Search results for: pathogen

Authors: C. A. Gilkeson, C. J. Noakes, P. A. Sleigh, M. A. I. Khan, M. A. Camargo-Valero

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Understanding how airborne pathogens are transported through hospital wards is essential for determining the infection risk to patients and healthcare workers. This study utilizes Computational Fluid Dynamics (CFD) simulations to explore possible pathogen transport within a six-bed partitioned Nightingalestyle hospital ward. Grid independence of a ward model was addressed using the Grid Convergence Index (GCI) from solutions obtained using three fullystructured grids. Pathogens were simulated using source terms in conjunction with a scalar transport equation and a RANS turbulence model. Errors were found to be less than 4% in the calculation of air velocities but an average of 13% was seen in the scalar field. A parametric study of variations in the pathogen release point illustrated that its distribution is strongly influenced by the local velocity field and the degree of air mixing present.

Keywords: Natural, Ventilation, Pathogen, Transport

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Authors: Jörg Linde, Ekaterina Buyko, Robert Altwasser, Udo Hahn, Reinhard Guthke

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Reverse engineering of full-genomic interaction networks based on compendia of expression data has been successfully applied for a number of model organisms. This study adapts these approaches for an important non-model organism: The major human fungal pathogen Candida albicans. During the infection process, the pathogen can adapt to a wide range of environmental niches and reversibly changes its growth form. Given the importance of these processes, it is important to know how they are regulated. This study presents a reverse engineering strategy able to infer fullgenomic interaction networks for C. albicans based on a linear regression, utilizing the sparseness criterion (LASSO). To overcome the limited amount of expression data and small number of known interactions, we utilize different prior-knowledge sources guiding the network inference to a knowledge driven solution. Since, no database of known interactions for C. albicans exists, we use a textmining system which utilizes full-text research papers to identify known regulatory interactions. By comparing with these known regulatory interactions, we find an optimal value for global modelling parameters weighting the influence of the sparseness criterion and the prior-knowledge. Furthermore, we show that soft integration of prior-knowledge additionally improves the performance. Finally, we compare the performance of our approach to state of the art network inference approaches.

Keywords: Pathogen, network inference, text-mining, Candida albicans, LASSO, mutual information, reverse engineering, linear regression, modelling.

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Authors: Arshad Javaid, Syeda F. Naqvi

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Macrophomina phaseolina is a devastating soil-borne fungal plant pathogen that causes charcoal rot disease in many economically important crops worldwide. So far, no registered fungicide is available against this plant pathogen. This study was planned to examine the antifungal activity of an allelopathic grass Cenchrus pennisetiformis (Hochst. & Steud.) Wipff. for the management of M. phaseolina isolated from cowpea [Vigna unguiculata (L.) Walp.] plants suffering from charcoal rot disease. Different parts of the plants viz. inflorescence, shoot and root were extracted in methanol. Laboratory bioassays were carried out using different concentrations (0, 0.5, 1.0, …, 3.0 g mL-1) of methanolic extracts of the test allelopathic grass species to assess the antifungal activity against the pathogen. In general, extracts of all parts of the grass exhibited antifungal activity. All the concentrations of methanolic extracts of shoot and root significantly reduced fungal biomass by 20–73% and 40–80%, respectively. Methanolic shoot extract was fractionated using n-hexane, chloroform, ethyl acetate and n-butanol. Different concentrations of these fractions (3.125, 6.25, …, 200 mg mL-1) were analyzed for their antifungal activity. All the concentrations of n-hexane fraction significantly reduced fungal biomass by 15–96% over corresponding control treatments. Higher concentrations (12.5–200 mg mL-1) of chloroform, ethyl acetate and n-butanol also reduced the fungal biomass significantly by 29–100%, 46–100% and 24–100%, respectively.

Keywords: Antifungal activity, Cenchrus pennisetiformis, Macrophomina phaseolina, natural fungicides

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Authors: Tin A. Khaing

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Aloe vera has been used worldwide both for pharmaceutical, food, and cosmetic industries due to the plethora of biological activities of some of its metabolites. The aim of this study was to evaluate the antifungal and antioxidant activities of the leaf extract. The antifungal activity was determined by the agar-well diffusion method against plant and human fungal pathogens. The methanol and ethanol portions of the extracts studied were more bioactive than ethyl acetate portion. It was also observed that the activity was more pronounced on plant pathogen than human pathogen except Candida albicans. This is an indication that the extract has the potential to treat plant fungal infections. The Aloe extract showed the significant antioxidant activity by the DPPH radical scavenging method. Therefore, the Aloe extract provided as natural antioxidant has been used in health foods for medical and preservative purposes.

Keywords: Aloe vera, antifungal, antioxidant, DPPH

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Authors: Umapriya.R., S.Shrihari

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Drinking water is one of the most valuable resources available to mankind. The presence of pathogens in drinking water is highly undesirable. Because of the Lateritic soil, the iron concentrations were high in ground water. High concentration of iron and other trace elements could restrict bacterial growth and modify their metabolic pattern as well. The bacterial growth rate reduced in the presence of iron in water. This paper presents the results of a controlled laboratory study conducted to assess the inhibition of micro-organism (pathogen) in well waters in the presence of dissolved iron concentrations. Synthetic samples were studied in the laboratory and the results compared with field samples. Predictive model for microbial inhibition in the presence of iron is presented. It was seen that the bore wells, open wells and the field results varied, probably due to the nature of micro-organism utilizing the iron in well waters.

Keywords: Disinfection, Disinfectant, Iron, Laterite.

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Authors: M. Naimi, M. B. Khaled

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The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: L_m14, L_m21, L_m23, L_m24, and L_m25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: L_m21: 0.56%, L_m23: 0%, L_m25: 1.67%, L_m14: 1.88%, L_m24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46 and 10.16± 0.24 8.5 ± 0.40 10 mm for, L_m21, L_m23, L_m25, L_m14and L_m24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for L_m21, L_m23, L_m25 and 2.08 AU/ml for L_m14, L_m24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for L_m21, L_m23, L_m25, L_m14, L_m24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

Keywords: Exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens.

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Authors: Boci J., Çabeli P., Shtylla T., Kumbe I.

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The development of the poultry industry in Albania is mainly based on the existence of intensive modern farms with huge capacities, which often are mixed with other forms. Colibacillosis is commonly displayed regardless of the type of breeding, delivering high mortality in poultry industry. The mechanisms with which pathogen enterobacters are able to cause the infection in poultry are not yet clear. The routine diagnose in the field, followed by isolation of E. coli and species of Salmonella genres in reference laboratories cannot lead in classification or full recognition of circulative strains in a territory, if it is not performed a differentiation among the present microorganisms in intensive farms and those in rural areas. In this study were isolated 1.496 strains of E. coli and 378 Salmonella spp. This study, presents distribution of poultry pathogenosity of E.coli and Salmonella spp., based on the usage of innovative diagnostic methods.

Keywords: poultry, E.coli, Salmonella spp., Enterobacter

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Authors: JiEun Jeong, Dong Hyun Kim, Bharat Bhusan Patnaik, Se Won Kang, HeeJu Hwang, Yong Hun Jo, Dae-Hyun Seog, YeonSooHan, Yong Seok Lee

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An expressed sequence tag (EST) analysis provideus portions of expressed genes. We have constructed cDNA library and determined randomly sequences from cDNA library clones of T. molitor injected with acholeplasma lysate. We identified the homologous to a galectin gene. As the result of cloning and characterization of novel, we found that the protein has an open reading frame (ORF) of 495 bp, with 164 amino acid residues and molecular weight of 18.5 kDa. To characterize the role of novel Tm-galectin in immune system, we quantified the mRNA level of galectin at different times after treatment with immune elicitors. The galectin mRNA was up-regulated about 7-folds within 18 hrs. This suggests that Tm-galectin is a novel member of animal lectins, and has a role in the process of pathogen recognition. Our study would be helpful for the study on immune defense system and signaling cascade.

Keywords: EST, Innate immunity, Tenebrio molitor, Galectin.

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Authors: Campbell Townsend, David Hamilton

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Previous research has demonstrated that negative attitudes towards people with physical disabilities and obesity are predicted by a component of perceived vulnerability to disease; germ aversion. These findings have been suggested as illustrations of an evolved but over-active mechanism which promotes the avoidance of pathogen-carrying individuals. To date, this interpretation of attitude formation has not been explored with regard to people with intellectual disability, and no attempts have been made to examine possible mediating factors. This study examined attitudes in 333 adults and demonstrated that the moderate positive relationship between germ aversion and negative attitudes toward people with intellectual disability is fully mediated by social dominance orientation, a general preference for hierarchies and inequalities among social groups. These findings have implications for the design of programs which attempt to promote community acceptance and inclusion of people with disabilities.

Keywords: avoidance, evolutionary psychology, intellectual disability, prejudice

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Authors: M. Grabowski, K. Macnar, J. Skrzyński

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Huge losses in apple production are caused by pathogens that cannot be seen shortly after harvest. After-harvest thermotherapy treatments can considerably improve control of storage diseases on apples and become an alternative to chemical pesticides. In the years 2010-2012 carried out research in this area. Apples of 'Topaz' cultivar were harvested at optimal maturity time for long storage and subject to water bath treatment at 45, 50, 52, 55°C for 60, 120, 180 and 240 seconds. The control was untreated fruits. After 12 and 24 weeks and during so called simulated trade turnover the fruits were checked for their condition and the originators of diseases were determined by using the standard phytopathological methods. The most common originator of 'Topaz' apple infection during storage were the fungi of genus Gloeosporium. In this paper it was proven that for effective protection of 'Topaz' apples against diseases, thermotherapy by using water treatments at temperature range of 50-52°C is quite sufficient.

Keywords: apple storage diseases, prolonged fruit storage, 'Topaz' apples, thermotherapeutic treatments.

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Authors: Jayashree Ramana

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An alarming emergence of multidrug-resistant strains of the tuberculosis pathogen Mycobacterium tuberculosis and continuing high worldwide incidence of tuberculosis has invigorated the search for novel drug targets. The enzyme glutamate racemase (MurI) in bacteria catalyzes the stereoconversion of L-glutamate to D-glutamate which is a component of the peptidoglycan cell wall of the bacterium. The inhibitors targeted against MurI from several bacterial species have been patented and are advocated as promising antibacterial agents. However there are none available against MurI from Mycobacterium tuberculosis, due to the lack of its threedimensional structure. This work accomplished two major objectives. First, the tertiary structure of MtMurI was deduced computationally through homology modeling using the templates from bacterial homologues. It is speculated that like in other Gram-positive bacteria, MtMurI exists as a dimer and many of the protein interactions at the dimer interface are also conserved. Second, potent candidate inhibitors against MtMurI were identified through docking against already known inhibitors in other organisms.

Keywords: Glutamate racemase, homology modeling, docking, drug resistance.

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Authors: Virupakshaiah DBM, Madiha Ahmed, Smita T. Patil, Chandrakanth Kelmani

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Tuberculosis (TB) is a bacterial infectious disease caused by the obligate human pathogen, Mycobacterium tuberculosis. Multidrug-resistant tuberculosis (MDR-TB) is a global reality that threatens tuberculosis control. Resistance to antibiotic Rifampicin, occurs in 95% of cases through nucleotide substitutions in an 81-bp core region of the rpoB i.e; beta subunit of DNA dependant RNA polymerase. In this paper, we studied the Rifampicin-rpoB receptor interactions In silico. First, homology modeling was performed to obtain the three dimensional structure of Mycobacterium rpoB. Sixty analogs of Rifampicin were prepared using Marvin sketch software. Both original Rifampicin and the analogs were docked with rpoB and energy values were obtained. Out of sixty analogs, 43 analogs had lesser energy values than conventional Rifampicin and hence are predicted to have greater binding affinity to rpoB. Thus, this study offers a route for the development of Rifampicin analogs against multi drug resistant Mycobacterium rpoB.

Keywords: Marvin Sketch, Mycobacterium tuberculosis, Rifampicin, rpoB.

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Authors: Hanan Moawad, Naglaa M. Abd EL-Rahman

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Erwinia carotovora var. carotovora is the main cause of soft rot in potatoes. Hyphaene thebaica was studied for biocontrol of E. carotovora which inhibited growth of E. carotovora on solid medium, a comparative study of classical and ultrasound-assisted extractions of Hyphaene thebaica fruit. The use of ultrasound decreased significant the total time of treatment and increase the total amount of crude extract. The crude extract was subjected to determine the in vitro, by a bioassay technique revealed that the treatment of paper disks with ultrasound extraction of Hyphaene thebaica reduced the growth of pathogen and produced inhibition zones up to 38mm in diameter. The antioxidant activity of ultrasound-ethanolic extract of Doum fruits (Hyphaene thebaica) was determined. Data obtained showed that the extract contains the secondary metabolites such as Tannins, Saponin, Flavonoids, Phenols, Steroids, Terpenoids, Glycosides and Alkaloids.

Keywords: Ultrasound, classical extract, Biological control, Erwinia carotovora, Hyphaene thebaica.

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Authors: A. Doosti, S. Moshkelani

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Brucellosis is a zoonotic disease; its symptoms and appearances are not exclusive in human and its traditional diagnosis is based on culture, serological methods and conventional PCR. For more sensitive, specific detection and differentiation of Brucella spp., the real time PCR method is recommended. This research has performed to determine the presence and prevalence of Brucella spp. and differentiation of Brucella abortus and Brucella melitensis in house mouse (Mus musculus) in west of Iran. A TaqMan analysis and single-step PCR was carried out in total 326 DNA of Mouse's spleen samples. From the total number of 326 samples, 128 (39.27%) gave positive results for Brucella spp. by conventional PCR, also 65 and 32 out of the 128 specimens were positive for B. melitensis, B. abortus, respectively. These results indicate a high presence of this pathogen in this area and that real time PCR is considerably faster than current standard methods for identification and differentiation of Brucella species. To our knowledge, this study is the first prevalence report of direct identification and differentiation of B. abortus and B. melitensis by real time PCR in mouse tissue samples in Iran.

Keywords: Differentiation, B. abortus, B. melitensis, TaqManprobe, Iran.

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Authors: F. O. Omoya

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Food borne illnesses have been reported to be a global health challenge. Annual incidences of food–related diseases involve 76 million cases, of which only 14 million can be traced to known pathogens. Poor hygienic practices have contributed greatly to this. It has been reported that in the year 2000 about 2.1 million people died from diarrheal diseases, hence, there is a need to ensure food safety at all level. This study focused on the sterility examination and inhibitory effect of honey samples on selected gram negative and gram positive food borne pathogen from South West Nigeria. The laboratory examinations revealed the presence of some bacterial and fungal contaminations of honey samples and that inhibitory activity of the honey sample was more pronounced on the gram negative bacteria than the gram positive bacterial isolates. Antibiotic sensitivity test conducted on the different bacterial isolates also showed that honey was able to inhibit the proliferation of the tested bacteria than the employed antibiotics.

Keywords: Food borne illness, gram positive and gram negative bacteria, honey, and inhibitory activity.

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Authors: Waleed S. Alwaneen

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In many countries, cow dung is used as farm manure and for biogas production. Several bacterial strains associated with cow dung such as Campylobacter, Salmonella sp. and Escherichia coli cause serious human diseases. The objective of the present study was to investigate the use of insect larvae including fruit beetle, waxworms and tiger worms to improve the breakdown of agricultural wastes and reduce their pathogen loads. Fresh cow faeces were collected from a cattle farm and distributed into plastic boxes (100 g/box). Each box was provided with 10 larvae of fruit beetle, Waxworms and Tiger worms, respectively. There were 3 replicates in each treatment including the control. Bacteria were isolated weekly from both control and cow faeces to which larvae were added to determine the bacterial populations. Results revealed that the bacterial load was higher in the cow faeces treated with fruit beetles than in the control, while the bacterial load was lower in the cow faeces treated with waxworms and tiger worms than in the control. The activities of the fruit beetle larvae led to the cow faeces being liquefied which provided a more conducive growing media for bacteria. Therefore, higher bacterial load in the cow faeces treated with fruit beetle might be attributed to the liquefaction of cow faeces.

Keywords: Fruit beetle, waxworms, tiger worms, waste conditioning, composting.

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Authors: M. Dahiru, O. I. Enabulele

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The research aims to investigate the occurrence of multidrug-resistant Acinetobacter, in carrot and estimate the role of carrot in its transmission in a rapidly growing urban population. Thus, 50 carrot samples were collected from Jakara wastewater irrigation farms and are analyzed on MacConkey agar and screened by Microbact 24E (Oxoid) and susceptibility of isolates is tested against 10 commonly used antibiotics. Acinetobacter baumannii and A. lwoffii were isolated in 22.00% and 16% of samples respectively. Resistance to ceporex and penicillin of 36.36% and 27.27% in A. baumannii, and sensitivity to ofloxacin, pefloxacin, gentimycin and co-trimoxazole were observed. However, for A. lwoffii apart from 37.50% resistance to ceporex, it was also resistant to all other drugs tested. There were similarities in the resistances shown by A. baumannii and A. lwoffii to fluoroquinolones and β- lactame drug families in addition to between sulfonamide and animoglycoside demonstrated by A. lwoffii. Significant correlation in similarities were observed at P < 0.05 to CPX to NA (46.2%), and SXT to AU (52.6%) A. baumannii and A. lwoffii respectively and high multi drug resistance (MDR) of 27.27% and 62.50% by A. baumannii and A. lwoffii respectively. The occurrence of multidrug-resistance pathogen in carrot is a serious challenge to public health care, especially in a rapidly growing urban population where subsistence agriculture contributes greatly to urban livelihood and source of vegetables.

Keywords: Urban agriculture, Public health, Fluoroquinolone, Sulfonamide, Multidrug-resistance.

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Authors: T. Azouaoui – Ait Kettout, R. Gaceb – Terrak, F. Rahmania

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Date production in North Africa is facing a worrying slowdown and a decline because of Fusarium wilt or bayoud date palm (Phoenix dactylifera L., caused by Fusarium oxysporum f. sp. albedinis (F. o. a). The objective of this work is to study the in vitro effect of flavonoid aglycones extracted from the roots of two cultivars of date palm (one sensitive to bayoud (Deglet Nour) and the other resistant (Takerboucht)) on the growth and production fusaric acid of the pathogen. Results show that during the first week of development of F. o. a on potato dextrose liquid medium, the flavonoid aglycones extracts of the susceptible cultivar roots stimulates mycelial growth as well as conidiogenesis of F.o.a, nevertheless it has no effect on the synthesis of fusaric acid. However, the flavonoid aglycones extract of resistant cultivar roots stimulates mycelial growth and decreases both the number of conidia production and fusaric acid. It therefore appears possible that the resistant cultivar aglycones have two types of action: they either inhibit the synthesis of fusaric acid, or they metabolize this toxin into hydrosoluble product, this is called detoxification.

Keywords: Flavonoid Aglycones, date palm, fusaric acid, Fusarium oxysporum f. sp. albedinis.

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Authors: Wilailak Siripornadulsil, Siriyanapat Tasaku, Jutamas Buahorm, Surasak Siripornadulsil

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The objectives of this study were to isolate LAB from various sources, dietary supplement, Thai traditional fermented food, and freshwater fish and to characterize their potential as probiotic cultures. Out of 1,558 isolates, 730 were identified as LAB based on isolation on MRS agar supplemented with a bromocresol purple indicator&CaCO₃ and Gram-positive, catalase- and oxidase-negative characteristics. Eight isolates showed the potential probiotic properties including tolerance to acid, bile salt & heat, proteolytic, amylolytic & lipolytic activities and oxalate-degrading capability. They all showed the antimicrobial activity against some Gram-negative and Gram-positive pathogenic bacteria. Based on 16S rDNA sequence analysis, they were identified as Enterococcus faecalis BT2 & MG30, Leconostoc mesenteroides SW64 and Pediococcus pentosaceous BD33, CF32, NP6, PS34 & SW5. The health beneficial effects and food safety will be further investigated and developed as a probiotic or protective culture used in Nile tilapia belly flap meat fermentation.

Keywords: Lactic acid bacteria, pathogen, probiotic, protective culture.

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Authors: Fatih Özogul, Nurten Toy, Yesim Özogul

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The influences of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on cadaverine and other biogenic amines production by Listeria monocytogenes and Staphylococcus aureus were investigated in lysine decarboxylase broth (LDB) using HPLC. Cell free solutions were prepared from Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus acidilactici and Streptococcus thermophiles. Two different concentrations that were 50% and 25% CFS and the control without CFSs were prepared. Significant variations on biogenic amine production were observed in the presence of L. monocytogenes and S. aureus (P < 0.05). The function of CFS on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine formation by L. monocytogenes and S. aureus in control were 500.9 and 948.1 mg/L, respectively while the CFSs of LAB induced 4-fold lower cadaverine production by L. monocytogenes and 7-fold lower cadaverine production by S. aureus. The CFSs resulted in strong decreases in cadaverine and putrescine production by L. monocytogenes and S. aureus, although remarkable increases were observed for histamine, spermidine, spermine, serotonin, dopamine, tyramine and agmatine in the presence of LAB in lysine decarboxylase broth.

Keywords: Cell-free solution, lactic acid bacteria, cadaverine, food borne-pathogen.

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Authors: M. A. Gad, A. Z. Al-Herrawy

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Microsporidia comprises various pathogenic species can infect humans by means of water. Moreover, chlorine disinfection of drinking-water has limitations against this protozoan pathogen. A total of 48 water samples were collected from two drinking water treatment plants having two different filtration systems (slow sand filter and rapid sand filter) during one year period. Samples were collected from inlet and outlet of each plant. Samples were separately filtrated through nitrocellulose membrane (142 mm, 0.45 µm), then eluted and centrifuged. The obtained pellet from each sample was subjected to DNA extraction, then, amplification using genus-specific primer for microsporidia. Each microsporidia-PCR positive sample was performed by two species specific primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis. The results of the present study showed that the percentage of removal for microsporidia through different treatment processes reached its highest rate in the station using slow sand filters (100%), while the removal by rapid sand filter system was 81.8%. Statistically, the two different drinking water treatment plants (slow and rapid) had significant effect for removal of microsporidia. Molecular identification of microsporidia-PCR positive samples using two different primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis showed the presence of the two pervious species in the inlet water of the two stations, while Encephalitozoon intestinalis was detected in the outlet water only. In conclusion, the appearance of virulent microsporidia in treated drinking water may cause potential health threat.

Keywords: Removal, efficacy, microsporidia, drinking water treatment plants, PCR.

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Authors: Zonglin Yang, Tatsuya Akiyama, Kerry S. Williamson, Michael J. Franklin, Thiruvarangan Ramaraj

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Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Keywords: Image informatics, Pseudomonas aeruginosa, biofilm, FISH, computer vision, data visualization.

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Authors: D. Boecker, R. Breves, F. Herth, Z. Zhang, C. Bulitta

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Pathogen-carrying aerosol particles are recognized as important infection carriers like those in the current Corona pandemic. This infection route is often underestimated yet represents the infection route that has been least systematically countered to date. Particularly, the transmission indoors is of the highest concern but current indoor safety measures (e.g.: distancing, masks, filters) provide only limited protection. Inhalation of hypochlorous acid (HOCl) containing aerosols may become an alternate route to attack the incubating microbes in-situ and so potentially lead to a reduction of symptoms of already infected individuals. We investigated a facility-wide air-disinfection concept utilizing the potential of vaporized HOCl to become a disinfecting agent for populated indoor atmospheres. Aerosolized bacterial microbes were used as surrogates for a viral contamination, particularly the enveloped coronavirus. For the room air purification tests we aerosolized bacterial suspensions into lab chambers preloaded with vaporized HOCl solutions. Concentration of ‘free active chlorine’ in the test chamber atmosphere was determined with a special gas sensor system (Draeger AG, Lübeck, Germany) controlling the amount of vaporized HOCl via an aerosolis® device (oji Europe GmbH, Nauen, Germany). We could confirm the disinfecting power of HOCl in suspensions and determined the high efficacy of vaporized HOCl to disinfect atmospheres of populated indoor places at safe and non-irritant levels.

Keywords: Hypochlorous acid, HOCl, indoor air cleaning, infection control, microbial air burden, protective atmosphere.

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: Trichoderma spp., Meloidogyne javanica, Fusarium oxysporum f.sp. radicis lycopersici, biocontrol.

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Authors: M. Naimi, M. B. Khaled

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The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physico-chemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: Biocontrol, in situ and ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

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Authors: Maria C. Proença, Maria T. Rebelo, Maria J. Alves, Sofia Cunha

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Vector-borne diseases are transmitted to humans by mosquitos, sandflies, bugs, ticks, and other vectors. Some are re-transmitted between vectors, if the infected human has a new contact when his levels of infection are high. The vector is infected for lifetime and can transmit infectious diseases not only between humans but also from animals to humans. Some vector borne diseases are very disabling and globally account for more than one million deaths worldwide. The mosquitoes from the complex Culex pipiens sl. are the most abundant in Portugal, and we dispose in this moment of a data set from the surveillance program that has been carried on since 2006 across the country. All mosquitos’ species are included, but the large coverage of Culex pipiens sl. and its importance for public health make this vector an interesting candidate to assess risk of disease amplification. This work focus on ports and airports identified as key areas of high density of vectors. Mosquitoes being ectothermic organisms, the main factor for vector survival and pathogen development is temperature. Minima and maxima local air temperatures for each area of interest are averaged by month from data gathered on a daily basis at the national network of meteorological stations, and interpolated in a geographic information system (GIS). The range of temperatures ideal for several pathogens are known and this work shows how to use it with the meteorological data in each port and airport facility, to focus an efficient implementation of countermeasures and reduce simultaneously risk transmission and mitigation costs. The results show an increased alert with decreasing latitude, which corresponds to higher minimum and maximum temperatures and a lower amplitude range of the daily temperature.

Keywords: Human health, risk assessment, risk management, vector-borne diseases.

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Authors: Mohammadjavad Mehrabanpour, Maryam Ranjbar Bushehri, Dorsa Mehrabanpour

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Respiratory diseases are the most important problems in the country’s poultry industry, particularly when it comes to broiler flocks. Ornithobacterium rhinotracheale (ORT) is a species that causes poor performance in growth rate, egg production, and mortality. This pathogen causes a respiratory infection including pulmonary alveolar inflammation, and pneumonia of birds throughout the world. Newcastle disease (ND) is a highly contagious disease in poultry, and also, it causes considerable losses to the poultry industry. The aim of this study was to evaluate the simultaneous occurrence of ORT and ND and NDV isolation by inoculation in embryonated eggs and confirmed by RT-PCR in broiler chicken flocks in Fars province. In this study, 318 blood and 85 tissue samples (brain, trachea, liver, and cecal tonsils) were collected from 15 broiler chicken farms. Survey serum antibody titers against ORT by using a commercial enzyme-linked immunosorbent assay (ELISA) kit performed. Evaluation of antibody titer against ND virus is performed by hemagglutination inhibition test. Virus isolation with chick embryo eggs 9-11 and RT-PCR method were carried out. A total of 318 serum samples, 135 samples (42.5%) were positive for antibodies to ORT and titer of HI antibodies against NDV in 122 serum samples (38/4%) were 7-10 (log2) and 61 serum samples (19/2%) had occurrence antibody titer against Newcastle virus and ORT. Results of the present study indicated that 20 tissue samples were positive in embryonated egg and in rapid hemagglutination (HA) test. HI test with specific ND positive serum confirmed that 6 of 20 samples. PCR confirmed that all six samples were positive and PCR products of samples indicated 535-base pair fragments in electrophrosis. Due to the great economic importance of these two diseases in the poultry industry, it is necessary to design and implement a comprehensive plan for prevention and control of these diseases.

Keywords: ELISA, Newcastle disease, Ornithobacterium rhinotracheale, seroprevalence.

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Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Keywords: Biofilm, dairy products, lactic acid bacteria, yeast.

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Authors: Ibrahim M. S. Shnawa, Bashar A. H. E. Alsadi, Kalida K. Alniaem

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In the immunologic sense, clinical infection is a state of failure of the immune system to combat the pathogenic weapon of the bacteria invading the host. A motile gram negative vibroid organism associated with marked mono and poly nuclear cell responses was traced during the examination of a clinical material from an infected common carp Cyprinus carpio. On primary plate culture, growth was shown to be pure, dense population of an Aeromonas-like colony morphotype. The pure isolate was found to be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and 37C, oxidase positive utilizes glucose through fermentative pathway, resist 0/129 and novobiocin, produces alanine and lysine decarboxylases but non-producing ornithine dehydrolases. Tests for the in vitro determinants of pathogenicity has shown to be; Betahaemolytic onto blood agar, gelatinase, casienase and amylase producer. Three in vivo determinants of pathogenicity were tested as, the lethal dose fifty, the pathogenesis and pathogenicity. It was evident that 0.1 milliliter of the causal bacterial cell suspension of a density 1 x 107 CFU/ml injected intramuscularly into an average of 100gms fish toke five days incubation period, then at the day six morbidity and mortality were initiated. LD50 was recorded at the day 12 post-infection. Use of an LD50 doses to study the pathogenicity, reveals mononuclear and polynuclear cell responses, on examining the stained direct films of the clinical materials from the experimentally infected fish. Re-isolation tests confirm that the reisolant is same. The course of the infection in natural case was shown manifestation of; skin ulceration, haemorrhage and descaling. On evisceration, the internal organs were shown; congestion in the intestines, spleen and, air sacs. The induced infection showed a milder form of these manifestations. The grading of the virulence of this organism was virulent causing chronic course of infections as indicated from the pathogenesis and pathogenicity studies. Thus the infectious bacteria were consistent with Aeromonas hydrophila, and the infection was chronic.

Keywords: Piscan, inflammatory respnonse, pure culture, pathogen, chronic, infection.

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Authors: M. I. Ribah, M. Jibir, Y. A. Bashar, S. S. Manga

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Food safety is a significant and growing public health problem in the world and Nigeria as a developing country, since food-borne diseases are important contributors to the huge burden of sickness and death of humans. In Nigeria, traditional ready-to-eat meat products (RTE-MPs) like balangu, tsire, guru and dried meat products like kilishi, dambun nama, banda, were reported to be highly appreciated because of their eating qualities. The consumption of these products was considered as safe due to the treatments that are usually involved during their production process. However, during processing and handling, the products could be contaminated by pathogens that could cause food poisoning. Therefore, a hazard identification for pathogenic bacteria on some traditional RTE-MPs was conducted in Kebbi and Sokoto States, Nigeria. A total of 116 RTE-MPs (balangu-38, kilishi-39 and tsire-39) samples were obtained from retail outlets and analyzed using standard cultural microbiological procedures in general and selective enrichment media to isolate the target pathogens. A six-fold serial dilution was prepared and using the pour plating method, colonies were counted. Serial dilutions were selected based on the prepared pre-labeled Petri dishes for each sample. A volume of 10-12 ml of molten Nutrient agar cooled to 42-45°C was poured into each Petri dish and 1 ml each from dilutions of 102, 104 and 106 for every sample was respectively poured on a pre-labeled Petri plate after which colonies were counted. The isolated pathogens were identified and confirmed after series of biochemical tests. Frequencies and percentages were used to describe the presence of pathogens. The General Linear Model was used to analyze data on pathogen presence according to RTE-MPs and means were separated using the Tukey test at 0.05 confidence level. Of the 116 RTE-MPs samples collected, 35 (30.17%) samples were found to be contaminated with some tested pathogens. Prevalence results showed that Escherichia coli, salmonella and Staphylococcus aureus were present in the samples. Mean total bacterial count was 23.82×106 cfu/g. The frequency of individual pathogens isolated was; Staphylococcus aureus 18 (15.51%), Escherichia coli 12 (10.34%) and Salmonella 5 (4.31%). Also, among the RTE-MPs tested, the total bacterial counts were found to differ significantly (P < 0.05), with 1.81, 2.41 and 2.9×104 cfu/g for tsire, kilishi, and balangu, respectively. The study concluded that the presence of pathogenic bacteria in balangu could pose grave health risks to consumers, and hence, recommended good manufacturing practices in the production of balangu to improve the products’ safety.

Keywords: Ready-to-eat meat products, retail outlets, safety assessment, public health.

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