) of the methanogenic community resulted in different band patterns: CYN presented some strong bands, which became weaker in the analyzed treatments. Some bands appeared weaker, especially in MIC and STD, but not in STA and LEU. MIC seemed to increase the total number of weak bands. Overall, the tannin-rich plants negatively affected the F. succinogenes population and caused changes in the structure of the methanogenic community.

scholarly journals Protective Effect of the Nramp1 BB Genotype against Brucella abortus in the Water Buffalo (Bubalus bubalis)

2006 ◽  
Vol 75 (2) ◽  
pp. 988-996 ◽  
Author(s):  
Rosanna Capparelli ◽  
Flora Alfano ◽  
Maria Grazia Amoroso ◽  
Giorgia Borriello ◽  
Domenico Fenizia ◽  
...  
Keyword(s):  
Brucella Abortus ◽  
Untranslated Region ◽  
Complement Fixation Test ◽  
Water Buffalo ◽  
Complement Fixation ◽  
Bubalus Bubalis ◽  
Lower Number ◽  
Agglutination Test ◽  
Intracellular Bacteria ◽  
Antibacterial Protein

ABSTRACT We tested 413 water buffalo cows (142 cases and 271 controls) for the presence of anti-Brucella abortus antibodies (by the skin test, the agglutination test, and the complement fixation test) and the Nramp1 genotype (by capillary electrophoresis). Four alleles (Nramp1A, -B, -C, and -D) were detected in the 3′ untranslated region of the Nramp1 gene. The BB genotype was represented among only controls, providing evidence that this genotype confers resistance to Brucella abortus. The monocytes from the BB (resistant) subjects displayed a higher basal level of Nramp1 mRNA and a lower number of viable intracellular bacteria than did the monocytes from AA (susceptible) subjects. The higher basal level of the antibacterial protein Nramp1 most probably provides the BB animals with the possibility of controlling bacteria immediately after their entry inside the cell.

Model plants for studying the interaction between Methylobacterium mesophilicum and Xylella fastidiosa

2006 ◽  
Vol 52 (5) ◽  
pp. 419-426 ◽  
Author(s):  
Fernando D Andreote ◽  
Paulo T Lacava ◽  
Cláudia S Gai ◽  
Welington L Araújo ◽  
Walter Maccheroni, Jr. ◽  
...  
Keyword(s):  
Bacterial Communities ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Xylella Fastidiosa ◽  
Specific Primers ◽  
Gradient Gel Electrophoresis ◽  
Polymerase Chain ◽  
Methylobacterium Mesophilicum ◽  
Biofilm Structure ◽  
Nicotiana Clevelandii

Over the last few years, endophytic bacterial communities associated with citrus have been studied as key components interacting with Xylella fastidiosa. In this study, we investigated the possible interaction between the citrus endophyte Methylobacterium mesophilicum SR1.6/6 and X. fastidiosa in model plants such as Catharanthus roseus (Madagaskar periwinkle) and Nicotiana clevelandii (Clevelands tobacco). The aim of this study was to establish the fate of M. mesophilicum SR1.6/6 after inoculation of C. roseus and N. clevelandii plants, using PCR–DGGE (polymerase chain reaction – denaturing gradient gel electrophoresis) and plating techniques. Shifts in the indigenous endophytic bacterial communities were observed in plants inoculated with strain SR1.6/6, using specific primers targeting α- and β-Proteobacteria. Cells of strain SR1.6/6 were observed in a biofilm structure on the root and hypocotyl surfaces of in vitro seedlings inoculated with M. mesophilicum SR1.6/6. This emphasizes the importance of these tissues as main points of entrance for this organism. The results showed that C. roseus and N. clevelandii could be used as model plants to study the interaction between M. mesophilicum and X. fastidiosa.Key words: endophytic, Methylobacterium, model plants, DGGE.

scholarly journals Gastrointestinal Colonization with a Cephalosporinase-Producing Bacteroides Species Preserves Colonization Resistance against Vancomycin-Resistant Enterococcus and Clostridium difficile in Cephalosporin-Treated Mice

2014 ◽  
Vol 58 (8) ◽  
pp. 4535-4542 ◽  
Author(s):  
Usha Stiefel ◽  
Michelle M. Nerandzic ◽  
Michael J. Pultz ◽  
Curtis J. Donskey
Keyword(s):  
Clostridium Difficile ◽  
Intestinal Tract ◽  
Denaturing Gradient Gel Electrophoresis ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Content Type ◽  
Antibiotic Effect ◽  
Gradient Gel Electrophoresis ◽  
Vancomycin Resistant

ABSTRACTAntibiotics that are excreted into the intestinal tract may disrupt the indigenous intestinal microbiota and promote colonization by health care-associated pathogens. β-Lactam, or penicillin-type, antibiotics are among the most widely utilized antibiotics worldwide and may also adversely affect the microbiota. Many bacteria are capable, however, of producing β-lactamase enzymes that inactivate β-lactam antibiotics. We hypothesized that prior establishment of intestinal colonization with a β-lactamase-producing anaerobe might prevent these adverse effects of β-lactam antibiotics, by inactivating the portion of antibiotic that is excreted into the intestinal tract. Here, mice with a previously abolished microbiota received either oral normal saline or an oral cephalosporinase-producing strain ofBacteroides thetaiotaomicronfor 3 days. Mice then received 3 days of subcutaneous ceftriaxone, followed by either oral administration of vancomycin-resistantEnterococcus(VRE) or sacrifice and assessment ofin vitrogrowth of epidemic and nonepidemic strains ofClostridium difficilein murine cecal contents. Stool concentrations of VRE and ceftriaxone were measured, cecal levels ofC. difficile24 h after incubation were quantified, and denaturing gradient gel electrophoresis (DGGE) of microbial 16S rRNA genes was performed to evaluate the antibiotic effect on the microbiota. The results demonstrated that establishment of prior colonization with a β-lactamase-producing intestinal anaerobe inactivated intraintestinal ceftriaxone during treatment with this antibiotic, allowed recovery of the normal microbiota despite systemic ceftriaxone, and prevented overgrowth with VRE and epidemic and nonepidemic strains ofC. difficilein mice. These findings describe a novel probiotic strategy to potentially prevent pathogen colonization in hospitalized patients.

Demethylation of a hypermethylated P15/INK4B gene in patients with myelodysplastic syndrome by 5-Aza-2′-deoxycytidine (decitabine) treatment

Blood ◽  
2002 ◽  
Vol 100 (8) ◽  
pp. 2957-2964 ◽  
Author(s):  
Michael Daskalakis ◽  
Tudung T. Nguyen ◽  
Carvell Nguyen ◽  
Per Guldberg ◽  
Gabriele Köhler ◽  
...  
Keyword(s):  
Bone Marrow ◽  
High Risk ◽  
Myelodysplastic Syndrome ◽  
Mononuclear Cells ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Methylation Status ◽  
Bone Marrow Biopsies ◽  
Methylation Inhibitor ◽  
Gradient Gel Electrophoresis

p16 and p15, 2 inhibitors of cyclin-dependent kinases, are frequently hypermethylated in hematologic neoplasias. Decitabine, or 5-Aza-2′-deoxycytidine, reverts hypermethylation of these genes in vitro, and low-dose decitabine treatment improves cytopenias and blast excess in ∼50% of patients with high-risk myelodysplastic syndrome (MDS). We examined p15and p16 methylation status in bone marrow mononuclear cells from patients with high-risk MDS during treatment with decitabine, using a methylation-sensitive primer extension assay (Ms-SNuPE) to quantitate methylation, and denaturing gradient gel electrophoresis (DGGE) and bisulfite-DNA sequencing to distinguish individually methylated alleles. p15 expression was serially examined in bone marrow biopsies by immunohistochemistry. Hypermethylation in the 5′ p15 gene region was detected in 15 of 23 patients (65%), whereas the 5′ p16 region was unmethylated in all patients. Among 12 patients with hypermethylation sequentially analyzed after at least one course of decitabine treatment, a decrease in p15 methylation occurred in 9 and was associated with clinical response. DGGE and sequence analyses were indicative of hypomethylation induction at individual alleles. Immunohistochemical staining for p15 protein in bone marrow biopsies from 8 patients with p15 hypermethylation revealed low or absent expression in 4 patients, which was induced to normal levels during decitabine treatment. In conclusion, frequent, selectivep15 hypermethylation was reversed in responding MDS patients following treatment with a methylation inhibitor. The emergence of partially demethylated epigenotypes and re-establishment of normal p15 protein expression following the initial decitabine courses implicate pharmacologic demethylation as a possible mechanism resulting in hematologic response in MDS.

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