M. L. Riskuwa-Shehu, H. Ismail, A. Y. Fardami, U. B. Ibrahim
{"title":"Pennisetum purpureum Improved Polycyclic Aromatic Hydrocarbons Removal in Weathered-Petroleum Contaminated Soil","authors":"M. L. Riskuwa-Shehu, H. Ismail, A. Y. Fardami, U. B. Ibrahim","doi":"10.24018/ejbio.2022.3.3.285","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.3.285","url":null,"abstract":"Petroleum hydrocarbons are among the major driving forces of advancement in the last Century. Some of the hydrocarbons especially polycyclic aromatics are however of health and environmental significance, due to their recalcitrance and persistence leading to adverse effects on health and ecosystem stability. A number of treatment technologies have been used to cleanup hydrocarbon contaminants and the use of phytoremediation technology have recently been described as promising. In this study, phytoremediation of weathered crude oil contaminated soil was carried out in a microcosm using Pennisetumpurpureum for 60 days. Pristine soil samples were collected and mixed with weathered petroleum contaminated soil to achieve 5%, 25%, 35% and 50% w/w contamination levels. Bacterial species in the rhizosphere were isolated and identified and residual oil was extracted and analyzed using GC-MS. Results showed that there was high bacterial population in rhizosphere (5.0×105 cfu/g to 6.4×105 cfu/g) than non-rhizosphere soil (2.4 ×105 cfu/g to 4.0×105 cfu/g); and Bacillus spp. (64.71%) were observed to be predominant in the rhizosphere followed by Micrococcus spp. (17.65%), Pseudomonas aeruginosa (5.88%), Klebsiella pneumoniae (5.88%) and Flavobacterium sp. (5.88%). Hydrocarbon concentration in the rhizosphere was reduced by 82.5%, 60.5%, 58.0% and 48.8% respectively. Complex polycyclic aromatic hydrocarbon compounds detected in the control using GC-MS were significantly reduced or completely degraded. Polycyclic aromatic hydrocarbons such as anthracene, naphthalene, fluorene, benzo (a) anthracene, pyrene and chrysene were significantly reduced at a rate ranging between 13.33% and 97.54%. Based on the rate of PAHs reduction observed in this study, it was evident that P. purpureumsupports cleanup of persistent hydrocarbon contaminants in soil environment. The use of this plant in large scale petroleum hydrocarbon cleanup under field conditions should be investigated.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48186191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C. Odu, J. Egbere, I. Onyimba, P. Ghamba, Stanley Godiya, T. M. Isyaka, Joy O. Collins-Odu, Mediatrix A. Idigo, A. Ngene
{"title":"Resistance Profiles of Bacteria Isolated from Wastewater in the State Specialist Hospital Maiduguri","authors":"C. Odu, J. Egbere, I. Onyimba, P. Ghamba, Stanley Godiya, T. M. Isyaka, Joy O. Collins-Odu, Mediatrix A. Idigo, A. Ngene","doi":"10.24018/ejbio.2022.3.3.362","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.3.362","url":null,"abstract":"Antibiotics being an effective antibacterial did not really last long as expected. This is because as soon as they were discovered, bacteria started developing resistance against it. Even though studies have shown that resistance genes existed long before the discovery of antibiotics, the reason why we have increased growth in antibiotics resistance is as a result of misuse and overuse of the agents in human and agricultural settings. The aim of this study is to identify the common bacterial species in wastewater and to determine their resistance pattern to regularly/commonly used antibiotics. Samples were collected from the general wastewater channel of the state specialist hospital, Maiduguri. The bacterial isolates identified were Enterobacter spp, Staphylococcus aureus, Pseudomonas spp, Proteus spp, Klebsiella spp, Salmonella spp, Bacillus spp, Coagulase -ve Staphylococcus and other Coliform spp. The antibiotics susceptibility of the bacterial isolates was assayed using the disc diffusion method; OPTUDISCs. The level of sensitivity was highest with Ofloxacin and Levofloxacin having 78.5% and 75%, while resistance was highest with Cloxacillin, Augumentin, Ampicillin and Tetracycline with 96.4%, 89.2, 88.9 and 85.1% respectively. This shows that except for Levofloxacin, all isolates have already been exposed to antibiotics, which resulted in them developing multi resistance. As a result of this multi resistance, there is need to develop other alternative therapy that will be able to tackle antibiotics resistance and surveillance on trends in antibiotic resistance.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46816937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
R. Aso, C. Hammuel, T. Adé, J. Briska, C. S. Hyelnaya
{"title":"Microbial Contamination of Applied Instrument in Female Hair-Dressers Salon in Wukari, Taraba State, Nigeria","authors":"R. Aso, C. Hammuel, T. Adé, J. Briska, C. S. Hyelnaya","doi":"10.24018/ejbio.2022.3.2.361","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.361","url":null,"abstract":"Hairdresser’s salons are public places that can contribute to the spread of viral, fungi and bacterial pathogens. However, little is known about the contamination of hairdressing tools by bacterial and fungal pathogens. Hence, this study was conducted to determine bacterial and fungal contaminants of tools used in hairdressing salons within Wukari metropolis, Taraba State. Eighty (80) different samples were collected from combs, brushes, rollers, and hairdryers used in hairdressing salons using sterile swab stick moistened with normal saline. Samples were cultured aerobically on nutrient agar, MacConkey agar, and sheep blood agar for bacterial isolation and potato dextrose agar for fungal isolation. Bacterial isolates were identified using conventional biochemical tests while fungal isolates were identified on the basis of their cell wall structure using the lactophenol cotton blue stain. Antibiotic sensitivity pattern of bacterial isolates was tested using the modified Kirby-Bauer disc diffusion method. Sixty-seven (83.75%) of the collected sample were positive for bacterial and/or fungal contamination, yielding twenty-two (22) and eighteen (18) isolates each of bacteria and fungi. The bacterial isolates were Staphylococcus aureus (81.82%), Staphylococcus epidermidis (13.64%), and Escherichia coli (4.64%) while the fungal isolates were Aspergillus fumigatus (31.25%), Aspergillus flavus (50%), Aspergillus niger (6.25%), Madurella grisea (6.25%), and Rhizopus stolonifera (6.25%). Bacterial isolates were generally sensitive to ciprofloxacin, gentamicin, rifampicin, ofloxacin, and streptomycin. The highest resistances were against cefuroxime, trimethoprim-sulfamethoxazole, ampicillin, and augmentin. The presence of these microorganisms on hairdressing tools is an indication of poor hygienic practices among hairstylists in Wukari and these tools can serve as vehicles for the transmission of bacterial pathogens. Hence, appropriate measures should be taken to reduce the microbial load from hairdressing salons instruments.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47348211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
I. Onyimba, M. S. Chomini, M. O. Job, A. Njoku, J. A. Onoja, D. Isaac, I. C. Isaac, A. Ngene
{"title":"Evaluation of the Suitability of Tigernut Milk and Tigernut-Cow Composite Milks for Yoghurt Production","authors":"I. Onyimba, M. S. Chomini, M. O. Job, A. Njoku, J. A. Onoja, D. Isaac, I. C. Isaac, A. Ngene","doi":"10.24018/ejbio.2022.3.2.366","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.366","url":null,"abstract":"The cost of animal milk used in yoghurt production in Nigeria has continued to rise, making the price of yoghurt to become prohibitive. This study investigated the suitability of tiger nut milk for yoghurt production. Five milk formulations prepared from tiger nut milk and cow milk were used to produce yoghurts. pH and titratable acidity (TA) of the milks and yoghurts were determined. The yoghurts were assessed for total plate count (TPC), total fungal count (TFC), fat content and sensory properties. Sensory properties were evaluated by a 10-man taste-panel using a 5-point hedonic scale. pH values of the yoghurts ranged from 3.94 – 4.68. TA values ranged from 0.56 – 0.64. TPC of the yoghurts ranged from 1.0 x 102 – 1.3 x 103 cfu/ml while TFC ranged from 0.1 x 101 – 0.3 x 101. The microbial counts of the yoghurt samples were within acceptable safety limits. The yoghurts were generally acceptable to the panelists. There were significant (P < .05) differences in the sensory scores for appearance, taste, texture, and overall acceptability. Tiger nut-cow milk (75:25) yoghurt had the highest appearance and taste scores while tiger nut milk (100) yoghurt had the highest texture and overall acceptability scores. Tiger nut milk (100) yoghurt was the most preferred yoghurt with an overall acceptability score of 4.8±0.42 followed by tiger nut-cow milk (75:25) yoghurt. Yoghurts analysed had fairly high fat contents. The study showed that tiger nut milk and tiger nut-cow milk composites could be used as alternatives to cow milk for yoghurt production.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42896886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jonas da Silva, I. P. G. do Amaral, Elisângela Afonso de Moura Kretzschmar, U. Vasconcelos
{"title":"Antifungal Coating Based on Pyocyanin Nanoparticles (Np-Pyo)","authors":"Jonas da Silva, I. P. G. do Amaral, Elisângela Afonso de Moura Kretzschmar, U. Vasconcelos","doi":"10.24018/ejbio.2022.3.2.360","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.360","url":null,"abstract":"Pyocyanin is a pigment produced by 95% of Pseudomonas aeruginosa strains and exhibits antimicrobial properties that can be used for different purposes. In this work, PMMA-based nanoparticles that were encapsulated into 200 µg/mL of pyocyanin (Np-Pyo) were produced by the nanoprecipitation method. They were evaluated with respect to antifouling activity against Aspergillus sp. and Penicillium sp. With an encapsulation efficiency of 56%, the NpPyo remained stable for 90 days. Their characteristics were satisfactory for the following parameters: average size (616.90±38.30 nm; blank: 282.58±22.89 nm), polydispersion index (0.51±0.01; blank: 0.45±0.78), zeta potential (-5.13±0.41 mV; blank: -6.44±1.12 mV) and pH (6.18±0.03; blank: 6.42±0.01). The in vitro biofilm formation assay was performed on dolomite coupons measuring 1 cm2, on which the formulation was applied. There were tested conditions with and without immersion for 72h at 30 ºC. In the tests with the immersed coupons, there was fungal colonization; this was, however, lower than that observed in the control. A. niger decreased by 3 log units. No growth was observed on the coupons that were not immersed. The results were promising and demonstrated viability as a means of antifouling protection, particularly on dry surfaces.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46477346","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Comparative Karyomorphological Analysis of In vivo and In vitro Grown Plants of Plumbago zeylanica: An Important Medicinal Plant","authors":"S. Majumder, A. Biswas, Md. Mahbubur Rahman","doi":"10.24018/ejbio.2022.3.2.333","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.333","url":null,"abstract":"The present report describes the comparative cytological analysis of in vivo and in vitro grown plant species of Plumbago zeylanica. Somatic chromosome number of in vivo and micro-propagated plants was confirmed to be 2n=28. Individual chromosome length of mother plant and micro-propagated plant was ranged from 2.26 to 5.54 µm and 2.26 to 5.46 µm, respectively. The total length of the haploid complement of in vivo and in vitro grown plants was 49.35 and 52.98 µm. The total form percent (TF%) of mother and in vitro grown plants was 45.11% and 45.58%, respectively and according to Stebbins classification (1971) both plants karyotype was fell into 2B symmetric type. The centromeric formula was for in vivo and in vitro grown plants were 3sm + 11m.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48331991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Priyanka J. Utgikar, Janvi H. Kadam, S. Rambhiya, Vrushali Inamdar, Preet D. Nagda, S. Barve, Pranjali P. Dhawal
{"title":"Titanium Dioxide and Its Effect on Human Health and Environment- An in vitro Study","authors":"Priyanka J. Utgikar, Janvi H. Kadam, S. Rambhiya, Vrushali Inamdar, Preet D. Nagda, S. Barve, Pranjali P. Dhawal","doi":"10.24018/ejbio.2022.3.2.352","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.352","url":null,"abstract":"This work focuses on understanding the in-vitro effects of Titanium Dioxide (TiO2) led in the environment on human health, algae, and agriculture. Human dermal fibroblasts (HDF) from juvenile foreskin and human red blood cells (HRBC) were taken as a model to study the effect of TiO2 on human health in vitro. The study intended to define the toxic effect of TiO2 at 50 ppm, 100 ppm, and 200 ppm. The effect of TiO2 on the algae cyanobacterium as a model to analyze the algal toxicity effect i.e. was checked for freshwater toxicity at the concentration of TiO2 at 100 and 200 ppm was studied. The effect of TiO2 on the growth of microorganisms in wastewater was studied to determine its biodegradation in-vitro for 5 days using dissolved oxygen determination and the biological oxidation demand (BOD). Finally, phytotoxicity was monitored by observing the effect of TiO2 on wheat seed germination. It was found that TiO2 had no effect on HDF and HRBC at the tested concentrations as no cell death and hemolysis were observed when the cells were treated with TiO2. However, a statistically significant algal toxicity of 32.14 % was observed at 100 ppm and a 42.86 % (p<0.01) decrease in biomass was observed at 200 ppm. Additionally, there was no effect found on BOD of wastewater in the presence or absence of TiO2. The TiO2 had a positive effect on wheat seed germination in a dose-dependent manner. There was an increase in root length from 3.4 cm to 4.3 cm and 4.6 cm at 100 and 200 ppm of TiO2, respectively. Also, a slight increase in shoot length was observed at 100 ppm and 200 ppm. However, visible root thinning was a drawback observed. Hence, the present study gives an elaborative insight into the effects of Titanium Dioxide on human health and the environment.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46372443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ketaki Walimbe, Pranjali P. Dhawal, Shruti A. Kakodkar
{"title":"Anticancer Potential of Biosynthesized Silver Nanoparticles: A Review","authors":"Ketaki Walimbe, Pranjali P. Dhawal, Shruti A. Kakodkar","doi":"10.24018/ejbio.2022.3.2.338","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.338","url":null,"abstract":"The field of nanotechnology has emerged as a promising course of study branching out into various biomedical fields such as therapeutics, imaging, and diagnostics. Metallic nanoparticles, specifically silver, are an important area of study due to their multifunctionality and diverse morphological characteristics. Amongst the various methods of synthesis of these multifunctional nanoparticles such as physical methods and chemical, green synthesis is the most suitable method due to its eco-friendly nature, cost-effectiveness, and ease of production. This article summarizes the broad spectrum of applications of green silver nanoparticles in the fields of cancer theranostics, imaging and diagnosis, and drug delivery. Bimetallic nanoparticles with silver as one of its major constituents are also explored to get a clear insight into the numerous prospective developments in the respective field.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44115944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Manasseh E. Atabor, T. O. Ndibe, B. Benjamin, V. Bakare, Chizoba K. Ejuama
{"title":"Assessment of Bioaccumulating Ability of Mariscus longibrateatus and Effects of Effluents from Kaduna Refinery and Petrochemical Company on the Plant","authors":"Manasseh E. Atabor, T. O. Ndibe, B. Benjamin, V. Bakare, Chizoba K. Ejuama","doi":"10.24018/ejbio.2022.3.2.344","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.344","url":null,"abstract":"The operations of Kaduna Refinery and Petrochemical Company (KRPC) lead to the generation of effluents. Plants growing in the drain are in constant exposure to these effluents. Mariscus longibrateatus, which is the most abundant plant species growing in the drain, was studied to determine the effects of the effluents on the plant. Plant samples and soil on which the plant grows, were analyzed for heavy metals. Manganese (1.30mg/g) and copper (1.30mg/g) had the highest concentration of heavy metals in the roots. The plant samples had thinner leaves than the control plant, which may be attributed to dehydration and some hidden injuries. The leaf whole vascular bundle of the studied plants ranged from 19085–20790µm2 whereas phloem and xylem area ranged from 3995 – 4290µm2 and 6584-7004µm2 respectively. Transfer and bioaccumulation factors revealed that heavy metals were not effectively transferred from the root to the stem, but the plant was able to survive in the drain containing KRPC effluents. It is concluded that the effluents caused some changes in the plant. It can be inferred that Mariscus longibrateatus are tolerant to the toxicants in the effluents, and therefore, this plant species is recommended for bioremediation study because of its tolerant ability to heavy metals.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46655005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Docking and Molecular Dynamics Simulations Reveal A Possible Site of Interaction of Olive Leaf Extract Hydroxytyrosol with Polyunsaturated Fatty Acid 5-Lipoxygenase Human Enzyme","authors":"M. Valentini, E. Pieroni, A. Concas, M. Pisu","doi":"10.24018/ejbio.2022.3.2.345","DOIUrl":"https://doi.org/10.24018/ejbio.2022.3.2.345","url":null,"abstract":"The polyphenol hydroxytyrosol (HT), a molecule easily extracted from olive oil production waste, has well known antioxidant and anti-inflammatory properties. In literature various bioassays points to a clear inhibitory effects on the polyunsaturated fatty acid 5-lipoxygenase enzyme (LOX5) which is a current target for pharmaceutical intervention for various inflammatory diseases. We have investigated the hypothesis of direct interaction of HT with LOX5 through blind docking and a 200 nanoseconds long molecular dynamics. Analysis of the results highlights the stability of the interaction of HT in the putative binding site with LOX5. This is in accord with the hypothesis of an allosteric way of action of HT to inhibit the activity of the LOX5 also suggesting the use of HT structure as a scaffold to design LOX5 inhibitors with improved activity and specificity.","PeriodicalId":72969,"journal":{"name":"European journal of biology and biotechnology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47423942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}