Journal of Advanced Biotechnology and Experimental Therapeutics

01-08

Authors: Bulla Humpy, Swetha Senthil Kumar, Sahabudeen Sheik Mohideen

Hermetia illuciens, commonly known as black soldier fly (BSF) can convert organic biomass into manure and insect biomass into protein and fat. Currently, in the waste management industry, BSF larvae are predominantly used to convert valorized organic substrates into fertilizer and to produce biogas. They are also alternatively used as viable protein substitutes for animal feed as well as human consumer products. The treatment of organic wastes before BSFL consumption indicated a positive stimulation in the biomass conversion rate, larval growth rate and overall larval performance parameters. The current study was designed to analyze the effect of the bio-stimulation-based pre-treatment strategy of the feed on performance parameters, such as larval weight, larval survival rate, substrate reduction by pre-treatment of biowaste for better growth, efficient performance, and good biomass composition in BSFL. In our study, we noticed significant up- and downregulation of several larval parameters by assessing substrate composition, larval growth parameters and substrate reduction rate. The bio-stimulated waste showed larval weight gain when compared to larvae grown in control feed. The heat pretreated waste was not suitable for larval growth as the parameters assessed were observed to be declined. Further investigations are needed to comprehend how BSFL reared on pretreated substrates enhances the nutritional composition of larvae.

17-24

Authors: Aghras Sabah Nawar, Zeena Hadi Obaid, Qaiser Ibrahim Sheikh

Polycystic ovary syndrome (PCOS) is one of the most common types of endocrine syndromes in women, where the disease is described as chronic anovulation and hyperandrogenism, which was characterized by the formation of small cysts (fluid-filled sacs) in the ovaries. This study was carried out to introduce knowledge when plasma and erythrocytes portions were chosen to compare the antioxidant and oxidant status of PCOS patients treated or without treatment with metformin. Samples included 75 women diagnosed with PCOS according to Rotterdam criteria. The control group included healthy volunteers with regular menstrual cycles and with no symptoms of hyperandrogenism. Total antioxidants capacity (TAC), total oxidant status (TOS), and Cu\Zn SOD enzyme activity were examined in both plasma and erythrocytes. The activity of the Cu\Zn SOD enzyme was determined based on the ability of the enzyme to inhibit the autoxidation of pyrogallol. Body mass index (BMI), hormonal tests (LH and FSH) showed significant differences in PCOS patients as compared to control. In both plasma and erythrocyte, TAC levels were significantly decreased in PCOS patients. Though the activity of superoxide dismutase was significantly higher in plasma and erythrocytes in patients. On the other hand, PCOS patients showed significantly higher levels of TOS in plasma, while erythrocytes did not show any significant differences. In addition, metformin treatment significantly decreased levels of TAC, TOS and SOD1 activity in PCOS patients. Considering all, it is concluded that both TOS and SOD1 activity introduce the same pattern in blood samples, TAC displays a different line of responses which can be due to the crucial role of SOD1 as an antioxidant factor.

35-43

Authors: May K. Ismael, Lubna M. Rasuol, Yasir Basim Qaddoori

SARS-CoV-2 stands for severe acute respiratory syndrome coronavirus 2. Matrix metalloproteinases- 9 (MMP-9) performs a crucial physiological role. In addition to its role in the molecular basis of lung fibrosis, this enzyme may also play a part in the “cytokine storm,” which may represent one of the potential scenarios during coronavirus infection. Tissue inhibitors of metalloproteinase (TIMPs) are well-known for their ability to regulate MMP activity during remodeling of the extracellular matrix. As cytokines, they are also thought of as signaling molecules that impact on a wide range of biological processes. This study aimed to investigate the link between each of MMP-9 and TIMP-1, and COVID19 disease. A total of 58 COVID-19 patients and 30 apparently healthy adults enrolled in this study. The ORF1ab, E and N genes of SARS-CoV-2 were detected using multiplex real-time PCR, while the ELISA technique was used to estimate the level of serum MMP-9, TIMP-1, and C-reactive protein (CRP). The study results demonstrated higher concentrations of MMP-9 in COVID-19 patients (2810 ± 1160 pg/ml) compared to controls (2110 ± 850 pg/ml), with non-significant differences (p=0.002). Unlike, TIMP-1, showed considerably higher levels in the patient’s group (541.53 ± 201.42 pg/ml) than in controls (276.33 ± 67.26 pg/ml) with high significant differences (p ≤ 0.001). Considering this study, TIMP-1 in COVID patients most likely play an important role in inflammatory response. Its clinical utility as a biomarker may be insufficient, but it provides a useful data in the diagnosis of COVID‐19.

58-66

Authors: Md Faruk Hasan, Jannatul Ferdous, Amit Kumar Dutta, Fahmida Begum Mina, Sumon Karmakar, Biswanath Sikdar

Anthracnose is one of the most devastating fungi causing twig dieback and postharvest fruit decay. Present study was aimed to identification of anthracnose fungus. Infected leaves of Citrus limon were collected and cultured on potato dextrose agar (PDA) media for pathogenic fungus isolation. The isolated fungal pure culture was characterized by physiological and morphological characterization methods. Biological control measures of the fungus were evaluated by disc diffusion methods. The highest growth and development of isolated fungus was detected in PDA media at pH 7 in fructose as the best carbon source and 0.05gm NaCl concentrations at 37°C. Pathogenicity potency of isolate was performed on lemon, orange and malta, and showed typical anthracnose symptoms after incubation at 25°C for 5 days. For antifungal activity, 200µgm/disc methanolic extract of Psidium guajava showed 14.33±0.66 mm inhibition zone against the isolated fungus. From the present investigations, identified anthracnose causing fungus and it’s controlling techniques may help for further research for the isolation of drugs related compound for controlling this disease.

84-93

Authors: Dwi Wahyuning Asih, Agung Dwi Wahyu Widodo, Rebekah Juniati Setiabudi, Sulistiawati, Wiwiek Tyasningsih, Manik Retno Wahyunitisari

Biofilms are composed of more than one species and are often called polymicrobial biofilms, so research is needed on the formation of polymicrobial biofilms, especially between fungi and bacteria. This study aimed to analyze the total biomass and metabolic activity of biofilms formed from the interaction of fungi (Candida tropicalis) with various bacteria, including Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii. Bacterial and fungal cultures were suspended in tryptic soy broth (TSB) medium, and biofilm was cultivated in two 96-well microplates for 48 hours at 37°C. The crystal violet assay was used to detect the total biomass biofilm, and the tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, or MTT) assay was used to obtain the metabolic activity of biofilm with the optical density (OD) value using an ELISA reader. The results of this study obtained significant data from both parameters. The highest average value was found in the C. tropicalis treatment group (2,412 ± 0.825 on the biomass test results; 1,525 ± 0.473 on the metabolic activity test results), while the lowest was seen in the S. aureus treatment group (0.512 ± 0.224 in the biomass test results; 0.978 ± 0.349 in the metabolic activity test results). In conclusion, it was confirmed that biofilm biomass could be generated strongly in each treatment in the total biomass parameter, while metabolic activity data suggested that each treatment could carry out cell proliferation.

109-122

Authors: Ayilya Bakthavatchalam Loganathan, Nazeer Rasool Abdul

Chronic obstructive pulmonary disease (COPD) is a progressive disease that is certainly preventable and treatable and is caused due to the continuous exposure to noxious substances and toxic gases and is characterized by airflow limitation and chronic inflammation in the lungs coupled with persistent symptoms in the respiratory tract leading to obstructive bronchiolitis and parenchymal emphysema. The incidence and progression of COPD is a complicated pathological phenomenon, and the overall severity is due to its exacerbations and comorbidities in individuals. Further, COPD is a major contributor to the global years of life lost and by 2030 it would be the third leading causes of mortality in the world. Also, knowledge on COPD, its associated conditions, and the clinical understanding of the disease date back to the 16th century. The prevalence, morbidity and mortality may vary across the globe based on their exposure to smoking, tobacco, occupational pollutants, indoor pollution, outdoor air pollution, gender, age, and genetic inheritance. Considering the continuous exposure to toxic substances and aging of the general population, the burden and prevalence of COPD are estimated to increase substantially in the coming years. Furthermore, COVID-19 patients with pre-existing COPD conditions suffer from severe disease progression and delay in recovery.

133-139

Authors: Yance Kaitana, Nia Kurnia, Max Tulung, Josef Sem Berth Tuda, Juliet Merry Eva Mamahit, Trina Ekawati Tallei

Aedes spp. are the primary vectors of arboviruses such as dengue, Zika, and chikungunya. To date, neither an approved vaccine nor a drug that can definitively prevent or treat these diseases. Vector control continues to be an essential method of disease prevention. Plant-based insecticides are an alternative to chemical insecticides because they are less toxic to non-target insects and degrade more quickly. The Papuan people have used Pangi leaves for generations as a potent natural chemical against head lice and mosquito larvae. This study aimed to evaluate the potential of pangi (Pangium edule) leaf extract as a biolarvicide and mosquito repellent. Fresh pangi leaves were extracted with water by using a blender to pulverize the leaves. One-way ANOVA was used to examine the variance in mortality rates, and the LC50 value was calculated for probit analysis. Dead larvae were identified and counted. In addition to testing the effectiveness of the extracts as biolarvicide, the extract was also tested as potential insect repellents. The findings demonstrated that pangi leaf extract can kill mosquito larvae. The concentration of the extract affected larval mortality. Furthermore, the extract demonstrated its effectiveness as an insect repellent. Pangi leaf extract contains alkaloids, flavonoids, tannins, saponins, phenolic compounds, and cyanide compounds, with alkaloids being the most abundant. The study found that there was a direct correlation between the concentration of the insecticide and the number of dead Aedes spp. larvae. The LC50 probit test revealed that pangi leaf extract belongs to the category of toxic substances. Several compounds, including alkaloids, flavonoids, tannins, saponins, and cyanide, may be responsible for the toxicity of pangi leaf extract. Thus, pangi leaf extract might be established as a means of overcoming numerous health issues caused by mosquito vectors.

149-160

Authors: Rana H. K. Al-Rubaye; Rakad M. Kh AL-Jumaily

Breast cancer (BC) is the most common malignant tumor in women and the leading cause of cancer deaths worldwide. This work was conducted to estimate the roles of oxidative stress, vitamin B12, homocysteine (HCY), and DNA methylation in BC disease progression. Sixty BC patients (age range 33–80 years) and 30 healthy controls were recruited for this study. Patients with BC were split to group 1 consisted of stage II BC women (low level), and group 2 consisted of patients in stages III and IV (high level). Malondialdehyde (MDA), glutathione peroxidase 3 (GPX3), HCY, and vitamin B12 levels in the study groups were measured. Also, the 5-methylcytosine (5mC) global DNA methylation levels were evaluated. The results showed a significant increase in HCY, and MDA in BC patients compared to healthy controls, with evident increases observed in those with advanced-stage BC (stages III and IV). They were accompanied by significantly reduced levels of 5mC, with a positive correlation between 5mC and the different stages of BC. Also, patients in advanced stages and those with a poor prognosis were exposed to low levels of vitamin B12 and GPX3 (except for the patients in stage IV, which showed increased GPX3 levels). The findings of this study suggest that the differences in global DNA methylation levels at the various phases may be used as a risk factor for developing BC, which indicates the involvement of GPX3 and HCY in BC progression.

172-183

Authors: Md. Hakimul Haque, Md. Zohurul Islam, Md. Lovelu Miah, Sunny Kumar Das, Subir Sarker

Urinary tract infection (UTI) is the leading cause of hospitalization due to bacterial infection, and the frequency of multidrug-resistant Escherichia coli isolates from these infections is increasing worldwide. The current study aims to isolate and characterize antibiotic-resistant Escherichia coli and their antibiogram typing from urine samples of humans. From April to December 2019, a total of 60 human urine samples were collected aseptically and treated to primary isolation by propagation in nutrient broth followed by culture on various agar media. Gram’s staining, string techniques, biochemical characterization, PCR, and Sanger sequencing were performed to confirm E. coli. The Kirby-Bauer disk diffusion technique was used to test the susceptibility of all bacterial strains to thirteen typically prescribed antibiotics. The overall prevalence of E. coli in UTI was 66.67%. Three variations were noted in E. coli, all of which were single substitutions (A>T, C>T, and T>A). Phylogenetic analysis of the 16S rRNA revealed that the E. coli discovered in this study belonged to the genus Escherichia but was distinct from those identified in other countries. The antibiograms revealed that all the isolates (100%) were resistant to penicillin, ampicillin, and amoxicillin; 94.87% to doxycycline; 79.16% to gentamycin; 75.48% to ciprofloxacin; 73.07% to erythromycin; 71.66% to levofloxacin; 47.36% to ceftriaxone; and 46.66% to tetracycline. In contrast, all E. coli strains were sensitive to amikacin (95%), vancomycin (92.50%), and azithromycin (92.50%). People with a UTI often have multidrug-resistant E. coli in their urine samples, which calls for a one-health strategy to deal with this rapidly changing condition.

198-209

Authors: Anindya Das, Ahmed Faisal Sumit

The food consumers and stakeholders of Bangladesh have limited knowledge and perception over genetically modified (GM) foods. As no studies have been done regarding this among the Bangladeshi educated people on a large scale, hence this study aims to determine the level of knowledge and attitude regarding GM foods; and to explore the related factors as well. A cross-sectional survey was done with a close-ended questionnaire in the four largest divisions of Bangladesh where respondents were above 18 years of age and completed at least higher secondary level education. It was observed that among the total 614 respondents, 24.8% had no acquaintance with GM food. Of the remaining 462 respondents, 41.8% had better knowledge and 30.7% had positive attitude. The level of knowledge significantly varied with respondents’ gender, occupation, monthly income, educational status, and discipline studied where male, pharmaceutical workers, respondents with monthly income of 50001-100000 BDT, science studied respondents had significantly better knowledge than their counterparts. Similarly, respondents’ attitude on GMOs significantly differed with gender, occupation, monthly income, and education in which positive attitude were shown by respondents with better knowledge. Finally, logistic regression analysis showed that female and business studied respondents had significantly less likely to have better knowledge and positive attitude than reference group, while respondents with monthly income between 50001-100000 BDT had more likely to have better knowledge and positive attitude, and NGO workers had more likely to have positive attitude than references. Thus, this study will help the policymakers of Bangladesh to perceive the current scenario of public demands on GM foods.

222-230

Authors: Sunao Wada, MH Mahbub, Natsu Yamaguchi, Ryosuke Hase, Yuki Nakagami, Hidekazu Takahashi, Hiroyuki Saito, Junki Shimokawa, Rie Watanabe, Tsuyoshi Tanabe

Exercise training has the potential for inducing enhancements in peripheral circulation, which can play important preventive and therapeutic roles in peripheral circulatory diseases. However, the relevant published studies show conflicting and inconclusive results. Furthermore, useful or optimum ambient temperature for this purpose has not yet been established. Therefore, we investigated the acute responses in peripheral circulation from exposure of healthy subjects to treadmill exercise under different ambient temperatures; A total of 12 young adult volunteers (males 6, females 6) randomly underwent three sessions of treadmill exercise for 30 min under three different ambient temperatures (10°C, 20°C, and 30°C), at a predetermined exercise intensity. Before and after the intervention, leg skin blood flow (SBF) was measured by laser speckle flowgraphy and hand skin temperature (ST), by digital thermometry; After the cessation of treadmill exercise, compared to the corresponding baseline values, a significant increase in SBF was observed under all ambient temperature conditions (P<0.005). During intervention, ST showed a significant decrease at 10th min of intervention under all ambient temperatures (P<0.005) with a subsequent increasing trend in it. After intervention, a significant increase in ST was observed under 20°C condition only (P<0.05). Also, after intervention, the observed increase in systolic blood pressure was less significant under 20°C condition; Treadmill exercise appears to be a useful intervention modality in inducing improvements in peripheral circulation. However, exposure to treadmill exercise at or near 20°C ambient temperature might be recommended for the purpose.

243-253

Authors: Supavinee Sukcharoen, Paphada Watcharapo, Kanokkorn Vichitsakul, Jiraporn Arunpanichlert, Pichayanoot Rotkrua, Boonchoy Soontornworajit

Platelet-derived growth factor-BB (PDGF-BB), a multifunctional growth factor secreted by platelets, endothelial and cancer cells, functions as an efficient and accurate biomarker. PDGF- BB has been employed in various therapeutic applications. Tuning its bioactivity via a specific binding ligand could enhance the PDGF-BB functionality. Aptamer, a single-stranded oligonucleotide that exhibits a selective binding to PDGF-BB, is the molecule of our interest. This research aims to evaluate the PDGF-BB aptamer binding by growth factor-coated particles. PDGF-BB-coated particles were confirmed by enzyme-linked immunosorbent assay (ELISA), and aptamer binding evaluation was investigated by flow cytometry. To optimize the condition for binding evaluation, the washing cycle and concentration of PDGF-BB, polystyrene particle (PS) and aptamer were studied using flow cytometer. Based on the flow cytometry results, the concentration of PDGF-BB showed an optimal level on the number of fluorescent particles. The percentage of fluorescent particles increased when the washing cycle was reduced, and the particle concentration was decreased. The percentage of fluorescent particles also increased proportionally with the aptamer concentration. Using a flow cytometer to detect fluorescent signal from growth factor-coated particle is a promising strategy to study biomolecular binding.

265-275

Authors: Sumaiya Islam, Md. Eftakhar Jahan Bhuiyan, Md. Golzar Hossain, Md. Eliusur Rahman Bhuiyan, Sharmin Akter

At relatively low concentrations in the diet, molasses is extensively used as a vital energy supplement to improve growth, meat quality, and general health condition. However, there is little information on feeding molasses to steers: that’s why the study sought to determine the effects of urea molasses supplementation on growth performance and hemato-biochemical parameters. Twenty Sahiwal steers between 1.5 and 2 years old were divided into two groups (n=10); steers of group A received untreated straw (Control), while steers of group B received urea molasses straw (UMS). The body weight of the steers was measured at every 15 days intervals, and after 90 days of the experimental period blood samples were collected and serum was prepared for hemato-biochemical analysis. The result revealed that body weight of the UMS-treated steers increased significantly from day 15 to day 90 of the experiment compared to the untreated control. UMS induced a significant increase in hematological parameters such as TEC (p<0.01), Hb (p<0.01), and PCV (p<0.05), whereas there were no alterations in TLC and DLC values. Further, UMS treated group showed no significant changes in biochemical parameters such as AST, ALT, and lipid profile (TC, TG, HDL, LDL) while urea and creatinine levels increased significantly (p<0.05). Therefore, it could be suggested that UMS had beneficial effects on growth performance and hemato-biochemical parameters in Sahiwal steers.

286-300

Authors: Md. Moniruzzaman, Mst. Merina Mostari, Shirmin Islam, Mst. Maskera Jinnah, Jui Biswas, Suvro Biswas, Shahriar Zaman, Md. Abu Saleh, Md. Salah Uddin

Rumex dentatus is a medicinal plant with a variety of bioactive components that can be used to treat infections and multidrug-resistant (MDR) microbes. Infections caused by MDR microbes may pose a real threat and the available antibiotics can no longer be used to control or kill those bacteria. Thus, the study aimed to ascertain the antibacterial activity of Rumex dentatus leaf extract and identify a potent compound that can be used as a drug developmental agent may against Staphylococcus aureus. Disc diffusion method, mortality assay of brine shrimp, and DPPH free radical scavenging assay were carried out for in vitro analysis. Standard computational tools and servers such as Pymol, PyRx, and Discovery Studio were used for carrying out the in-silico studies. Staphylococcus aureus was isolated from the infected area of eczema sufferers and identified through morphological, biochemical, and 16SrRNA sequence analysis. Rumex dentatus leaf extract was diluted with methanol and it showed the maximum zone of inhibition (14.33 ± 0.68 mm) after applying it against Staphylococcus aureus at the dose of 150µg/disc. Furthermore, the extract revealed remarkable antioxidant activity and the death rate of brine shrimp and leaf extract concentration were positively connected. Finding pharmacological targets with a high affinity for the FosB (4nb2) protein, which controls antibiotic resistance in Staphylococcus aureus, was done via molecular docking. 1-alpha-18O-1,25-dihydroxycholecalciferol (CID-5280453), androstan-3-ol, 9-methyl-(3 beta,5 alpha) (CID-22215820), phenylacetaldehyde (CID-998), and benzenepropanoic acid, 3,5-bis(1,1-dimethylethyl)-4-hydroxy- methyl ester (CID-62603) substances had binding energies of -6.8 Kcal/mol, -6.6 Kcal/mol, -5.6 kcal/mol, and -5.5 kcal/mol, respectively. As an organic source of medicines, Rumex dentatus leaf extract could be used to combat the antimicrobial-resistant pathogen.