Molecular Detection of Bartonella quintana among Long-Tailed Macaques (Macaca fascicularis) in Thailand

Bartonella quintana is a zoonotic pathogen with a worldwide distribution. Humans and non-human primates are considered to be natural reservoir hosts for B. quintana. However, information on the molecular epidemiology of this organism is very limited in regard to long-tailed macaques (Macaca fascicularis) in Thailand. Therefore, this study aimed to investigate the occurrence and genetic diversity of Bartonella spp. among long-tailed macaques in Thailand. In total, 856 blood samples were collected from long-tailed macaques in Thailand. All specimens were screened for Bartonella spp. using a polymerase chain reaction (PCR) assay targeting the 16S rRNA, gltA and ftsZ genes. All positive samples were further analyzed based on nucleotide sequencing, phylogenetic analysis and multiple sequence alignment analysis. Only one macaque showed a positive result in the PCR assays based on the 16S rRNA, gltA and ftsZ genes. Nucleotide sequencing and phylogenetic analysis revealed that the obtained sequences were closely related to B. quintana previously detected in non-human primates. Single-nucleotide polymorphisms (SNPs) were detected in the gltA and ftsZ gene sequences. This study revealed that long-tailed macaques in Thailand carried B. quintana. Despite the low infection rate detected, long-tailed macaques may be a reservoir of B. quintana.

PENGELOMPOKKAN SEPULUH VARIETAS TEMBAKAU (Nicotiana tabacum) BERDASARKAN KERAGAMAN RUNUTAN BASA PARSIAL GEN PMT(PUTRESCINE N-METHYLTRANSFERASE) Clustering of ten tobacco (Nicotiana tabacum) varieties based on the partial PMT (putrescine N-methyltranfera

<p align="center">Abstrak</p><p>Gen <em>PMT</em> adalah gen penyandi enzim putresina N-metiltransferase (PMT) yang berperan dalam lintasan biosintesis nikotin pada tanaman tembakau (<em>Nicotiana tabacum</em>). Sepuluh varietas tembakau yang memiliki perbedaan tingkat kadar nikotin diuji untuk mempelajari: (1) keragaman runutan basa parsial gen <em>PMT</em> dari masing-masing varietas, dan (2) kekerabatan antara sepuluh varietas tembakau yang diuji berdasarkan keragaman runutan basa parsial gen <em>PMT</em>. Keragaman runutan basa dianalisis dengan mensejajarkan data runutan basa dari sepuluh varietas tembakau yang diuji dengan runutan basa dari <em>Ntpmt_Sindoro1</em> (JQ438825) yang telah tersimpan dalam <em>database</em><em> </em><em>genbank NCBI</em>. Hasil pensejajaran digunakan untuk menghitung matriks jarak, yang selanjutnya digunakan untuk menganalisis hubungan kekerabatan diantara sepuluh varietas tembakau. Hasil analisis memperlihatkan adanya variasi ukuran dan jumlah runutan basaparsial gen <em>PMT</em> asal sepuluh varietas tembakau yang dianalisis. Hasil analisis juga memperlihatkan bahwa runutan basa parsial gen <em>PMT</em> tersebut berasal/diturunkan dari sumber (<em>ancestor</em>) yang sama dan terkait dengan biosintesis nikotin pada tembakau. Runutan basaparsial gen <em>PMT</em> dari sepuluh varietas yang dianalisis memisahkan antara kelompok tembakau introduksi (kadar nikotin rendah-sedang) dengan kelompok tembakau lokal (kadar nikotin sedang-tinggi). Dua kelompok memisah berdasarkan level kadar nikotin, danperbedaan/perubahan susunan basa pada situs-situs tertentu dari runutan basaparsial gen <em>PMT</em> yang dianalisis. Informasi tentang mutasi yang terjadi pada situs-situs runutan basa dari parsial gen <em>PMT</em><em> </em>dapat digunakan untuk mempelajari keterkaitan antara perubahan basa pada fragmen gen <em>PMT</em> dengan kandungan nikotin total tembakau yang terjadi selama proses evolusi.</p><p>Kata kunci: Analisis pengelompokkan, gen <em>PMT,</em>Nikotin, <em>Nicotiana tabacum</em></p><p align="center"><strong> </strong></p><p align="center">Abstract</p><p><strong> </strong><em>PMT</em> gene is the gene encoded putrescine N-methiltransferase which is related to nicotine biosinthesis in tobacco (<em>Nicotiana tabacum</em>). Ten tobacco varieties with different nicotine level were used inthis study. The aims of this study were: (1) to analyze thepartial <em>PMT</em> gene sequence diversity among ten tobacco varieties, and (2) to evaluate the closed-relationship amongten tobacco varieties based on their partial<em>PMT</em> gene sequences diversity.Sequence diversity was analyzed by multiple sequence alignment between the partial<em>PMT</em> gene sequence of the ten tobacco varietiesand <em>Ntpmt_Sindoro1 </em>sequence deposited in the NCBI gene-bank database.The phylogenetic relationship amongthe sequences was inferred by genetic distancebetween pairs of sequences using the pairwise and multiple sequence alignment analysis. Analysis of the sequences showed that all varieties analyzed had varied in size and number of the <em>PMT</em> gene fragments yielded. The analysis also revealed that thepartial<em>PMT</em> gene sequencesarecoming from the same ancestor which related to nicotine biosynthesis in tobacco. Phylogenetic analysis separated the partial<em>PMT</em> gene sequences into two different branches significantly (bootstrap value = 100), and clustered together based on tobacco types with different nicotine level in whichcould be due to some baseschanged on the specific sites of the<em>PMT</em> gene sequences. This information could be used to study the relationship between some bases changed on the specific sites of the<em>PMT</em> gene sequences and the nicotine content variation yielded by the ten tobacco varieties that is happened during evolution time.</p><p>Key words: Clustering analysis, <em>PMT</em> gene, nicotine, <em>Nicotiana tabacum</em></p>

Sequence and analysis of the mitochondrial DNA control region in the sugarcane borer Diatraea saccharalis (Lepidoptera: Crambidae)

This study aimed at the sequence and analysis of the mtDNA control region (CR) of the Diatraea saccharalis. The genome PCR amplification was performed using the complementary primers to the flanking regions of Bombyx mori CR mitochondrial segment. The sequencing revealed that the amplified product was 568 bp long, which was smaller than that observed for B. mori (725 bp). Within the amplified segment, a sequence with 338 nucleotides was identified as the control region, which displayed a high AT content (93.5%). The D. saccharalis mtDNA CR multiple sequence alignment analysis showed that this region had high similarity with the Lepidoptera Cydia pomonella.