Characterization and growth profile of proteolytic strain PK-4 isolated from local slaughterhouse wastewater

This study aims to characterize and to determine the growth profile of the proteolytic bacteria isolated from Giwangan slaughterhouse wastewater in Yogyakarta City, and to observe the optimum temperature as well as the pH condition for growing in the nutrient medium. Isolation of bacteria from slaughterhouse wastewater was done with a sample from 4 different locations. The isolates were then grown on a medium with a skim (at 0;0.5;1%;1.5 and 2%) and pH condition (at 7;8;9;10 and 11). The bacterial growth profile was measured based on the number of cells (CFU/mL), the size of the bacterial colony diameter, the diameter of the clear zone, and the proteolytic index. Strain PK4 was proved to have proteolytic activity. The characterization of Strain PK4 has shown for colony morphology with a circle shape, white color, flat edges, and convex elevation. The cell morphology was a cocci-shaped, red color, Gram-negative, and having a catalase-positive. The bacterial colony diameter, halo diameter, and proteolytic index were increased significantly (P<0.05) with the increase of skim milk addition. The optimal growth at the medium has reached by the addition of 1–2 % Skim and pH alkaline (>7). It could be concluded that Strain PK4 was classified as alkalophilic and had the potency as alkaline protease producing bacteria.

Characterization of Tn6349, a novel mosaic transposon carrying poxtA, cfr and other resistance determinants, inserted in the chromosome of an ST5-MRSA-II strain of clinical origin

ObjectivesTo characterize the genetic element carrying the poxtA oxazolidinone resistance gene found in the poxtA index strain Staphylococcus aureus AOUC-0915 isolated from a cystic fibrosis patient.MethodsThe genetic context of poxtA was investigated by bioinformatics analysis of WGS data of strain AOUC-0915, followed by PCR and confirmatory Sanger sequencing for repetitive regions. Conjugation and electrotransformation experiments were carried out to assess horizontal transferability using S. aureus and Enterococcus faecalis recipients. Production of phage particles was evaluated by PCR using DNA preparations obtained after phage induction. Excision of the transposon carrying poxtA was evaluated by inverse PCR experiments for detection of circular intermediates.ResultspoxtA was found to be associated with a 48 kb composite transposon of original structure, named Tn6349, inserted into a φN315-like prophage. The transposon was bounded by two IS1216 insertion sequences, carried several resistance genes [erm(B), cfr, poxtA and fexB] and exhibited a mosaic structure made by a derivative of plasmid pE35048-oc (previously described in an Enterococcus faecium clinical isolate) and Tn6657, a novel composite transposon carrying the poxtA and fexB genes. Excision ability of Tn6349 as a circular intermediate was demonstrated. Transferability of Tn6349 or modules thereof to S. aureus or E. faecalis by either conjugation or electrotransformation was not detected. Induction of the φN315-like prophage carrying Tn6349 was not observed.ConclusionsThis study describes the structure of Tn6349, a novel composite transposon carrying several resistance determinants to anti-ribosomal drugs, including cfr and poxtA, from an oxazolidinone-resistant MRSA strain. Analysis of Tn6349 revealed a modular structure that could favour the mobilization of its resistance determinants.

Evaluation of Cervical Elastography Strain Pattern to Predict Preterm Birth

Purpose To evaluate cervical elastography strain pattern as a predictive marker for spontaneous preterm delivery (SPTD). Materials and Methods In this case-control study cervical length (CL) and elastographic data (strain ratio, elastography index, strain pattern score) were acquired from 335 pregnant women (20th – 34th week of gestation) by transvaginal ultrasound. Data of 50 preterm deliveries were compared with 285 normal controls. Strain ratio and elastography index were calculated by placing two regions of interest (ROIs) in parallel on the anterior cervical lip. The strain ratio was determined by dividing the higher strain value by the lower one. The elastography index was defined as the maximum of the strain ratio curve. Elastographic images were assigned a new established strain pattern (SP) score between 0 and 2 according to the distribution of strain induced by compression. Results Elastography index, SP score and CL differed between preterm and normal pregnancies (1.61 vs. 1.27, p < 0.001; SP score value of “2”: n = 31 (62 %) vs. n = 36 (12.6 %), p < 0.001; CL 30.7 vs. 41.0 mm, p < 0.001; respectively). The elastography index and SP score were associated with a higher predictive potential than CL measurement alone (AUC 0.8059 (area under the curve); AUC 0.7716; AUC 0.7631; respectively). A combination of all parameters proved more predictive than any single parameter (AUC 0.8987; respectively). Conclusion Higher elastography index and SP scores were correlated with an elevated risk of SPTD and are superior to CL measurement as a predictive marker. A combination of these parameters could be used as a “Cervical Index” for the prediction of SPTD.

The importance of compression elastography in the evaluation of thyroid nodule malignancy

Introduction/Objective. Compression, also called strain elastography imaging techniques, represent new echotomographic modality, which is a promising method for the differentiation of benign from malignant lesions, not only in the thyroid gland but also in other organs. The objective of this study is to evaluate the importance of compression elastography in the differentiation of benign and malignant thyroid nodules. Methods. We performed echotomographic examinations in B mode, and examinations using compression elastography in a total of 186 persons (152 females and 34 males, with the average age of 45.3 ? 13.5 years), with 264 nodules in the thyroid gland. Elastography was done in two steps: the first one through scoring elastographic figures, and the second one through the determination of the resistance index (strain ratio ? SR). Results. Using elastography scores by Fukunari, 44 of 60 malignant nodules had a score of 3?4, while 152 of the 204 benign nodules had a score of 1?2. Using the receiver operating characteristic (ROC) analysis, the best cut-off point obtained using elastography scores was 2, with a sensitivity of 73.3% and specificity of 74.5%. Using the software-calculated SR we found that out of 89 nodules with SR ? 2.5, 52 were malignant nodules, while out of 175 nodules with SR < 2.5, 167 were benign nodules. Using the ROC analysis, the best cut-off point obtained using SR was > 2.5, with a sensitivity of 86.7%, and specificity of 81.9%. Conclusion. As a follow-up of standard echotomographic examination in B mode, compression elastography is a newly developed and promising technique in the differentiation of benign from malignant lesions.

Tn6249, a New Tn6162Transposon Derivative Carrying a Double-Integron Platform and Involved with Acquisition of theblaVIM-1Metallo-β-Lactamase Gene in Pseudomonas aeruginosa

ABSTRACTThe In70.2 integron platform appears to be a conserved structure involved in the dissemination of theblaVIM-1metallo-β-lactamase gene inPseudomonas aeruginosa. The genetic context of the In70.2 integron platform fromP. aeruginosaVR-143/97, the VIM-1-producing index strain isolated in Italy in 1997, was fully characterized by a next-generation sequencing approach refined by conventional sequencing. The In70.2 integron platform from VR-143/97 was found to be associated with a defective Tn402-like transposon inserted into theurf2gene of a Tn3family transposon of an original structure, named Tn6249, which also carried a partially deletedmeroperon and an In90 integron platform in a tail-to-tail orientation. Tn6249was inserted into a PACS171b-like genomic island, which was in turn inserted into theendAgene of thePseudomonaschromosomal backbone. Tn6249showed a similar structure and a conserved location with respect to that of Tn6060, a Tn3family transposon associated with In70.2 and carrying a double-integron platform, which was detected in a VIM-1-producingP. aeruginosastrain isolated in Australia in 2008. Both Tn6249and Tn6060are apparently derived from Tn6162, a mercury resistance transposon carrying an integron platform, which was found inP. aeruginosaisolates from different geographic locations. The conservation of the genetic context of Tn6249and Tn6060suggests anin situevolution of these elements after the insertion of a Tn6162-like ancestor into the PACS171b-like genomic island (GI) present in the genome of a successful widespreadP. aeruginosaclonal lineage.

Dissemination of a pSCFS3-Likecfr-Carrying Plasmid in Staphylococcus aureus and Staphylococcus epidermidis Clinical Isolates Recovered from Hospitals in Ohio

ABSTRACTNineteen linezolid-resistantStaphylococcus epidermidisand twoStaphylococcus aureusisolates recovered from two medical institutions in northeast Ohio and anS. aureuscfrindex strain previously collected in the same facilities during the 2007 SENTRY Antimicrobial Surveillance Program were investigated for the genetic basis of oxazolidinone resistance and the location ofcfr. S. aureusisolates were typed by pulsed-field gel electrophoresis (PFGE),spatyping, and multilocus sequence typing (MLST). The location ofcfrwas determined by Southern blotting and hybridization. Plasmid sequencing was performed using the 454 Life Sciences (Roche) GS-FLX DNA platform. The twoS. aureusisolates showed unique PFGE patterns but were multilocus sequence type 5 (ST5) andspatype t002, whereas theS. aureusindex strain was ST239 and t037. Southern blot and hybridization experiments showed thatcfrwas plasmid located and that theS. epidermidisisolates, one of theS. aureusisolates, and theS. aureusindex strain shared an identicalcfr-carrying plasmid (39.3 kb). Sequencing results confirmed these findings. A 10-kb fragment containingcfrshowed the highest identity (99.9%) to a 9.5-kb fragment of plasmid pSCFS3 from a bovineStaphylococcus lentusisolate from Germany. In addition, these 39.3-kb plasmids from humanS. epidermidisandS. aureusexhibited BglII restriction profiles very similar to that observed for plasmid pSCFS3. Thecfr-carrying plasmid detected in the remainingS. aureusisolate (7.9 kb) was distinct and showed the highest identity to the chromosomalcfrintegrate found in the chromosomal DNA of aProteus vulgarisisolate from a pig in China.