Evaluasi Program Penyediaan Air Minum Dan Sanitasi Berbasis Masyarakat (Pamsimas) Di Kecamatan Tembalang

Pembangunan tidak lain merupakan suatu proses Perubahan yang berlangsung secara sadar, terencana dan berkelanjutan dengan sasaran utamanya adalah untuk meningkatkan kesejahteraan hidup manusia. Salah satu pembangunan yang menjadi perhatian adalah kebutuhan air bersih dan sanitasi. Pamsimas merupakan salah satu bentuk solusi dari kurangnya air bersih dan sanitasi di Indonesia. Tetapi pelaksanaan Pamsimas masih belum optimal, tidak terkecuali pelaksanaan di Kecamatan Tembalang Kota Semarang. Tujuan dari penelitian ini adalah untuk mengevaluasi Program Penyediaan Air Minum dan Sanitasi Berbasis Masyarakat (Pamsimas) di Kecamatan Tembalang. Dalam evaluasi ini digunakan enam kriteria evaluasi yaitu efisiensi, efektivitas, kecukupan, perataan, responsivitas dan ketepatan. Hasil penelitian ini menunjukkan bahwa Pamsimas di Kecamatan Tembalang sudah efektif dalam memenuhi kebutuhan air bersih dan sanitasi. Tetapi masih ditemui kekurangan, seperti kurang meratanya pembangunan tower air dan perpipaan Pamsimas, selain tidak merata masih banyak warga miskin yang masih belum dapat mendapatkan Pamsimas. Rekomendasi untuk meningkatkan perataan pembangunan Pamsimas dapat dilakukan pada saat musyawarah penentuan prioritas pembangunan dilakukan oleh seluruh elemen pelaksana Pamsimas, hal ini agar tercipta keadilan dan pemerataan pada pembangunan karena diawasi langsung oleh semua pihak yang terkait

Characteristics of oligonucleotides used for <i>Echinococcus granulosus</i> complex multiplex PCR.

a<p>) Strict specific bases in each primer are written in bold. Tiny characters mark additional polymorphic sites (but not strict).</p

Specificity of the mPCR approach based on number of cycles.

<p>(<b>A–C</b>) Different quantities of <i>E. granulosus s.s.</i> (G1) DNA were used as templates in the mPCR: 5 ng (lane 1), 25 ng (lane 2), 50 ng (lane 3), 100 ng (lane 4) and 250 ng (lane 5). The mPCR was run with 25 cycles (<b>A</b>), 30 cycles (<b>B</b>) or 35 cycles (<b>C</b>) of amplification. For the <i>E. granulosus s.s.</i> (G1) template, the genotype was clearly detectable in all setups, but performing the mPCR with 30 or 35 cycles resulted in a visible background smear and some very light additional bands. A reduced setup was performed for the other genotypes (<b>D</b>). The mPCR was run with 5 ng (lanes 1, 3, 5, 7 and 9) or with 250 ng (lanes 2, 4, 6, 8 and 10) and 30 cycles of amplification. In contrast to <i>E. granulosus s.s.</i> (G1/G2/G3) (lanes 1 and 2) which showed only minor unspecific products, the mPCR amplified unspecific products for <i>E. equinus</i> (G4) (lanes 3 and 4), <i>E. ortleppi</i> (G5) (lanes 5 and 6), <i>E. canadensis</i> (G6/G7) (lanes 7 and 8) and <i>E. canadensis</i> (G8/G10) (lanes 9 and 10). Thus, additional numbers of PCR cycles result in unspecific PCR products hampering the readout. M: 100-bp DNA ladder (Promega).</p

Geographical origin, hosts and numbers of <i>E. granulosus</i> isolates and their corresponding species/strains based on multiplex-PCR results.

*<p><i>cox1</i>-sequenced samples: 14 ovine and 39 human Tunisian samples as well as the 6 ovine Argentinean samples were identified as <i>E. granulosus s.s.</i> (G1) and the 7 Argentinean porcine samples were identified as <i>E. canadensis</i> (G7).</p

Genotype profile of the <i>E. granulosus</i> complex by mPCR.

<p>(<b>A</b>) Schematic representation of the genotype specific banding patterns amplified by mPCR: (lane 1) <i>E. granulosus s.s.</i> (G1/G2/G3), (lane 2) <i>E. equinus</i> (G4), (lane 3) <i>E. ortleppi</i> (G5), (lane 4) <i>E. canadensis</i> (G6/G7), (lane 5) <i>E. canadensis</i> (G8/G10), (lane 6) <i>E. multilocularis</i> and (lane 7) <i>E. vogeli</i>. The product sizes are specified in bp and the corresponding genes are shown in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002017#pntd-0002017-t001" target="_blank">Table 1</a>. (<b>B</b>) Result of a mPCR using 5 ng of purified template DNA of the known <i>Echinococcus</i> species described above (lanes 1–7) visualized on a 2% agarose gel. The target of 1232 bp is specific for the <i>Echinococcus</i> genus and is also amplified for <i>E. multilocularis</i> (lane 6) and <i>E. vogeli</i> (lane 7). The 110 bp band allows specific detection of <i>E. granulosus</i> complex members (lanes 1–5). All bands between 1232 bp and 110 bp specifically detected one <i>E. granulosus</i> complex species/genotype and showed no cross-reactivity with other members. (<b>C</b>) Specificity test of the mPCR for the genus <i>Echinococcus</i> and other closely related cestodes of the family; <i>E. granulosus</i> (G1/G2/G3) (lane 1), <i>E. multilocularis</i> (lane 2), <i>E. vogeli</i> (lane 3), <i>T. saginat</i>a (lane 4), <i>T. solium</i> (lane 5), <i>T. crassiceps</i> (lane 6), <i>T. taeniaformis</i> (lane 7) and <i>T. pisiformis</i> (lane 8). The expected banding pattern was observed for <i>E. granulosus</i> (G1/G2/G3) (lane 1), <i>E. multilocularis</i> (lane 2) and <i>E. vogeli</i> (lane 3) and no PCR products were detected for the <i>Taenia</i> samples. N: PCR-negative control (ddH₂O). M: 100-bp DNA ladder (Promega).</p

Direct mPCR on frozen and fixed <i>E. granulosus</i> complex material.

<p>(<b>A</b>) 1 µl (lane 1) or 2 µl (lane 2) of previously frozen hydatid fluid aspirated from an equid cyst was used directly in the mPCR without resulting in genotype specific PCR products. In parallel, 1 ml of the hydatid fluid was boiled for 30 min followed by a centrifugation step. Different volumes of the resulting supernatant were used in the mPCR (0.25 µl lane 3, 0.5 µl lane 4, 1 µl lane 5, 1.5 µl lane 6, 2 µl lane 7, 2.5 µl lane 8, 3 µl lane 9, 10 µl lane 10). Note that using 1–3 µl resulted in the detection of <i>E. equinus</i> (G4), although with some minor additional background amplicons. Frozen (<b>B</b>) and EtOH-fixed (<b>C</b>) <i>E. granulosus s.s.</i> (G1) protoscoleces were treated by alkaline lysis and the supernatant was used without (lanes 1 and 2) or with dilution (1∶1 lane 3, 1∶2 lane 4, 1∶4 lane 5, 1∶6 lane 6, 1∶8 lane 7, 1∶10 lane 8, 1∶25 lane 9) for mPCR. Undiluted supernatant (1 µl lane 1 and 2 µl lane 2) resulted in failed mPCR in both setups. If 2 µl of diluted supernatant was used for mPCR, genotyping was successfully performed for frozen protoscoleces on dilution ratios of 1∶8 to 1∶10 and for EtOH-fixed protoscoleces on ratios between 1∶2 to 1∶4. M: 100-bp DNA ladder (Promega).</p

Detection limit of one species in a dual-species DNA mixture.

<p><b>A</b>) To mimic a mixture of different <i>Echinococcus granulosus</i> complex members, as it can occur in egg-derived samples, DNA from <i>E. granulosus s.s.</i> (G1) and <i>E. canadensis</i> (G6) was mixed in different ratios and the mPCR was performed using 250 ng (lanes 1–5), 50 ng (lanes 6–9) or 5 ng (lanes 10–16) DNA template. The detection limit of one species in a dual-species DNA mixture was measured at 5% (lane 4, 250 ng template DNA), 2.5% (lane 6, 50 ng template DNA) and 20% (lanes 11 and 15, each 5 ng template DNA). <b>B</b>) To test if all 11 targets can be amplified in parallel, 5 ng template DNAs from <i>E. granulosus s.s.</i> (G1), <i>E. equinus</i> (G4), <i>E. ortleppi</i> (G5), <i>E. canadensis</i> (G6) and <i>E. canadensis</i> (G10) were mixed and used together in one single mPCR. All targets were successfully amplified and no missing or non-specific amplicon was detected (lane 1). Lane 2 shows the virtual banding pattern. Amplicon sizes and genotype specificities are marked on the left side. M: 100-bp DNA ladder (Promega).</p