Biotechnology Journal International <p style="text-align: justify;"><strong>Biotechnology Journal International (ISSN:&nbsp;2456-7051)</strong> publishes original research papers, review articles and short communications on all areas of Biotechnology including cell biology, genetics, microbiology, immunology, molecular biology, biochemistry, embryology,&nbsp; immunogenetics, cell and tissue culture, molecular ecology, genetic engineering and biological engineering, bioremediation and biodegradation, bioinformatics, biotechnology regulations, pharmacogenomics, gene therapy, plant, animal, microbial and environmental biotechnology.&nbsp;The journal also encourages the submission of useful reports of negative results. This is a quality controlled,&nbsp;OPEN&nbsp;peer reviewed, open access INTERNATIONAL journal.</p> en-US (Biotechnology Journal International) (Biotechnology Journal International) Thu, 17 Sep 2020 11:11:39 +0000 OJS 60 Effects of Cadmium Stress on Photosynthetic Physiological Characteristics of Two Rorippa Species <p>The photosynthetic physiological characteristics of <em>Rorippa amphilia</em> and <em>Rorippa sylvestris</em> were studied by pot experiment under Cd stress, and 7 Cd treatments were 0, 1, 5, 10, 50, 100, 200(mg·kg<sup>-1</sup>)respectively. The results showed that, (1) The Chla, chlb, and Ch(la+b) changed little when the Cd concentration in the soil was less than 100 mg·kg<sup>-1</sup>. There was no significant difference among the treatments. When Cd concentration was more than or equal to 100 mg·kg<sup>-1</sup>, all indexes decreased significantly, Chla/b increased gradually with the increase of Cd stressful concentration; (2) Net photosynthetic rate (<em>P<sub>n</sub></em>), stomatal conductance (<em>G<sub>s</sub></em>) and transpiration rate (<em>T<sub>r</sub></em>) first increased and then decreased with the increase of Cd stressful concentration, while intercellular carbon dioxide concentration (<em>C<sub>i</sub></em>) increased significantly; (3) The photochemical quenching coefficient (qP) and electron transfer rate (ETR)&nbsp; decreased gradually with the increase of Cd stressful concentration, while the non-photochemical quenching coefficient (NPQ) increased with the increase of Cd stress concentration. It indicated that the photosynthetic mechanism of leaves was damaged; the maximum photochemical quantum yield (Fv/Fm) and potential activity (Fv/Fo) of PSII decreased gradually with the increase of Cd stress concentration. When Cd concentration was more than 50 mg·kg<sup>-1</sup>, the decrease was very obvious, showing photoinhibition. The concentration of Cd below 50 mg·kg<sup>-1</sup> had little effect on photosynthesis. These results provide a theoretical application by using these two species of <em>Rorippa</em> to ecological restore the Cd contaminated farmland and abandoned mines.</p> Yuan Yun- Ning, Wang Lin- Yu, Chen Hao, Xu Yu- Feng ##submission.copyrightStatement## Wed, 04 Nov 2020 00:00:00 +0000 Selection and Validation of Reference Genes for Quantitative Real-Time PCR Expression Analysis of Candidate Genes in Carposina sasakii (Lepidoptera: Carposinidae) <p><em>Carposina sasakii</em> is one of the most important pests on the quality of stone and pome fruits. Investigation of a gene expression level in the species is hampered because of the gap of validated reference genes. The expression variation in the transcription levels of eight candidate reference genes, Actin (<em>ACT</em>), Tubulinbeta-1 (<em>TUB</em>), Ribosomal protein 49 (<em>RP49</em>), Elongation factor1-alpha (<em>EF-1a</em>), Elongation factor1-b (<em>EF-1b</em>), Elongation factor1-d (<em>EF-1d</em>), Ribosomal proteinL13 (<em>RPL13</em>) and Glyceraldehyde-3-phosphate dehydrogenase (<em>GAPDH</em>), were analyzed by quantitative real-time PCR (qPCR). The stability and ranking of these gene expression profiles in three organ types (head, thorax and abdomen), three developmental stages (larva, pupa and moth), and five diapause states (non-diapause, pre-diapause, diapause 0 d, diapause 20 d and diapause 60 d) were assessed using two algorithm-based methods, geNorm and NormFinder. <em>EF-1a</em>, <em>ACT</em> and <em>GAPDH</em> were evaluated to be the three stable reference genes based on the important observations and comprehensive analysis, whereas <em>TUB</em> and <em>EF-1b </em>showed low expression stability. Best gene combinations for different qPCR analysis in <em>C.</em> <em>sasakii </em>could be chosen from the three stable reference genes, the using of two reference genes is sufficient to effectively normalize qPCR data in <em>C. sasakii</em>. The study laid the foundation for gene expression analysis in <em>C.</em> <em>sasakii </em>and provided new information for the selection of reference genes.</p> Zuobing Yan, Yongli Li, Zhou Zhou, Yongan Zhang, Liangjian Qu ##submission.copyrightStatement## Thu, 31 Dec 2020 00:00:00 +0000 Impact of Drifts Resulting from Pesticide Application on Soil Microorganisms around Waste Receptacles in Port Harcourt City, Nigeria <p>Pesticides are toxic substance used to reduce or kill pests but the deposits on soil environment can remain there for long period of time causing adverse effects on soil microorganisms which are responsible for soil health conditions. This study was carried out to determine the impact of pesticide drifts on soil microorganisms in a waste receptacle around Port Harcourt city. Soil samples were obtained from various depths around waste receptacles with hand auger using standard analytical procedures. Microbial analysis was done according to prescribed standard methods. Characterization and identification of the isolates were based on their cultural, morphological, and cellular characteristics. Results obtained showed that the bacterial isolates were identified as <em>Staphylococus aureus</em>, <em>Bacillus subtilis</em>, <em>Bacillus megaterium</em>, <em>Pseudomonas </em>sp and <em>Micrococcus </em>sp while fungal isolates include <em>Aspergillus niger, Penicillium </em>sp, <em>Fusarium siculi, and Aspergillus fumigatus,</em> <em>Aspergillus nidulas, Microsporium canis </em>and <em>Yeast. </em>The results of the microbial counts revealed that Total Heterotrophic Bacteria (THB) had 2.08 ×10<sup>9</sup> cfu/g at a depth of 30-45 cm while Total Heterotrophic Fungi (THF) had 6.0×10<sup>6 </sup>cfu/g before application with a mean value of 1.02×10<sup>9</sup><sub>&nbsp;&nbsp; </sub>and&nbsp; <sub>&nbsp;&nbsp;</sub>2.8 × 10<sup>6 </sup>cfu/g respectively while after application THB had 4.1 × 10<sup>8</sup> and 4.6 × 10<sup>8 </sup>cfu/g for 0-15 and 30-45 cm respectively while the THF recorded 1.0 × 10<sup>6 </sup>and 0.6 × 10<sup>6 </sup>cfu/g for 0-15 cm and 30-45 cm respectively. However there was a drastic decrease in the number of microbes in the soils after pesticide application especially at the surface soil depth of 0-15 cm. This shows that the pesticides application affected microbial population by reducing their numbers in the soil and this may in turn affect soil health and physiological status of their habitat. It is therefore recommended that proper surveillance during pesticide application should be considered to avoid drift to non-target organisms and that concentrations of pesticides to be used should be taken into account to avoid reduction in the number of microorganisms in soils because of the vital roles they play in maintaining soil health.</p> D. Momoh, C. L. Eze, D. N. Ogbonna ##submission.copyrightStatement## Thu, 17 Sep 2020 00:00:00 +0000 RT-PCR Mediated Identification of SARS-CoV-2 Patients from Particular Regions of Bangladesh and the Multi-factorial Analysis Considering Their Pre and Post Infection Health Conditions <p><strong>Background: </strong>SARS-CoV-2 is a type of coronavirus shows bizarre features with fatality rates at 34.4%. The clinical and non-clinical features of COVID-19 might be causative factors for more rapid community spreading compared to MERS and SARS-CoV-1.</p> <p><strong>Objectives: </strong>To study the efficacy of RT-PCR based detection of SARS-CoV-2 patients. Besides, the patients' pre and post-infection health conditions depending on selective clinical and nonclinical parameters were analyzed.</p> <p><strong>Materials and Methods:&nbsp; </strong>Clinical and non- clinical data were collected from 205 randomly selected COVID-19 patients, identified through RT-PCR from different areas of Bangladesh. The data were analyzed using statistical and analytical tools to illustrate the impact of the pandemic situation on the sufferers.&nbsp;</p> <p><strong>Results: </strong>Molecular identification of the patients through RT-PCR has been observed as the most efficient option for detecting SARS-CoV-2 patients. Patients with the smoking habit have been reported to be more prone to the COVID-19 infection; surprisingly the non-smoker female was the worse suffers. The patients aged 60-69 years were the worst sufferers with every adverse health conditions mentioned. Professionally, 28% self-employed male and 8.19% female were infected. The most infected individuals were house wife (32.029%), as next to the health care workers (22.1%). With the increased age group, a decreased number of smokers were found. &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Nebulization of 5% home and 12% hospital treated patients were provided, with Oxygen for 26% of hospitalized patients. After recovery, combined physical and psychological complications were observed in 19% of male and 11% female. The socioeconomic, environmental and geographical annotations exposed a clear relationship with the rate of infection as revealed from the nonclinical data analysis.</p> <p><strong>Conclusion: </strong>RT-PCR proved its specialty in COVID-19 detection. In addition, impacts of different clinical and nonclinical factors on the physical conditions of the nCoV patients were found significant in the research.</p> Kazi Mahmuda Akter, Tanjum Tushi, Sabrina Jahan Mily, Rashenda Aziz Mohona, Samin Anis, Adittya Kishor Chakraborty, Elmee Tabassum, Tahmid Ul Islam, Ozifatun Jannat Akhi, Israt Sultana Nishe, Badrun Nahar Laxy, Sanjida Sultana Zerin, Abdiqani Jama Roble, Md. Imran Hossain, Sharmin Ahmed, Salauddin Al Azad ##submission.copyrightStatement## Fri, 11 Dec 2020 00:00:00 +0000 Effects of Surface Sterilization Protocol and Haustorium Removal on Germination and Growth of Hybrid PB121 Coconut Palm (Cocos nucifera L.) Zygotic Embryos Cultured in vitro <p><strong>Aims:</strong> This study aims to identify the best surface sterilization and evaluate the effect of haustorium suppression on <em>in vitro</em> germination of coconut palm (<em>Cocos nucifera</em> L.) zygotic embryos.</p> <p><strong>Study Design:</strong> Survival rate and contamination rate of zygotic embryos after different surface sterilization treatments, regeneration rate and organogenesis through the number of leaves and the length of shoots after haustorium suppression were determined. For data processing, the Analysis of Variance was used to compare the means which were separated according to Tukey test &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(P = 0.05).</p> <p><strong>Place and Duration of Study:</strong> Coconut fruits (hybrid PB121) were collected 12 to 14 months after controlled pollination from CRAPP (Centre de Recherches Agricoles Plantes Pérennes), station of Sèmè-kpodji in Benin. Experiments were done in Central Laboratory of Plant Biotechnology and Plant Improvement, University of Abomey-Calavi and conducted from june to december in 2019.</p> <p><strong>Methodology:</strong> For the zygotic embryos surface sterilization, four treatments combining three concentrations (3%, 6% and 15%) of commercial bleach (Javel la Croix© containing 12° active chlorine) and immersion durations (5 min, 10 min and 20 min) were tested and the survival rate were determined for each treatment after two months culture. The zygotic embryos were then divided in two sets (haustorium excised embryos set and the whole embryos set) and cultured in modified Y3 medium supplemented with 7 g L<sup>-1</sup> agar, 2.5 g L<sup>-1</sup> activated charcoal, 5% sucrose, &nbsp;6.10<sup>-3</sup> mM 2.4 D (2.4-dichlorophonoxyacetic acid), gibberellic acid and 0.3 mM BAP(6-benzylaminopurine). After five months culture, the regeneration rate, the number of leaves and the length of shouts were recorded.</p> <p><strong>Results:</strong> The high survival rate (80%) was obtained with 6% of bleach and 20 min for the immersion duration without pre-disinfection. The suppression of haustorium have significantly increased the number of leaves (4.3 ± 0.02) and the length of shoots (16.2 ±0.7cm) compared to the whole zygotic embryos.</p> <p><strong>Conclusion:</strong> This protocol can help to ensure better surface sterilization of zygotic embryos before their <em>in vitro</em> culture and the development of vigorous plantlets in order to improve the slow growth of plantlets, when transferred to the greenhouse or field.&nbsp;</p> Arnaud Agbidinoukoun, Euloge Rimson Somakpe, Serge Sètondji Houedjissin, Florent Engelmann, Corneille Ahanhanzo ##submission.copyrightStatement## Thu, 31 Dec 2020 00:00:00 +0000 Recombinant Co-Expression of Collagen A1 (I) Fragment with the Prolyl 4-Hydroxylases (P4H) Subunits in Komagataella phaffii <p>Recombinant collagen and collagen-like products are increasingly replacing animal-sourced collagen that is difficult to produce in safe and standard quality. In this study to produce hydroxylated collagen, a 400 base pair collagen fragment of the bovine COL1A1 gene was co-expressed with prolyl-4-hydroxylase subunit α (P4Hα) and prolyl-4-hydroxylase subunit β(P4Hβ) encoding the P4H enzyme in <em>Komagataella phaffii</em>. For this purpose, each target gene was inserted into the pPICZαA vector and then cloned in <em>E. coli</em> DH5α cells. Subsequently, co-expression vectors were constructed using recombinant vectors isolated from positive clones according to the <em>in vitro</em> multimer ligation method. All recombinant expression and co-expression vectors were transformed into <em>K. phaffii</em> X33 cells by electroporation. The results of reverse transcriptase-polymerase chain reaction (PCR) proved that all target genes were transcribed by recombinant strains. The expression of recombinant proteins was performed for 96 hours by methanol-fed cultivation, and the concentration of the purified proteins from the culture medium was measured by the His-Tag enzyme-linked immunosorbent&nbsp;assay (ELISA) method. The concentrations of rP4Hα and rP4Hβ, and rCol1 proteins expressed individually by recombinant strains were determined to be 10.69 µg/L, 10.74 µg/L, and 8.61 µg/L, respectively, while the concentrations of co-expressed rP4Hα/β and rP4Hα/β/rCol1 proteins were 7.82 µg/L and 5.02 µg/L, respectively. These results showed that the target genes were successfully expressed and co-expressed in the recombinant <em>K. phaffii</em> cell.</p> Esra Avci, Zülal Kesmen ##submission.copyrightStatement## Thu, 31 Dec 2020 00:00:00 +0000 Antioxidant, Hypoglycemic and Neuroprotective Activities of Extracts from Fruits Native to the Amazon Region: A Review <p>The Amazon forest has the largest biome on the planet, and it is estimated that only 16 to 20% of the identified animal, and plant biodiversity. Considering plant diversity, we will highlight the biological properties of the fruits extracts of Arecaceae, Caryocaraceae, Malvaceae, Myrtaceae, Sapindaceae, and Solanaceae’s families due to their significant biological actions. This review presents the antioxidant, glycemic control, and neuroprotective activities from ten fruit extracts distributed in six botanical families in the Amazon region. We obtained 801 publications (described from 2010 to 2020), of which 64 articles were selected by the benchmark previously chosen. The antioxidant effect was the dominant effect observed in the studies used for this review, followed by glycemic control and protective actions in neurons. This review provides a synopsis of the recent literature exploring the extracts from native fruits to the Amazon region that could efficiently prevent pathologies associated with oxidative stress, and cellular maintenance mechanisms.&nbsp;</p> Klenicy Kazumy de Lima Yamaguchi, Anderson de Oliveira Souza ##submission.copyrightStatement## Thu, 22 Oct 2020 00:00:00 +0000