An Advanced Approach Based on Artificial Neural Networks to Identify Environmental Bacteria

Environmental micro-organisms include a large number of taxa and some species that are generally considered nonpathogenic, but can represent a risk in certain conditions, especially for elderly people and immunocompromised individuals. Chemotaxonomic identification techniques are powerful tools for environmental micro-organisms, and cellular fatty acid methyl esters (FAME) content is a powerful fingerprinting identification technique. A system based on an unsupervised artificial neural network (ANN) was set up using the fatty acid profiles of standard bacterial strains, obtained by gas-chromatography, used as learning data. We analysed 45 certified strains belonging to Acinetobacter, Aeromonas, Alcaligenes, Aquaspirillum, Arthrobacter, Bacillus, Brevundimonas, Enterobacter, Flavobacterium, Micrococcus, Pseudomonas, Serratia, Shewanella and Vibrio genera. A set of 79 bacteria isolated from a drinking water line (AMGA, the major water supply system in Genoa) were used as an example for identification compared to standard MIDI method. The resulting ANN output map was found to be a very powerful tool to identify these fresh isolates.

Visfatin and Apelin Are New Interrelated Adipokines Playing Role in the Pathogenesis of Type 2 Diabetes Mellitus Associated Coronary Artery Disease in Postmenopausal Women

Visfatin and apelin are two new adipokines that recently gained a special interest in diabetes research. This study was conducted to study the interplay between these two adipokines and their correlation with other inflammatory and biochemical parameters in type 2 diabetic (T2D) postmenopausal women with CAD. Visfatin and apelin were measured by enzyme-linked immunoassay (ELISA). Visfatin was found to be significantly higher in the following groups: T2D patients without CAD, non-obese and obese T2D patients with CAD when compared to control group. Apelin was found to be significantly lower in non-obese and obese T2D patients with CAD when compared to control group. Visfatin and apelin were found to be significantly associated with each other and with other biochemical parameters. The current study provides evidence for the interplay between visfatin and apelin through the inflammatory milieu characteristic of T2D and their possible role in the pathogenesis of CAD complication of T2D. 

Antibacterial Activity of Lactic Acid Bacteria Isolated from Table Olives against Skin Pathogens

The aim of this study was to assess the effect of LAB isolated from Iranian native olives on the opportunistic skin pathogens, Pseudomonas aeruginosa and Staphylococcus aureus. Lactic Acid Bacteria were isolated from the brine of each sample in the prior of time. The samples were spread on MRS agar for isolation of lactobacillus and for lactococcus. 28 strains of labs were isolated. The labs were centrifuged, the supernatant was strewed and pellet was used to inoculation in wells or at blank disks. 20μl of each pellet was inoculated to blank disks and 40μl of each pellet was inoculated to each well. The result of disk and well diffusion agar against these pathogens were confirmed each other. The size of inhibition zone was different according to the type of bacteria, the method and the concentrations of labs.

New Effective Strains of Bacteria Bacillus thuringiensis ssp. israelensis for Bloodsucking Mosquito Control

Five original strains of entomopathogenic bacteria with insecticidal activity against mosquito larvae of the genera Aedes, Culex and Anopheles have been isolated from natural conditions in Armenia and characterized. According to morphological, physiological and biochemical parameters, all isolates were identified as Bacillus thuringiensis spp. israelensis (Bti). High larvicidal activity has been showed by three strains Bti. These strains can be recommended for industrial production of bacterial preparations.

Wasp Venom Peptides may play a role in the Pathogenesis of Acute Disseminated Encephalomyelitis in Humans: A Structural Similarity Analysis

Acute disseminated encephalomyelitis (ADEM) has been reported to develop after a hymenoptera sting, but its pathogenesis is not known in detail. Myelin basic protein (MBP)- specific T cells have been detected in the blood of patients with ADEM, and a proportion of these patients develop multiple sclerosis (MS). In an attempt to understand the mechanisms underlying ADEM, molecular mimicry between hymenoptera venom peptides and the human immunodominant MBP peptide was scrutinized, based on the sequence and structural similarities, whether it was the root of the disease. The results suggest that the three wasp venom peptides have low sequence homology with the human immunodominant MBP residues 85-99. Structural similarity analysis among the three venom peptides and the MS-related HLA-DR2b (DRA, DRB1*1501)-associated immunodominant MHC binding/TCR contact residues 88-93, VVHFFK showed that hyaluronidase residues 7-12, phospholipase A1 residues 98-103, and antigen 5 residues 109-114 showed a high degree of similarity 83.3%, 100%, and 83.3% respectively. In conclusion, some wasp venom peptides, particularly phospholipase A1, may potentially act as the molecular motifs of the human 3HLA-DR2b-associated immunodominant MBP88-93, and possibly present a mechanism for induction of wasp sting-associated ADEM.

The Presence of Enterobacters (E.Coli and Salmonella spp.) in Industrial Growing Poultry in Albania

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.

Role of Oxidative DNA Damage in Pathogenesis of Diabetic Neuropathy

Oxidative stress is considered to be the cause for onset and the progression of type 2 diabetes mellitus (T2DM) and complications including neuropathy. It is a deleterious process that can be an important mediator of damage to cell structures: protein, lipids and DNA. Data suggest that in patients with diabetes and diabetic neuropathy DNA repair is impaired, which prevents effective removal of lesions. Objective: The aim of our study was to evaluate the association of the hOGG1 (326 Ser/Cys) and XRCC1 (194 Arg/Trp, 399 Arg/Gln) gene polymorphisms whose protein is involved in the BER pathway with DNA repair efficiency in patients with diabetes type 2 and diabetic neuropathy compared to the healthy subjects. Genotypes were determined by PCR-RFLP analysis in 385 subjects, including 117 with type 2 diabetes, 56 with diabetic neuropathy and 212 with normal glucose metabolism. The polymorphisms studied include codon 326 of hOGG1 and 194, 399 of XRCC1 in the base excision repair (BER) genes. Comet assay was carried out using peripheral blood lymphocytes from the patients and controls. This test enabled the evaluation of DNA damage in cells exposed to hydrogen peroxide alone and in the combination with the endonuclease III (Nth). The results of the analysis of polymorphism were statistically examination by calculating the odds ratio (OR) and their 95% confidence intervals (95% CI) using the ¤ç2-tests. Our data indicate that patients with diabetes mellitus type 2 (including those with neuropathy) had higher frequencies of the XRCC1 399Arg/Gln polymorphism in homozygote (GG) (OR: 1.85 [95% CI: 1.07-3.22], P=0.3) and also increased frequency of 399Gln (G) allele (OR: 1.38 [95% CI: 1.03-1.83], P=0.3). No relation to other polymorphisms with increased risk of diabetes or diabetic neuropathy. In T2DM patients complicated by neuropathy, there was less efficient repair of oxidative DNA damage induced by hydrogen peroxide in both the presence and absence of the Nth enzyme. The results of our study suggest that the XRCC1 399 Arg/Gln polymorphism is a significant risk factor of T2DM in Polish population. Obtained data suggest a decreased efficiency of DNA repair in cells from patients with diabetes and neuropathy may be associated with oxidative stress. Additionally, patients with neuropathy are characterized by even greater sensitivity to oxidative damage than patients with diabetes, which suggests participation of free radicals in the pathogenesis of neuropathy.

Antimicrobial, Antiplasmid and Cytotoxicity Potentials of Marine Algae Halimeda opuntia and Sarconema filiforme Collected from Red Sea Coast

The antimicrobial, antiplasmid and cytotoxic activities of marine algae Halimeda opuntia and Sarconema filiforme were investigated. Antimicrobial bioassay against some human pathogenic bacteria and yeast were conducted using disc diffusion method. Halimeda extract exhibited antibacterial activity against six species of microrganisms, with significant inhibition against Staphylococcus aureus. While Sarconema extract was better potent as antifungal against Candida albicans. Comparative antibacterial studies showed that Halimeda extract showed equivalent or better activity as compared with commercial antibiotic when tested against Staphylococcus aureus. Further tests conducted using dilution method showed both extracts as having bacteriostatic mode of action against the tested microorganisms. Methanol extract of two species showed significant cytotoxicity (LC50

Mycoflora of Activated Sludge with MBRs in Berlin, Germany

Thirty six samples from each (aerobic and anoxic) activated sludge were collected from two wastewater treatment plants with MBRs in Berlin, Germany. The samples were prepared for count and definition of fungal isolates; these isolates were purified by conventional techniques and identified by microscopic examination. Sixty tow species belonging to 28 genera were isolated from activated sludge samples under aerobic conditions (28 genera and 58 species) and anoxic conditions (26 genera and 52 species). The obtained data show that, Aspergillus was found at 94.4% followed by Penicillium 61.1 %, Fusarium (61.1 %), Trichoderma (44.4 %) and Geotrichum candidum (41.6 %) species were the most prevalent in all activated sludge samples. The study confirmed that fungi can thrive in activated sludge and sporulation, but isolated in different numbers depending on the effect of aeration system. Some fungal species in our study are saprophytic, and other a pathogenic to plants and animals.

An Example of Post-Harvest Thermotherapy as a Non-Chemical Method of Pathogen Control on Apples of Topaz Cultivar in Storage

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.

Prevalence of Epstein-Barr Virus Latent Membrane Protein-1 in Jordanian Patients with Hodgkin's Lymphoma and Non- Hodgkin's Lymphoma

The aim of this study was to estimate the frequency of EBV infection in Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL) occurring in Jordanian patients. A total of 55 patients with lymphoma were examined in this study. Of 55 patients, 30 and 25 were diagnosed as HL and NHL, respectively. The four HL subtypes were observed with the majority of the cases exhibited the mixed cellularity (MC) subtype followed by the nodular sclerosis (NS). The high grade was found to be the commonest subtype of NHL in our sample, followed by the low grade. The presence of EBV virus was detected by immunostating for expression of latent membrane protein-1 (LMP-1). The frequency of LMP-1 expression occurred more frequent in patients with HL (60.0%) than in patients with NHL (32.0%). The frequency of LMP-1 expression was also higher in patients with MC subtype (61.11%) than those patients with NS (28.57%). No age or gender difference in occurrence of EBV infection was observed among patient with HL. By contrast, the prevalence of EBV infection in NHL patients aged below 50 was lower (16.66%) than in NHL patients aged 50 or above (46.15%). In addition, EBV infection was more frequent in females with NHL (38.46%) than in male with NHL (25%). In NHL cases, the frequency of EBV infection in intermediate grade (60.0%) was high when compared with frequency of low (25%) or high grades (25%). In conclusion, analysis of LMP-1 expression indicates an important role for this viral oncogene in the pathogenesis of EBV-associated malignant lymphomas. These data also support the previous findings that people with EBV may develop lymphoma and that efforts to maintain low lymphoma should be considered for people with EBV infection.

Regulatory Effects of Carbon Sources on Tabtoxin Production (A β-lactam Phytotoxin of Pseudomonas syringae pv. tabaci)

The effects of divers carbon substrates were investigated for the tabtoxin production of an isolated pathogenic Pseudomonas syringae pv. tabaci, the causal agent of wildfire of tobacco and are discussed in relation to the bacterium growth. The isolated organism was grown in batch culture on Woolley's medium (28°C, 200 rpm, during 5 days). The growth has been measured by the optical density (OD) at 620 nm and the tabtoxin production quantified by Escherichia coli (K-12) bioassay technique. The growth and the tabtoxin production were both influenced by the substrates (sugars, amino acids, organic acids) used, each, as a sole carbon source and as a supplement for the same amino acids. The most significant quantities of tabtoxin were obtained in presence of some amino acids used as sole carbon source and/or as supplement.

Detection of Pathogenic Escherichia coli Strains Pollution in Red Deer Meat in Latvia and Determination the Compatibility of VT1, VT2, eae A Genes in their Isolate

Tasks of the work were study the possible E.coli contamination in red deer meat, identify pathogenic strains from isolated E.coli, determine their incidence in red deer meat and determine the presence of VT1, VT2 and eaeA genes for the pathogenic E.coli. 8 (10%) samples were randomly selected from 80 analysed isolates of E.coli and PCR reaction was performed on them. PCR was done both on initial materials – samples of red deer meat - and for already isolated liqueurs. Two of analysed venison samples contain verotoxin-producing strains of E. coli. It means that this meat is not safe to consumer. It was proven by the sequestration reaction of E. coli and by comparison of the obtained results with the database of microorganism genome available on the internet that the isolated culture corresponds to region 16S rDNS of E. coli thus presenting correctness of the microbiological methods.

Full-genomic Network Inference for Non-model organisms: A Case Study for the Fungal Pathogen Candida albicans

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.

The Effect of Sowing Time on Phytopathogenic Characteristics and Yield of Sunflower Hybrids

The field research was carried out at the Látókép AGTC KIT research area of the University of Debrecen in Eastern-Hungary, on the area of the aeolain loess of the Hajdúság. We examined the effects of the sowing time on the phytopathogenic characteristics and yield production by applying various fertilizer treatments on two different sunflower genotypes (NK Ferti, PR64H42) in 2012 and 2013. We applied three different sowing times (early, optimal, late) and two different treatment levels of fungicides (control = no fungicides applied, double fungicide protection). During our investigations, the studied cropyears were of different sowing time optimum in terms of yield amount (2012: early, 2013: average). By Pearson’s correlation analysis, we have found that delaying the sowing time pronouncedly decreased the extent of infection in both crop years (Diaporthe: r=0.663**, r=0.681**, Sclerotinia: r=0.465**, r=0.622**). The fungicide treatment not only decreased the extent of infection, but had yield increasing effect too (2012: r=0.498**, 2013: r=0.603**). In 2012, delaying of the sowing time increased (r=0.600**), but in 2013, it decreased (r= 0.356*) the yield amount.

Analysis of Plasmids and Restriction Fragment Length Polymorphisms of Acinetobacter baumannii Isolated from Hospitals- AL Jouf Region- KSA

Abstract–The objectives of the current study are to determine the prevalence, etiological agents, drug susceptibility pattern and plasmid profile of Acinetobacter baumannii isolates from Hospital-Acquired Infections (HAI) at Community Hospital, Al Jouf Province, Saudi Arabia. A total of 1890 patients had developed infection during hospital admission and were included in the study. Among those who developed nosocomial infections, 15(9.4), 10(2.7) and 118 (12.7) had respiratory tract infection (RTI), blood stream infections (BSI) and urinary tract (UTI) respectively. A total of 268 bacterial isolates were isolated from nosocomial infection. S. aureus was reported in 23.5% for of the total isolates followed by Klebsiella pneumoniae (17.5%), E. coli (17.2%), P. aeruginosa (11.9%), coagulase negative staphylococcus (9%), A. baumannii (7.1%), Enterobacter spp. (3.4%), Citrobacter freundii (3%), Proteus mirabilis (2.6%), and Proteus vulgaris and Enterococcous faecalis (0.7%). Isolated organisms are multi-drug resistant, predominantly Gram-positive pathogens with a high incidence of methicillin-resistant S. aureus, extended spectrum beta lactamase and vancomycin resistant enterococci organisms. The RFLP (Fragment Length Polymorphisms) patterns of plasmid preparations from isolated A. baumannii isolates had altered RFLP patterns, possibly due to the presence of plasmid(s). Five A. baumannii isolates harbored plasmids all of which were not less than 2.71kbp in molecular weight. Hence, it showed that the gene coding for the isolates were located on the plasmid DNA while the remaining isolates which have no plasmid might showed gene coding for antibiotic resistance being located on chromosomal DNA. Nosocomial infections represent a current problem in Community Hospital, Al Jouf Province, Saudi Arabia. Problems associated with SSI include infection with multidrug resistant pathogens which are difficult to treat and are associated with increased mortality.

A Review on Application of Chitosan as a Natural Antimicrobial

In recent years application of natural antimicrobials instead of conventional ones, due to their hazardous effects on health, has got serious attentions. On the basis of the results of different studies, chitosan, a natural bio-degradable and non-toxic biopolysaccharide derived from chitin, has potential to be used as a natural antimicrobial. Chitosan has exhibited high antimicrobial activity against a wide variety of pathogenic and spoilage microorganisms, including fungi, and Gram-positive and Gramnegative bacteria. The antimicrobial action is influenced by intrinsic factors such as the type of chitosan, the degree of chitosan polymerization and extrinsic factors such as the microbial organism, the environmental conditions and presence of the other components. The use of chitosan in food systems should be based on sufficient knowledge of the complex mechanisms of its antimicrobial mode of action. In this article we review a number of studies on the investigation of chitosan antimicrobial properties and application of them in culture and food mediums.

The Effectiveness of Tebuconazole and Chitosan in Inhibiting the Growth of Fusarium Species on Winter Wheat Grain under Field Conditions

A three-year field experiment (2010-2012) was conducted to determine the abundance of epiphytic and endophytic filamentous fungi colonizing the grain of winter wheat cv. Bogatka. Wheat spikes were protected with tebuconazole or chitosan at the watery ripe stage. Untreated plants served as control. Tebuconazole exerted an inhibitory effect primarily on F. culmorum and F. graminearum, and its effectiveness was determined by the pressure from pathogens that infected wheat spikes during the growing season. Chitosan did not suppress the growth of Fusarium species and Alternaria alternata.

Recent Outbreaks of Highly Pathogenic Avian Influenza Virus in Chickens and Ducks in Egypt: Pathological Study

Clinically, chickens showed progressively developed clinical signs represented by sever cyanosis of the comb and wattles with hemorrhage at the shanks, depression, and ruffling feathers with loss of appetite and high daily mortalities. The ducks showed severe neurological signs as torticollus, seizures and inability to stand with mild signs of diarrhea and depression. Grossly, chickens showed hemorrhages and congestion in most of the organs particularly lung, liver, spleen, trachea and kidney. The examined ducks showed multiple petechial hemorrhages, multifocal hemorrhagic necrosis in the pancreas, pulmonary edema, congestion and hemorrhage in meninges and congestion in the skeletal muscles. Histopathology revealed severe congestion and hemorrhages in most of the organs particularly lung, liver and kidney. Microscopic erosive tracheitis, sever pulmonary congestion and perivascular oedema and lymphogranulocytic pneumonia were constant. The liver showed hepatocyts necrosis and lympho-granulocytic infiltration. The kidney showed renal tubular necrosis and diffuse congestion. Multifocal, neuronal necrosis, hemorrhages, multifocal glial nodules, lympho- histiocytic perivascular cuffing, and occasional neuronophagia were observed in the cerebrum. Other organs showed moderate changes.

Study on Phytochemical Properties, Antibacterial Activity and Cytotoxicity of Aloe vera L.

The aim of the study was to investigate phytochemical properties, antimicrobial activity and cytotoxicity of Aloe vera. The phytochemical screening of the extracts of leaves of A. vera revealed the presence of bioactive compounds such as alkaloids, tannins, flavonoids phenolic compounds, and etc. with absence of cyanogenic glycosides. Three different solvents such as methanol, ethanol and Di-Methyl sulfoxide were used to screen the antimicrobial activity of A. vera leaves against four human clinical pathogens by agar well diffusion method. The maximum antibacterial activities were observed in methanol extract followed by ethanol and Di-Methyl sulfoxide. It was also found that remarkable antibacterial activities with methanolic and ethanolic extracts of A. vera compared with the standard antibiotic, tetracycline that was not active against E. coli and S. boydii and supported the view that A. vera is a potent antimicrobial agent compared with the conventional antibiotic. Moreover, the brine shrimps (Artemia salina) toxicity test exhibited LC50 value was 569.52 ppm. The resulting data indicated that the A. vera plant have less toxic effects on brine shrimp. Hence, it is signified that Aloe vera plant extract is safe to be used as an antimicrobial agent.