Showing papers in "International Journal of Microbiology in 2013"
TL;DR: Guava leaf-extract might be a good candidate in the search for a natural antimicrobial agent as it showed inhibitory activity against gram-positive bacteria, whereas the gram-negative bacteria were resistant to all the solvent extracts.
Abstract: Aim. To determine the antimicrobial potential of guava (Psidium guajava) leaf extracts against two gram-negative bacteria (Escherichia coli and Salmonella enteritidis) and two gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) which are some of foodborne and spoilage bacteria. The guava leaves were extracted in four different solvents of increasing polarities (hexane, methanol, ethanol, and water). The efficacy of these extracts was tested against those bacteria through a well-diffusion method employing 50 μL leaf-extract solution per well. According to the findings of the antibacterial assay, the methanol and ethanol extracts of the guava leaves showed inhibitory activity against gram-positive bacteria, whereas the gram-negative bacteria were resistant to all the solvent extracts. The methanol extract had an antibacterial activity with mean zones of inhibition of 8.27 and 12.3 mm, and the ethanol extract had a mean zone of inhibition of 6.11 and 11.0 mm against B. cereus and S. aureus, respectively. On the basis of the present finding, guava leaf-extract might be a good candidate in the search for a natural antimicrobial agent. This study provides scientific understanding to further determine the antimicrobial values and investigate other pharmacological properties.
221 citations
TL;DR: PrestoBlue and Alamar Blue are resazurin based reagents that resulted in a quick and easily distinguishable colour change that allowed for visual readings of the minimum inhibitory concentration of various positive drug controls on various microbial strains.
Abstract: This study compared different commercially available viability reagents. The growth indicator reagents include p-iodonitrotetrazolium violet (INT), PrestoBlue, and Alamar Blue which were used for antimicrobial analysis against Streptococcus mutans, Prevotella intermedia, Propionibacterium acnes, and Mycobacterium tuberculosis. PrestoBlue and Alamar Blue are resazurin based reagents that resulted in a quick and easily distinguishable colour change that allowed for visual readings. INT and Sodium 3′-[1-(phenyl amino-carbonyl)-3,4-tetrazolium]-bis-[4-methoxy-6-nitro] benzene sulfonic acid hydrate (XTT) are tetrazolium based reagents which are converted to a formazan dye in the presence of metabolically active mitochondria enzyme. For cell viability analysis, reagents XTT and PrestoBlue were compared. PrestoBlue was able to clearly indicate the minimum inhibitory concentration (MIC) of various positive drug controls on various microbial strains. PrestoBlue was also a good indicator of the 50% inhibitory concentration (IC50) of positive drug controls on various cell lines.
136 citations
TL;DR: Short term pot culture experiment with maize revealed that seed bacterization with P29 @ 10 g·kg−1 significantly enhanced total dry mass and uptake of N, K, Mn, and Zn and uptake in broth was almost acidic in all the cases.
Abstract: Zinc (Zn) is one of the essential micronutrients required for optimum plant growth. Substantial quantity of applied inorganic zinc in soil is converted into unavailable form. Zinc solubilising bacteria are potential alternates for zinc supplement. Among 10 strains screened for Zn solubilisation, P29, P33, and B40 produced 22.0 mm clear haloes on solid medium amended with ZnCO3. Similarly, P17 and B40 showed 31.0 mm zone in ZnO incorporated medium. P29 and B40 showed significant release of Zn in broth amended with ZnCO3 (17 and 16.8 ppm) and ZnO (18 and 17 ppm), respectively. The pH of the broth was almost acidic in all the cases ranging from 3.9 to 6.1 in ZnCO3 and from 4.1 to 6.4 in ZnO added medium. Short term pot culture experiment with maize revealed that seed bacterization with P29 @ 10 g·kg−1 significantly enhanced total dry mass (12.96 g) and uptake of N (2.268%), K (2.0%), Mn (60 ppm), and Zn (278.8 ppm).
130 citations
TL;DR: The potential of the bacteriophage-derived peptidase, CHAPK, as a biocidal agent for the rapid disruption of biofilm-forming staphylococci, commonly associated with bovine mastitis, is demonstrated.
Abstract: New antibacterial agents are urgently needed for the elimination of biofilm-forming bacteria that are highly resistant to traditional antimicrobial agents. Proliferation of such bacteria can lead to significant economic losses in the agri-food sector. This study demonstrates the potential of the bacteriophage-derived peptidase, , as a biocidal agent for the rapid disruption of biofilm-forming staphylococci, commonly associated with bovine mastitis. Purified applied to biofilms of Staphylococcus aureus DPC5246 completely eliminated the staphylococcal biofilms within 4 h. In addition, was able to prevent biofilm formation by this strain. The lysin also reduced S. aureus in a skin decolonization model. Our data demonstrates the potential of as a biocidal agent for prevention and treatment of biofilm-associated staphylococcal infections or as a decontaminating agent in the food and healthcare sectors.
83 citations
TL;DR: Two native strains BF11 and BF21 degrading keratin completely were characterized and showed a significant increase in the amounts of cysteine, cystine, methionine, and total free amino acids during the fermentation period suggesting the hydrolysis of disulphide by the isolates.
Abstract: Keratinase producing microorganisms are being increasingly utilized for degradation and recycling of poultry feather waste. Two native strains BF11 (Bacillus subtilis) and BF21 (Bacillus cereus) degrading keratin completely were characterized. The native strains produced more than 10 KU/mL of enzyme. Strain improvement resulted in isolation of MBF11 and MBF21 from BF11 and BF21 isolates, respectively. Optimization of nutritional and physical parameters of these MBF isolates at laboratory scale increased the overall keratinase activity by 50-fold resulting in a yield of 518–520 KU/mL. Fermentation media designed with starch as carbon source and soya bean meal as nitrogen source supported high levels of enzyme production. The optimum conditions for enzyme production were determined to be pH 8.5 and temperatures of 45–55°C for MBF11 and 37°C for MBF21, respectively. Culture filtrate showed a significant increase in the amounts of cysteine, cystine, methionine, and total free amino acids during the fermentation period. The ratio of organic sulphur concentration was also considerably higher than that of the inorganic sulphate in the culture filtrate suggesting the hydrolysis of disulphide by the isolates.
61 citations
TL;DR: Evaluation of in vitro interactions between plant extracts and oxacillin described in terms of fractional inhibitory concentration (FIC) indices revealed synergistic or additive effects of plant Extracts and clearly synergistic effects of essential oil from Salvia officinalis with Oxacillin in methicillin-resistant Staphylococcus epidermidis.
Abstract: The crude extracts of plants from Asteraceae and Lamiaceae family and essential oils from Salvia officinalis and Salvia sclarea were studied for their antibacterial as well as antibiotic resistance modifying activity. Using disc diffusion and broth microdilution assays we determined higher antibacterial effect of three Salvia spp. and by evaluating the leakage of 260 nm absorbing material we detected effect of extracts and, namely, of essential oils on the disruption of cytoplasmic membrane. The evaluation of in vitro interactions between plant extracts and oxacillin described in terms of fractional inhibitory concentration (FIC) indices revealed synergistic or additive effects of plant extracts and clearly synergistic effects of essential oil from Salvia officinalis with oxacillin in methicillin-resistant Staphylococcus epidermidis.
58 citations
TL;DR: A statistical model was developed in this study to describe bioethanol production through a batch fermentation process of sugarcane molasses by locally isolated Saccharomyces cerevisiae Y-39 and a maximum ethanol production of 255 g/L was obtained.
Abstract: A statistical model was developed in this study to describe bioethanol production through a batch fermentation process of sugarcane molasses by locally isolated Saccharomyces cerevisiae Y-39. Response surface methodology RSM based on central composite face centered design CCFD was employed to statistically evaluate and optimize the conditions for maximum bioethanol production and study the significance and interaction of incubation period, initial pH, incubation temperature, and molasses concentration on bioethanol yield. With the use of the developed quadratic model equation, a maximum ethanol production of 255 g/L was obtained in a batch fermentation process at optimum operating conditions of approximately 71 h, pH 5.6, 38°C, molasses concentration 18% wt.%, and 100 rpm.
52 citations
TL;DR: Replacement of seawater with a defined salt formulation in production medium and subsequent statistical media optimization to ensure consistent as well as improved antibiotic production by Streptomyces sp.
Abstract: Streptomyces sp. JAJ06 is a seawater-dependent antibiotic producer, previously isolated and characterised from an Indian coastal solar saltern. This paper reports replacement of seawater with a defined salt formulation in production medium and subsequent statistical media optimization to ensure consistent as well as improved antibiotic production by Streptomyces sp. JAJ06. This strain was observed to be proficient to produce antibiotic compound with incorporation of chemically defined sodium-chloride-based salt formulation instead of seawater into the production medium. Plackett-Burman design experiment was applied, and three media constituents, starch, KBr, and CaCO3, were recognised to have significant effect on the antibiotic production of Streptomyces JAJ06 at their individual levels. Subsequently, Response surface methodology with Box-Behnken design was employed to optimize these influencing medium constituents for the improved antibiotic production of Streptomyces sp. JAJ06. A total of 17 experiments were conducted towards the construction of a quadratic model and a second-order polynomial equation. Optimum levels of medium constituents were obtained by analysis of the model and numerical optimization method. When the strain JAJ06 was cultivated in the optimized medium, the antibiotic activity was increased to 173.3 U/mL, 26.8% increase as compared to the original (136.7 U/mL). This study found a useful way to cultivate Streptomyces sp. JAJ06 for enhanced production of antibiotic compound.
40 citations
TL;DR: Strain, origin, and environmental conditions can determine the level of biofilm production by L. monocytogenes isolates and significant differences between isolates origin, temperature, and sublethal acidic stress were observed concerning the ability to form biofilms.
Abstract: Objective. A total of 725 Listeria monocytogenes isolates, 607 from various foods and 118 from clinical cases of listeriosis, were investigated concerning their ability to form biofilms, at 4°C during 5 days and at 37°C during 24 h. Methods. Biofilm production was carried out on polystyrene tissue culture plates. Five L. monocytogenes isolates were tested for biofilm formation after being exposed to acidic and osmotic stress conditions. Results. Significant differences (
38 citations
TL;DR: From 194 faecal dropping samples of common house geckos collected from offices, houses, integrated farm units, and hostels, guest houses, and dining rooms of different canteen/mess, 326 bacterial isolates of enteric bacteria belonging to 17 genera and 34 species were detected.
Abstract: From 194 faecal dropping samples of common house geckos collected from offices (60), houses (88), integrated farm units (IFS,18) and hostels, guest houses, and dining rooms of different canteen/mess (HGM, 28), 326 bacterial isolates of enteric bacteria belonging to 17 genera and 34 species were detected. Escherichia coli were the most frequently (39) isolated followed by Citrobacter freundii (33), Klebsiella pneumonia (27), Salmonella indica (12), Enterobacter gergoviae (12), and Ent. agglomerans (11). Other important bacteria isolated from gecko droppings were Listonella damsela (2), Raoultella terrigena (3), S. salamae (2), S. houtenae (3), Edwardsiella tarda (4), Edwardsiella hoshinae (1), and Klebsiella oxytoca (2). Of the 223 isolates tested for antimicrobial drug sensitivity, 27 (12.1%) had multiple drug resistance (MDR). None of the salmonellae or edwardsiellae had MDR however, MDR strains were significantly more common among Escherichia spp. (P = 1.9 × 10−5) and isolates from IFS units (P = 3.58 × 10−23). The most effective herbal drug, Ageratum conyzoides extract, inhibited growth of only 27.8% of strains tested followed by ethanolic extract of Zanthoxylum rhetsa (13.9%), eucalyptus oil (5.4%), patchouli oil (5.4%), lemongrass oil (3.6%), and sandalwood oil (3.1%), and Artemisia vulgaris essential oil (3.1%).
31 citations
TL;DR: The Malaysian isolates were highly susceptible to the current antibiotics used for the treatment of melioidosis in Malaysia, and multiple resistances to the antibiotics used in the maintenance therapy are the cause for a concern.
Abstract: Acute melioidosis may present as localised or septicaemic infections and can be fatal if left untreated. Burkholderia pseudomallei resistant to antibiotics used for the treatment of melioidosis had been reported. The aim of this study was to determine the in vitro antibiotic susceptibility patterns of Burkholderia pseudomallei isolated in Malaysia to a panel of antibiotics used for the treatment of melioidosis and also to potential alternative antibiotics such as tigecycline, ampicillin/sulbactam, and piperacillin/tazobactam. A total of 170 Burkholderia pseudomallei isolates were subjected to minimum inhibitory concentration determination using -test method to eleven antibiotics. All isolates were sensitive to meropenem and piperacillin/tazobactam. For ceftazidime, imipenem, amoxicillin/clavulanic acid, and doxycycline resistance was observed in 1 isolate (0.6%) for each of the antibiotics. Trimethoprim/sulfamethoxazole resistance was observed in 17 (10%) isolates. For other antibiotics, ampicillin/sulbactam, chloramphenicol, tigecycline, and ciprofloxacin resistance were observed in 1 (0.6%), 6 (3.5%), 60 (35.3%) and 98 (57.7%) isolates respectively. One isolate B170/06 exhibited resistance to 4 antibiotics, namely, ciprofloxacin, chloramphenicol, trimethoprim/sulfamethoxazole, and tigecycline. In conclusion, the Malaysian isolates were highly susceptible to the current antibiotics used in the treatment of melioidosis in Malaysia. Multiple resistances to the antibiotics used in the maintenance therapy are the cause for a concern.
TL;DR: Four nontyphoidal Salmonella strains with resistance to extended-spectrum cephalosporins and nonclassical quinolone resistance phenotype were studied and the circulation of qnrB19 associated with bla TEM-1, bla SHV-12, and bla CTX-M-2 is highlighted.
Abstract: Four nontyphoidal Salmonella strains with resistance to extended-spectrum cephalosporins and nonclassical quinolone resistance phenotype were studied. Two S. Give were isolated from pediatric patients with acute gastroenteritis, and two S. Heidelberg were recovered from raw chicken meat. Phenotypic characterization included antimicrobial susceptibility testing and detection of extended-spectrum β -lactamases (ESBLs) by the double-disc synergy method. The detection of quinolone resistance-determining regions (QRDR) of gyrA, gyrB, and gyrC genes, bla ESBLs genes, and plasmid-mediated quinolone resistance (PMQR) determinants was carried out by molecular methods. Plasmid analysis included Southern blot and restriction patterns. Transferability of resistance genes was examined by transformation. bla TEM-1 + bla SHV-12 genes were detected in S. Give SG9611 and bla TEM-1 + bla CTX-M-2 in the other three strains: S. Give SG9811, S. Heidelberg SH7511, and SH7911. Regardless of origin and serovars, the qnrB19 gene was detected in the 4 strains studied. All determinants of resistance were localized in plasmids and successfully transferred by transformation. This study highlights the circulation of qnrB19 associated with bla TEM-1, bla SHV-12, and bla CTX-M-2 in S. Give and S. Heidelberg in Venezuela. The recognition of factors associated with increasing resistance and the study of the molecular mechanisms involved can lead to a more focused use of antimicrobial agents.
TL;DR: A screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-resistant Staphylococcus aureus, MRSA, and Pseudomonas aeruginosa is developed and suggested that the screen used is sufficient and appropriate to detect secreted antibacterial fungal compounds resulting from mutagenesis of A.oryzae.
Abstract: Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-resistant Staphylococcus aureus (MRSA) in vitro. To randomly generate A. oryzae mutant strains, spores were treated with ethyl methanesulfonate (EMS). Over 3000 EMS-treated A. oryzae cultures were tested in the screen, and one isolate, CAL220, exhibited altered morphology and antibacterial activity. Culture supernatant from this isolate showed antibacterial activity against Methicillin-sensitive Staphylococcus aureus, MRSA, and Pseudomonas aeruginosa, but not Klebsiella pneumonia or Proteus vulgaris. The results of this study support our hypothesis and suggest that the screen used is sufficient and appropriate to detect secreted antibacterial fungal compounds resulting from mutagenesis of A. oryzae. Because the genome of A. oryzae has been sequenced and systems are available for genetic transformation of this organism, targeted as well as random mutations may be introduced to facilitate the discovery of novel antibacterial compounds using this system.
TL;DR: Using the polyphasic taxonomical approach and phenotypic characteristic studies, the isolate BTSS 1001 was characterized as marine actinomycete Streptomyces rochei.
Abstract: Actinomycetes isolated from marine sediments along the southeast coast of Bay of Bengal were investigated for amylolytic activity. Marine actinomycete BTSS 1001 producing an alkaline amylase was identified from marine sediment of Diviseema coast, Bay of Bengal. The isolate produced alkaline amylase with maximum amylolytic activity at pH 9.5 at 50°C. The organism produced white to pale grey substrate mycelium and grayish aerial mycelium with pinkish brown pigmentation. A comprehensive study of morphological, physiological parameters, cultural characteristics, and biochemical studies was performed. The presence of iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0, and anteiso-C17 : 0 as the major cellular fatty acids, LL-diaminopimelic acid as the characteristic cell wall component, and menaquinones MK-9H(6) and MK-9H(8) as the major isoprenoid quinones is attributed to the strain BTSS 1001 belonging to the genus Streptomyces. Comparison of 16S rRNA gene sequences showed that strain BTSS 1001 exhibited the highest similarities to the type strains of Streptomyces rochei (99%), Streptomyces plicatus (99%), and Streptomyces enissocaesilis (99%). Using the polyphasic taxonomical approach and phenotypic characteristic studies, the isolate BTSS 1001 was characterized as marine actinomycete Streptomyces rochei.
TL;DR: These new regulatory mechanisms which could be exploited to prevent MDR development in C. albicans are summarized, namely, ROS, iron, hypoxia, lipids, morphogenesis, and transcriptional and signaling networks.
Abstract: Continuous deployment of antifungals in treating infections caused by dimorphic opportunistic pathogen Candida albicans has led to the emergence of drug resistance resulting in cross-resistance to many unrelated drugs, a phenomenon termed multidrug resistance (MDR). Despite the current understanding of major factors which contribute to MDR mechanisms, there are many lines of evidence suggesting that it is a complex interplay of multiple factors which may be contributed by still unknown mechanisms. Coincidentally with the increased usage of antifungal drugs, the number of reports for antifungal drug resistance has also increased which further highlights the need for understanding novel molecular mechanisms which can be explored to combat MDR, namely, ROS, iron, hypoxia, lipids, morphogenesis, and transcriptional and signaling networks. Considering the worrying evolution of MDR and significance of C. albicans being the most prevalent human fungal pathogen, this review summarizes these new regulatory mechanisms which could be exploited to prevent MDR development in C. albicans as established from recent studies.
TL;DR: Under normal growth conditions, expression of SaNOS was highest during early exponential phase of the bacterial growth and deletion led to increased susceptibility of the mutant cells compared to wild-type S. aureus, while the addition of a nitric oxide donor, sodium nitroprusside, restored oxidative stress tolerance of the SaN OS mutant.
Abstract: Nitric oxide and its derivative peroxynitrites are generated by host defense system to control bacterial infection. However certain Gram positive bacteria including Staphylococcus aureus possess a gene encoding nitric oxide synthase (SaNOS) in their chromosome. In this study it was determined that under normal growth conditions, expression of SaNOS was highest during early exponential phase of the bacterial growth. In oxidative stress studies, deletion of SaNOS led to increased susceptibility of the mutant cells compared to wild-type S. aureus. While inhibition of SaNOS activity by the addition of L-NAME increased sensitivity of the wild-type S. aureus to oxidative stress, the addition of a nitric oxide donor, sodium nitroprusside, restored oxidative stress tolerance of the SaNOS mutant. The SaNOS mutant also showed reduced survival after phagocytosis by PMN cells with respect to wild-type S. aureus.
TL;DR: The prevalence of MRSA carriage among black college students at a university setting is found to be low, highlighting the need of future study which involves multiinstitutions and other ethnic group to assess the association of black race withMRSA carriage.
Abstract: Background. Black people in the USA is afflicted with a higher rate of methicillin-resistant Staphylococcus aureus (MRSA) infection. This study determined the prevalence of MRSA carriage among black college students at a university setting. Methods. Hand and nasal swabs were collected and screened for MRSA by mannitol fermentation, coagulase, and DNase activities and their resistance to oxacillin. MRSA isolates were analyzed for antimicrobial resistance pattern, genetic profile for staphylococcal cassette chromosome mec (SCCmec) type, pulsed-field type, multilocus sequence type (ST), and the presence of Panton-Valentine leukocidin (PVL) gene. Results. MRSA was isolated from 1 of the 312 (0.3%) hand swabs and 2 of the 310 (0.65%) nasal swabs, respectively. All isolates lack multidrug resistance and have type IV SCCmec, characteristic of community-associated MRSA. These isolates were a ST8-MRSA-IVa-PVL(+) (USA300 strain), a ST8-MRSA-IVb-PVL(−), and a new MLST, ST2562-MRSA-IV-PVL(−), identified in this study. These isolates were thus not transmitted among students. Conclusion. We found a low rate of MRSA carriage among students in a black university. Our finding highlights the need of future study which involves multiinstitutions and other ethnic group to assess the association of black race with MRSA carriage.
TL;DR: A cell-based assay utilizing a fluorescence reporter construct expressed in a neuronal cell model to study toxin activity in situ is described, which can detect as little as 100 pM BoNT/A activity within living cells.
Abstract: The standard procedure for definitive detection of BoNT-producing Clostridia is a culture method combined with neurotoxin detection using a standard mouse bioassay (MBA). The mouse bioassay is highly sensitive and specific, but it is expensive and time-consuming, and there are ethical concerns due to use of laboratory animals. Cell-based assays provide an alternative to the MBA in screening for BoNT-producing Clostridia. Here, we describe a cell-based assay utilizing a fluorescence reporter construct expressed in a neuronal cell model to study toxin activity in situ. Our data indicates that the assay can detect as little as 100 pM BoNT/A activity within living cells, and the assay is currently being evaluated for the analysis of BoNT in food matrices. Among available in vitro assays, we believe that cell-based assays are widely applicable in high-throughput screenings and have the potential to at least reduce and refine animal assays if not replace it.
TL;DR: It is concluded that a mode of transmission of environmental microbes to patients was present in the Neonatal Unit of RSCM and thus needed to be overcome.
Abstract: Acinetobacter baumannii (A. baumannii) is Gram-negative coccobacilli that has emerged as a nosocomial pathogen. Several reports in Indonesia showed the continuous presence of A. baumannii. This study aimed to determine the incidence of A. baumannii bacteremia in neonates in the Neonatal Unit Dr. Cipto Mangunkusumo Hospital (RSCM), Jakarta, Indonesia, and assess its role in blood stream infection using antibiogram and genotyping by pulsed-field gel electrophoresis (PFGE). Subjects were neonates with clinical sepsis. Blood specimens from the neonates and samples of suspected environment within the Neonatal Unit were cultivated. Antimicrobial resistance profiles were classified for analysis purpose. A. baumannii isolates were genotyped by PFGE to determine their similarity. A total of 24 A. baumannii were isolated from 80 neonates and the environment during this period of study. Seven isolates from the neonates showed multiple antimicrobial resistance (MDR), and 82% (n = 17) of the environment isolates were also MDR. Antibiotype "d" seemed to be predominant (62.5%). PFGE analysis showed a very close genetic relationship between the patients and environment isolates (Dice coefficient 0.8-1.0). We concluded that a mode of transmission of environmental microbes to patients was present in the Neonatal Unit of RSCM and thus needed to be overcome.
TL;DR: Bacteriovorax were quantified in US Atlantic, Gulf, and Pacific seawater to determine baseline levels of these predatory bacteria and possible seasonal fluctuations in levels and there was a relatively strong negative correlation between temperature and Bacter Giovorax levels for Gulf seawater.
Abstract: Bacteriovorax were quantified in US Atlantic, Gulf, and Pacific seawater to determine baseline levels of these predatory bacteria and possible seasonal fluctuations in levels. Surface seawater was analyzed monthly for 1 year from Kailua-Kona, Hawaii; the Gulf Coast of Alabama; and four sites along the Delaware Bay. Screening for Bacteriovorax was performed on lawns of V. parahaemolyticus host cells. Direct testing of 7.5 mL portions of seawater from the Atlantic, Pacific, and Gulf coasts gave mean annual counts ≤12.2 PFU. Spikes in counts were observed at 3 out of 4 sites along the Delaware Bay 1 week after Hurricane Sandy. A comparison of summer versus winter counts showed significantly more Bacteriovorax (P ≤ 0.0001) in the Delaware Bay during the summer and significantly more (P ≤ 0.0001) in the Gulf during the winter, but no significant seasonal differences (P > 0.05) for Hawaiian seawater. Bacteriovorax counts only correlated with seawater salinity and temperature at one Delaware site (r = 0.79 and r = 0.65, resp.). There was a relatively strong negative correlation between temperature and Bacteriovorax levels (r = -0.585) for Gulf seawater. Selected isolates were sequenced and identified by phylogenetic analysis as Bacteriovorax clusters IX, X, XI, and XII.
TL;DR: Examples from the own experiences and the literature are focused on to provide insight into the limitations of single target testing in molecular biology, and multitargeted testing in microbiological molecular assays should be a rule.
Abstract: Advantages of PCR assays over more conventional culture-based diagnostics include significantly higher sensitivities and shorter turnaround times They are particularly useful when patient treatment has already been initiated or for specimens that may contain microorganisms that are slow-growing, difficult to culture, or for which culture methods do not exist However, due to genome variability, single target testing might lead to false-negative results This paper focuses on examples from our own experiences and the literature to provide insight into the limitations of single target testing in molecular biology Lessons learned from these experiences include the careful design of diagnostic assays, preferably multitargeted, the importance of investigating the incidence and epidemiology of infection in detail, the frequent participation in appropriate quality assurance schemes, and the importance of continuous attentiveness by investigators when confronted with inconsistent results In conclusion, multitargeted testing in microbiological molecular assays should be a rule
TL;DR: Liquid egg is a significant Salmonella vehicle, showing a need to continue the vaccination of chickens to prevent S. Enteritidis contamination in Japanese eggs, and further study is needed to evaluate Salmoneella contamination in seed sprouts with more sampling from retailers there.
Abstract: The study aimed to evaluate the prevalence of Salmonella in retail and wholesale foods in Fukuoka Prefecture, Japan. A total of 2,021 samples collected between 1999 and 2010 were tested using a culture method. Samples consisted of liquid eggs (), meat (beef and pork) (), offal (), processed meats (), seafood (), processed seafood (dried fish) (), vegetables (), processed vegetables (), fruits (), and herbs () from 574 outlets and wholesale agents in 15 areas (five samples were undocumented regarding outlets). Overall, liquid egg showed significantly () higher frequencies of Salmonella contamination (13.3%) than beef (1/423, 0.2%) and pork (3/235, 1.3%). Salmonella enterica subsp. enterica serovar Enteritidis, the most common serovar as a human pathogen, were isolated from two liquid egg samples. No Salmonella were isolated from seafood and vegetable-related samples including seed sprouts (). In conclusion, liquid egg is a significant Salmonella vehicle, showing a need to continue the vaccination of chickens to prevent S. Enteritidis contamination in Japanese eggs. Moreover, further study is needed to evaluate Salmonella contamination in seed sprouts with more sampling from retailers there.
TL;DR: Interestingly, cloned folP with three mutations derived from Streptococcus mutans 8 conferred substantial resistance against sulfonamide to folP-deficient bacteria, suggesting that folP point mutations only partially explain bacterial resistance to sulf onamide.
Abstract: Cotrimoxazole inhibits dhfr and dhps and reportedly selects for drug resistance in pathogens. Here, Streptococcus mutans isolates were obtained from saliva of HIV/AIDS patients taking cotrimoxazole prophylaxis in Uganda. The isolates were tested for resistance to cotrimoxazole and their folP DNA (which encodes sulfonamide-targeted enzyme dhps) cloned in pUC19. A set of recombinant plasmids carrying different point mutations in cloned folP were separately transformed into folP-deficient Escherichia coli. Using sulfonamide-containing media, we assessed the growth of folP-deficient bacteria harbouring plasmids with differing folP point mutations. Interestingly, cloned folP with three mutations (A37V, N172D, R193Q) derived from Streptococcus mutans 8 conferred substantial resistance against sulfonamide to folP-deficient bacteria. Indeed, change of any of the three residues (A37V, N172D, and R193Q) in plasmid-encoded folP diminished the bacterial resistance to sulfonamide while removal of all three mutations abolished the resistance. In contrast, plasmids carrying four other mutations (A46V, E80K, Q122H, and S146G) in folP did not similarly confer any sulfonamide resistance to folP-knockout bacteria. Nevertheless, sulfonamide resistance (MIC = 50 μM) of folP-knockout bacteria transformed with plasmid-encoded folP was much less than the resistance (MIC = 4 mM) expressed by chromosomally-encoded folP. Therefore, folP point mutations only partially explain bacterial resistance to sulfonamide.
TL;DR: DGGE profiling is a molecular method to describe the microbial population associated with ticks and demonstrate some of the complexity and variety of tick-borne microorganisms.
Abstract: Ticks acquire a wide range of microorganisms as a natural part of their lifecycle. Bacteria, viruses, and protozoa can be transmitted to ticks during feeding and free-living phases. DGGE profiling is a molecular method to describe the microbial population associated with ticks and demonstrate some of the complexity and variety of tick-borne microorganisms. The present study profiled a total of 120 I. ricinus ticks, which were divided into three equally sized groups. We found that B. burgdorferi s.l.-infected ticks presented a pattern consisting of bacterial Pseudomonas spp. (67.5%), Bacillus spp. (50%), and Sphingomonas spp. (77.5%), while A. phagocytophilum-infected ticks were associated with Pseudomonas spp. (82.5%) and Sphingomonas spp. (57.5%). All profiles had one or more Pseudomonas species present, and the intramitochondrial endosymbiont Candidatus Midichloria mitochondrii was present in more than 25% of the samples. Statistical analysis demonstrated that the microbial communities were not significantly different between the groups and that the groups could not be characterised by a specific microbial population.
TL;DR: Abiotic and biotic factors mimicking conditions that might induce resting spores in temperate and tropical regions were tested with isolates from Norway and Brazil and no combination of conditions induced the formation of a high number of resting spores.
Abstract: Neozygites floridana is an obligate mite pathogenic fungus in the Entomophthoromycota. It has been suggested that resting spores of this fungus are produced as a strategy to survive adverse conditions. In the present study, possible mechanisms involved in the regulation of resting spore formation were investigated in the hosts Tetranychus urticae and Tetranychus evansi. Abiotic and biotic factors mimicking conditions that we, based on earlier field studies, thought might induce resting spores in temperate and tropical regions were tested with isolates from Norway and Brazil. A total of 42 combinations of conditions were tested, but only one induced the formation of a high number of resting spores in only one isolate. The Brazilian isolate ESALQ1420 produced a large number of resting spores (51.5%) in T. urticae at a temperature of 11°C, photoperiod of 10L:14D, and light intensity of 42–46 (μmol m−2 s−1) on nonsenescent plants (nondiapausing females). Resting spores of the Brazilian N. floridana isolate ESALQ1421 were found at very low levels (up to 1.0%). Small percentages of T. urticae with resting spores (0–5.0%) were found for the Norwegian isolate NCRI271/04 under the conditions tested. The percentages of resting spores found for the Norwegian isolate in our laboratory studies are similar to the prevalence reported in earlier field studies.
TL;DR: Data in combination with the phenotypic distinctiveness of AK-1T clearly indicate that the strain represents a novel species, for which the name Agromyces arachidis sp.
Abstract: A Gram-positive, yellowish bacterium strain AK-1(T) was isolated from soil sample collected from peanut (Arachis hypogaea) crop field and studied by using a polyphasic approach. The organism had morphological and chemotaxonomic properties consistent with its classification in the genus Agromyces. Phylogenetic analysis of the 16S rRNA gene sequence showed that strain AK-1(T) was closely related to Agromyces aurantiacus (98.6%) followed by Agromyces soli (98.3%), Agromyces tropicus (97.6%), Agromyces ulmi (97.3%), Agromyces flavus (97.2%), and Agromyces italicus (97.0%), whereas the sequence similarity values with respect to the other Agromyces species with validly published names were between 95.3 and 96.7 %. However, the DNA-DNA hybridization values obtained between strain AK-1(T) and other related strains were well below the threshold that is required for the proposal of a novel species. The DNA G + C content of the strain is 71.8 mol%. The above data in combination with the phenotypic distinctiveness of AK-1(T) clearly indicate that the strain represents a novel species, for which the name Agromyces arachidis sp. nov. is proposed. The type strain is AK-1(T) (=MTCC 10524(T) = JCM 19251(T)).
TL;DR: It has been shown that no trend to emerging species from 1998 to 2008 could be detected, and the frequently isolated non-Candida albicans species isolated in the DGP departments have already been detected in or before 1997.
Abstract: From 1997 to 2009, 1,862 dermatology, gynaecology, and paediatrics (DGP) associated clinical yeast isolates were analysed for species occurrence, specimen origin and type, (multi-) resistance pattern, and testing period. The top seven of the isolated DGP-associated species remained the same as compared to total medical wards, with Candida albicans (45%) as most frequent pathogen. However, the DGP wards and DGP ICUs showed species-specific profiles; that is, the species distribution is clinic-specific similar and however differs in their percentage from ward to ward. By applying the “one fungus one name” principle, respectively, the appropriate current taxonomic species denominations, it has been shown that no trend to emerging species from 1998 to 2008 could be detected. In particular the frequently isolated non-Candida albicans species isolated in the DGP departments have already been detected in or before 1997. As yeasts are part of the cutaneous microbiota and play an important role as opportunistic pathogens for superficial infections, proper identification of the isolates according to the new nomenclature deems to be essential for specific and calculated antifungal therapy for yeast-like DGP-related infectious agents.
TL;DR: Results suggest that variations exist in the survival of these serotypes of STEC within mixed ruminal microorganism fluid media when supplemented with CBP, and further research is needed to determine why CBPs affect STEC serotypes differently.
Abstract: Citrus byproducts (CBPs) are utilized as a low cost nutritional supplement to the diets of cattle and have been suggested to inhibit the growth of both Escherichia coli O157:H7 and Salmonella. The objective of this study was to examine the effects in vitro that varying concentrations of CBP in the powdered or pelleted variety have on the survival of Shiga-toxin Escherichia coli (STEC) serotypes O26:H11, O103:H8, O111:H8, O145:H28, and O157:H7 in bovine ruminal microorganism media. The O26:H11, O111:H8, O145:H28, and O157:H7 serotypes did not exhibit a change in populations in media supplemented with CBP with either variety. The O103:H8 serotype displayed a general trend for an approximate
TL;DR: Evidence suggests that phages with ctxAB are genetically diverse and can infect not only V. cholerae and V. mimicus but also other species and genera in the form of a pseudolysogen.
Abstract: We initially attempted to isolate a Vibrio cholerae O1 El Tor biotype that carries a novel variant of the cholera toxin gene (ctxAB) from environmental waters of Indonesia, where the seventh cholera pandemic by V. cholerae O1 El Tor biotype began. Nested PCR targeting the gene revealed that a total of eight strains were found to carry ctxAB. However, sequencing of the 16S rRNA genes of these isolates showed they were not V. cholerae but were either Klebsiella, Enterobacter, Pantoea, or Aeromonas. Subsequent nested PCR assays targeting all genes known to be encoded on the CTX phage (i.e., zot, ace, orfU, cep, rstB, rstA, and rstR) showed that one isolate belonged to the Enterobacter genus carried all the genes tested, while the other isolates lacked either 2, 3, or 5 of the genes. This evidence suggests that phages with ctxAB are genetically diverse and can infect not only V. cholerae and V. mimicus but also other species and genera in the form of a pseudolysogen.