Search results for: opportunistic pathogens
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 616

Search results for: opportunistic pathogens

346 Effect of Metarhizium robertsii in Rhipicephalus microplus hemocytes

Authors: Jessica P. Fiorotti, Maria C. Freitas, Caio J. B. Coutinho-Rodrigues, Mariana G. Camargo, Emily S. Mesquita, Amanda R. C. Corval, Ricardo O. B. Bitencourt, Allan F. Marciano, Diva D. Spadacci-Morena, Patricia S. Golo, Isabele C. Angelo, Vania R. E. P. Bittencourt

Abstract:

The bovine tick, Rhipicephalus microplus, is an arthropod of great importance in veterinary medicine leading to anemia, weight loss, animals' leather depreciation and also acting as a vector of many pathogens. In this way, the parasitism causes a loss of 3.24 billion dollars per year in Brazil. Knowingly, entomopathogenic fungi act as natural controller of some arthropods, acting mainly by active penetration through the cuticle. However, it can also act on the hemolymph and through the production of mycotoxins. Hemocytes are responsible for the cellular immune response and participate in the processes of phagocytosis, nodulation and encapsulation and may undergo changes when challenged by pathogens. The aim of the present study was to evaluate changes in R. microplus hemocytes after inoculation of Metarhizium robertsii using transmission electron microscopy. The isolate ARSEF 2575 and 200 engorged R. microplus females were used. The groups were divided into control, in which the females were inoculated with 5 μL of sterile distilled water solution and 0.1% Tween 80, and a group inoculated with 5 μL of fungal suspension at the concentration of 10⁷ conidia mL⁻¹. The experiment was performed in duplicate and each group contained 50 females. Twenty-four hours after fungal inoculation, hemolymph was collected through the cuticle dorsal surface perforation of the tick females. After collection, the hemolymph samples were centrifuged at 500 x g for 3 minutes at 4 °C, the plasma was discarded and the hemocyte pellet was resuspended in 50 μl PBS. The suspension material was fixed in 2% glutaraldehyde in Millonig buffer for three hours. After fixation, the material was centrifuged at 500 x g for 3 minutes, the supernatant was discarded and the cells were resuspended in a wash solution. Subsequently, the cells were post-fixed with 1% osmium tetroxide in phosphate buffer for one hour at room temperature and dehydrated in increasing concentrations of ethanol, and then embedded in Epon resin. The ultrathin sections were examined under the LEO EM 906E transmission electron microscopy at 80kV. The ultrastructural results revealed that.in control group, the cells were considered intact, in which the granulocytes were observed with granules of different electrodensities, intact mitochondria and cytoplasm without vacuolization. In addition, granulocytes showed plasma membrane projections similar to pseudopodia. Plasmatocytes presented as irregularly shaped cells, with the eccentric nucleus, agranular cytoplasm and some cells presented pseudopodia. Nevertheless, in the group exposed to the fungus, most of the cells presented in degeneration. The granulocytes found had fewer granules in the cytoplasm and more vacuoles. Plasmatocytes, after treatment, presented many vacuoles also in the cytoplasm and the lysosomes presented great amount of electrodense material in their interior. Thus, the results suggest that the fungus has a depressant action in the immune system of the tick, not only by the cell degranulation, but also suggesting that this leads to morphological changes in the hemocytes and may even trigger processes such as phagocytosis.

Keywords: bovine tick, cellular defense, entomopathogenic fungi, immune response

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345 Recreation and Environmental Quality of Tropical Wetlands: A Social Media Based Spatial Analysis

Authors: Michael Sinclair, Andrea Ghermandi, Sheela A. Moses, Joseph Sabu

Abstract:

Passively crowdsourced data, such as geotagged photographs from social media, represent an opportunistic source of location-based and time-specific behavioral data for ecosystem services analysis. Such data have innovative applications for environmental management and protection, which are replicable at wide spatial scales and in the context of both developed and developing countries. Here we test one such innovation, based on the analysis of the metadata of online geotagged photographs, to investigate the provision of recreational services by the entire network of wetland ecosystems in the state of Kerala, India. We estimate visitation to individual wetlands state-wide and extend, for the first time to a developing region, the emerging application of cultural ecosystem services modelling using data from social media. The impacts of restoration of wetland areal extension and water quality improvement are explored as a means to inform more sustainable management strategies. Findings show that improving water quality to a level suitable for the preservation of wildlife and fisheries could increase annual visits by 350,000, an increase of 13% in wetland visits state-wide, while restoring previously encroached wetland area could result in a 7% increase in annual visits, corresponding to 49,000 visitors, in the Ashtamudi and Vembanad lakes alone, two large coastal Ramsar wetlands in Kerala. We discuss how passive crowdsourcing of social media data has the potential to improve current ecosystem service analyses and environmental management practices also in the context of developing countries.

Keywords: coastal wetlands, cultural ecosystem services, India, passive crowdsourcing, social media, wetland restoration

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344 Inhibition of Mixed Infection Caused by Human Immunodeficiency Virus and Herpes Virus by Fullerene Compound

Authors: Dmitry Nosik, Nickolay Nosik, Elli Kaplina, Olga Lobach, Marina Chataeva, Lev Rasnetsov

Abstract:

Background and aims: Human Immunodeficiency Virus (HIV) infection is very often associated with Herpes Simplex Virus (HSV) infection but HIV patients are treated with a cocktail of antiretroviral drugs which are toxic. The use of an antiviral drug which will be active against both viruses like ferrovir found in our previous studies is rather actual. Earlier we had shown that Fullerene poly-amino capronic acid (FPACA) was active in case of monoinfection of HIV-1 or HSV-1. The aim of the study was to analyze the efficiency of FPACA against mixed infection of HIV and HSV. Methods: The peripheral blood lymphocytes, CEM, MT-4 cells were simultaneously infected with HIV-1 and HSV-1. FPACA was added 1 hour before infection. Cells viability was detected by MTT assay, virus antigens detected by ELISA, syncytium formation detected by microscopy. The different multiplicity of HIV-1/HSV-1 ratio was used. Results: The double viral HIV-1/HSV-1 infection was more cytopathic comparing with monoinfections. In mixed infection by the HIV-1/HSV-1 concentration of HIV-1 antigens and syncytium formations increased by 1,7 to 2,3 times in different cells in comparison with the culture infected with HIV-1 alone. The concentration of HSV-1 increased by 1,5-1,7 times, respectively. Administration of FPACA (1 microg/ml) protected cells: HIV-1/HSV-1 (1:1) – 80,1%; HIV-1/HSV-1 (1:4) – 57,2%; HIV-1/HSV-1 (1:8) – 46,3 %; HIV-1/HSV-1 (1:16) – 17,0%. Virus’s antigen levels were also reduced. Syncytium formation was totally inhibited in all cases of mixed infection. Conclusion: FPACA showed antiviral activity in case of mixed viral infection induced by Human Immunodeficiency Virus and Herpes Simplex Virus. The effect of viral inhibition increased with the multiplicity of HIV-1 in the inoculum. The mechanism of FPACA action is connected with the blocking of the virus particles adsorption to the cells and it could be suggested that it can have an antiviral activity against some other viruses too. Now FPACA could be considered as a potential drug for treatment of HIV disease complicated with opportunistic herpes viral infection.

Keywords: antiviral drug, human immunodeficiency virus (hiv), herpes simplex virus (hsv), mixed viral infection

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343 Isolation and Probiotic Characterization of Lactobacillus plantarum and Lactococcus lactis from Gut Microbiome of Rohu (Labeo rohita)

Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

Abstract:

Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

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342 Chemicals to Remove and Prevent Biofilm

Authors: Cynthia K. Burzell

Abstract:

Aequor's Founder, a Marine and Medical Microbiologist, discovered novel, non-toxic chemicals in the ocean that uniquely remove biofilm in minutes and prevent its formation for days. These chemicals and over 70 synthesized analogs that Aequor developed can replace thousands of toxic biocides used in consumer and industrial products and, as new drug candidates, kill biofilm-forming bacteria and fungi Superbugs -the antimicrobial-resistant (AMR) pathogens for which there is no cure. Cynthia Burzell, PhD., is a Marine and Medical Microbiologist studying natural mechanisms that inhibit biofilm formation on surfaces in contact with water. In 2002, she discovered a new genus and several new species of marine microbes that produce small molecules that remove biofilm in minutes and prevent its formation for days. The molecules include new antimicrobials that can replace thousands of toxic biocides used in consumer and industrial products and can be developed into new drug candidates to kill the biofilm-forming bacteria and fungi -- including the antimicrobial-resistant (AMR) Superbugs for which there is no cure. Today, Aequor has over 70 chemicals that are divided into categories: (1) Novel natural chemicals. Lonza validated that the primary natural chemical removed biofilm in minutes and stated: "Nothing else known can do this at non-toxic doses." (2) Specialty chemicals. 25 of these structural analogs are already approved under the U.S. Environmental Protection Agency (EPA)'s Toxic Substances Control Act, certified as "green" and available for immediate sale. These have been validated for the following agro-industrial verticals: (a) Surface cleaners: The U.S. Department of Agriculture validated that low concentrations of Aequor's formulations provide deep cleaning of inert, nano and organic surfaces and materials; (b) Water treatments: NASA validated that one dose of Aequor's treatment in the International Space Station's water reuse/recycling system lasted 15 months without replenishment. DOE validated that our treatments lower energy consumption by over 10% in buildings and industrial processes. Future validations include pilot projects with the EPA to test efficacy in hospital plumbing systems. (c) Algae cultivation and yeast fermentation: The U.S. Department of Energy (DOE) validated that Aequor's treatment boosted biomass of renewable feedstocks by 40% in half the time -- increasing the profitability of biofuels and biobased co-products. DOE also validated increased yields and crop protection of algae under cultivation in open ponds. A private oil and gas company validated decontamination of oilfield water. (3) New structural analogs. These kill Gram-negative and Gram-positive bacteria and fungi alone, in combinations with each other, and in combination with low doses of existing, ineffective antibiotics (including Penicillin), "potentiating" them to kill AMR pathogens at doses too low to trigger resistance. Both the U.S. National Institutes for Health (NIH) and Department of Defense (DOD) has executed contracts with Aequor to provide the pre-clinical trials needed for these new drug candidates to enter the regulatory approval pipelines. Aequor seeks partners/licensees to commercialize its specialty chemicals and support to evaluate the optimal methods to scale-up of several new structural analogs via activity-guided fractionation and/or biosynthesis in order to initiate the NIH and DOD pre-clinical trials.

Keywords: biofilm, potentiation, prevention, removal

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341 Hanta Virus Infection in a Child and Sequelae

Authors: Vijay Samuel, Tina Thekkekkara, Shoma Ganguly

Abstract:

There is no reported Hanta Seoul virus infection in children in the UK so far, making it quite challenging for clinicians in diagnosing, predicting and prognosticating the outcome of the infection to patients and parents. We report a case of a ten-year-old girl who presented with pyrexia associated with headache, photophobia and abdominal pain. The family had recently acquired two pet rats six weeks ago. She appeared flushed with peri-oral pallor, coated the strawberry tongue, inflamed tonsils and bilateral cervical lymphadenopathy. Her liver and splenic edges were palpable. Investigations showed that she was thrombocytopenic with deranged renal and liver functions. An ultrasound abdomen demonstrated a mildly enlarged spleen, peripancreatic lymph node and an acalculous cholecystitis. In view of her clinical presentation, a diagnosis of leptospirosis was considered and she was commenced on intravenous benzylpenicillin. The following day she became oliguric, developed significant proteinuria and her renal function deteriorated. Following conservative management, her urine output gradually improved along with her renal function, proteinuria and thrombocytopaenia. Serology for leptospirosis and various other viruses were negative. Following discussion with the Rare and Imported Pathogens Laboratory at Porton hanta virus serology was requested and found to be strongly positive for Seoul hanta virus. Following discharge she developed palpitations, fatigue, severe headache and cognitive difficulties including memory loss and difficulties in spelling, reading and mathematics. Extensive investigations including ECG, MRI brain and CSF studies were performed and revealed no significant abnormalities. Since 2012, there have been six cases of acute kidney injury due to Hantavirus infection in the UK. Two cases were from the Humber region and were exposure to wild rats and the other four were exposed to specially bred pet fancy rats. Hanta virus infections can cause mild flu like symptoms but two clinical syndromes are associated with severe disease including haemorrhagic fever with renal syndrome, which may be associated with thrombocytopenia and Hantavirus cardiopulmonary syndrome. Neuropsychological impairments reported following hantavirus pulmonary syndrome and following Puumala virus infection have been reported. Minor white matter lesions were found in about half of the patients investigated with MRI brain. Seoul virus has a global distribution owing to the dispersal of its carrier host rats, through global trade. Several ports in the region could explain the possible establishment of Seoul virus in local populations of rats in the Yorkshire and Humber region. The risk of infection for occupationally exposed groups is 1-3% compared to 32.9% for specialist pet rat owners. The report highlight’s the importance of routinely asking about pets in the family. We hope to raise awareness of the emergence of hantavirus infection in the UK, particularly in the Yorkshire and Humber region. Clinicians should consider hantavirus infection as a potential cause of febrile illness causing renal impairment in children. Awareness of the possible neuro-cognitive sequele would help the clinicians offer appropriate information and support to children and their families. Contacting Rare and Imported Pathogens Laboratory at Porton is a useful resource for clinicians in UK when they consider unusual infections.

Keywords: Seoul hantavirus in child Porton, UK Acute kidney injury

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340 Interaction of Steinernema Glaseri, an Entomopathogenic Nematode with a Predatory Fungus Arthrobotrys Superba on Different Nutrient Media

Authors: Varsha Baweja

Abstract:

Steinernema glaseri is known to be the most potent biocontrol agent against a number of insect pests of various orders and of diverse habitats under laboratory conditions. But in nature many micro pathogens may affect the efficacy of such entomopathogenic nematodes. Keeping this in view, the interaction of Steinernema glaseri with a predatory fungus Arthrobotrys superba was assessed on eight different nutrient media. The activity of A.superba was evaluated in terms of trap formation, conidiophore formation, and number of adhesive cells formed in the presence and absence of nematodes. The fungus failed to form any trap on any of the culture media in the absence of nematodes. However, in the presence of nematodes, the trap formation by the test fungus was increased but the number of conidiophores decreased with increase in dilution of Corn Meal Agar from 5% to 2%. Higher number of chlamydospores were observed in phenylalanine treated medium which indicates the inhibiting effect of phenylalanine on the growth of A. superba. Our results suggest that care should be taken during release of entomopathogenic nematodes in an agroecosystem for managing various insect pests in a more efficient manner.

Keywords: Entomopathogenic Nematode , Steinernema Glaseri, Predatory Fungus, Arthrobotrys Superba

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339 Beijerinckia indica Extracellular Extract Mediated Green Synthesis of Silver Nanoparticles with Antioxidant and Antibacterial Activities against Clinical Pathogens

Authors: Gopalu Karunakaran, Matheswaran Jagathambal, Nguyen Van Minh, Evgeny Kolesnikov, Denis Kuznetsov

Abstract:

This work investigated the use of Beijerinckia indica extracellular extract for the synthesis of silver nanoparticles using AgNO3. The formation of nanoparticles was confirmed by different methods, such as UV-Vis absorption spectroscopy, XRD, FTIR, EDX, and TEM analysis. The formation of silver nanoparticles (AgNPs) was confirmed by the change in color from light yellow to dark brown. The absorbance peak obtained at 430 nm confirmed the presence of silver nanoparticles. The XRD analysis showed the cubic crystalline phase of the synthesized nanoparticles. FTIR revealed the presence of groups that acts as stabilizing and reducing agents for silver nanoparticles formation. The synthesized silver nanoparticles were generally found to be spherical in shape with size ranging from 5 to 20 nm, as evident by TEM analysis. These nanoparticles were found to inhibit pathogenic bacterial strains. This work proved that the bacterial extract is a potential eco-friendly candidate for the synthesis of silver nanoparticles with promising antibacterial and antioxidant properties. 

Keywords: antioxidant activity, antimicrobial activity, Beijerinckia indica, characterisation, extracellular extracts, silver nanoparticles

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338 Research Cooperation between of Ukraine in Terms of Food Chain Safety Control in the Frame of MICRORISK Project

Authors: Kinga Wieczorek, Elzbieta Kukier, Remigiusz Pomykala, Beata Lachtara, Renata Szewczyk, Krzysztof Kwiatek, Jacek Osek

Abstract:

The MICRORISK project (Research cooperation in assessment of microbiological hazard and risk in the food chain) was funded by the European Commission under the FP7 PEOPLE 2012 IRSES call within the International Research Staff Exchange Scheme of Marie Curie Action and realized during years from 2014 to 2015. The main aim of the project was to establish a cooperation between the European Union (EU) and the third State in the area important from the public health point of view. The following organizations have been engaged in the activity: National Veterinary Research Institute (NVRI) in Pulawy, Poland (coordinator), French Agency for Food, Environmental and Occupational Health & Safety (ANSES) in Maisons Alfort, France, National Scientific Center Institute of Experimental and Clinical Veterinary Medicine (NSC IECVM), Kharkov and State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise (SSRILDVSE) Kijev Ukraine. The results of the project showed that Ukraine used microbiological criteria in accordance with Commission Regulation (EC) No 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs. Compliance concerns both the criteria applicable at the stage of food safety (retail trade), as well as evaluation criteria and process hygiene in food production. In this case, the Ukrainian legislation also provides application of the criteria that do not have counterparts in the food law of the European Union, and are based on the provisions of Ukrainian law. Partial coherence of the Ukrainian and EU legal requirements in terms of microbiological criteria for food and feed concerns microbiological parameters such as total plate count, coliforms, coagulase-positive Staphylococcus spp., including S. aureus. Analysis of laboratory methods used for microbiological hazards control in food production chain has shown that most methods used in the EU are well-known by Ukrainian partners, and many of them are routinely applied as the only standards in the laboratory practice or simultaneously used with Ukrainian methods. The area without any legislation, where the EU regulation and analytical methods should be implemented is the area of Shiga toxin producing E. coli, including E. coli O157 and staphylococcal enterotoxin detection. During the project, the analysis of the existing Ukrainian and EU data concerning the prevalence of the most important food-borne pathogens on different stages of food production chain was performed. Particularly, prevalence of Salmonella spp., Campylobacter spp., L. monocytogenes as well as clostridia was examined. The analysis showed that poultry meat still appears to be the most important food-borne source of Campylobacter and Salmonella in the UE. On the other hand, L. monocytogenes were seldom detected above the legal safety limit (100 cfu/g) among the EU countries. Moreover, the analysis revealed the lack of comprehensive data regarding the prevalence of the most important food-borne pathogens in Ukraine. The results of the MICRORISK project are networking activities among researches originations participating in the tasks will help with a better recognition of each other regarding very important, from the public health point of view areas such as microbiological hazards in the food production chain and finally will help to improve food quality and safety for consumers.

Keywords: cooperation, European Union, food chain safety, food law, microbiological risk, Microrisk, Poland, Ukraine

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337 Monitoring and Evaluation of the Reverse Osmosis Reject Wastewater from the Sulaibiya Wastewater Treatment Plant in Kuwait

Authors: Mishari Khajah, Mohd. Elmuntasir Ahmed, Abdullah Al-Matouq, Farah Al-Ajeel, Fatemah Dashti, Ahmed Shishter

Abstract:

The overall aim of this study was to monitor and evaluate the effluent quality of a reverse osmosis (RO) reject wastewater from the biggest wastewater treatment plant in the world that is using RO and ultrafiltration membranes in their processes to reclaim water for indirect potable water reuse from municipal wastewaters. The RO reject wastewater or brine included various contaminants that could harm the human health and the environment such as trace organics, organic matters, heavy metals, nutrients and pathogens. Unfortunately, there are no legally binding regulatory guidelines for brine management in Kuwait as many countries around the world. This study monitors and evaluate the RO reject wastewater (brine) generated from the Sulaibiya Wastewater Treatment Plant. Samples were collected and analyzed about 37 parameters for one-year period, twice a month, and compare it to Kuwait Environment Public Authority, KEPA. Results showed that the heavy metals parameters were above KEPA standards, which needs to be treated.

Keywords: domestic wastewater, management, potable water, RO reject wastewater, Sulaibiya wastewater treatment plant

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336 Developing a Set of Primers Targeting Chondroitin Ac Lyase Gene for Specific and Sensitive Detection of Flavobacterium Columnare, a Causative Agent of Freshwater Columnaris

Authors: Mahmoud Mabrok, Channarong Rodkhum

Abstract:

Flavobacterium columanre is one of the devastating pathogen that causes noticeable economic losses in freshwater cultured fish. Like other filamentous bacteria, F. columanre tends to aggregate and fluctuate to all kind of media, thus revealing obstacles in recognition of its colonies. Since the molecular typing is the only fundamental tool for rapid and precise detection of this pathgen. The present study developed a species-specific PCR assay based on cslA unique gene of F. columnare. The cslA gene sequences of 13 F. columnare, strains retrieved from gene bank database, were aligned to identify a conserved homologous segment prior to primers design. The new primers yielded amplicons of 287 bp from F. columnare strains but not from relevant or other pathogens, unlike to other published set that showed no specificity and cross-reactivity with F. indicum. The primers were sensitive and detected as few as 7 CFUs of bacteria and 3 pg of gDNA template. The sensitivity was reduced ten times when using tissue samples. These primers precisely defined all field isolates in a double-blind study, proposing their applicable use for field detection.

Keywords: Columnaris infection, cslA gene, Flavobacterium columnare, PCR

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335 Central Retinal Venous Occlusion Associated O Bilateral Optic Nerve Infiltration Revealing Relapse Of An Acute Lymphoblastic Leukemia

Authors: Fendouli Ines, Zaafrane Nesrine, Mhamdi Hana, Knani Leila, Ghorbel Mohamed

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Introduction: Ocular infiltration of leukemia can involve orbit, uveal tract, retina and optic nerve. It may result from direct ocular infiltration by leukemic cells or indirect ocular involvement resulting from secondary hematologic changes, opportunistic infections and complications of various modalities of therapy. We here in report a case of central venous retinal occlusion associated to optic nerve infiltration as presenting signs of a relapse of acute lymphoblastic leukemia. Case Report: A twelve-year-old male -patient of acute B lymphoblastic leukemia presented with headaches and bilateral blurred vision in the left ee. Ophthalmic examination showed a visual acuity reduced to counting fingers in the right eye and no light perception in the left eye. Funduscopy revealed a voluminous disc edema surrounded by retinal haemorrhages in the right eye, and venous tortusities, papillary edema, and hemorrages suggesting central retinal venous occlusion in the LE. Swept source optical coherence tomography revealed a serous retinal detachment in the RE and .hyperreflective inner layers with macular edema in the left eye. Cerebro-orbital MRI showed bilateral thickened left optic nerve. There were no radiological signs of true papillary edema due to intracranial hypertension secondary to central nervous system involvement. Myelogram and lumbar punction demonstrated blast infiltration and confirmed ocular relapse of the leukemia. Conclusion: Ocular involvement lymphoblastic acute leukemias decreased since the introduction of a systematic prophylactic treatment of central nervous system. Periodic ophthalmic examination is necessary to allow early diagnosis and treatment.

Keywords: acute leukemia, optic nerve, infiltration, relapse

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334 Colorful Textiles with Antimicrobial Property Using Natural Dyes as Effective Green Finishing Agents

Authors: Shahid-ul-Islam, Faqeer Mohammad

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The present study was conducted to investigate the effect of annatto, teak and flame of the forest natural dyes on color, fastness, and antimicrobial property of protein based textile substrate. The color strength (K/S) of wool samples at various concentrations of dyes were analysed using a Reflective Spectrophotometer. The antimicrobial activity of natural dyes before and after application on wool was tested against common human pathogens Escherichia coli, Staphylococcus aureus, and Candida albicans, by using micro-broth dilution method, disc diffusion assay and growth curve studies. The structural morphology of natural protein fibre (wool) was investigated by Scanning Electron Microscopy (SEM). Annatto and teak natural dyes proved very effective in inhibiting the microbial growth in solution phase and after application on wool and resulted in a broad beautiful spectrum of colors with exceptional fastness properties. The results encourage the search and exploitation of new plant species as source of dyes to replace toxic synthetic antimicrobial agents currently used in textile industry.

Keywords: annatto, antimicrobial agents, natural dyes, green textiles

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333 The Potential of Rhizospheric Bacteria for Mycotoxigenic Fungi Suppression

Authors: Vanja Vlajkov, Ivana PajčIn, Mila Grahovac, Marta Loc, Dragana Budakov, Jovana Grahovac

Abstract:

The rhizosphere soil refers to the plant roots' dynamic environment characterized by their inhabitants' high biological activity. Rhizospheric bacteria are recognized as effective biocontrol agents and considered cardinal in alternative strategies for securing ecological plant diseases management. The need to suppress fungal pathogens is an urgent task, not only because of the direct economic losses caused by infection but also due to their ability to produce mycotoxins with harmful effects on human health. Aspergillus and Fusarium species are well-known producers of toxigenic metabolites with a high capacity to colonize crops and enter the food chain. The bacteria belonging to the Bacillus genus has been conceded as a plant beneficial species in agricultural practice and identified as plant growth-promoting rhizobacteria (PGPR). Besides incontestable potential, the full commercialization of microbial biopesticides is in the preliminary phase. Thus, there is a constant need for estimating the suitability of novel strains to be used as a central point of viable bioprocess leading to market-ready product development. In the present study, 76 potential producing strains were isolated from the rhizosphere soil, sampled from different localities in the Autonomous Province of Vojvodina, Republic of Serbia. The selective isolation process of strains started by resuspending 1 g of soil samples in 9 ml of saline and incubating at 28° C for 15 minutes at 150 rpm. After homogenization, thermal treatment at 100° C for 7 minutes was performed. Dilution series (10-1-10-3) were prepared, and 500 µl of each was inoculated on nutrient agar plates and incubated at 28° C for 48 h. The pure cultures of morphologically different strains indicating belonging to the Bacillus genus were obtained by the spread-plate technique. The cultivation of the isolated strains was carried out in an Erlenmeyer flask for 96 h, at 28 °C, 170 rpm. The antagonistic activity screening included two phytopathogenic fungi as test microorganisms: Aspergillus sp. and Fusarium sp. The mycelial growth inhibition was estimated based on the antimicrobial activity testing of cultivation broth by the diffusion method. For the Aspergillus sp., the highest antifungal activity was recorded for the isolates Kro-4a and Mah-1a. In contrast, for the Fusarium sp., following 15 isolates exhibited the highest antagonistic effect Par-1, Par-2, Par-3, Par-4, Kup-4, Paš-1b, Pap-3, Kro-2, Kro-3a, Kro-3b, Kra-1a, Kra-1b, Šar-1, Šar-2b and Šar-4. One-way ANOVA was performed to determine the antagonists' effect statistical significance on inhibition zone diameter. Duncan's multiple range test was conducted to define homogenous groups of antagonists with the same level of statistical significance regarding their effect on antimicrobial activity of the tested cultivation broth against tested pathogens. The study results have pointed out the significant in vitro potential of the isolated strains to be used as biocontrol agents for the suppression of the tested mycotoxigenic fungi. Further research should include the identification and detailed characterization of the most promising isolates and mode of action of the selected strains as biocontrol agents. The following research should also involve bioprocess optimization steps to fully reach the selected strains' potential as microbial biopesticides and design cost-effective biotechnological production.

Keywords: Bacillus, biocontrol, bioprocess, mycotoxigenic fungi

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332 Effect of Temperature and Relative Humidity on Aerosol Spread

Authors: Getu Hailu, Catelynn Hettick, Niklas Pieper, Paul Kim, Augustine Hamner

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Airborne transmission is a problem that all viral respiratory diseases have in common. In late 2019, a disease outbreak, now known as SARS-CoV-2, suddenly expanded across China and the rest of the world in a matter of months. Research on the spread and transmission of SARS-CoV-2 airborne particles is ongoing, as well as the development of strategies for the prevention of the spread of these pathogens using indoor air quality (IAQ) methods. By evaluating the surface area of pollutants on the surface of a mannequin in a mock-based clinic room, this study aims to better understand how altering temperature and relative humidity affect aerosol spread and contamination. Four experiments were carried out at a constant temperature of 70 degrees Fahrenheit but with four different humidity levels of 0%, 30%, 45 percent, and 60%. The mannequin was placed in direct aerosol flow since it was discovered that this was the position with the largest exposed surface area. The findings demonstrate that as relative humidity increased while the temperature remained constant, the amount of surface area infected by virus particles decreased. These findings point to approaches to reduce the spread of viral particles, such as SARS-CoV-2 and emphasize the significance of IAQ controls in enclosed environments.

Keywords: IAQ, ventilation, COVID-19, humidity, temperature

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331 Epigenetic Mechanisms Involved in the Occurrence and Development of Infectious Diseases

Authors: Frank Boris Feutmba Keutchou, Saurelle Fabienne Bieghan Same, Verelle Elsa Fogang Pokam, Charles Ursula Metapi Meikeu, Angel Marilyne Messop Nzomo, Ousman Tamgue

Abstract:

Infectious diseases are one of the most important causes of morbidity and mortality worldwide. These diseases are caused by micro-pathogenic organisms, such as bacteria, viruses, parasites, and fungi. Heritable changes in gene expression that do not involve changes to the underlying DNA sequence are referred to as epigenetics. Emerging evidence suggests that epigenetic mechanisms are important in the emergence and progression of infectious diseases. Pathogens can manipulate host epigenetic machinery to promote their own replication and evade immune responses. The Human Genome Project has provided new opportunities for developing better tools for the diagnosis and identification of target genes. Several epigenetic modifications, such as DNA methylation, histone modifications, and non-coding RNA expression, have been shown to influence infectious disease outcomes. Understanding the epigenetic mechanisms underlying infectious diseases may result in the progression of new therapeutic approaches focusing on host-pathogen interactions. The goal of this study is to show how different infectious agents interact with host cells after infection.

Keywords: epigenetic, infectious disease, micro-pathogenic organism, phenotype

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330 Chemical Composition, Antioxidant and Antimicrobial Activities of the Essential Oils of Different Pinus Species from Kosovo

Authors: Fatbardhë Kurti, Giangiacomo Beretta, Behxhet Mustafa, Fabrizio Gelmini, Avni Hajdari

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Chemical profile, antioxidant and antimicrobial activity of total and fractionated essential oils (EOs) (F1 – hexane, F2 – hexane/diethyl ether, F3 – diethyl ether) derived from five Pinus species (Pinus heldreichii, P. peuce, P. mugo, Pinus nigra, P. sylvestris), were investigated. The hydrodistilled EOs and their chromatographic fractions (direct solid phase extraction, SPE) were analysed by GC-MS and 112 compounds separated and identified. The main constituents were α-pinene, β-pinene, D-limonene, β-caryophyllene, germacrene D, bornyl acetate and 3-carene. The antioxidant activities of total EOs were lower than those of the corresponding fractions, with F2 the strongest in all cases. EOs and fractions showed different degrees of antibacterial efficacy against different microbial pathogens (moderately strong antimicrobial activity against C. albicans and C. krusei ,while low or no activity against E. faecalis and E. coli strains). The detected inhibition zones and MICs for the EOs and fractions were in the range of 14 -35 mm and 0.125 - 1% (v/v), respectively. The components responsible for the antioxidant and antimicrobial activity were oxygenated monoterpenes and sesquiterpenes recovered in the polar EO fractions. These activities seem to be regulated by reciprocal interactions among the different subclasses of phytochemical species present in the EOs.

Keywords: antagonism, antioxidant activity, antibacterial activity, essential oil, fractions, GC-MS, pinus

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329 Efficacy Enhancement of Hydrophobic Antibiotics Employing Rhamnolipid as Biosurfactant

Authors: Abdurrahim A. Elouzi, Abdurrauf M. Gusbi, Ali M. Elgerbi

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Antibiotic resistance has become a global public-health problem, thus it is imperative that new antibiotics continue to be developed. Major problems are being experienced in human medicine from antibiotic resistant bacteria. Moreover, no new chemical class of antibiotics has been introduced into medicine in the past two decades. The aim of the current study presents experimental results that evaluate the capability of bio surfactant rhamnolipid on enhancing the efficacy of hydrophobic antibiotics. Serial dilutions of azithromycin and clarithromycin were prepared. A bacterial suspension (approximately 5 X 105 CFU) from an overnight culture in MSM was inoculated into 20 ml sterile test tube each containing a serial 10-fold dilution of the test antibiotic(s) in broth with or without 200 mgL-1 rhamnolipid. The tubes were incubated for 24 h with vigorous shaking at 37°C. Antimicrobial activity in multiple antibiotic-resistant gram-negative bacteria pathogens and gram-positive bacteria were assessed using optical density technique. The results clearly demonstrated that the presence of rhamnolipid significantly improved the efficiency of both antibiotics. We hypothesized that the addition of rhamnolipid at low concentration, causes release of LPS which results in an increase in cell surface hydrophobicity. This allows increased association of cells with hydrophobic antibiotics resulting in increased cytotoxicity rates.

Keywords: hydrophobic antibiotics, biosurfactant, rhamnolipid, azithromycin, clarithromycin

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328 Homology Modelling of Beta Defensin 3 of Bos taurus and Its Docking Studies with Molecules Responsible for Formation of Biofilm

Authors: Ravinder Singh, Ankita Gurao, Saroj Bandhan, Sudhir Kumar Kashyap

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The Bos taurus Beta defensin 3 is a defensin peptide secreted by neutrophils and epithelial that exhibits anti-microbial activity. It is one of the crucial components forming an innate defense against intra mammary infections in livestock. The beta defensin 3 by virtue of its anti-microbial activity inhibits major mastitis pathogens including Staphylococcus aureus and Pseudomonas aeruginosa etc, which are also responsible for biofilm formation leading to antibiotic resistance phenomenon. Therefore, the defensin may prove as a non-conventional option to treat mastitis. In this study, computational analysis has been performed including sequence comparison among species and homology modeling of Bos taurus beta defensin 3 protein. The assessments of protein structure were done using the protein structure and model assessment tools integrated in Swiss Model server, which employs various local and global quality evaluation parameters. Further, molecular docking was also carried out between the defensin peptide and the components of biofilm to gain insight into various interactions and structural differences crucial for functionality of this protein.

Keywords: beta defensin 3, bos taurus, docking, homology modeling

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327 Assessment of the Possible Effects of Biological Control Agents of Lantana camara and Chromolaena odorata in Davao City, Mindanao, Philippines

Authors: Cristine P. Canlas, Crislene Mae L. Gever, Patricia Bea R. Rosialda, Ma. Nina Regina M. Quibod, Perry Archival C. Buenavente, Normandy M. Barbecho, Cynthia Adeline A. Layusa, Michael Day

Abstract:

Invasive plants have an impact on global biodiversity and ecosystem function, and their management is a complex and formidable task. Two of these invasive plant species, Lantana camara and Chromolaena odorata, are found in the Philippines. Lantana camara has the ability to suppress the growth of and outcompete neighboring plants. Chromolaena odorata causes serious agricultural and economical damage and causes fire hazards during dry season. In addition, both species has been reported to poison livestock. One of the known global management strategies to control invasive plants is the introduction of biological control agents. These natural enemies of the invasive plants reduce population density and impacts of the invasive plants, resulting in the balance of the nature in their invasion. Through secondary data sources, interviews, and field validation (e.g. microhabitat searches, sweep netting, opportunistic sampling, photo-documentation), we investigated whether the biocontrol agents previously released by the Philippine Coconut Authority (PCA) in their Davao Research Center to control these invasive plants are still present and are affecting their respective host weeds. We confirm the presence of the biocontrol agent of L. camara, Uroplata girardi, which was introduced in 1985, and Cecidochares connexa, a biocontrol agent of C. odorata released in 2003. Four other biocontrol agents were found to affect L. camara. Signs of damage (e.g. stem galls in C. odorata, and leaf mines in L. camara) signify that these biocontrol agents have successfully established outside of their release site in Davao. Further investigating the extent of the spread of these biocontrol agents in the Philippines and their damage to the two weeds will contribute to the management of invasive plant species in the country.

Keywords: invasive alien species, biological control agent, entomology, worst weeds

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326 Analysis of Microbiological Quality and Detection of Antibiotic Residue in Bovine Raw Milk Produced in Blida State, Algeria

Authors: M. N. Boukhatem, M. A. Ferhat, K. Mansour

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Bovine raw milk represents a favorable environment for the growth of several food-spoilage strains and some pathogens. It must meet stringent standards to ensure the highest microbiological and toxicological qualities.In order to assess the microbiological risks associated with the consumption of this food, we conducted this study to determine the microbiological quality of bovine raw milk (54 samples) commercialized at the state of Blida (Algeria). The samples analyzed were unsatisfactory in terms of total flora where 61.11% of samples were considered as non acceptable in terms of quality standards, fecal coliforms (40.74%), fecal streptococci (55.55%) and staphylococci (74.07%). Salmonella and Clostridium strains were not detected in all the samples. Furthermore, antibiotic residues were found in 26% of analysed samples. These results reflect non-compliance with the rules of good hygiene practices at milking, storage, transportatio, and sale of milk. Bovine raw milk consumed presents a serious health risk to the population of the study areas.The livestock coaching actors and dissemination of good hygiene practices throughout the production chain are needed to improve the quality of local milk.

Keywords: bovine raw milk, microbiological quality, fecal coliforms, antibiotic residue, Blida state

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325 Bacterial Contamination of Kitchen Sponges and Cutting Surfaces and Disinfection Procedures

Authors: Hayyan I Al Taweil

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Background: The most common of bacterium in kitchen sponges and cutting surfaces which can play a task within the cross-contamination of foods, fomites and hands by foodborne pathogens. Aims and Objectives: This study investigated the incidence of bacterium in kitchen Sponge, and cutting surfaces. Material and methods: a complete of twenty four kitchen Sponges were collected from home kitchens and therefore the numbers of mesotrophic microorganism, coliform microorganism, E. coli, Salmonella, genus {pseudomonas|bacteria genus} and staphylococci in every kitchen Sponges were determined. Microbiological tests of all sponges for total mesophilic aerobic microorganism, S. aureus, Pseudomonas, Salmonella spp., and E. coli were performed on days 3, 7, and 14 by sampling. The sponges involved in daily use in kitchens countenosely with the dishwasher detergent a minimum of doubly daily Results: Results from the overall mesophilic aerobic microorganism, indicate a major increase within the variety of log CFU/ml. the amount of E. coli was reduced, Salmonella spp. was stabled, S. aureus was enhanced from the sponges throughout fourteen days. Genus Pseudomonas was enhanced and was the dominant micro flora within the sponges throughout fourteen days.

Keywords: Kitchen Sponges, Microbiological Contamination, Disinfection; cutting surface; , Cross-Contamination

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324 A Review on Climate Change and Sustainable Agriculture in Southeast Nigeria

Authors: Jane O. Munonye

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Climate change has both negative and positive effects in agricultural production. For agriculture to be sustainable in adverse climate change condition, some natural measures are needed. The issue is to produce more food with available natural resources and reduce the contribution of agriculture to climate change. The study reviewed climate change and sustainable agriculture in southeast Nigeria. Data from the study were from secondary sources. Ten scientific papers were consulted and data for the review were collected from three. The objectives of the paper were as follows: to review the effect of climate change on one major arable crop in southeast Nigeria (yam; Dioscorea rotundata); evident of climate change impact and methods for sustainable agricultural production in adverse weather condition. Some climatic parameter as sunshine, relative humidity and rainfall have negative relationship with yam production and significant at 10% probability. Crop production was predicted to decline by 25% per hectare by 2060 while livestock production has increased the incidence of diseases and pathogens as the major effect to agriculture. Methods for sustainable agriculture and damage of natural resources by climate change were highlighted. Agriculture needs to be transformed as climate changes to enable the sector to be sustainable. There should be a policy in place to facilitate the integration of sustainability in Nigeria agriculture.

Keywords: agriculture, climate change, sustainability, yam

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323 Absolute Lymphocyte Count as Predictor of Pneumocystis Pneumonia in Patients With Unknown HIV Status at a Private Tertiary Hospital

Authors: Marja A. Bernardo, Coreena A. Bueser, Cybele Lara R. Abad, Raul V. Destura

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Pneumocystis jirovecii pneumonia (PCP) is the most common opportunistic infection among people with HIV. Early consideration of PCP should be made even in patients whose HIV status is unknown as delay in treatment may be fatal. The use of absolute lymphocyte count (ALC) has been suggested as an alternative predictor of PCP especially in resource limited settings where PCR testing is costly or delayed. Objective: To determine whether the absolute lymphocyte count (ALC) can be used as a screening tool to predict Pneumocystis pneumonia in patients with unknown HIV status admitted at a private tertiary hospital. Methods: A retrospective cross-sectional study was conducted at a private tertiary medical center. Inpatient medical records of patients aged 18 years old and above from January 2012 to May 2014, in whom a clinical diagnosis of Pneumocystis jirovecii pneumonia was made were reviewed for inclusion. Demographic data, clinical features, hospital course, PCP PCR and HIV results were recorded. Independent t-test and chi-square analysis was used to determine any statistical difference between PCP-positive and PCP-negative groups. Mann-Whitney U-test was used for comparison of hospital stay. Results: There were no statistically significant differences in baseline characteristics between PCP positive and negative groups. While both the percent lymphocyte count (0.14 ± 0.13 vs 0.21 ± 0.16) and ALC (1160 ± 528.67 vs 1493.70 ± 988.61) were lower for the PCP-positive group, only the percent lymphocyte count reached a statistically significant difference (p= 0.067 vs p= 0.042). Conclusion: A quick determination of the ALC may be useful as an additional parameter to help screen for and diagnose pneumocystis pneumonia. In our study, the ALC of patients with PCP appear to be lower than in patients without PCP. A low ALC (e.g. below 1200) may help with the decision regarding empiric treatment. However, it should be used in conjunction with the patient’s clinical presentation, as well as other diagnostic tests. Larger, prospective studies incorporating the ALC with other clinical predictors are necessary to optimally predict those who would benefit from empiric or expedited management for potential PCP.

Keywords: Pneumocystis carinii pneumonia, Absolute Lymphocyte Count, infection, PCP

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322 Forest Policy and Its Implications on Private Forestry Development: A Case Study in Rautahat District, Nepal

Authors: Dammar Bahadur Adhikari

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Community forestry in Nepal has got disproportionately high level of support from government and other actors in forestry sector. Even though master plan for forestry sector (1989) has highlighted community and private forestry as one component, the government policies and other intervention deliberately left out private forestry in its structure and programs. The study aimed at providing the pathway for formulating appropriate policies to address need of different kind of forest management regimes in Rautahat district, Nepal. The key areas the research focused were assessment of current status of private forestry, community forest users' understanding on private forestry; criteria for choosing species of private forestry and factors affecting establishment of private forestry in the area. Qualitative and quantitative data were collected employing questionnaire survey, rapid forest assessment and key informant interview. The study found out that forest policies are imposed due to intense pressure of exogenous forces than due to endogenous demand. Most of the local people opine that their traditional knowledge and skills are not sufficient for private forestry and hence need training on the matter. Likewise, local use, market value and rotation dictate the choice of species for plantation in private forests. Currently district forest office is the only government institution working in the area of private forestry all other governmental and non-governmental organizations have condoned. private forestry. Similarly, only permanent settlers in the area are found to establish private forests other forest users such as migrants and forest encroachers follow opportunistic behavior to meet their forest product need from community and national forests. In this regard, the study recommends taking appropriate step to support other forest management system including private forestry provide community forestry the benefits of competition as suggested by Darwin in 18th century, one and half century back and to help alleviate poverty by channelizing benefits to household level.

Keywords: community forest, forest management, poverty, private forest, users’ group

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321 Unconventional Strategies for Combating Multidrug Resistant Bacterial Biofilms

Authors: Soheir Mohamed Fathey

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Biofilms are complex biological communities which are hard to be eliminated by conventional antibiotic administration and implemented in eighty percent of humans infections. Green remedies have been used for centuries and have shown obvious effects in hindering and combating microbial biofilm infections. Nowadays, there has been a growth in the number of researches on the anti-biofilm performance of natural agents such as plant essential oil (EOs) and propolis. In this study, we investigated the antibiofilm performance of various natural agents, including four essential oils (EOs), cinnamon (Cinnamomum cassia), tea tree (Melaleuca alternifolia), and clove (Syzygium aromaticum), as well as propolis versus the biofilm of both Gram-positive pathogenic bacterium Staphylococcus aureus and Gram-negative pathogenic bacterium Pseudomonas aeruginosa which are major human and animal pathogens rendering a high risk due to their biofilm development ability. The antibiofilm activity of the tested agents was evaluated by crystal violet staining assay and detected by scanning electron and fluorescent microscopy. Antibiofilm performance declared a potent effect of the tested products versus the tested bacterial biofilms.

Keywords: biofilm, essential oils, electron microscopy, fluorescent

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320 Contribution to the Study of the Microbiological Quality of Chawarma Sold in Biskra

Authors: Sara Boulmai̇z

Abstract:

In order to study the microbiological quality of chawarma sold in Biskra, a sampling through some fastfoods of the city was done, the parameters studied are highlighted according to the criteria required by the country's trade management. Microbiological analyzes revealed different levels of contamination by microorganisms. The 10 samples were of an overall view of unsatisfactory quality, and according to the standards, no sample was satisfactory. The range of total aerobic mesophilic flora found is between 105 and 1.2 × 10 7 CFU / g, that of fecal coliforms is 104 to 2.4 × 10 5 CFU / g. The suspected pathogenic staphylococci were between 3.103 and 2.7.106 CFU / g. Salmonellae were absent in all samples, whereas sulphite-reducing anaerobes were present in a single sample. The rate of E. cloacae was between 103 and 6.104 CFU / g. As for fungi and safe mice, their rate was 103 to 107 CFU / g. The study of the sensitivity of antibiotics showed multi-resistance to all the antibiotics tested, although there is a sensitivity towards others. All strains of Staphylococcus aureus tested demonstrated resistance against erythromycin, 30% against streptomycin, and 10% against tetracycline. While the strains of E. cloacae were resistant in all strains to amoxicillin, ceftazidime, cefotaxime, and erythromycin, while they were sensitive to fosfomycin, sulfamethoxazole trimethoperine, ciprofloxacin, and tetracycline. While against chlorophenicol and ofloxacin, the sensitivity was dominant, although there was intermediate resistance. In this study demonstrates that foodborne illnesses remain a problem that arises in addition to the increasingly observed bacterial resistance and that, after all, healthy eating is a right.

Keywords: chawarma, microbiological quality, pathogens., street food

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319 Electrochemical Bioassay for Haptoglobin Quantification: Application in Bovine Mastitis Diagnosis

Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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318 Nanoderma: Ecofriendly Nano Biofungicides for Controlling Plant Pathogenic Fungi

Authors: Kamel A. Abd-Elsalam, Alexei R. Khokhlov

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Studies on bioefficacy (in vitro and in vivo) and mode of action of the nanocides against the most important plant diseases in Egypt and Russia might assist in the goal of sustainable agriculture. To our knowledge, few researchers have evaluated the combined antimicrobial effect of inorganic nanoparticles (NPs) with bioorganic pesticides for controlling plant pathogens in the greenhouse and open field, decontrol investigated synergistic effect. In the current project, we will develop eco-friendly alternative management strategies including the use of heavy nanometal-tolerant Trichoderma strains and the main effective material in conventional fungicides (curpic, sulfur, phosphorus and zinc) for controlling plant diseases. Studies on bioefficacy and the mechanism of the nanocides against the most important plant diseases in Egypt were evaluated. There is a growing need to establish mechanisms of action for nano bio and/or fungicides to assist the design of new compounds or combinations of compounds, in order to understand resistance mechanisms and to provide a focus for toxicological attention. Nanofungicides represent an emerging technological development that could offer a range of benefits including increased efficacy, durability, and a reduction in the amounts of active ingredients that need to be used.

Keywords: biohybrids, biocides, bioagent, plant pathogenic fungi

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317 An Update on Linezolid against Methicillin-Resistant Staphylococcus Aureus Clinical Isolates from Pakistan

Authors: Tayaba Dastgeer, Farhan Rasheed, Muhammad Saeed, Maqsood Ahmad, Zia Ashraf, Abdul Waheed, Muhammad Kamran, Mohsin Khurshid

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Objectives: The study aimed to determine the efficacy of linezolid against clinical isolates of methicillin-resistant staphylococcus aureus (MRSA). Methodology: This cross-sectional study was conducted in the microbiology department of Allama Iqbal Medical College Lahore from August 2017 to September 2019. Isolates were confirmed as MRSA via the presence of the mec-A gene. Confirmed MRSA isolates were processed for susceptibility testing against different antimicrobials, especially linezolid, via the disc diffusion method. Zone sizes were interpreted according to CLSI guidelines. Results: Various types of clinical samples were included in the study; however, the highest frequency of MRSA isolates was found in pus samples, followed by other clinical samples. Among hospitalized patients, most MRSA isolates were obtained from patients in the surgical ward. Of 243 mec-A gene detected isolates, Vancomycin and linezolid showed 100% susceptibility, chloramphenicol showed declining resistance 78 (32.09%), and emerging sensitivity 165 (67.90%) against MRSA. Conclusion: Linezolid is a very efficient drug against MRSA, but the use of this novel drug must be conserved for vancomycin-resistant Staphylococcus aureus or when more resistant pathogens are suspected.

Keywords: MRSA, chloramphenicol, linezolid, nosocomial infections

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