scholarly journals Molecular Identification and Classification of Strawberry Phylloid Fruit Phytoplasma in Group 16SrI, New Subgroup

Plant Disease ◽  
2002 ◽  
Vol 86 (8) ◽  
pp. 920-920 ◽  
Author(s):  
R. R. Jomantiene ◽  
J. L. Maas ◽  
F. Takeda ◽  
R. E. Davis
Keyword(s):  
West Virginia ◽  
16S Rdna ◽  
Dna Amplification ◽  
Universal Primer ◽  
Greenhouse Production ◽  
Chain Reactions ◽  
Group I ◽  
16S Rdna Pcr ◽  
Restriction Sites ◽  
Clover Phyllody

Plants of commercial strawberry (Fragaria × ananassa Duch., cv. Camarosa) exhibiting extensive fruit phyllody (development of leafy structures from achenes) were observed in a winter greenhouse production facility in West Virginia. In July 2001, 95 dormant, cold-stored plants were purchased from a California strawberry nursery, potted and grown in this West Virginia facility. Five of the plants developed fruits with phylloid growths. These fruits were assessed for phytoplasma infection using nested polymerase chain reactions (PCRs) in which initial ribosomal (r) DNA amplification was primed by phytoplasma-universal primer pair P1/P7 (2), and rDNA reamplification was primed by primer pair R16F2n/R16R2 (1). Amplification of phytoplasma-characteristic 1.2-kbp 16S rDNA in the nested reactions primed by R16F2n/R16R2 confirmed that the symptomatic plants were infected by a phytoplasma, termed strawberry phylloid fruit (StrawbPhF) phytoplasma. No phytoplasma DNAs were amplified from healthy plants. Restriction fragment length polymorphism (RFLP) patterns of 16S rDNA digested with AluI, KpnI, HhaI, HaeIII, HpaII, MseI, RsaI, and Sau3A1 restriction endonucleases indicated that StrawbPhF phytoplasma belonged to group 16SrI (group I, aster yellows phytoplasma group) according to the phytoplasma classification system of Lee et al. (4). However, the collective patterns distinguished StrawbPhF from its closest known relative, clover phyllody (CPh) phytoplasma, and from all other phytoplasmas classified in group 16SrI. On the basis of the RFLP patterns of 16S rDNA, the StrawbPhF was classified in group 16SrI, new subgroup R. The StrawbPhF phytoplasma 1.2-kbp 16S rDNA PCR product was cloned in Escherichia coli using TOPO TA Cloning Kit (Invitrogen, Carlsbad, CA), sequenced, and the sequence deposited in GenBank under Accession No. AY102275. The StrawbPhF 16S rDNA sequence shared 99.9 and 99.8% similarity with the two sequence heterogeneous operons, rrnA and rrnB, respectively, of CPh phytoplasma, and shared 99.9% similarity with 16S rDNA of the unclassified cirsium yellows (CirY) phytoplasma (GenBank Accession No. AF200431) reported in Cirsium arvense L. in Lithuania (3). The restriction sites in 16S rDNA of StrawbPhF were identical to those in 16S rDNA of CPh rrnA and CirY. Three restriction sites (AluI, HaeIII, and MseI) and three base substitutions distinguished StrawbPhF 16S rDNA from rrnB of CPh phytoplasma. No evidence was obtained for the presence of a second (sequence heterogeneous) rRNA operon in StrawbPhF phytoplasma, as reported in CPh phytoplasma (4), which clearly distinguishes this phytoplasma from CPh phytoplasma. Future studies on StrawbPhF phytoplasma may provide important information on the evolution of phytoplasmas. References: (1) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (2) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (3) R. Jomantiene et al. Phytopathology 90:S39, 2000. (4) I.-M. Lee et al. Int J. Syst. Bacteriol. 48:1153, 1998.

scholarly journals Single and Mixed Phytoplasma Infections in Phyllody- and Dwarf-Diseased Clover Plants in Lithuania

Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1061-1066 ◽  
Author(s):  
Juozas B. Staniulis ◽  
Robert E. Davis ◽  
Rasa Jomantiene ◽  
Audrone Kalvelyte ◽  
Ellen L. Dally
Keyword(s):  
16S Rdna ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Restriction Sites ◽  
Polymerase Chain ◽  
Dwarf Disease ◽  
Clover Phyllody

Naturally diseased plants of clover (Trifolium spp.) exhibiting symptoms of clover phyllody (virescence and phyllody of flowers) or of clover dwarf (abnormally small leaves, shortened internodes, proliferation of shoots, and dwarf growth habit) were observed in fields in Lithuania. Phytoplasma group-specific polymerase chain reactions (PCRs) and restriction fragment length polymorphism (RFLP) analysis of 16S rDNA revealed that the plants were infected by two mutually distinct phytoplasmas. Clover phyllody-diseased plants were infected by a subgroup 16SrI-C (subgroup I-C) phytoplasma (CPh-L) related to clover phyllody (CPh-C) phytoplasma in Canada. Clover dwarf-diseased plants were infected by both CPh-L and a phytoplasma (CYE-L) related to clover yellow edge (CYE-C) phytoplasma (subgroup 16SrIII-B = III-B) in Canada. A 1.8-kbp fragment of rRNA operon from CYE-L was amplified, cloned, and sequenced, and putative restriction sites mapped. This sequence shared high similarity (99.7%) with that of CYE-C and exhibited no differences from CYE-C in RFLP patterns of 16S rDNA; therefore, we tentatively classified CYE-L in subgroup 16SrIII-B (type strain, CYE = CYE-C phytoplasma) of the X-disease phytoplasma group. These findings extend the known geographical ranges of subgroup I-C and subgroup III-B taxa to the region of northern Europe including Lithuania and suggest a role of the subgroup III-B phytoplasma in clover dwarf disease.

scholarly journals Simultaneous Discrimination between 15 Fish Pathogens by Using 16S Ribosomal DNA PCR and DNA Microarrays

2004 ◽  
Vol 70 (7) ◽  
pp. 4216-4221 ◽  
Author(s):  
Adelaide E. Warsen ◽  
Melissa J. Krug ◽  
Stacey LaFrentz ◽  
Danielle R. Stanek ◽  
Frank J. Loge ◽  
...  
Keyword(s):  
16S Rdna ◽  
Ribosomal Dna ◽  
Bacterial Species ◽  
Simultaneous Detection ◽  
Detection Sensitivity ◽  
Fish Pathogens ◽  
Universal Primer ◽  
16S Ribosomal Dna ◽  
Glass Slides ◽  
16S Rdna Pcr

ABSTRACT We developed a DNA microarray suitable for simultaneous detection and discrimination between multiple bacterial species based on 16S ribosomal DNA (rDNA) polymorphisms using glass slides. Microarray probes (22- to 31-mer oligonucleotides) were spotted onto Teflon-masked, epoxy-silane-derivatized glass slides using a robotic arrayer. PCR products (ca. 199 bp) were generated using biotinylated, universal primer sequences, and these products were hybridized overnight (55°C) to the microarray. Targets that annealed to microarray probes were detected using a combination of Tyramide Signal Amplification and Alexa Fluor 546. This methodology permitted 100% specificity for detection of 18 microbes, 15 of which were fish pathogens. With universal 16S rDNA PCR (limited to 28 cycles), detection sensitivity for purified control DNA was equivalent to <150 genomes (675 fg), and this sensitivity was not adversely impacted either by the presence of competing bacterial DNA (1.1 × 106 genomes; 5 ng) or by the addition of up to 500 ng of fish DNA. Consequently, coupling 16S rDNA PCR with a microarray detector appears suitable for diagnostic detection and surveillance for commercially important fish pathogens.

scholarly journals First Report of Barley as Host of a Phytoplasma Belonging to Group 16SrI, Subgroup B, and Ribosomal Protein Subgroup rpI-B in Lithuania

Plant Disease ◽  
2005 ◽  
Vol 89 (3) ◽  
pp. 339-339 ◽  
Author(s):  
L. Urbanaviciene ◽  
R. Jomantiene ◽  
R. E. Davis
Keyword(s):  
Ribosomal Protein ◽  
16S Rdna ◽  
Rflp Analysis ◽  
Gene Sequences ◽  
Universal Primer ◽  
Group I ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Template Dna ◽  
Aster Yellows Phytoplasma

Numerous diseased plants of barley (Hordeum vulgaris L.) exhibiting twisted, abnormally thin and yellowed awns, reduced spikelets, and general stunting and yellowing were observed in fields in the Vilnius and Kaisiadorys regions of Lithuania. The possible association of a phyto-plasma with the disease, termed barley deformation (BaDef), was assessed using polymerase chain reaction (PCR). Three phytoplasma universal primer pairs (P1/P7, R16F2n/R16R2, and rpF1/rpR1) (1,2,4) were employed to amplify ribosomal (r) RNA gene (rDNA) and ribosomal protein (rp) gene sequences. Template DNA extractions and PCR (direct and nested) were conducted as previously described (4). Although DNA was amplified in PCRs containing template extracted from diseased plants, no amplification was observed in PCRs containing DNA from symptomless plants sampled from the same fields. The BaDef phytoplasma was identified and classified according to Lee et al. (4) through restriction fragment length polymorphism (RFLP) analysis of 1.2-kbp 16S rDNA amplified in the PCR primed by primer pair R16F2n/R16R2 and analysis of the 1.2-kbp rp gene sequences amplified in PCR primed by primer pair rpF1/rpR1. On the basis of collective RFLP patterns of amplified 16S rDNA and rp gene sequences, the BaDef phytoplasma was classified as a member of group 16SrI (group I, aster yellows phytoplasma group), subgroup B (16SrI-B), and rp subgroup rpI-B. Ribosomal protein subgroup B was distinguished from other rp subgroups on the basis of the presence of a recognition site for HpaII. The 1.8-kbp rDNA product of PCR primed by P1/P7 and the 1.2-kbp rpF1/rpR1 PCR product were cloned and sequenced, and the sequences were deposited in GenBank under Accession No. AY734453 for the BaDef 16S rDNA and Accession No. AY735448 for the BaDef rp gene sequence. Previously, only oat proliferation (OatP) phytoplasma, a member of subgroup 16SrI-A, had been characterized in a cereal crop (Avena sativa L.) in Europe (3); BaDef is another phytoplasmal disease threatening cereal crops in the region. References: (1) S. Deng and D. Hiruki. J. Microbiol. Methods 14:53, 1991. (2) D. E. Gundersen and I. M. Lee. Phytopathol. Mediterr. 35:144, 1996. (3) R. Jomantiene et al. Plant Dis. 86:443, 2002. (4) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

scholarly journals Molecular Evidence for the Presence of Maize Bushy Stunt Phytoplasma in Corn in Brazil

Plant Disease ◽  
1997 ◽  
Vol 81 (8) ◽  
pp. 957-957 ◽  
Author(s):  
I. P. Bedendo ◽  
R. E. Davis ◽  
E. L. Dally
Keyword(s):  
16S Rdna ◽  
Dna Sequences ◽  
Dna Amplification ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Chromosomal Dna ◽  
Chain Reactions ◽  
Leaf Reddening ◽  
Corn Plants ◽  
Maize Bushy Stunt Phytoplasma

Previously, electron microscopy revealed that the corn (Zea mays L.) disease characterized by stunting and leaf reddening and commonly known as “red stunt” in Brazil is associated with plant infection by an unidentified phytoplasma (formerly mycoplasmalike organism) (1). During recent years, corn production in Brazil has been seriously affected by the increasing prevalence of a disease exhibiting symptoms similar to those of “red stunt.” The present investigation was initiated to determine the possible association of a phytoplasma with the current disease problem and to attain definitive molecular identification of any associated phytoplasma. To detect the possible presence of a phytoplasma in diseased corn in Brazil, plants exhibiting symptoms of stunting and leaf reddening in the field in 1995 and 1996 were assayed for the presence of phytoplasma DNA sequences by the use of polymerase chain reactions (PCR). We used primer pairs R16mF2/R16mR1 and R16F2n/R16R2 in nested PCRs (2) to prime phytoplasma-universal amplification of 16S ribosomal (r) DNA. Oligonucleotide pair rpF1/rpR1 (3) was used to prime phytoplasma-universal amplification of ribosomal protein (rp) gene operon sequences. Phytoplasma identification was accomplished by restriction fragment length polymorphism (RFLP) analysis of amplified 16S rDNA and rp gene operon sequences. Primer pair MBS-F1/MBS-R1 (4) was used to prime amplification of a maize bushy stunt (MBS)-specific chromosomal DNA sequence. Preparation of template DNAs, PCR conditions, and RFLP analyses of PCR products were as previously described (2–4). DNA amplification was observed in all PCRs containing template DNAs derived from symptomatic plants, indicating phytoplasmal infection of corn in Brazil. No DNA amplification was observed in PCR containing template DNA from healthy control corn plants. Polymorphisms in amplified 16S rDNA were those characteristic of phytoplasmas classified in 16S rRNA gene group 16SrI, subgroup I-B, of which MBS phytoplasma is a member (3). Collective RFLP patterns of amplified rp gene operon sequences were similar or identical to those observed in parallel tests for a known reference strain of MBS phytoplasma, indicating that the Brazilian corn plants were infected by MBS phytoplasma. Amplification of MBS-characteristic DNA was observed in PCRs containing MBS-specific primer pair MBS-F1/MBS-R1 and DNA from diseased corn, confirming infection of the plants by MBS phytoplasma. This work provides the first firm evidence for association of maize bushy stunt phytoplasma with the current disease problem of corn in Brazil. References: (1) A. S. Costa et al. Rev. Soc. Bras. Fitopatol. Piracicaba 4:39, 1971. (2) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35: 144,1996. (3) D. E. Gundersen et al. Int. J. Syst. Bacteriol. 46:64, 1996. (4) N. A. Harrison et al. Plant Dis. 80:263, 1996.

scholarly journals First Report of Oat as Host of a Phytoplasma Belonging to Group 16SrI, Subgroup A

Plant Disease ◽  
2002 ◽  
Vol 86 (4) ◽  
pp. 443-443 ◽  
Author(s):  
R. Jomantiene ◽  
R. E. Davis ◽  
A. Alminaite ◽  
D. Valiunas ◽  
R. Jasinskaite
Keyword(s):  
16S Rdna ◽  
Plant Species ◽  
Nested Pcr ◽  
Sequence Similarity ◽  
Intergenic Spacer ◽  
Rflp Analysis ◽  
23S Rrna ◽  
Universal Primer ◽  
Intergenic Spacer Region ◽  
Group I

Diseased plants of oat (Avena sativa L.) exhibiting abnormal proliferation of spikelets were observed in the field in Raseniai, Lithuania. The possible association of a phytoplasma with the disease, termed oat proliferation (OatP), was determined using polymerase chain reaction (PCR) for amplification of phytoplasmal ribosomal (r) RNA gene (rDNA) sequences from template DNA extracted from the diseased oats. DNA extractions and nested PCRs were conducted as previously described (2). In the nested PCRs, the first reaction was primed by phytoplasma-universal primer pair P1/P7, and the second (nested) PCR was primed by primer pair R16F2n/R16R2 (F2n/R2). Phytoplasmal rDNA was amplified in the nested PCR, indicating that the plants contained a phytoplasma, designated oat proliferation (OatP) phytoplasma. The OatP phytoplasma was identified and classified according to the system of Lee et al. (2) through restriction fragment length polymorphism (RFLP) analysis of 16S rDNA amplified in the PCR primed by F2n/R2. On the basis of collective RFLP patterns of the 16S rDNA, the OatP phytoplasma was classified as a member of group 16SrI (group I, aster yellows phytoplasma group). The RFLP patterns of the 16S rDNA were indistinguishable from those of 16S rDNA from tomato big bud (BB) phytoplasma and other phytoplasmas classified in group I, subgroup A (subgroup I-A, tomato big bud phytoplasma subgroup). The 1.8-kbp rDNA product of PCR primed by primer pair P1/P7 was cloned, and its nucleotide sequence was determined. The sequence was deposited in GenBank under Accession No. AF453416. Results from putative restriction site analysis of the cloned and sequenced rDNA were in excellent agreement with the results from enzymatic RFLP analysis of uncloned rDNA from OatP-diseased oat plants. Sequence similarity between the 1.8-kbp rDNA of OatP phytoplasma and that of BB phytoplasma (GenBank No. AF222064) was 99.2%; 9 of the 14 base changes were in the 16S-23S rRNA intergenic spacer region. The base differences in rDNA may signal that the OatP and BB phytoplasmas are mutually distinct in their biologies. Phytoplasmas classified in subgroup I-A have previously been reported in a broad range of plant species in North America and Europe, although there are no previous definitive reports of oat as a host of a subgroup I-A phytoplasma (3,4). In 1977, Fedotina (1) reported electron microscopy of a mycoplasma-like organism (phytoplasma) in pseudorosette-diseased oat plants in Siberia, but the identity of that phytoplasma remains unknown. Subgroup I-A phytoplasma strains are geographically widespread and have been found in numerous plant species (3,4). The discovery reported here, of a subgroup I-A phytoplasma in diseased oats in Lithuania, provokes questions concerning possible impacts of this phytoplasma on oat cultivation in central Europe and other regions. References: (1) V. L. Fedotina. Arch. Phytopathol. Pflanzenschutz 13:177, 1977. (2) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (3) C. Marcone et al. Int. J. Syst. Evol. Microbiol. 50:1703, 2000. (4) D. Valiunas et al. Plant Dis. 85:804, 2001.

scholarly journals First Report of Elm Yellows Phytoplasma Infecting Clover in China

Plant Disease ◽  
2009 ◽  
Vol 93 (3) ◽  
pp. 321-321 ◽  
Author(s):  
Z. N. Li ◽  
X. Zheng ◽  
H. J. Wei ◽  
X. Q. Yu ◽  
W. J. Wu ◽  
...  
Keyword(s):  
16S Rdna ◽  
Polymorphism Analysis ◽  
Universal Primer ◽  
Thin Sections ◽  
First Report ◽  
Ctab Method ◽  
Plant Pathol ◽  
Group Relationships ◽  
Elm Yellows ◽  
Clover Phyllody

In the summer of 2008, phyllody and enlarged petioles resembling symptoms of phytoplasma infection were observed on clover (Trifolium repens) plants in lawns on the campus of Northwest A&F University. Typical phytoplasma-like bacteria were observed in the phloem cells when ultra-thin sections from leaf midrib tissues were examined with transmission electron microscopy. Nested PCR assays were used to verify the association of phytoplasma with the disease. Total DNA was extracted from the phloem of leaf midribs from 20 symptomatic plants and six symptomless plants using the modified CTAB method (1). Using the phytoplasma universal primer pair R16mF2/R16mR1 followed by specific primers R16F2n/R16R2 (4), PCR products of 1.4 and 1.2 kb were amplified, respectively, from symptomatic plants only. Jujube witches'-broom (JWB) and paulownia witches'-broom (PaWB) phytoplasma DNA samples served as controls and were used to study group relationships. After sequencing of the 16S rDNA fragment (GenBank Accession No. FJ436792), a BLAST search determined that the clover phytoplasma shared closest homology (99.6%) with JWB (GenBank Accession No. FJ154846) phytoplasmas compared with lesser identity (90.4%) with PaWB (GenBank Accession No. EF199937). Subsequent restriction fragment length polymorphism analysis of the PCR-amplified 1.2-kb 16S rDNA R16(1)F1/R1 fragment indicated that the phytoplasma associated with the disease belongs to subgroup 16SrV-B of the elm yellows phytoplasma group. Clover phyllody phytoplasma were previously reported to infect clover in Canada (GenBank Accession No. L33762) (3) and Italy (GenBank Accession No. X77482) (2). The phytoplasma reported here shared 86.7 and 90.0% identity with the clover phyllody phytoplasma above, respectively, much lower than that with Elm yellows phytoplasma group. To our knowledge, this is the first report of Elm yellows phytoplasma infecting clover in China. References:(1) E. Angelini et al. Vitis 40:79, 2001. (2) G. Firrao et al. Eur. J. Plant Pathol. 102:817, 1996. (3) N. A. Harrison et al. Plant Pathol. 52:147, 2003. (4) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

DETECTING MYCOBACTERIUM TUBERCULOSIS RAPIDLY AND MYCOBACTERIUM TUBERCULOSIS STRAINS WITHOUT IS6110 SEQUENCE BY PCR ASSAY

2012 ◽  
pp. 15-19
Author(s):  
Thi Chau Anh Nguyen ◽  
Hoang Bach Nguyen ◽  
Hai Duong Huynh ◽  
Nu Xuan Thanh Le ◽  
Xuan Cuong Le ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
16S Rdna ◽  
False Negative ◽  
Clinical Samples ◽  
Tuberculosis Complex ◽  
Pcr Assay ◽  
Complex Objects ◽  
Method Performance ◽  
Gene Coding ◽  
16S Rdna Pcr

Background: The Nested IS6110 PCR is used for detecting tuberculosis, however IS6110 sequence is not present in the genome of all strains of M.tuberculosis, the result may be false negative. The gene coding 16S ribosome always contains a short sequence specific to M. tuberculosis complex. Objects: Performance of the 16S Real-time PCR to detect M. tuberculosis and combining to the nested IS6110 PCR to determine the rate of Mtb strains without IS6110 from clinical samples. Materials and method: Performance of 16S rDNA PCR by commercial kit of Viet A Inc. for all 480 samples, the samples which were positive with the 16S rDNA PCR were retested in IS6110 PCR assay by in-house kit. Results: The Realtime 16S rDNA PCR detected 258 cases (53.8%) of tuberculosis. There were 3 (1.2 %) M. tuberculosis strains which do not harbor IS6110 sequence in genome. Conclusion: The IS6110 nested PCR can be applied more widely than the 16S rDNA realtime PCR. In case of using IS6110 PCR assay, results may show a low proportion of false negative. Combining 16S rDNA PCR with the IS6110 based PCR allowed detection of deletion of IS6110 sequence in M. tuberculosis isolates.

Bacterial Species Identification after DNA Amplification with a Universal Primer Pair

1999 ◽  
Vol 66 (3) ◽  
pp. 205-211 ◽  
Author(s):  
Kevin M. McCabe ◽  
Yao-Hua Zhang ◽  
Bing-Ling Huang ◽  
Elizabeth A. Wagar ◽  
Edward R.B. McCabe
Keyword(s):  
Species Identification ◽  
Bacterial Species ◽  
Dna Amplification ◽  
Universal Primer

Isolation of Enterobacter sakazakii from Stable Flies, Stomoxys calcitrans L. (Diptera: Muscidae)†

2006 ◽  
Vol 69 (3) ◽  
pp. 671-673 ◽  
Author(s):  
F. MRAMBA ◽  
A. BROCE ◽  
L. ZUREK
Keyword(s):  
16S Rdna ◽  
Foodborne Pathogen ◽  
Animal Manure ◽  
Stomoxys Calcitrans ◽  
Fecal Coliforms ◽  
Stable Fly ◽  
Enterobacter Sakazakii ◽  
Stable Flies ◽  
16S Rdna Pcr ◽  
Glucose Agar

Enterobacter sakazakii is an opportunistic foodborne pathogen that causes meningitis, enterocolitis, and sepsis, primarily in immunocompromised infants. Previously, it was suggested that stable flies, Stomoxys calcitrans, were a vector or reservoir of this pathogen. In our study, by means of a culturing approach combined with 16S rDNA PCR–restriction fragment length polymorphism genotyping and sequencing, we screened 928 individual stable flies collected in Kansas and Florida. Two stable flies (0.2%) were positive for E. sakazakii. In addition, 411 (44%) stable flies carried bacteria-forming red colonies (presumably enterics) on a violet red bile glucose agar (mean count = 6.4 × 104 CFU per fly), and 120 (13%) stable flies carried fecal coliforms (mean count = 8.7 × 103 CFU per fly). Sequencing of 16S rDNA showed that enterics from violet red bile glucose agar were represented by several genera, including Escherichia, Shigella, Providencia, Enterobacter, Pantoea, Proteus, Serratia, and Morganella. Our study shows that stable flies carry bacteria typically present in animal manure (a developmental site of stable fly larvae), which indicates that the natural reservoir of E. sakazakii is the digestive tract or manure of domestic animals. The low prevalence of E. sakazakii associated with stable flies suggests that stable flies do not play a major role as a reservoir or vector of this pathogen.

scholarly journals Bakteri Simbion Gastropoda Pleuroploca trapesium Dari Perairan Ternate, Sebagai Alternatif Antibakteri MDR (Bacterial Symbiont Gastropoda Pleuroploca trapezium from Ternate, as Alternative Antibacterial MDR)

2014 ◽  
Vol 19 (1) ◽  
pp. 55
Author(s):  
Delianis Pringgenies ◽  
Person Pesona Renta
Keyword(s):  
16S Rdna ◽  
Pathogenic Bacteria ◽  
Dna Amplification ◽  
Sensitivity Test ◽  
Symbiotic Bacteria ◽  
Drug Resistant ◽  
16S Rdna Sequence Analysis ◽  
Close Relationship ◽  
Antibiotic Compounds ◽  
Bacterial Sensitivity

Bakteri yang resisten terhadap beberapa jenis antibakteri ini dikenal dengan bakteri multi drug resistant (MDR).Untuk mengatasi permasalahan tersebut, perlu dilakukan pencarian senyawa antibiotik baru yang lebih efektif dan efisien dalam mengatasi permasalahan bakteri MDR. Penelitian bertujuan untuk mengetahui potensi bakteri yang bersimbiosis dengan gastropoda Pleuroploca trapezium sebagai sumber antibakteri MDR. Sampel Moluska dikoleksi dari perairan Ternate, Maluku. Tahapan penelitian meliputi isolasi bakteri, skrining  bakteri simbion yang potensi sebagai anti bakteri MDR, uji antibakteri, isolasi bakteri patogen klinis MDR; uji sensitivitas anti-bakteri, ekstraksi, amplifikasi dan sekuensing DNA. Hasil 16S urutan r-DNA dianalisis dan diedit menggunakan program Genetix dan diikuti dengan analisis urutan 16S rDNA. Hasil menunjukkan bahwa terdapat 19 isolat bakteri dengan 5 bakteri aktif yang berasosiasi dengan Pleuroploca trapezium. Berdasarkan besarnya zona hambat yang dibentuk dan konsistensi munculnya zona hambatan, isolat terbaik adalah TPT 4.7. Isolat ini memiliki hubungan yang dekat dengan Paracoccus  sp. MBIC4019 dengan homologi sebesar 95% yang menunjukkan kekerabatan ditingkat genus. Hasil penelitian ini memberikan harapan adanya potensi besar sebagai bahan antibakteri baru. Kata kunci: antibakteri, simbion, Pleuroploca trapezium, multi drugs resistantThe bacteria resistant to some antibiotics are known as multi drug resistant (MDR). To overcome the problem, it is needed to search for a new antibiotic compounds more effectively and efficiently. This study aims to identify potential from symbionts of Pleuroploca trapezium as a source of antibacteria MDR and identifying the bacteria that were active against the MDR. Samples were collected from Ternate, Maluku. Isolation of symbiotic bacteria, screening for bacteria which producing secondary metabolites as anti-MDR bacteria, antibacterial test, isolation of clinical pathogenic bacteria of MDR. Conducting anti-bacterial sensitivity test,  sensitivity test for antibacterial,  DNA exctraction, DNA amplification based on PCR method, DNA sequencing.  Result of 16S r-DNA sequence was then analyzed and edited using GENETYX program and followed by 16S rDNA sequence analysis. Screening of bacteria associated with P. trapezium resulted in 19 isolates with 5 active bacteria. Based on the size of the zone forming and the consistency of zone, so the best isolate is TPT 4.7. The identification shows that TPT 4.7 has a close relationship with the Paracoccus sp. MBIC4019 with homologi of 95%, which shows the relationship at the genus level. Its suggest that these results are very promising as a new antibacterial material. Keywords: antibacterial, symbiotic bacteria, Pleuroploca trapezium, multi drugs resistant

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