Microbiology

A small proportion of human T-cell leukemia virus type-1 (HTLV-1)-infected individuals develop adult T-cell leukemia/lymphoma, a chemotherapy-resistant lymphoproliferative disease with a poor prognosis. HTLV-1-specific cytotoxic T lymphocytes (CTLs), potential anti-tumor/virus effectors, are impaired in adult T-cell leukemia/lymphoma patients. Here, using Japanese monkeys naturally infected with simian T-cell leukemia/T-lymphotropic virus type-1 (STLV-1) as a model, we demonstrate that short-term-cultured autologous peripheral blood mononuclear cells (PBMCs) can serve as a therapeutic vaccine to activate such CTLs. In a screening test, STLV-1-specific CTL activity was detectable in 8/10 naturally STLV-1-infected monkeys. We conducted a vaccine study in the remaining two monkeys with impaired CTL responses. The short-term-cultured PBMCs of these monkeys spontaneously expressed viral antigens, in a similar way to PBMCs from human HTLV-1 carriers. The first monkey was subcutaneously in...

Asymptomatic infections often proceed undetected, yet can still prime the host to be sensitive to secondary environmental stress. While the mechanisms underlying disease caused by asymptomatic infections are unknown, it is believed that productive pathogen replication is required. We report that the environmental stress of carbon dioxide (CO2) anesthesia converts an asymptomatic rhabdovirus infection in Drosophila to one that is lethal. This lethality results from a pool of infectious virus in glial cells and is regulated by the antiviral RNAi pathway of the host. CO2 sensitivity is caused by the fusogenic activity of the viral glycoprotein, which results in fusion of neurons and glia. Expression of the viral glycoprotein, but not a fusion defective mutant, is sufficient to cause CO2 sensitivity, which can occur even in the absence of productive viral replication. These findings highlight how viral proteins, independent of pathogen replication, may predispose hosts to life-threateni...

Organogenesis involves an initial surge of cell proliferation, leading to differentiation. This is followed by cell death in order to remove extra cells. During early development, there is little or no cell death. However,there is a lack of information concerning the genes required for survival during the early cell-proliferation phase. Here, we show that Lobe(L) and the Notch (N) ligand Serrate (Ser), which are both involved in ventral eye growth, are required for cell survival in the early eye disc. We observed that the loss-of-ventral-eye phenotype in L or Ser mutants is due to the induction of cell death and the upregulation of secreted Wingless (Wg). This loss-of-ventral-eye phenotype can be rescued by (i) increasing the levels of cell death inhibitors, (ii)reducing the levels of Hid-Reaper-Grim complex, or (iii) reducing canonical Wg signaling components. Blocking Jun-N-terminal kinase (JNK) signaling, which can induce caspase-independent cell death, significantly rescued vent...

A fundamental feature of developing tissues and organs is generation of planar polarity of cells in an epithelium with respect to the body axis. The Drosophila compound eye shows two-tier dorsoventral (DV) planar polarity. At the individual ommatidium level, the eight photoreceptors in each unit eye form a dorsoventrally asymmetric cluster. At the level of eye field, hundreds of ommatidia in the upper and lower halves of an eye are uniformly polarized dorsally or ventrally, respectively. This results in DV mirror symmetries about the equator. The uniform orientations of photoreceptor clusters over long distance in the eye field provide an excellent model for studying the genetic basis of long-range planar polarity. Ommatidial DV polarity can be detected in third instar eye imaginal disc during the early stage of retinal diVerentiation. Recent studies have strongly suggested that the foundation for this DV polarity pattern is laid much earlier in undiVerentiated eye disc. The eye disc primordium is partitioned into the DV compartments of independent cell lineages. The Iroquois-Complex genes specify the dorsal fate whereas their absence in the ventral compartment results in the ventral fate with expression of the fringe gene. Genetic evidence suggests that the interaction of dorsal and ventral cells at the DV boundary in early eye disc generates an organizing center for growth and patterning of the eye field. In this chapter, we outline key genetic events involved in early DV patterning and growth of eye disc, and its potential role in organizing long-range signaling for DV planar polarity during later diVerentiation of the eye.

Fungal chitinases play important roles in the decomposition of wastes, mycoparasitism, and biocontrol of nematodes and plant pathogens through chitin biodegradation. This study was conducted during 2013-2017 to investigate the presence of chitinase genes in Trichoderma and Clonostachys species from the Birjand plain, and to evaluate their ability to degrade chitin. Fungal spores and soil suspensions were cultured on minimal medium containing 1% colloidal chitin from crab bodies to isolate chitinolytic fungi. Chitinolytic ability of the isolates was evaluated on this medium by staining with 1% Lugol's iodine solution and screening for the production of a bright halo around the colonies. Fifty-two isolates capable of degrading chitin were recovered. DNA extracted from the isolates was amplified using Chit2 or DECH degenerative primers that are related to the chitinase gene, and their sequences were aligned using the NCBI GenBank database. The Chit2 and DECH primers amplified 600-bp and 250-bp fragments, respectively, and according to sequence alignment, the isolates had sequences similar to that of the chi18 chitinase genes. Morphological and molecular characterization allowed identifying the isolates as belonging to the species Trichoderma harzianum (n = 41), T. longibrachiatum (n = 1), T. virens (n = 3), T. brevicompactum (n = 1), Clonostachys rosea (n = 5), and C. rogersoniana (n = 1), some of which may potentially be used as biocontrol agents of pathogenic nematodes and fungi. This is the first report of isolation of fungi capable of chitin biodegradation from the South Khorasan Province in Eastern Iran.

Background: Although a dramatic decrease in AIDS progression has been observed after Highly Active Anti Retroviral Therapy (HAART) in both low-and high-resource settings, few data support that fact in low-resource settings. This study describes the incidence of AIDS-defining illnesses (ADI) after HAART initiation and analyzes their risk factors in a low-resource setting. A focus was put on CD4 cell counts and viral load measurements. Methods: 404 HIV-1-infected Senegalese adult patients were enrolled in a prospective observational cohort and data censored as of April 2008. A Poisson regression was used to model the incidence of ADIs over two periods and to assess its association with baseline variables, current CD4, current viral load, CD4 response, and virological response. Results: ADI incidence declined from 20.5 ADIs per 100 person-years, 95% CI = [16.3;25.8] during the first year to 4.3, 95% CI = [2.3;8.1] during the fourth year but increased afterwards. Before 42 months, the decrease was greater in patients with clinical stage CDC-C at baseline and with a viral load remaining below 1000 cp/mL but was uniform across CD4 strata (p = 0.1). After 42 months, 293 patients were still at risk. The current CD4 and viral load were associated with ADI incidence (decrease of 21% per 50 CD4/mm 3 and of 61% for patients with a viral load < 1000 cp/ mL). During the first four years, a uniform decline of ADI incidence was observed even in patients with low CD4-cell counts at HAART initiation as long as the viral load remained undetectable. An increase was noted later in patients with immunologic and virological failures but also in patients with only virological failure.

Objective: Increasing antifungal resistance and biofilm formation among Candida species is an intimidating public health concern, especially at the hospital settings. In light of this, the current study was designed to assess the biofilm-forming ability of clinically isolated Candida albicans and determine their antifungal susceptibility against both the planktonic and sessile forms. Methods: A total of 58 Candida isolates from different clinical samples received at the Microbiology laboratory of KIST Medical College and Teaching Hospital, Lalitpur, Nepal in between April to October 2018 were included in the study. Isolation and identification of C. albicans was done following standard microbiological procedures that comprised of microscopic observations along with germ tube formation and biochemical tests. Besides qualitative investigation of biofilm by tube method, it was also investigated quantitatively by crystal violet staining method and metabolic activity of the biofilm was a...

Staphylococcus aureus (MSSA) and methicillin resistant S. aureus (MRSA) infections. So, it is utmost important to perform the susceptibility test for erythromycin and clindamycin. And, there is concern on use of this antibiotic in the presence of erythromycin resistance because of the possibility of induction of cross-resistance among members of macrolide, lincosamide and streptogramin B (MLS B ) group. During August 2011 to May 2012, a total of 207 isolates of S. aureus were isolated and among which 29.47% (61) isolates were confirmed as MRSA by cefoxitin (30 µg) disc. All the isolates were further processed for MLSB resistance test by double disc diffusion test of erythromycin (2 µg) and clindamycin (15 µg) at a distance of 15 and 22 mm between them. This study result show 12.56% (26) and 14.49% (30) of inducible macrolide-lincosamide-streptogramin B phenotype (iMLS B ) resistance type at 22 and 15 mm disc distance, respectively, showing 15 mm disc distance is potential than 22 mm and 17.39% (36) of cMLS B resistance type. Similarly, both iMLSB and cMLSB are greater in MRSA than MSSA and constitutes 18.05 (11) and 36.06% (22), respectively. Thus, this study concludes that D-test should be used as a mandatory method and is more potential in 15 mm disc apart.

This thesis contains no material which has been accepted for a degree or diploma by the University or any other institution , except by way of background information and duly acknowledged in the thesis , and to the best of my knowledge and belief no material previously published or written by another person except where due acknowledgment is made in the text of the thesis.

In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity. Mixed infection with the different A. actinomycetemcomitans serotypes is frequent in periodontitis patients; accordingly, the role of this bacterial species in the pathogenesis of periodontitis may differ depending whether patients or periodontal lesions harbour one or more of the A. actinomycetemcomitans serotypes. We hypothesized that different combinations of these serotypes could be associated with distinct host responses and hence different inflammatory patterns. This investigation was aimed to assess whether the increased immuno-stimulatory potential attributed to the serotype b of A. actinomycetemcomitans on immune cells is able to be modified during co-infection with other A. actinomycetemcomitans serotypes. Dendritic cells (DCs) were obtained from healthy subjects and stimulated with the different A. actinomycetemcomitans serotypes or their purified LPS using the fol...

Fresh cut-fruits produce is one of the quickest developing food industries ever. Food poisoning has been connected with the utilization of poor quality of fruits. The present study was conducted to isolate bacteria from cantaloupe skin, characterize and identify the bacteria from cantaloupe fruit skins using biochemical tests and 16S rRNA gene analysis and to investigate the effect of antimicrobial agents on the growth of bacteria and all of these objective were successfully achieved. Four unique bacteria based on different morphology were isolated by using spread plate method. A total of ten biochemical tests were performed. For bacterial identification, the PCR products of the 16S rRNA gene were amplified, purified and sent for sequencing. Bacterium CG1, CG2, CG3 and CG4 bacteria were identified as Exigubactirum sp CG1, Exigubactirum sp. CG2, Pseudomonas sp. CG3, and Microbacterium sp. CG4. The XY-12 antimicrobial agent with two different concentrations (0.6 mL/ L and 0.3 mL/ L) w...

Studies on the distribution, epidemiology and pathogenesis of Cryptococcus neoformans on the island of Puerto Rico are few. We have studied mouse virulence and in vitro antifungal susceptibility of 133 isolates of C. neoformans: 121 environmental and 12 clinical (9 from AIDS patients), that were isolated in Puerto Rico. In experimental CD-1 mice infected intravenously, the mean lethal dose 50~o values (28 days) were > 5.2 • 106 and 1.1 x 105 cells/mouse for environmental and clinical isolates, respectively. Using an agar dilution assay, the minimum inhibitory concentrations of amphotericin B, ketoconazole and 5-fluorocytosine were comparable for environmental and clinical isolates in both yeast nitrogen dextrose base agar and Kimmig's agar. These data suggest a difference in lethality for mice, but no difference in antifungal susceptibility of environmental and clinical isolates of C. neoformans obtained in Puerto Rico.

Reports on the antimicrobial potentials of Euphorbia heterophylla are scanty globally. This study investigated the antimicrobial effects of E. heterophylla against microorganisms of clinical importance. Cold water, hot water, chloroform and methanol extracts of the leaf, stem and fruit of E. heterophylla were obtained. The phytochemical properties of the plant parts were determined, and antimicrobial analyses of extracts investigated against sixteen clinical isolates, in accordance with standard procedures. The microorganisms tested were nine clinical bacterial strains which included Pseudomonas aeruginosa, Pseudomonas aeruginosa NCIB 950, Salmonella typhi, Escherichia coli, Escherichia coli NCIB 86, Staphylococcus aureus, Staphylococcus aureus NCIB 8588, Klebsiella pneumoniae and Serratia marcescens, and seven fungal strains which were Candida albicans, Aspergillus fumigatus, Aspergillus flavus, Aspergillus parasiticus, Trichoderma viride, Trichophyton rubrum, and Malassezia furfur...

Mangifera indica, commonly called mango, is an edible fruit that are also used for juice and wine production. Fresh leaves of M. indica were collected from the premises of Adekunle Ajasin University, Akungba Akoko Ondo State, Nigeria. Materials used, including Gram staining reagents, for the study are properly sterilized. Similarly, the culture media used, including Nutrient agar, MacConkey agar, Eosin methylene blue agar, Salmonella-Shigella agar, Mannitol salt agar, Mueller-Hinton agar and Nutrient broth, were properly sterilized. The bacterial isolates identified and used for the study were Staphylococcus aureus, Salmonella typhi Klebsiella spp., Pseudomonas aeruginosa, Escherichia coli, and Citrobacter spp. Antimicrobial susceptibility test of the Mango (Mangifera indica) leaves extract against the test bacteria shows that 200 mg/ml and 175 mg/ml extract of the plant source had observable effect against test organisms, particularly Escherichia coli, Salmonella typhi, and to a va...

Background: Platelet transfusion therapy poses many challenges in veterinary clinical practice. Lack of readily available blood donors, short shelf-life, and inability to administer a sufficient number of platelets to meet a dog's transfusion need are the major difficulties encountered. Platelet additive solutions are already in use at American and European human blood banks, showing to be a realistic alternative. This study compares the in vitro platelet function in plasma, Composol, or SSP+ during storage for 13 days. Platelet rich plasma-platelet concentrate with 35% plasma and 65% platelet additive solutions (Composol or SSP+) and a control group (100% plasma) were prepared. Swirling, platelet count, blood gases, metabolic variables, platelet activation markers, and apoptosis markers were analyzed on days 1, 5, 9 and 13. Results: Swirling was well preserved and pH was acceptable (> 6.2) during storage for all platelet additive solutions units until day 9. SSP + units showed more stable pH and metabolic variables until day 13. Platelets in plasma showed higher glucose consumption than in Composol or in SSP+. The platelet additive solutions units showed better platelet metabolism maintenance, reduced glucose consumption and lactate production. The apoptotic markers were still low for 9 days in platelet concentrates with platelet additive solutions, suggesting the possibility to extend the shelf life with the use of SSP+ or Composol. Conclusions: Our findings suggest that the uses of Composol and SSP+ in canine platelet concentrates are potential alternatives in veterinary blood banks.

Gram-negative bacteria and bacteria in biofilms are very difficult to eradicate and are at the origin of the most antibiotic-resistant bacteria. Therapeutic alternatives less susceptible to mechanisms of resistance are urgently needed to respond to an alarming increase of resistant nosocomial infections. Antibacterial photodynamic inactivation (PDI) generates oxidative stress that triggers multiple cell death mechanisms more difficult to counteract by bacteria. We explore PDI of multidrug-resistant bacterial strains collected from patients and show how positive charge distribution in the photosensitizer drug impacts on the efficacy of inactivation. We demonstrate the relevance of size for drug diffusion in biofilms. Designed meso-imidazolyl porphyrins of small size with positive charges surrounding the macrocycle enabled the inactivation of bacteria in biofilms by 6.9 log units at 5 nM photosensitizer concentration and 5 J cm -2 , which offers new opportunities to treat biofilm infections.

Massarina ingoldiana occurs worldwide on a variety of dead plant substrates in aquatic habitats. This species has been accommodated in Massarina or Lophiostoma in Pleosporales, Dothideomycetes, but the validity of either of these taxonomic placements has not been confirmed with molecular data. In addition morphological variations occur among different populations of this species causing problems in identification. To evaluate the generic placement and monophyly of M. ingoldiana and the taxonomic usefulness of variable morphological features, phylogenetic analyses based on SSU and LSU sequences of ribosomal DNA were conducted for 10 putative strains of this species and its relatives. Phylogenies revealed that M. ingoldiana sensu lato is polyphyletic and comprises two distinct lineages within Pleosporales. Neither lineage was congeneric with either Massarina or Lophiostoma. Based on molecular data and a reevaluation of morphology, two new genera, Lindgomyces and Tingoldiago, are established for the two lineages of M. ingoldiana sensu lato. Lindgomyces includes four species, L. ingoldianus comb. nov. (5 M. ingoldiana sensu stricto), L. rotundatus sp. nov. (5 M. ingoldiana sensu lato), L. cinctosporae sp. nov. and L. breviappendiculatus comb. nov. (5 Lophiostoma breviappendiculatum). A new aquatic family, Lindgomycetaceae, is proposed for Lindgomyces and its sister taxon, Massariosphaeria typhicola. Isolates of a fungus from submerged Phragmites, with ascospores similar to those of M. ingoldiana, occurred in an additional single species lineage distant from that of M. ingoldiana (Lindgomyces). This fungus is described as Tingoldiago graminicola gen. & sp. nov. The discovery that Tingoldiago, which occurs in a lineage distantly related to Lindgomyces but has morphologically similar ascospores and ascospore sheaths, suggests that the elaborate ascospore sheath in M. ingoldiana has arisen in two separate lineages as a result of convergent evolution in response to the aquatic environment. The large gelatinous sheath previously was considered one of the most distinctive and stable features for species identification of M. ingoldiana.

A new ascomycete species, Natipusilla bellaspora, collected from submerged woody debris in a freshwater stream at Los Amigos Biological Station, Madre De Dios in the Peruvian Amazon is described and illustrated. This fungus is characterized by small, globose to subglobose, hyaline ascomata; small, globose to subglobose, eight-spored fissitunicate asci; one-septate, multiguttulate ascospores with two different gelatinous sheaths, an outer amorphous sheath that enlarges in water and an inner sheath that has a distinctive persistent shape and is attached to the ascospore apex. Morphologically N. bellaspora differs from other Natipusilla species in having larger ascospores and two ascospore sheaths. A second Natipusilla species, N. limonensis, is reported for the first time from Peru. Based on the unique morphological characters of taxa in Natipusilla and results of previous molecular phylogenetic analyses with other members of the Dothideomycetes, we establish Natipusillaceae fam. nov. for this unique tropical freshwater clade.

Massarina ingoldiana occurs worldwide on a variety of dead plant substrates in aquatic habitats. This species has been accommodated in Massarina or Lophiostoma in Pleosporales, Dothideomycetes, but the validity of either of these taxonomic placements has not been confirmed with molecular data. In addition morphological variations occur among different populations of this species causing problems in identification. To evaluate the generic placement and monophyly of M. ingoldiana and the taxonomic usefulness of variable morphological features, phylogenetic analyses based on SSU and LSU sequences of ribosomal DNA were conducted for 10 putative strains of this species and its relatives. Phylogenies revealed that M. ingoldiana sensu lato is polyphyletic and comprises two distinct lineages within Pleosporales. Neither lineage was congeneric with either Massarina or Lophiostoma. Based on molecular data and a reevaluation of morphology, two new genera, Lindgomyces and Tingoldiago, are established for the two lineages of M. ingoldiana sensu lato. Lindgomyces includes four species, L. ingoldianus comb. nov. (5 M. ingoldiana sensu stricto), L. rotundatus sp. nov. (5 M. ingoldiana sensu lato), L. cinctosporae sp. nov. and L. breviappendiculatus comb. nov. (5 Lophiostoma breviappendiculatum). A new aquatic family, Lindgomycetaceae, is proposed for Lindgomyces and its sister taxon, Massariosphaeria typhicola. Isolates of a fungus from submerged Phragmites, with ascospores similar to those of M. ingoldiana, occurred in an additional single species lineage distant from that of M. ingoldiana (Lindgomyces). This fungus is described as Tingoldiago graminicola gen. & sp. nov. The discovery that Tingoldiago, which occurs in a lineage distantly related to Lindgomyces but has morphologically similar ascospores and ascospore sheaths, suggests that the elaborate ascospore sheath in M. ingoldiana has arisen in two separate lineages as a result of convergent evolution in response to the aquatic environment. The large gelatinous sheath previously was considered one of the most distinctive and stable features for species identification of M. ingoldiana.

A coupled analysis of the reaction kinetics and microstructural changes of Fe 2 O 3 /Al 2 O 3 oxygen carriers during redox reaction cycles for chemical looping systems was conducted. The microstructural changes in the oxygen carrier particles were investigated using an image analysis method with cross-sectional backscattered electron images, and microstructural information such as the particle size, porosity, and two-phase boundary between the iron oxide and pores was obtained. The microstructural changes and the degradation kinetics during redox cycles were investigated under various operating conditions (reaction temperatures, reduction times, oxygen partial pressures during the oxidation process, and weight ratios of the oxygen carriers). The degradation coefficient and the reaction enthalpy of the particles in the oxidation process are linearly related, implying that increasing the local temperature of the oxygen carriers causes the coarsening of iron oxide particles via sintering and microstructural changes. Our analysis contributes to the design of highly stable oxygen carrier particles and the improvement of the operating conditions for chemical looping systems.

A coupled analysis of the reaction kinetics and microstructural changes of Fe 2 O 3 /Al 2 O 3 oxygen carriers during redox reaction cycles for chemical looping systems was conducted. The microstructural changes in the oxygen carrier particles were investigated using an image analysis method with cross-sectional backscattered electron images, and microstructural information such as the particle size, porosity, and two-phase boundary between the iron oxide and pores was obtained. The microstructural changes and the degradation kinetics during redox cycles were investigated under various operating conditions (reaction temperatures, reduction times, oxygen partial pressures during the oxidation process, and weight ratios of the oxygen carriers). The degradation coefficient and the reaction enthalpy of the particles in the oxidation process are linearly related, implying that increasing the local temperature of the oxygen carriers causes the coarsening of iron oxide particles via sintering and microstructural changes. Our analysis contributes to the design of highly stable oxygen carrier particles and the improvement of the operating conditions for chemical looping systems.

Numerosi studi scientifici in campo nutrizionale umano sottolineano l'importanza di ridurre il consumo di acidi grassi saturi (AGS), responsabili di numerose patologie e di incrementare l'utilizzo di carni ricche di acidi grassi polinsaturi (PUFA), più salutari. Tuttavia le diete contenenti PUFA, se non integrate con antiossidanti, rendono le carni più suscettibili a fenomeni di ossidazione e meno conservabili. L'obiettivo di questo studio è stato quello di valutare l'effetto di diverse diete (dieta contenente olio di palma, OP, ricca in AGS; dieta contenente olio di mais, OM, ricca in PUFA e dieta contenente olio di mais con aggiunta di un fitocomplesso antiossidante, OML) sulla digeribilità delle diete e sulle variazioni del profilo acidico, sulla qualità e sulla conservabilità (metodo TBARS) della carne suina. Lo studio, effettuato su 18 suini, suddivisi in 3 gruppi di 6 animali ciascuno, è durato 8 settimane precedute da 7 giorni di adattamento. Nessuna differenza statisticamente significativa è stata evidenziata per i parametri chimico-fisici (pH, colore, perdite di cottura, forza di taglio) delle carni, per la digeribilità delle diete, per la composizione centesimale e per i livelli di TBARS della carne fresca. La composizione in acidi grassi delle carni ha evidenziato differenze statisticamente significative (p<0.05) per l'acido palmitico (C16:0) e l'indice aterogenico che sono risultati superiori nelle carni del gruppo di suini OP rispetto ai gruppi OM ed OML. Dal punto di vista nutrizionale si può quindi affermare che l'inserimento di olio di mais nelle diete dei suini sia in grado di modificare il profilo degli acidi grassi delle carni aumentando il grado di insaturazione, ma non predisponendo la carne fresca a fenomeni perossidativi. Suini, digeribilità, qualità della carne, vino rosso liofilizzato, Liofenol.

 A bromovirus isolated from sunflower (Helianthus annuus L.) was characterised and shown to be highly related to Pelargonium zonate spot virus (PZSV), a virus reported in Italy, Spain, France, USA and Israel. Sunflower plants showing chlorotic concentric ring and linear pattern symptoms were observed near Paraná city (Entre Ríos Province) in commercial sunflower crops. This was the first observation of this type of symptoms in sunflower in Argentina, but similar ones have been reported in Africa and Mexico. The aim of this study was to characterize this new sunflower disease.  The characterization was based on virus transmission, host range, electron microscopy and comparison of its sequence with those of related viruses.  Virus transmission efficiency by mechanical inoculation to sunflower plants at the V2 vegetative stage was nearly 100%. Seed transmission was negative as plants derived from seeds of systemically infected plants showed no symptoms. A host range including 27 spe...

 Two potyviruses that infect sunflower (Helianthus annuus L.) crops have been reported in Argentina. Sunflower chlorotic mottle virus (SuCMoV) is the most widely distributed one, with two strains: common and ring spot, which have been completely characterized. Sunflower mild mosaic virus (SuMMoV) that produces a chlorotic mild mosaic on sunflower commercial crops was detected in the rural area of Paraná (Entre Ríos Province). This virus has been biologically and serologically characterized but not fully characterized at the molecular level. The aim of this work was to determine the complete sequence of SuMMoV and study its genomic organization.  A total RNA extraction was made from a preparation enriched with this virus. The quality of preparation was confirmed through electron microscopy and RT-PCR with specific primers for capsid protein gene. Total RNA obtained was pyrosequenced and the sequences obtained were analyzed.  Electron microscopy from rapid preparations of samples e...

This is the first survey to determine the occurrence, prevalence, and distribution of peanut (Arachis hypogaea) viral diseases in Argentina. It was conducted in the province of Córdoba, which has 92% of the country&#39;s peanut production. It included the main peanut viruses Peanut mottle virus (PeMoV), Peanut stripe virus (PStV), Cucumber mosaic virus (CMV), Peanut stunt virus (PSV), Tomato spotted wilt virus (TSWV), and Groundnut ringspot virus (GRSV). Leaf samples from 1,028 individual peanut plants with virus-like symptoms and 986 samples from asymptomatic plants were collected in six counties of Córdoba over 3 years and serologically tested for the presence of viruses. PeMoV was the most frequently detected virus, found in 58.8, 34.2, and 23.4% of samples from the 2003–04, 2004–05, and 2005–06 growing seasons, respectively, and it was found in all sampled counties. Also, it was the only virus detected in asymptomatic plants. Less than 4% of symptomatic plants were infected with...

Outbreaks in fish of motile Aeromonad septicemia (MAS) caused by Aeromonas hydrophila have caused a great concern worldwide. Here, for the first time, we provide two complete genomes of epidemic A. hydrophila strains isolated in China. To gain an insight into the pathogenicity of epidemic A. hydrophila, we performed comparative genomic analyses of five epidemic strains belonging to sequence type (ST) 251, together with the environmental strain ATCC 7966 T . We found that the known virulence factors, including a type III secretion system, a type VI secretion system and lateral flagella, are not required for the high virulence of the ST251 clonal group. Additionally, our work identifies three utilization pathways for myo-inositol, sialic acid and L-fucose providing clues regarding the factors that underlie the epidemic and virulent nature of ST251 A. hydrophila. Based on the geographical distribution and biological resources of the ST251 clonal group, we conclude that ST251 is a high-risk clonal group of A. hydrophila which may be responsible for the MAS outbreaks in China and the southeastern United States.

An arsenic-resistant bacterium, AsRB1, was isolated from the fronds of Pteris vittata grown in a site contaminated with copper chromium arsenate. The bacterium exhibited resistance to arsenate, arsenite, and antimony in the culture medium. AsRB1, like Pseudomonas putida, grew on MacConkey and xylose-lactose-desoxycholate agars and utilized citrate but, unlike P. putida, was positive for indole test and negative for oxidase test. A phylogenetic analysis of the 16S rRNA gene showed that AsRB1 is a proteobacterium of the beta subclass, related to Pseudomonas saccharophila and Variovorax paradoxus. Following an exogenous supply of arsenate, most arsenic occurred as arsenite in the medium and the cell extracts, suggesting reduction and extrusion of arsenic as the mechanism for arsenic resistance in AsRB1.

Most illnesses affecting the oral cavity are proven to have infectious origin. Several categories of chemical agents have been used in the chemical control of dental biofilm through strategies that aim at reducing bacterial adhesion and inhibiting the growth and the proliferation of microorganisms on the tooth surface. The use of plants in folk medicine and in Dentistry, as well as the spread of successful cases, has led to scientific exploration, resulting in chemical-pharmacological knowledge of thousands of plants. The present study aimed to evaluate the anti-adherence activity of Lippia sidoides Cham., comparing the results with those of 0.12% chlorhexidine by means of an in vitro simulation of dental biofilm. The studied bacterial strains were Streptococcus mutans, Streptococcus sanguinis and Lactobacillus casei, main responsible for the biofilm adherence. The studied extract was effective in inhibiting the adherence of Streptococcus mutans up to a concentration of 1:16, compar...

Candida albicans is a dimorphic fungus that can grow in yeast morphology or hyphal form depending on the surrounding environment. This ubiquitous fungus is present in skin and mucus membranes as a potential pathogen that under opportunistic conditions causes a series of systemic and superficial infections known as candidiasis, moniliasis or simply candidiasis. There has been a steady increase in the prevalence of candidiasis that is expressed in more virulent forms of infection. Although candidiasis is commonly manifested as mucocutaneous disease, life-threatening systemic invasion by this fungus can occur in every part of the body. The severity of candidal infections is associated with its morphological shift such that the hyphal morphology of the fungus is most invasive. Of importance, aberrant multiplication of Candida yeast is also associated with the pathogenesis of certain mucosal diseases. In this study, we assessed the anti-candidal activity of the volatile anesthetic isoflu...

Ability of Bacillus sp., Pseudomonas fluorescens, and Trichoderma sp. to Control Tomato Bacterial Wilt DiseaseRalstonia solanacearum is known as the causal agent of bacterial wilt on tomato.  The bacteria may infect all stadia of plant growth and decrease tomato production. Biological control using antagonistic microbes is considered as a potential control alternative for the disease. This research was aimed to assay the ability of combination treatment of Bacillus sp., Pseudomonas fluorescens, and Trichoderma sp. in controlling R. solanacearum and its effect on growth and yield of tomato in the field. The treatments consisted of control (without antagonistic microbes), mixed combination of Bacillus sp. B8 + Bacillus sp. B11 + Trichoderma sp.; Bacillus sp. B8 + Pseudomonas flourescens P8 + Trichoderma sp.; and Streptomisin sulfat 20%. Antagonistic microbes was applied at planting time, as much as 100 mL per plant. The result showed that mixed combination of Bacillus sp. B8 + Bacillu...

Research aiming for knowing ability of biological agents in remeding contaminated soil with Fusarium oxysporum f.sp. lycopersici and in controlling Fusarium wilt was carried out at the screen house for five months. Completely Randomized Block Design was used with 12 treatments and repeated three times. The agents tested were Trichoderma harzianum isoleted from ginger, ginseng, or banana, Bacillus subtilis, and Pseudomonas fluorescens P60. Result of the research showed that all the agents could decrease population density of F. oxysporum f.sp. lycopersici ranging from 21.01 to 40.03% applied two weeks before planting to the soil. The most effective agent was P. fluorescens P60 by decreasing the population of 35.08%, suppressing the disease of 28.15%, and increasing the agent itself for 50%.

Traditional materials from new sourcesconflicts in analytical methods for calcium carbonate. Calcium carbonate, (E170) is a common food and pharmaceutical additive/ingredient. In addition to a source of calcium, the carbonate has uses including as a colour, acidity regulator and bulking agent. Globally, a range of regulatory agencies and pharmacopoeia control the analyses and specification of additives in food, supplements, pharmaceutical substances and excipients. Accordingly, a range of specifications and analyses exist for calcium carbonate depending on the application and market of the product. In this contribution, we analyse calcium carbonates from geological, synthetic and biogenic sources, focussing on acid insoluble impurities, a test required by current monographs. Analysis of calcium carbonate from different origins may require modification of existing tests to comply with regulatory bodies, due to the variation of impurities specific to the source of the material. We suggest an analytical approach involving centrifugation that improves analytical efficiency (up to 85% time reduction), especially for calcium carbonate of biological origin.

Objectives: Aspergillus spp. are the most frequently isolated filamentous fungi in the sputum of patients with cystic fibrosis (CF). Resistance to the azoles, the mainstay of current antifungal therapy, has been increasingly observed worldwide, but few data are available on the resistance of Aspergillus spp. in German CF patients. This study investigated the epidemiology of Aspergillus spp. and the molecular origin of azole resistance in a large German CF centre. Methods: In total, 2677 respiratory samples from 221 CF patients collected between April 2010 and April 2013 were analysed; of these, 573 yielded Aspergillus spp., which were screened for azole resistance. Isolates with reduced susceptibility to itraconazole and/or voriconazole were tested according to the EUCAST reference procedure. Sequencing of cyp51A, the target of azole antifungals, was performed in all resistant isolates. Results: Six isolates obtained from four patients were highly resistant to itraconazole (all identified as Aspergillus fumigatus sensu stricto); five of them were pan-azole resistant. The TR34/L98H mutation was the most frequent mutation identified in azole-resistant isolates (n¼4), followed by M220L and TR46/Y121F/T289A, a mutation previously reported from Belgium and the Netherlands only. Three of four patients harbouring azole-resistant A. fumigatus had not received any prior azole treatment. Conclusions: Resistance to azoles in Aspergillus spp. is still infrequent in German CF patients and is mainly caused by the TR34/L98H mutation. Worryingly, pan-azole-resistant TR46/Y121F/T289A has spread to Germany. Azole resistance has to be considered also in azole-naive CF patients and susceptibility testing of Aspergillus spp. isolates should be performed in all patients requiring treatment.

Several promising candidate drugs that target bacterial adherence and biofilm formation are being developed. Such hopeful drugs cannot be studied in chronic rhinosinusitis (CRS) without the evaluation of such virulence criteria in different forms of the disease with and without nasal polyposis (CRSwNP and CRSsNP). The aim of this study was to evaluate bacterial adherence, response to antibiotics and degree of accumulation of bacterial biofilms as new targets of treatment in CRSwNP and CRSsNP. Twenty CRS patients and 10 normal subjects with airway obstructing concha bullosa were prospectively enrolled in the present study. Scanning electron microscopy and cultures were performed on paranasal sinus tissue samples. Bacterial adherence tests using the tissue culture plate method were measured quantitatively. Strongly adherent bacteria were identified significantly in 6/9 (77%) cases of CRSsNP in comparison to 1/7 (14%) cases of CRSwNP. Strongly adherent bacteria that were sensitive to ciprofloxacin, vancomycin, and impenim were identified in 75% of the cultured coagulase positive staphylococci. A significant difference (P = 0.007) in the degree of accumulation of bacterial biofilms existed between the two groups. In CRSsNP, a more advanced stage of bacterial biofilms with strong bacterial adherence was observed which make them attractive targets for new drugs. In CRSwNP, lower stage bacterial biofilms with low bacterial adhesion were identified, which may help explain the low bacterial virulence in an environment of suboptimal, organizational arrangements.

Background: Common bean is the most important staple grain legume for direct human consumption and nutrition. It complements major sources of carbohydrates, including cereals, root crop, or plantain, as a source of dietary proteins. It is also a significant source of vitamins and minerals like iron and zinc. To fully play its nutritional role, however, its robustness against stresses needs to be strengthened. Foremost among these is drought, which commonly affects its productivity and seed quality. Previous studies have shown that photosynthate remobilization and partitioning is one of the main mechanisms of drought tolerance and overall productivity in common bean. Results: In this study, we sought to determine the inheritance of pod harvest index (PHI), a measure of the partitioning of pod biomass to seed biomass, relative to that of grain yield. We evaluated a recombinant inbred population of the cross of ICA Bunsi and SXB405, both from the Mesoamerican gene pool, to determine the effects of intermittent and terminal drought stresses on the genetic architecture of photosynthate allocation and remobilization in pods of common bean. The population was grown for two seasons, under well-watered conditions and terminal and intermittent drought stress in one year, and well-watered conditions and terminal drought stress in the second year. There was a significant effect of the water regime and year on all the traits, at both the phenotypic and QTL levels. We found nine QTLs for pod harvest index, including a major (17% of variation explained), stable QTL on linkage group Pv07. We also found eight QTLs for yield, three of which clustered with PHI QTLs, underscoring the importance of photosynthate remobilization in productivity. We also found evidence for substantial epistasis, explaining a considerable part of the variation for yield and PHI. Our results highlight the genetic relationship between PHI and yield and confirm the role of PHI in selection of both additive and epistatic effects controlling drought tolerance. These results are a key component to strengthen the robustness of common bean against drought stresses.

Background Phosphorus (P) is an essential macronutrient for plant growth that participates in a series of biological processes. Thus, P deficiency limits crop growth and yield. Although Stylosanthes guianensis (stylo) is an important tropical legume that displays adaptation to low phosphate (Pi) availability, its adaptive mechanisms remain largely unknown. Results In this study, differences in low-P stress tolerance were investigated using two stylo cultivars (‘RY2’ and ‘RY5’) that were grown in hydroponics. Results showed that cultivar RY2 was better adapted to Pi starvation than RY5, as reflected by lower values of relative decrease rates of growth parameters than RY5 at low-P stress, especially for the reduction of shoot and root dry weight. Furthermore, RY2 exhibited higher P acquisition efficiency than RY5 under the same P treatment, although P utilization efficiency was similar between the two cultivars. In addition, better root growth performance and higher leaf and root APas...

Introduction Anthracnose caused by CoUetotrichum lindemuthianum (Sacc. and Magn.) Scrib. is one of the most widespread and economically important fungal diseases of common bean {Phaseolus vulgaris LO« Genetic resistance is the most effective method of control of anthracnose in common bean where ten major resistance genes have been characterized. Inheritance of anthracnose resistance in the differential cultivar Widusa has not been fully characterized. To facilitate the use of Widusa as a source of resistance to anthracnose, we investigated its mode of inheritance in a series of allelism tests with previously characterized resistance genes.