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Pakistan Journal of Biological Sciences
eISSN: 1812-5735
pISSN: 1028-8880

Editor-in-Chief:  Hakan Ulukan
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Research Article
Phenotypic and Molecular Characterization of Multidrug Resistant Klebsiella pneumoniae Isolated from Different Clinical Sources in Al-Najaf Province-Iraq
Ahmed Abduljabbar Jaloob Aljanaby and Haneen Mohammed Reda Jaber Alhasnawi
Background and Objective: Burns infections and urinary tract infections are the most important prevalent diseases in Asian countries, such as Iraq. Klebsiella pneumoniae is one of the most important bacteria cause this type of infections especially in hospitals. Therefore, the aim of this study was to investigate the prevalence of multi-drug resistance K. pneumoniae and extended-spectrum beta-lactamases producing K. pneumoniae isolates from inpatients with urinary tract infection and burns infections in Al-Kufa hospital in Al-Najaf province, Iraq. Materials and Methods: A total of 285 clinical samples were collected from in-patients infected with urinary tract infection (141 urine samples) and burns infections (144 burns swabs). Fourteen different antibiotics were used by disc diffusion method and 13 antimicrobials resistance genes were used by PCR technique. Results: A total of 43 K. pneumoniae strains were isolated. The highest resistance rate was observed for amoxicillin 25 μg and amoxicillin+clavulanic acid 20+10 μg (97.67%) while the lowest resistance rate was observed for imipenem 10 μg (9.30%). The most common resistance associated-genes were blaSHV (86.04%) and at lower prevalence were IMP (9.30%). Conclusion: Klebsiella pneumoniae strains isolated from burns infections were more virulent than those isolated from urinary tract infections.
Research Article
Cloning and Expression Analysis of HbPR-1b and HbPR-3 in Hevea brasiliensis During Inoculation with Rigidoporus microporus
Natthakorn Woraathasin, Korakot Nakkanong and Charassri Nualsri
Background and Objective: Pathogenesis-related (PR) proteins are dramatically accumulated after pathogen infection. Innate defense response through increasing PR-proteins is important for rubber rootstock selection that is tolerant to the white root disease caused by Rigidoporus microporus. This study was aimed to investigate the expression levels of PR-1 and PR-3 genes in tolerant (PB5/51) and susceptible (BPM24 and RRIM600) rubber clones after R. microporus infection. Materials and Methods: The mRNA of HbPR-1b and HbPR-3 was isolated and characterized from rubber leaves. Gene expression levels of HbPR-1b and HbPR-3 were compared among three rubber clones (PB5/51, BPM24 and RRIM600) after R. microporus infection at 0, 12, 24, 48, 72 and 96 h using quantitative real-time PCR. The relative transcript abundances between inoculated and control plants were compared using the means of gene expression between time points and by Tukey’s HSD test. A probability value (p<0.05) was used to determine the significance of difference between time points. Results: The open reading frame of HbPR-1b is 492 bp with deduced 163 amino acid residues and the phylogenetic analysis showed it shared significant evolutionary history and clustering into group I of PR-protein. Moreover, the partial HbPR-3 was isolated with 390 bp. Gene expression levels of HbPR-1b and HbPR-3 showed marked differences in both transcripts depending on the rubber clones. Two genes demonstrated up-regulation of both tolerance and susceptibility in response to attack by R. microporus. The highest expression levels were found in seedlings of PB5/51 after inoculation. In RRIM600, low expression levels of HbPR-1b and HbPR-3 were initially observed but gradually increased at 24 h post inoculation. The transcription profile of HbPR-1b was stable expression in BPM24. Conclusion: The results demonstrated that the level ofHbPR-1b and HbPR-3 transcription can distinguish between tolerant and susceptible clones. The candidate defense genes to the white root disease were observed in PB5/51 seedlings, particularly HbPR-1b.
Research Article
Callus Induction from Various Organs of Dragon Fruit, Apple and Tomato on some Mediums
Rumiyati , Sismindari , Endang Semiarti, Asri Fajar Milasari, Dheatika Karina Sari, Nia Fitriana and Sekar Galuh
Background and Objective: Dragon fruit (Hylocereus spp.), apple (Malus sylvestris Mill.) and tomato (Solanum lycopersicum L.) are high potential sources of antioxidant compounds such as phenolics. The compounds have the capability of protecting cells and tissues against free radicals. Secondary metabolite produced by callus cell culture from plant organs also acts as a source of antioxidants. This study aimed to determine the optimal ratio of sucrose and 2,4-D in Murashige and Skoog (MS) medium for callus induction from different plant organ explants. With all of characteristic, callus can be used further for the development of natural cell regeneration agent. Methodology: This study was conducted using analytical technique. Suitable explants were obtained. They were developed in various concentrations of combination between MS medium and 2,4-D. Callus growth, including their weight and surface was then measured and analyzed by using one-way analysis of variance (ANOVA). Results: Callus was able to grow from its explants in 5-7 days after induction process. They were clear in color and had friable texture. The highest value of fresh weight of dragon fruit callus was obtained through MS supplemented with 1 μL L–1 2,4-D and 30 g sucrose. However, apple and tomato callus induction and growth maintenance reached optimal medium on MS supplemented with 30 g sucrose and 2 μL L–1 2,4-D. Conclusion: Callus of apple, dragon fruit and tomato was maintained upon MS supplemented with 30-40 g sucrose and 1-2 μL L–1 2,4-D for optimum induction and growth. The optimization of growth medium will give advantages for further development of natural cell regeneration agent.
Research Article
Influence of Light Intensity and Photoperiod on the Seed Germination of Four Rhododendron Species in Taiwan
Lei-Chen Lin and Chang-Sheng Wang
Background and Objective: There are 15 native Rhododendron species in Taiwan, among which 11 species are endemic and compose 73% of these native species. Although researchers predominantly use cuttings to propagate Rhododendron shrubs, there are no studies on the seed germination of Rhododendron species. The objective of this study was to evaluate the seed germination of four Rhododendron species in Taiwan under different light intensities and photoperiods. Materials and Methods: Two experiments on the seed germination percentage of R. breviperulatum, R. kanehirai, R. ovatum and R. simsii were conducted in this study. The first experiment was to identify the seed germination percentage of these four Rhododendron species using different light intensities (0, 700, 1400 and 3200 lux). The second experiment was to clarify the seed germination percentage of these four Rhododendron species using different photoperiods (0, 1, 4 and 16 h). All statistical analyses were performed using Statistical Package for the Social Science (SPSS12.0) for Windows software program. The data were analyzed using Tukey’s multiple range test at the p<0.05 significance level. Results: After 30 days, no seed germination occurred in darkness. The highest average seed germination percentages were all observed at 700 lux: R. breviperulatum (83.3%), R. kanehirai (68.9%), R. ovatum (85.6%) and R. simsii (92.2%). The highest average germination percentages of seeds were observed in R. breviperulatum at 16 h (83.3%), R. kanehirai at 1 h (60.0%), R. ovatum at 16 h (84.4%) and R. simsii at 16 h (85.6%). According to the results, these four Rhododendron species required light for germination. There were significant differences (p<0.05) in the seed germination of these four Rhododendron species for light intensity greater than 700 lux. Similar results were observed with photoperiods. Conclusion: The seed germination percentage of R. breviperulatum, R. ovatum and R. simsii increased with increasing photoperiod.
Research Article
Evaluation of Autothermal Thermophilic Aerobic Digester Performance for the Stabilization of Municipal Wastewater Sludge
Reza Shokoohi, Alireza Rahmani, Ghorban Asgari, Abdollah Dargahi, Yaser Vaziri and Mohammad Attar Abbasi
Background and Objective: Sludge stabilization process in terms of operational, environmental and economic indexes is the most important stage of treatment and its disposal. This study was aimed to determine the performance of Autothermal Thermophilic Aerobic Digestion (ATAD) system as one of the low-cost and biocompatible methods of sludge treatment. Materials and Methods: This study has been done using a laboratory scale Autothermal Thermophilic Aerobic Digestion (ATAD). The reactor was consisted of two polyethylene tanks with a final capacity of 100 L for each tank. Both tanks with all fittings were installed on a metal frame. The variables of study were temperature, dissolved oxygen, pH, volatile organic compounds, total solids, COD and the number of Ascaris eggs and fecal coliforms per gram of dry matter of the sludge. The temperature was measured hourly and the pH and dissolved oxygen were measured and controlled twice per day. One-way ANNOVA was applied to analyze reasults. Results: According to the results, the temperature of sludge increased from 11.7-61.2°C by biological reactions. Pathogen organisms were reduced from 80×106 to 503 in number during 72 h. After 6 days pathogen organisms and Ascaris eggs were removed completely. Volatile organic compounds and COD were reduced 42 and 38.3% respectively during the 6 days. Conclusion: It is concluded that the performance of ATAD in removing organic compounds from wastewater sludge were desirable. Resulted sludge from stabilization process were appropriate for use in agriculture as a soil supplement and met the indexes of class A sludge according to EPA’s standards (CFR 40 Part 503).

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