Jumlah Bakteri Staphylococcus Aureus Dan Skor California Mastitis Test (CMT) Pada Susu Kambing Peranakan Etawa Akibat Dipping Ekstrak Daun Babadotan (Ageratum Conyzoides L.)

The aim of this research is to determine the effect of teat dipping of Ettawa crossbred goat using babadotan leaves (Ageratum conyzoides Linn.) extract on the number of Staphylococcus aureusin milk. The udder inflammation degree also was determined using California Mastitis Test (CMT). The treatments were post milking teat dipping using antiseptic solutions containing 1%, 3%, and 5% of babadotan leaves extract (T1, T2 and T3, respectively). Milk samples were collected at before treatment (H0) and on the day 3, 6 and 9 day of the treatments (H3, H6 and H9, respectively). Commercially antiseptic povidone iodine was used as positive control (K+). Experimental research design was completely randomized design (CRD) split plot types, with the different extract concentration as the main plot and the day of treatment as subplot. CMT scores was analyzed using Kruskal-Wallis test. The results showed that babadotan leaves extract 5% had the same effectiveness (p>0,05) with povidone iodine to reduce the number of Staphylococcus aureusin milk. All extract concentrations (1%, 3% and 5%) had the same effectiveness (H>c0,05(3)) to decrease the CMT scores by postmilking teat dip treatments for 9 days

Use of alternate TSSs at LSP compared to ESP.

<p>Letters A to H annotate the different categories of arrangements of TSSs found at LSP compared to ESP that we were able to identify. Letters X, Y, Z and W represent TSS positions at ESP and LSP. The number of genes for which each type of rearrangement occurs is shown. For nucleotide positions of TSSs and genes to which the different types of rearrangements apply see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092690#pone.0092690.s008" target="_blank">Table S2</a>. Red arrows indicate TSS positions that were identified at LSP but not ESP. Rearrangements which apply to one or two genes are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092690#pone.0092690.s002" target="_blank">Fig. S2</a>.</p

Transcriptional architecture of the entire SPI1 and SPI2 loci at ESP and LSP in parental and Δ<i>relA</i>Δ<i>spoT</i> strains.

<p>Enriched (+) and non-enriched (−) cDNAs of <i>S</i>. Typhimurium parental (black) or ppGpp⁰ (<i>ΔrelAΔspoT</i>) (red) strains mapped onto the SPI1 and SPI2 loci. Operons are indicated by horizontal arrows and annotated according to the first gene. The Y axis in each lane represents 0–50 mapped reads per genome position. The dRNA-seq data for ESP was from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092690#pone.0092690-Ramachandran1" target="_blank">[19]</a>.</p

Promoter architecture and ppGpp-dependency of SPI2.

<p>Enriched (+) and non-enriched (−) cDNAs of <i>S</i>. Typhimurium SL1344 parent (black) or ppGpp⁰ (<i>ΔrelAΔspoT</i>) (red) strains mapped onto selected regions of the SPI2 locus. Operons are indicated by horizontal arrows and annotated according to the first gene. Transcript start sites are indicated by black arrows (A) Map of <i>ssaB</i> and <i>sseA</i> operons. The Y axis in each lane represents 0–50 mapped reads per genome position. (B) Map of <i>ssaR</i> operon. The Y axis in each lane represents 0–150 reads. (C) Map of the <i>ssrA</i> operon showing transcriptional read through into <i>ssrB</i> and <i>orf242</i>. Y axis in each lane represents 0–30 reads. (D) Location of transcript start sites identified previously and in this study; N.D.  =  not determined.</p

Correlation of ppGpp-repression of LSP TSSs of annotated genes with %GC content of discriminator regions.

<p>The analysis of GC content within the discriminator region was calculated using a custom Perl script. The inset shows the GC rich conserved region identified using MEME within the discriminator regions of promoters that were ppGpp-repressed by 20-fold or greater.</p

Annotation of TSSs.

<p>(A) TSSs were defined as primary (P), secondary (S) or internal (I). Primary TSSs were identified as having higher mapped read counts relative to secondary TSSs. Internal TSSs were located within the coding region (CDS) of a gene where a TSS was annotated for the gene immediately upstream. Primary and secondary internal TSSs (P,I and S,I respectively) were located within the CDS of a gene where there were no TSSs annotated for the gene immediately downstream. (B) Venn diagram showing overlap of <i>S</i>. Typhimurium LSP TSSs categories for all annotated genes.</p

Expression of an <i>S</i>. Typhimurium <i>sipC</i>::<i>lacZ</i> fusion in parental, Δ<i>relA</i>Δ<i>spoT</i> (ppGpp⁰), Δ<i>dksA</i> and Δ<i>rpoS</i> backgrounds during growth in aerobic LB batch cultures.

<p>Data is from 3 biological replicate experiments.</p

Effect of loss of ppGpp, Rpos and/or DksA on SPI1 and SPI2 expression at late-log phase (LLP), early stationary phase (ESP), mid-stationary phase (MSP) and late stationary phase (LSP).

<p>(A, E) SPI1 and SPI2 transcript levels in parent strain (SL1344); 1, 2, and 3 indicate biological replicate cultures. (B, F) SPI1 and SPI2 transcript levels in Δ<i>relA</i>Δ<i>spoT</i> (P) and Δ<i>rpoS</i> (R) strains; transcript levels are normalised to parental (WT) SPI1 transcript levels. (C, G) SPI1 and SPI2 transcript levels in a Δ<i>dksA</i> (D) strain normalised to parent (WT) strain. (D, H) Late stationary phase SPI1 and SPI2 transcript levels in Δ<i>relA</i>Δ<i>spoT</i> (P), Δ<i>rpoS</i> (R), and Δ<i>relA</i>Δ<i>spoT</i>Δ<i>rpoS</i> (RP) and Δ<i>relA</i>Δ<i>spoT</i>Δ<i>dksA</i> (DP) strains normalised to transcript levels in the SL1344 parent strain. Data from which the figure was compiled and statistical analysis is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127523#pone.0127523.s002" target="_blank">S2 Fig</a> and also deposited at Gene Expression Omnibus (GEO), superseries accession number GSE63715.</p

ChIP-chip profiles of RNAP distribution at the (A) SPI1 and (B) SPI2 loci in parental and Δ<i>dksA</i> and Δ<i>relA</i>Δ<i>spoT</i> strains at ESP and LSP viewed on the integrated genome browser (IGB) [65].

<p>Resolution of each peak is 100 nt.</p

Glycolysis is important but not essential for the invasion and intracellular replication of HeLa cells with <i>S</i>. Typhimurium.

<p>(A) Invasion assay of <i>S</i>. Typhimurium 4/74 parental and Δ<i>pfkAB</i> (JH3486) strains in HeLa cells (B). Intracellular replication assays of <i>S</i>. Typhimurium 4/74 parental and Δ<i>pfkAB</i> (JH3486) strains during infection of HeLa cells. The chart shows the percentage replication of bacteria between 2 h and 6 h. (C) Complementation of invasion of the <i>S</i>. Typhimurium Δ<i>pfkAB</i> strain in HeLa cells. (D) Complementation of intracellular replication of the <i>S</i>. Typhimurium Δ<i>pfkAB</i> strain in HeLa cells. Each bar represents the statistical mean from three biological replicates and the error bars represent the standard deviation. (The significant differences between the parental 4/74 strain (A, B), or the 4/74 (pWKS30) strain (C, D) and the mutant strains are shown by asterisks, *<i>p</i><0.05. **<i>p</i><0.01, and ***<i>p</i><0.001).</p