International Journal of Microbiology The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. Activity of Medicinal Plant Extracts on Multiplication of Mycobacterium tuberculosis under Reduced Oxygen Conditions Using Intracellular and Axenic Assays Sun, 31 Jan 2016 11:21:02 +0000 Aim. Test the activity of selected medicinal plant extracts on multiplication of Mycobacterium tuberculosis under reduced oxygen concentration which represents nonreplicating conditions. Material and Methods. Acetone, ethanol and aqueous extracts of the plants Acorus calamus L. (rhizome), Ocimum sanctum L. (leaf), Piper nigrum L. (seed), and Pueraria tuberosa DC. (tuber) were tested on Mycobacterium tuberculosis H37Rv intracellularly using an epithelial cell (A549) infection model. The extracts found to be active intracellularly were further studied axenically under reducing oxygen concentrations. Results and Conclusions. Intracellular multiplication was inhibited ≥60% by five of the twelve extracts. Amongst these 5 extracts, in axenic culture, P. nigrum (acetone) was active under aerobic, microaerophilic, and anaerobic conditions indicating presence of multiple components acting at different levels and P. tuberosa (aqueous) showed bactericidal activity under microaerophilic and anaerobic conditions implying the influence of anaerobiosis on its efficacy. P. nigrum (aqueous) and A. calamus (aqueous and ethanol) extracts were not active under axenic conditions but only inhibited intracellular growth of Mycobacterium tuberculosis, suggesting activation of host defense mechanisms to mediate bacterial killing rather than direct bactericidal activity. Purva D. Bhatter, Pooja D. Gupta, and Tannaz J. Birdi Copyright © 2016 Purva D. Bhatter et al. All rights reserved. Comparison of Procleix Ultrio Elite and Procleix Ultrio NAT Assays for Screening of Transfusion Transmitted Infections among Blood Donors in India Tue, 19 Jan 2016 15:06:17 +0000 Background. Introduction of nucleic acid testing (NAT) has helped in decreasing window period donations, resulting in increased safety of blood supplies. NAT combines the advantages of direct and highly sequence-specific detection of viral genomes. We analysed the performance of newer Procleix Ultrio Elite (PUE) and Procleix Ultrio assay (PUA) for the screening of the viral markers in our donor population. Material and Methods. 10,015 donor samples were screened by routine immunoassays and both versions of NAT. NAT yields detected were subjected to viral load estimation and to other serological markers. Results. A total of 21 NAT yields were detected; three were positive by both NAT systems, whereas 18 samples were reactive by PUE only. NAT yields include 18 HBV and 3 HCV yields, of which 17 HBV yields were occult infections and 1 was window period (WP) infection. All 3 HCV yields were WP infections. No HIV-1/HIV-2 yield was found. Conclusion. Efficient target capture chemistry in the new TMA assay version significantly improved sensitivity. NAT is superior to serological immunoassays for screening of the viral markers; and the efficient target capture system in the newer TMA assay, namely, the PUE system, has significantly improved sensitivity over the earlier versions. Rahul Chaurasia, Diptiranjan Rout, Shamsuz Zaman, Kabita Chatterjee, Hem Chandra Pandey, and Abhishek Kumar Maurya Copyright © 2016 Rahul Chaurasia et al. All rights reserved. Antibiotic-Resistant Bacteria Are Major Threats of Otitis Media in Wollo Area, Northeastern Ethiopia: A Ten-Year Retrospective Analysis Tue, 19 Jan 2016 12:30:21 +0000 Antibiotic resistance is an increasingly serious threat to human health that needs an urgent action. The aim of this study was to determine the prevalence and antibiotic susceptibility profiles of bacteria isolated from patient ear discharges suspected of otitis media. A retrospective analysis was performed using culture and antibiotic susceptibility test results of 1225 patients who visited Dessie Regional Health Research Laboratory from 2001 to 2011. Results showed a strong association () between age and the risk of acquiring middle ear infection. The predominant bacterial isolates were Proteus spp. (28.8%), Staphylococcus aureus (23.7%), and Pseudomonas spp. (17.2%). Most of the isolated bacteria showed high resistance to ampicillin (88.5%), ceftriaxone (84.5%), amoxicillin (81.9%), and tetracycline (74.5%). About 72.5% of Proteus spp. and 62.2% of Pseudomonas spp. have developed resistance to one and more antibiotics used to treat them. This retrospective study also revealed the overall antibiotic resistance rate of bacterial isolates was increased nearly twofold () over the last decade. Relatively, ciprofloxacin and gentamicin were the most effective antibiotics against all the isolates. In conclusion, antibiotic-resistant bacteria are alarmingly increasing in Wollo area, northeastern Ethiopia, and becoming a major public health problem in the management of patients with middle ear infection. Ayele Argaw-Denboba, Asrat Agalu Abejew, and Alemayehu Gashaw Mekonnen Copyright © 2016 Ayele Argaw-Denboba et al. All rights reserved. Eucalyptus Tree: A Potential Source of Cryptococcus neoformans in Egyptian Environment Wed, 13 Jan 2016 11:28:48 +0000 In Egypt, the River Red Gum (Eucalyptus camaldulensis) is a well-known tree and is highly appreciated by the rural and urban dwellers. The role of Eucalyptus trees in the ecology of Cryptococcus neoformans is documented worldwide. The aim of this survey was to show the prevalence of C. neoformans during the flowering season of E. camaldulensis at the Delta region in Egypt. Three hundred and eleven samples out of two hundred Eucalyptus trees, including leaves, flowers, and woody trunks, were collected from four governorates in the Delta region. Thirteen isolates of C. neoformans were recovered from Eucalyptus tree samples (4.2%). Molecular identification of C. neoformans was done by capsular gene specific primer CAP64 and serotype identification was done depending on LAC1 gene. This study represents an update on the ecology of C. neoformans associated with Eucalyptus tree in Egyptian environment. Mahmoud Elhariri, Dalia Hamza, Rehab Elhelw, and Mohamed Refai Copyright © 2016 Mahmoud Elhariri et al. All rights reserved. Modulation of mecA Gene Expression by Essential Oil from Salvia sclarea and Synergism with Oxacillin in Methicillin Resistant Staphylococcus epidermidis Carrying Different Types of Staphylococcal Chromosomal Cassette mec Wed, 06 Jan 2016 14:01:08 +0000 The essential oil (EO) from Salvia sclarea was shown to increase the susceptibility of methicillin resistant Staphylococcus epidermidis (MRSE) isolates to oxacillin. The purpose of this study was to investigate the effect of EO from S. sclarea on expression of mecA gene of MRSE carrying different types of staphylococcal chromosomal cassette (SCCmec) and to evaluate potential synergistic effect of EO with oxacillin. Using real-time PCR we found that EO alone inhibited the expression of the resistant genes mecA, mecR1, and mecI and blaZ, blaR1, and blaI. The use of the combination of EO with oxacillin resulted in significantly inhibited expression of mecA gene in all tested strains with different types of SCCmec. Using time-kill assay and checkerboard assay we confirmed synergistic effect of EO from S. sclarea and oxacillin in MRSE. Romana Chovanová, Mária Mikulášová, and Štefánia Vaverková Copyright © 2016 Romana Chovanová et al. All rights reserved. Spectrum and the In Vitro Antifungal Susceptibility Pattern of Yeast Isolates in Ethiopian HIV Patients with Oropharyngeal Candidiasis Tue, 05 Jan 2016 13:42:29 +0000 Background. In Ethiopia, little is known regarding the distribution and the in vitro antifungal susceptibility profile of yeasts. Objective. This study was undertaken to determine the spectrum and the in vitro antifungal susceptibility pattern of yeasts isolated from HIV infected patients with OPC. Method. Oral pharyngeal swabs taken from oral lesions of study subjects were inoculated onto Sabouraud Dextrose Agar. Yeasts were identified by employing conventional test procedures and the susceptibility of yeasts to antifungal agents was evaluated by disk diffusion assay method. Result. One hundred and fifty-five yeast isolates were recovered of which 91 isolates were from patients that were not under HAART and 64 were from patients that were under HAART. C. albicans was the most frequently isolated species followed by C. glabrata, C. tropicalis, C. krusei, C. kefyr, Cryptococcus laurentii, and Rhodotorula species. Irrespective of yeasts isolated and identified, 5.8%, 5.8%, 12.3%, 8.4%, 0.6%, and 1.3% of the isolates were resistant to amphotericin B, clotrimazole, fluconazole, ketoconazole, miconazole, and nystatin, respectively. Conclusion. Yeast colonization rate of 69.2% and 31% resistance to six antifungal agents was documented. These highlight the need for nationwide study on the epidemiology of OPC and resistance to antifungal drugs. Birhan Moges, Adane Bitew, and Aster Shewaamare Copyright © 2016 Birhan Moges et al. All rights reserved. In Vitro Antimicrobial Activity of Spices and Medicinal Herbs against Selected Microbes Associated with Juices Mon, 04 Jan 2016 12:49:29 +0000 In the present investigation, comparison of antimicrobial activities of different spices, Curcuma longa, Zingiber officinale, and Mentha arvensis, and medicinal herbs, such as Withania somnifera, Rauvolfia serpentina, Emblica officinalis, Terminalia arjuna, and Centella asiatica, was evaluated. Different extraction solvents (acetone, methanol, ethanol, and water) were used and extracts were examined against Bacillus cereus, Serratia sp., Rhodotorula mucilaginosa, Aspergillus flavus, and Penicillium citrinum isolated from juices. Extracts from the medicinal herb and spices have significant activity. B. cereus was the most sensitive and R. mucilaginosa was the most resistant among the microorganisms tested. Ethanolic and methanolic extract of C. asiatica displayed maximum diameter of inhibition zone against bacteria and yeast and percentage mycelial inhibition against moulds. This study confirmed the potential of selected extracts of spices as effective natural food preservative in juices. Romika Dhiman, Neeraj Aggarwal, Kamal Rai Aneja, and Manpreet Kaur Copyright © 2016 Romika Dhiman et al. All rights reserved. In Vitro Antimicrobial Activity of Essential Oil of Thymus schimperi, Matricaria chamomilla, Eucalyptus globulus, and Rosmarinus officinalis Mon, 04 Jan 2016 11:10:38 +0000 In this study, the in vitro antimicrobial activities of four plant essential oils (T. schimperi, E. globulus, R. officinalis, and M. Chamomilla) were evaluated against bacteria and fungi. The studies were carried out using agar diffusion method for screening the most effective essential oils and agar dilution to determine minimum inhibitory concentration of the essential oils. Results of this study revealed that essential oils of T. schimperi, E. globulus, and R. officinalis were active against bacteria and some fungi. The antimicrobial effect of M. chamomilla was found to be weaker and did not show any antimicrobial activity. The minimum inhibitory concentration values of T. schimperi were 15.75 mg/mL for most of the bacteria and fungi used in this study. The minimum inhibitory concentration values of the other essential oils were in the range of 15.75–36.33 mg/mL against tested bacteria. This study highlighted the antimicrobial activity of the essential oil of E. globulus, M. chamomilla, T. Schimperi, and R. officinalis. The results indicated that T. schimperi have shown strong antimicrobial activity which could be potential candidates for preparation of antimicrobial drug preparation. Awol Mekonnen, Berhanu Yitayew, Alemnesh Tesema, and Solomon Taddese Copyright © 2016 Awol Mekonnen et al. All rights reserved. Rapid Accumulation of Total Lipid in Rhizoclonium africanum Kutzing as Biodiesel Feedstock under Nutrient Limitations and the Associated Changes at Cellular Level Thu, 31 Dec 2015 09:35:29 +0000 Increase of total lipid and the proportion of the favorable fatty acids in marine green filamentous macroalga Rhizoclonium africanum (Chlorophyceae) was studied under nitrate and phosphate limitations. These stresses were given by both eliminating and doubling the required amounts of nitrate and phosphate salts in the growth media. A significant twofold increase in total lipid (193.03 mg/g) was achieved in cells in absence of nitrate in the culture medium, followed by phosphate limitation (142.65 mg/g). The intracellular accumulation of neutral lipids was observed by fluorescence microscopy. The scanning electron microscopic study showed the major structural changes under nutrient starvation. Fourier transform infrared spectroscopy (FTIR) revealed the presence of ester (C-O-C stretching), ketone (C-C stretching), carboxylic acid (O-H bending), phosphine (P-H stretching), aromatic (C-H stretching and bending), and alcohol (O-H stretching and bending) groups in the treated cells indicating the high accumulation of lipid hydrocarbons in the treated cells. Elevated levels of fatty acids favorable for biodiesel production, that is, C16:0, C16:1, C18:1, and C20:1, were identified under nitrate- and phosphate-deficient conditions. This study shows that the manipulation of cultural conditions could affect the biosynthetic pathways leading to increased lipid production while increasing the proportion of fatty acids suitable for biodiesel production. Gour Gopal Satpati, Sanjit Kanjilal, Rachapudi Badari Narayana Prasad, and Ruma Pal Copyright © 2015 Gour Gopal Satpati et al. All rights reserved. Carbon Sources for Yeast Growth as a Precondition of Hydrogen Peroxide Induced Hormetic Phenotype Wed, 30 Dec 2015 09:18:26 +0000 Hormesis is a phenomenon of particular interest in biology, medicine, pharmacology, and toxicology. In this study, we investigated the relationship between H2O2-induced hormetic response in S. cerevisiae and carbon sources in yeast growth medium. In general, our data indicate that (i) hydrogen peroxide induces hormesis in a concentration-dependent manner; (ii) the effect of hydrogen peroxide on yeast reproductive ability depends on the type of carbon substrate in growth medium; and (iii) metabolic and growth rates as well as catalase activity play an important role in H2O2-induced hormetic response in yeast. Ruslana Vasylkovska, Natalia Petriv, and Halyna Semchyshyn Copyright © 2015 Ruslana Vasylkovska et al. All rights reserved. Isolation of Bacillus sphaericus from Lombok Island, Indonesia, and Their Toxicity against Anopheles aconitus Mon, 14 Dec 2015 09:15:47 +0000 Malaria is endemic to Lombok Island, Indonesia. One approach to suppress malaria spread is to eliminate anopheline larvae in their habitat and the environmentally safe agent is bacteria, that is, Bacillus sphaericus. However, there is no information regarding local isolate of B. sphaericus that is toxic to mosquito larvae from Lombok. The aims of the study were to isolate B. sphaericus from soil in areas close to beach surrounding Lombok Island and to test their toxicity against 3rd instar Anopheles aconitus larvae. Soil samples were collected from 20 different sampling locations from Lombok Island and homogenized with sterile physiological salt solution. Suspension was heat-shocked at 80°C for 30 minutes and then spread onto antibiotic-supplemented NYSM solid medium. Colonies grown were characterized and subjected to initial toxicity test against anopheline larvae. Isolates with more than 50% killing percentage were subjected to bioassay testing against anopheline larvae. From 20 locations, 1 isolate showed mild toxicity (namely, isolate MNT) and 2 isolates showed high toxicity (namely, isolates SLG and TJL2) against An. aconitus. Those 3 isolates were potentially useful isolates, as they killed almost all larvae in 24 hours. The discovery of toxic indigenous isolates of B. sphaericus from Lombok Island opens opportunity to develop a biopesticide from local resources. Bambang Fajar Suryadi, Bagyo Yanuwiadi, Tri Ardyati, and Suharjono Copyright © 2015 Bambang Fajar Suryadi et al. All rights reserved. In Vitro Evaluation of Antimicrobial Efficacy of Extracts Obtained from Raw and Fermented Wild Macrofungus, Lenzites quercina Thu, 29 Oct 2015 11:00:38 +0000 In recent time, there is a major concern about antibiotic resistance displayed by some pathogenic microorganisms and this had involved a continuous search for natural antimicrobial products. The phytochemistry as well as antimicrobial activity of extracts obtained from Lenzites quercina was investigated. The extracts and purified fractions were, respectively, tested against indicator organisms using agar well diffusion and disc diffusion methods. The quantity of phytochemicals found in the extracts of L. quercina ranged from 14.4 to 20.7 mg/g for alkaloids, 6.1 to 12.8 mg/g for steroids, 4.5 to 10.6 mg/g for saponins, 2.8 to 17.2 mg/g for terpenoids, and 0.41 to 17.1 mg/g for flavonoids. The gas chromatography mass spectrophotometry (GCMS) analysis of the extract reveals the presence of caprylic acid, stearic acid, tetradecanoic acid, methyl-11-octadecenoate, oleic acid, and 4-methyl-2-propyl-1-pentanol. Extracts of L. quercina and its purified fractions exhibited wider range of inhibition (4 mm to 26 mm) on Staphylococcus aureus (ATCC 29213), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 35218), Methicillin Resistant Staphylococcus aureus (MRSA), Salmonella typhi, Bacillus cereus, Enterococcus faecalis, Candida albicans, and Aspergillus niger. The antimicrobial effects of L. quercina extracts indicate that this wild macrofungus contains significant amount of pharmacological agents, which could be extracted to curb the menace of antibiotic resistances by pathogenic organisms. Olusola Clement Ogidi, Victor Olusegun Oyetayo, and Bamidele Juliet Akinyele Copyright © 2015 Olusola Clement Ogidi et al. All rights reserved. Nosocomial Isolates and Their Drug Resistant Pattern in ICU Patients at National Institute of Neurological and Allied Sciences, Nepal Mon, 26 Oct 2015 06:17:58 +0000 Multidrug resistant organisms are increasing day by day and the cause is poorly known. This study was carried out from June 2011 to May 2012 at National Institute of Neurological and Allied Sciences Kathmandu, Nepal, with a view to determining drug resistant pathogens along with detection of extended spectrum β-lactamase (ESBL), AmpC β-lactamase (ABL), and metallo-β-lactamase (MBL) producing bacteria causing infection to ICU patients. A standard methodology was used to achieve these objectives as per recommendation of American Society for Microbiology. ESBL was detected by combined disc assay using cefotaxime and cefotaxime clavulanic acid, ABL by inhibitor based method using cefoxitin and phenylboronic acid, and MBL by imipenem-EDTA combined disk method. Two hundred and ninety-four different clinical samples such as tracheal aspirates, urine, pus, swabs, catheter tips, and blood were processed during the study. Most common bacteria were Acinetobacter spp. Of the total 58 Acinetobacter spp., 46 (79%) were MDR, and 27% were positive for ABL and 12% were for MBL. Of the 32 cases of Staphylococcus aureus, 18 (56%) were MDR. Findings of this study warrant routine β-lactamase testing in clinical isolates. Pashupati Bhandari, Ganesh Thapa, Bharat Mani Pokhrel, Dwij Raj Bhatta, and Upendra Devkota Copyright © 2015 Pashupati Bhandari et al. All rights reserved. Characterization of Staphylococcus aureus Isolates That Colonize Medical Students in a Hospital of the City of Cali, Colombia Thu, 01 Oct 2015 08:18:57 +0000 Introduction. Nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) represents a risk for the spread of bacteria. This study characterized the S. aureus isolated from medical students, who were in their clinical rotation at a hospital in the city of Cali. Materials and Methods. 216 students participated in the study and 63 isolates of S. aureus were evaluated for susceptibility and PCR amplification of agr and mecA genes. The origin of MRSA isolates was established by analyzing agr polymorphisms. Results. A total of 29.2% of students were colonized by S. aureus and nasal carriage rate was 23.6% and 14.3% MRSA. Three agr groups (agr II, and agr III) were identified; the agr I group was the most common, with a 35% prevalence; this group is from community origin. Conclusion. The present study demonstrates that medical students carry S. aureus strains, with the threat of spreading them both to community and hospital environments. Luis Fernando Collazos Marín, Gina Estupiñan Arciniegas, and Monica Chavez Vivas Copyright © 2015 Luis Fernando Collazos Marín et al. All rights reserved. Regulation of Expression of Oxacillin-Inducible Methionine Sulfoxide Reductases in Staphylococcus aureus Mon, 21 Sep 2015 12:04:12 +0000 Cell wall-active antibiotics cause induction of a locus that leads to elevated synthesis of two methionine sulfoxide reductases (MsrA1 and MsrB) in Staphylococcus aureus. To understand the regulation of this locus, reporter strains were constructed by integrating a DNA fragment consisting of the msrA1/msrB promoter in front of a promoterless lacZ gene in the chromosome of wild-type and MsrA1-, MsrB-, MsrA1/MsrB-, and SigB-deficient methicillin-sensitive S. aureus strain SH1000 and methicillin-resistant S. aureus strain COL. These reporter strains were cultured in TSB and the cellular levels of β-galactosidase activity in these cultures were assayed during different growth phases. β-galactosidase activity assays demonstrated that the lack of MsrA1, MsrB, and SigB upregulated the msrA1/msrB promoter in S. aureus strain SH1000. In S. aureus strain COL, the highest level of β-galactosidase activity was observed under the conditions when both MsrA1 and MsrB proteins were absent. The data suggest that the msrA1/msrB locus, in part, is negatively regulated by MsrA1, MsrB, and SigB in S. aureus. Kyle R. Baum, Zulfiqar Ahmad, and Vineet K. Singh Copyright © 2015 Kyle R. Baum et al. All rights reserved. Prevalence of Dermatophytic Infection and the Spectrum of Dermatophytes in Patients Attending a Tertiary Hospital in Addis Ababa, Ethiopia Sun, 13 Sep 2015 06:27:55 +0000 Background. Dermatophytosis is common worldwide and continues to increase. Objective. This study was undertaken to determine the prevalence of dermatophytosis and the spectrum of ringworm fungi in patients attending a tertiary hospital. Methods. Samples were collected from 305 patients. A portion of each sample was examined microscopically and the remaining portion of each sample was cultured onto plates of Sabouraud’s dextrose agar containing chloramphenicol with and without cycloheximide. Dermatophyte isolates were identified by studying macroscopic and microscopic characteristics of their colonies. Result. Of 305 samples, fungi were detected in 166 (54.4%) by KOH of which 95 were dermatophytes while 242 (79.4%) samples were culture positive of which 130 isolates were dermatophytes. Among dermatophyte isolates T. violaceum was the most common (37.7%) cause of infection. Tinea unguium was the predominant clinical manifestation accounting for 51.1% of the cases. Patients with age group 25–44 and 45–64 years were more affected. T. violaceum was the most common pathogen in tinea unguium and tinea capitis, whereas T. mentagrophytes was the most common pathogen in tinea pedis. Conclusion. Further intensive epidemiological studies of ring worm fungus induced dermatophytosis which have public health significance are needed. Gebreabiezgi Teklebirhan and Adane Bitew Copyright © 2015 Gebreabiezgi Teklebirhan and Adane Bitew. All rights reserved. Isolation of Fungi and Bacteria Associated with the Guts of Tropical Wood-Feeding Coleoptera and Determination of Their Lignocellulolytic Activities Wed, 26 Aug 2015 07:56:06 +0000 The guts of beetle larvae constitute a complex system where relationships among fungi, bacteria, and the insect host occur. In this study, we collected larvae of five families of wood-feeding Coleoptera in tropical forests of Costa Rica, isolated fungi and bacteria from their intestinal tracts, and determined the presence of five different pathways for lignocellulolytic activity. The fungal isolates were assigned to three phyla, 16 orders, 24 families, and 40 genera; Trichoderma was the most abundant genus, detected in all insect families and at all sites. The bacterial isolates were assigned to five phyla, 13 orders, 22 families, and 35 genera; Bacillus, Serratia, and Pseudomonas were the dominant genera, present in all the Coleopteran families. Positive results for activities related to degradation of wood components were determined in 65% and 48% of the fungal and bacterial genera, respectively. Our results showed that both the fungal and bacterial populations were highly diverse in terms of number of species and their phylogenetic composition, although the structure of the microbial communities varied with insect host family and the surrounding environment. The recurrent identification of some lignocellulolytic-positive inhabitants suggests that particular microbial groups play important roles in providing nutritional needs for the Coleopteran host. Keilor Rojas-Jiménez and Myriam Hernández Copyright © 2015 Keilor Rojas-Jiménez and Myriam Hernández. All rights reserved. Mycobiota and Natural Incidence of Aflatoxins, Ochratoxin A, and Citrinin in Indian Spices Confirmed by LC-MS/MS Thu, 02 Jul 2015 08:48:49 +0000 Nine different Indian spices (red chilli, black pepper, turmeric, coriander, cumin, fennel, caraway, fenugreek, and dry ginger) commonly cultivated and highly used in India were analysed for natural occurrence of toxigenic mycoflora and aflatoxins (AFs), ochratoxin A (OTA), and citrinin (CTN) contamination. Aspergillus flavus and Aspergillus niger were the most dominant species isolated from all types of spices. Red chilli samples were highly contaminated with aflatoxins (85.4%) followed by dry ginger (77.7%). 56% Aspergillus flavus from red chilli and 45% Aspergillus ochraceus from black pepper were toxigenic and produced aflatoxins and ochratoxin A, respectively. Qualitative detection and quantitative detection of mycotoxins in spices were analyzed by ELISA and further confirmed by LC-MS/MS. Penicillium citrinum produced citrinin in red chilli, black pepper, coriander, cumin, fenugreek, and dry ginger samples. The highest amount of AFs was found in red chilli (219.6 ng/g), OTA was in black pepper (154.1 ng/g), and CTN was in dry ginger samples (85.1 ng/g). The results of this study suggest that the spices are susceptible substrate for growth of mycotoxigenic fungi and further mycotoxin production. This is the first report of natural occurrence of citrinin in black pepper and dry ginger from India. Punam Jeswal and Dhiraj Kumar Copyright © 2015 Punam Jeswal and Dhiraj Kumar. All rights reserved. Antimicrobial Activities of a Plethora of Medicinal Plant Extracts and Hydrolates against Human Pathogens and Their Potential to Reverse Antibiotic Resistance Wed, 27 May 2015 09:56:18 +0000 Microbial infections till date remain a scourge of humanity due to lack of vaccine against some infections, emergence of drug resistant phenotypes, and the resurgence of infections amongst others. Continuous quest for novel therapeutic approaches remains imperative. Here we (i) assessed the effects of extracts/hydrolates of some medicinal plants on pathogenic microorganisms and (ii) evaluated the inhibitory potential of the most active ones in combination with antibiotics. Extract E03 had the highest DZI (25 mm). Extracts E05 and E06 were active against all microorganisms tested. The MICs and MBCs of the methanol extracts ranged from 16.667 × 103 μg/mL to 2 μg/mL and hydrolates from 0.028 to 333333 ppm. Extract E30 had the highest activity especially against S. saprophyticus (MIC of 6 ppm) and E. coli (MIC of 17 ppm). Combination with conventional antibiotics was shown to overcome resistance especially with E30. Analyses of the extracts revealed the presence of alkaloids, flavonoids, triterpenes, steroids, phenols, and saponins. These results justify the use of these plants in traditional medicine and the practice of supplementing decoctions/concoctions with conventional antibiotics. Nauclea pobeguinii (E30), the most active and synergistic of all these extracts, and some hydrolates with antimicrobial activity need further exploration for the development of novel antimicrobials. Dieudonné Lemuh Njimoh, Jules Clement N. Assob, Seraphine Ebenye Mokake, Dinga Jerome Nyhalah, Claude Kwe Yinda, and Bertrand Sandjon Copyright © 2015 Dieudonné Lemuh Njimoh et al. All rights reserved. Viral Agents of Diarrhea in Young Children in Two Primary Health Centers in Edo State, Nigeria Thu, 30 Apr 2015 07:32:40 +0000 Enteric viruses have been shown to be responsible for diarrhea among children during their early childhood. This study was carried out to determine the prevalence of rotavirus, adenovirus, and norovirus infection in young children with diarrhea in two primary health centers in Edo State, Nigeria. A total of 223 stool specimens were collected from children aged 0–36 months with clinical signs of diarrhea and 59 apparently healthy age-matched children as control. These specimens were investigated for three viral agents using immunochromatographic technique (ICT). The overall results showed that at least one viral agent was detected in 95/223 (42.6%) of the children with diarrhea while the control had none. The prevalence of rotavirus was 28.3%, adenovirus 19.3%, and norovirus 3.6%. There was a significant association between age group and infection (). Seasonal pattern of enteric viruses was not statistically significant (). The overall coinfection rate was 7.6% and rotavirus-adenovirus coinfection had the highest with 5.4%. Rotavirus was the most prevalent viral agent. Coinfections are not uncommon among the population studied. The most commonly associated clinical symptom of viral diarrhea in this study was vomiting. Viral diagnostic tests are advocated for primary health care facilities in this locality. Paul Erhunmwunse Imade and Nosakhare Odeh Eghafona Copyright © 2015 Paul Erhunmwunse Imade and Nosakhare Odeh Eghafona. All rights reserved. Rapid Identification of Pathogens in Positive Blood Culture of Patients with Sepsis: Review and Meta-Analysis of the Performance of the Sepsityper Kit Mon, 27 Apr 2015 13:59:11 +0000 Sepsis is one of the leading causes of deaths, and rapid identification (ID) of blood stream infection is mandatory to perform adequate antibiotic therapy. The advent of MALDI-TOF Mass Spectrometry for the rapid ID of pathogens was a major breakthrough in microbiology. Recently, this method was combined with extraction methods for pathogens directly from positive blood cultures. This review summarizes the results obtained so far with the commercial Sepsityper sample preparation kit, which is now approved for in vitro diagnostic use. Summarizing data from 21 reports, the Sepsityper kit allowed a reliable ID on the species level of 80% of 3320 positive blood culture bottles. Gram negative bacteria resulted consistently in higher ID rates (90%) compared to Gram positive bacteria (76%) or yeast (66%). No relevant misidentifications on the genus level were reported at a log(score)cut-off of 1.6. The Sepsityper kit is a simple and reproducible method which extends the MALDI-TOF technology to positive blood culture specimens and shortens the time to result by several hours or even days. In combination with antibiotic stewardship programs, this rapid ID allows a much faster optimization of antibiotic therapy in patients with sepsis compared to conventional workflows. Nils G. Morgenthaler and Markus Kostrzewa Copyright © 2015 Nils G. Morgenthaler and Markus Kostrzewa. All rights reserved. Effects of Two Salts Compounds on Mycelial Growth, Sporulation, and Spore Germination of Six Isolates of Botrytis cinerea in the Western North of Algeria Thu, 26 Mar 2015 14:06:37 +0000 Six isolates of Botrytis cinerea were isolated from leaves and stems of different tomato varieties taken from four areas in the northwest of Algeria where tomato is mostly grown in greenhouses and high tunnels. The purpose of this research was to determine the effect of two salts, NaCl and CaCl2, on three stages of Botrytis cinerea’s life cycle. All isolates tested were stimulated in 50 to 150 ppm; NaCl was the most effective treatment to increase mycelial growth at two tested concentrations. However, at 300 ppm concentration, CaCl2 completely inhibited the growth of mycelium; they reach 34.78% for the isolate TR46 and 26.72% for isolate F27. The sodium and calcium salts stimulated conidia production in liquid culture. We noticed that the effect of calcium chloride on sporulation was average while sodium chloride. In the medium containing 50 ppm, calcium chloride and sodium chloride increased the germination capacity of most isolates compared with the control. Other calcium salts, at 100 or 300 ppm, decreased the germination percentage of the conidia. With the exception of sodium salts, the inhibitions of germination reduce at 150 or 300 compared with the control. Conidial germination was slightly inhibited by sodium chloride only when the concentration was over 300 ppm. Boualem Boumaaza, Mohamed Benkhelifa, and Moulay Belkhoudja Copyright © 2015 Boualem Boumaaza et al. All rights reserved. Ebola Virus Disease 2013-2014 Outbreak in West Africa: An Analysis of the Epidemic Spread and Response Tue, 17 Mar 2015 07:33:29 +0000 The Ebola virus epidemic burst in West Africa in late 2013, started in Guinea, reached in a few months an alarming diffusion, actually involving several countries (Liberia, Sierra Leone, Nigeria, Senegal, and Mali). Guinea and Liberia, the first nations affected by the outbreak, have put in place measures to contain the spread, supported by international organizations; then they were followed by the other nations affected. In the present EVD outbreak, the geographical spread of the virus has followed a new route: the achievement of large urban areas at an early stage of the epidemic has led to an unprecedented diffusion, featuring the largest outbreak of EVD of all time. This has caused significant concerns all over the world: the potential reaching of far countries from endemic areas, mainly through fast transports, induced several countries to issue information documents and health supervision for individuals going to or coming from the areas at risk. In this paper the geographical spread of the epidemic was analyzed, assessing the sequential appearance of cases by geographic area, considering the increase in cases and mortality according to affected nations. The measures implemented by each government and international organizations to contain the outbreak, and their effectiveness, were also evaluated. Orlando Cenciarelli, Stefano Pietropaoli, Andrea Malizia, Mariachiara Carestia, Fabrizio D’Amico, Alessandro Sassolini, Daniele Di Giovanni, Silvia Rea, Valentina Gabbarini, Annalaura Tamburrini, Leonardo Palombi, Carlo Bellecci, and Pasquale Gaudio Copyright © 2015 Orlando Cenciarelli et al. All rights reserved. Enteropathogens Associated with Acute Diarrhea in Children from Households with High Socioeconomic Level in Uruguay Thu, 12 Mar 2015 08:18:15 +0000 Infectious diarrhea, a common disease of children, deserves permanent monitoring in all social groups. To know the etiology and clinical manifestations of acute diarrhea in children up to 5 years of age from high socioeconomic level households, we conducted a descriptive, microbiological, and clinical study. Stools from 59 children with acute community-acquired diarrhea were examined, and their parents were interviewed concerning symptoms and signs. Rotavirus, adenovirus, and norovirus were detected by commercially available qualitative immunochromatographic lateral flow rapid tests. Salmonella, Campylobacter, Yersinia, and Shigella were investigated by standard bacteriological methods and diarrheagenic E. coli by PCR assays. We identified a potential enteric pathogen in 30 children. The most frequent causes of diarrhea were enteropathogenic E. coli (EPEC), viruses, Campylobacter, Salmonella, and Shiga-toxin-producing E. coli (STEC). Only 2 patients showed mixed infections. Our data suggest that children with viral or Campylobacter diarrhea were taken to the hospital earlier than those infected with EPEC. One child infected with STEC O26 developed “complete” HUS. The microbiological results highlight the importance of zoonotic bacteria such as atypical EPEC, Campylobacter, STEC, and Salmonella as pathogens associated with acute diarrhea in these children. The findings also reinforce our previous communications about the regional importance of non-O157 STEC strains in severe infant food-borne diseases. Gustavo Varela, Lara Batthyány, María Noel Bianco, Walter Pérez, Lorena Pardo, Gabriela Algorta, Luciana Robino, Ramón Suárez, Armando Navarro, María Catalina Pírez, and Felipe Schelotto Copyright © 2015 Gustavo Varela et al. All rights reserved. Edible Mushrooms: Improving Human Health and Promoting Quality Life Tue, 20 Jan 2015 08:31:39 +0000 Mushrooms have been consumed since earliest history; ancient Greeks believed that mushrooms provided strength for warriors in battle, and the Romans perceived them as the “Food of the Gods.” For centuries, the Chinese culture has treasured mushrooms as a health food, an “elixir of life.” They have been part of the human culture for thousands of years and have considerable interest in the most important civilizations in history because of their sensory characteristics; they have been recognized for their attractive culinary attributes. Nowadays, mushrooms are popular valuable foods because they are low in calories, carbohydrates, fat, and sodium: also, they are cholesterol-free. Besides, mushrooms provide important nutrients, including selenium, potassium, riboflavin, niacin, vitamin D, proteins, and fiber. All together with a long history as food source, mushrooms are important for their healing capacities and properties in traditional medicine. It has reported beneficial effects for health and treatment of some diseases. Many nutraceutical properties are described in mushrooms, such as prevention or treatment of Parkinson, Alzheimer, hypertension, and high risk of stroke. They are also utilized to reduce the likelihood of cancer invasion and metastasis due to antitumoral attributes. Mushrooms act as antibacterial, immune system enhancer and cholesterol lowering agents; additionally, they are important sources of bioactive compounds. As a result of these properties, some mushroom extracts are used to promote human health and are found as dietary supplements. María Elena Valverde, Talía Hernández-Pérez, and Octavio Paredes-López Copyright © 2015 María Elena Valverde et al. All rights reserved. Childhood Septicemia in Nepal: Documenting the Bacterial Etiology and Its Susceptibility to Antibiotics Thu, 25 Dec 2014 13:08:08 +0000 Introduction. Children are among the most vulnerable population groups to contract illnesses. The varying microbiological pattern of septicemia warrants the need for an ongoing review of the causative organisms and their antimicrobial susceptibility pattern. Therefore, the objective of this study was to document the bacterial etiology of childhood septicemia and its antibiotic susceptibility profile. Methods. Cross-sectional type of study in 1630 suspected patients was conducted at CMCTH from January 2012 to December 2013. Blood samples were collected aseptically for culture. The organisms grown were identified by standard microbiological methods recommended by American Society for Microbiology (ASM) and subjected to antibiotic susceptibility testing by modified Kirby-Bauer disk diffusion method. Methicillin resistance was confirmed using cefoxitin and oxacillin disks methods. Results. Septicemia was detected in 172 (10.6%) cases. Among Gram-positive organisms, coagulase negative staphylococci (CoNS) were leading pathogen and Acinetobacter spp. were leading pathogen among Gram-negative isolates. Vancomycin, teicoplanin, and clindamycin were the most effective antibiotics against Gram-positive isolates while amikacin was effective against Gram-positive as well as Gram-negative isolates. Methicillin resistance was detected in 44.4% of Staphylococcus aureus. Conclusions. This study has highlighted the burden of bacterial etiology for septicemia among children in a tertiary care center of central Nepal. Shamshul Ansari, Hari Prasad Nepal, Rajendra Gautam, Sony Shrestha, Puja Neopane, Brihaspati Rimal, Fuleshwar Mandal, Safiur Rahman Ansari, and Moti Lal Chapagain Copyright © 2014 Shamshul Ansari et al. All rights reserved. Global Phenotypic Characterization of Effects of Fluoroquinolone Resistance Selection on the Metabolic Activities and Drug Susceptibilities of Clostridium perfringens Strains Sun, 21 Dec 2014 00:10:15 +0000 Fluoroquinolone resistance affects toxin production of Clostridium perfringens strains differently. To investigate the effect of fluoroquinolone resistance selection on global changes in metabolic activities and drug susceptibilities, four C. perfringens strains and their norfloxacin-, ciprofloxacin-, and gatifloxacin-resistant mutants were compared in nearly 2000 assays, using phenotype microarray plates. Variations among mutant strains resulting from resistance selection were observed in all aspects of metabolism. Carbon utilization, pH range, osmotic tolerance, and chemical sensitivity of resistant strains were affected differently in the resistant mutants depending on both the bacterial genotype and the fluoroquinolone to which the bacterium was resistant. The susceptibilities to gentamicin and erythromycin of all resistant mutants except one increased, but some resistant strains were less susceptible to amoxicillin, cefoxitin, ceftriaxone, chloramphenicol, and metronidazole than their wild types. Sensitivity to ethidium bromide decreased in some resistant mutants and increased in others. Microarray analysis of two gatifloxacin-resistant mutants showed changes in metabolic activities that were correlated with altered expression of various genes. Both the chemical structures of fluoroquinolones and the genomic makeup of the wild types influenced the changes found in resistant mutants, which may explain some inconsistent reports of the effects of therapeutic use of fluoroquinolones on clinical isolates of bacteria. Miseon Park and Fatemeh Rafii Copyright © 2014 Miseon Park and Fatemeh Rafii. All rights reserved. Microbiological Analysis of Surfaces of Leonardo Da Vinci’s Atlantic Codex: Biodeterioration Risk Wed, 10 Dec 2014 00:10:38 +0000 Following the discovery of discoloration on some pages of the Atlantic Codex (AC) of Leonardo da Vinci kept in the Biblioteca Ambrosiana in Milan, some investigations have been carried out to verify the presence of microorganisms, such as bacteria and fungi. To verify the presence of microorganisms a noninvasive method of sampling has been used that was efficient and allowed us to highlight the microbial facies of the material that was examined using conventional microbiological techniques. The microclimatic conditions in the storage room as well as the water content of the volume were also assessed. The combined observations allowed the conclusion that the discoloration of suspected biological origin on some pages of AC is not related to the presence or current attack of microbial agents. Gianfranco Tarsitani, Catia Moroni, Francesca Cappitelli, Giovanna Pasquariello, and Oriana Maggi Copyright © 2014 Gianfranco Tarsitani et al. All rights reserved. Candida glabrata Esophagitis: Are We Seeing the Emergence of a New Azole-Resistant Pathogen? Wed, 03 Dec 2014 07:17:51 +0000 Background. Candida glabrata (C. glabrata) has become a recognized pathogen in fungal esophagitis. A proportion of these isolates are azole-resistant which may have treatment implications. Variability in the prevalence of this organism exists in the limited data available. Objective. To determine the incidence of C. glabrata esophagitis in a North American hospital setting and to highlight factors that may predispose patients to this condition. Methods. Patient charts were collected from January 1, 2009 to July 30, 2011. Any charts of patients identified as having esophagitis with a positive fungal culture were reviewed for the species of Candida and the presence of factors that would predispose them to esophageal candidiasis. Results. The prevalence of Candida esophagitis based on culture was 2.2% (37 subjects). C. glabrata was the 2nd most prevalent pathogen identified (24.3% or 9 subjects). Of the C. glabrata cohort, all patients had at least one factor predisposing them to candidiasis. Conclusion. C. glabrata esophagitis makes up a large portion of the candidal esophagitis seen in hospital. C. glabrata infections were associated with at least one risk factor for candidal infection. Given its resistance to azole-based therapy, this may have treatment implications for how candidal esophagitis is approached by the clinician. Aze Wilson, Johan Delport, and Terry Ponich Copyright © 2014 Aze Wilson et al. All rights reserved. Assessment of the Contamination of Some Foodstuffs by Escherichia coli O157 in Benin, West Africa Mon, 24 Nov 2014 00:00:00 +0000 Escherichia coli O157 is a pathogenic bacterium causing haemorrhagic colitis. It represents a serious public health problem in Northern America and Europe, which can plague Africa. Most cases of mentioned poisoning were related to contaminated meat products and vegetables. The present work aimed to estimate the prevalence of E. coli O157 in meat and vegetables in Benin. For this purpose, 6 lots of faeces samples from pigs and 8 from cattle were collected at the farms on the outskirts of Cotonou. Similarly, 20 samples of carcasses, 20 samples of intestines and stomach, and 20 surfaces samples of slaughtering equipment were taken. Vegetables and environment materials in gardens have also been sampled for 84 samples. Bacteriological analyses revealed a percentage of contamination of 50% for pig faeces and 25% for cattle ones. All the meats from stalling parks have been contaminated by this bacterium. For vegetables, 14.6% of samples were contaminated by E. coli O157. The presence of this pathovar in animal breeding and slaughtering environment and in the gardens shows that Benin is not aware of the risks of foodborne illness associated with the consumption of contaminated products. Therefore, it urges including that germ in a systematic search during safety control of food products in Benin. Honoré Sourou Bankole, Victorien Tamègnon Dougnon, Roch Christian Johnson, T. J. Dougnon, Boniface Yehouenou, Sylvain Kougblenou, Maxime Agonsa, Magloire Legonou, Thomas Dadie, and Lamine Baba-Moussa Copyright © 2014 Honoré Sourou Bankole et al. All rights reserved.