A disease of grasshoppers caused by the bacterium Pseudomonas aeruginosa (Schroeter) Migula.

Rhamnus purshiana, or cascara, is a deciduous tall shrub or small tree as much as 9 m high with thin, smooth, silver-gray bark. It is often present in shady sites in redwood and mixed evergreen forests of the North Ameri-can west coast, from British Columbia to northern California. In July 2005, symptomatic leaves with irregular, black spots, 2 to 5 mm in diameter and concentrated toward the tips, were collected from four cascara plants in the Samuel P. Taylor State Park, Marin County, California. There was no evidence of defoliation. Pieces of necrotic tissue were plated on selective medium (PARP) and maintained at 19°C for 2 weeks. A Phytophthora sp. was consistently isolated and it was identified as P. ramorum on the basis of morphological and molecular traits published previously (3,4). The P. ramorum isolate Pr-418 has been deposited in the American Type Culture Collection (ATCC MYA-3676) and a portion of the internal transcribed sequence (ITS) of rDNA has been deposited in the NCBI database (GenBank Accession No. DQ168874). Koch's postulates were completed using the leaf-dip method (2) on detached leaves collected from three cascara plants growing at the University of California Botanical Garden at Berkeley. Zoospore inoculum was prepared by flooding a 2-week-old culture growing on V8 agar with sterile water for 4 days. The liquid was filtered after cold shocking at 4°C for 30 min and incubated at room temperature for 1 h. Fifteen leaves were dipped in the resulting zoospore suspension (1.6 × 104 zoospores per ml) for either 1 min (experiment 1) or overnight (experiment 2). Leaves used as negative controls were dipped in sterile water. After removal from the inoculum, excess liquid was allowed to drain. Leaves were maintained in a moist chamber at 19°C with 13 h of natural light for 1 week. After 3 days of incubation, necrotic spots similar to those observed in the field had developed on leaves in experiment 2, while no symptoms were observed in experiment 1. Necrotic lesions were observed on 12 and 15 of 15 leaves in experiments 1 and 2, respectively, after 7 days of incubation. For each leaf, the necrotic area and percent necrosis was determined by placing the leaves in a flatbed scanner and processing the images with Assess (Version 1.01; The American Phytopathological Society, St. Paul, MN). Lesions extended from the tip of the leaves and covered 3 ± 1% of the total leaf area in experiment 1 and 33 ± 3% in experiment 2. Reisolation of P. ramorum on PARP was successful for all inoculated leaves. P. ramorum was never isolated from negative controls and no symptoms of infection were observed. The leaf-dip inoculation method is a rapid and reliable indicator of host susceptibility to P. ramorum, with many species developing necrosis when exposed to high concentrations of zoospores (3). Our results show that exposure time to the pathogen can play an important role in the development of symptoms. R. purshiana has been previously reported as a host in Oregon (1,2), but to our knowledge, this is the first report of cascara as a natural host of P. ramorum in the state of California. Our results confirm those from Oregon (2). The impact of infection by P. ramorum on cascara is unknown.

The production of patulin and griseofulvin by 49 different isolates of Penicillium griseofulvum Dierckx was analyzed by high-performance liquid chromatography. Eleven isolates were obtained from pistachio nuts, 37 were obtained from wheat seeds, and 1 was obtained from the American Type Culture Collection. Activities of 19 enzymes were also assayed by the API ZYM system. From these results it may be deduced that there are two different groups among the strains tested which cannot be distinguished by morphological and cultural characteristics. One group of isolates did not produce detectable amounts of patulin and griseofulvin when grown in sucrose-yeast extract and Wickerham media, while enzymatic activities were quantitatively distinct from the other group, which produced patulin and griseofulvin in variable proportions. Leucine arylamidase, phosphoamidase, and beta-D-glucosidase are the main enzymes with differing activities between the two groups. Differences in physiological characteristics among isolates of a single species reveal shortcomings in the classification of the penicillia based only on morphological criteria. Thus, determination of the ability to yield mycotoxins and antibiotics as well as determination of enzymatic activities appear to be very valuable tools in the taxonomy of these fungi and for food toxicology.

A survey was conducted in April 2003 to identify viruses infecting faba bean (Vicia faba L.) in six regions (Beja, Bizerte, Cap-bon, Le Kef, Siliana, and Zaghouan) in Tunisia. A total of 292 faba bean samples with symptoms of viral infection (leaf rolling, yellowing, and mosaic) were collected. The samples were tested at the virology laboratory of the International Center for Agricultural Research in the Dry Areas (ICARDA), Syria, for 11 viruses using the tissue-blot immunoassay procedure (3). Specific rabbit polyclonal antisera were used to test for Chickpea chlorotic dwarf virus (CpCDV) (provided by H. J. Vetten, BBA, Braunschweig, Germany), Alfalfa mosaic virus (AMV), Bean yellow mosaic virus (BYMV), Broad bean mottle virus (BBMV), Broad bean stain virus (BBSV), Cucumber mosaic virus (CMV), and Pea seedborne mosaic virus (PSbMV) (ICARDA, Aleppo, Syria). In addition, four specific monoclonal antibodies were used to detect Bean leaf roll virus (BLRV) (4B10) (2), Beet western yellows virus (BWYV) (ATCC PVAS-647; American Type Culture Collection, Manassas, VA), Faba bean necrotic yellows virus (FBNYV) (3-2E9) (1), and Soybean dwarf virus (SbDV) (ATCC PVAS-650). Serological tests showed that BBMV, a beetle-transmitted and seedborne virus identified in 23.3% (68 samples) of the samples tested, was the most common. BLRV, FBNYV, BWYV, BYMV, SbDV, and PSbMV were detected in 56, 33, 31, 10, 5, and 1 sample(s) of 292 samples tested, respectively. AMV, BBSV, CMV, and CpCDV were not detected in any samples tested. In Tunisia, BLRV, BWYV, BYMV, FBNYV, and PSbMV have previously been reported in faba bean (4), but to our knowledge, this is the first record of SbDV affecting faba bean in Tunisia, where it was detected in two fields in the Cap-bon Region. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by western blots, extracts from SbDV-infected plants were observed to contain 23-kDa structural proteins, which reacted strongly with SbDV monoclonal antibodies. Transmission tests showed that the samples, which reacted with SbDV monoclonal antibodies, were transmitted to faba bean plants by the pea aphid (Acyrthosiphon pisum Harris) in a persistent manner. To our knowledge, this is the first report of SbDV naturally infecting faba bean in Tunisia and it could cause a serious problem to other leguminous crops grown in Tunisia, such as French bean and peas, which are hosts for the virus.

A field visit was conducted in March 2002 to identify viruses infecting the faba bean ( Vicia faba ) crop in four governorates (Fayoum, Beni Suef, El-Nobareia and Bihera) in Egypt. A total of 71 faba bean samples with symptoms of viral infection (leaf rolling, yellowing and mosaic) were collected. These were tested at the Virology Laboratory of ICARDA, Syria, for seven viruses using the tissue-blot immunoassay procedure (Makkouk & Comeau, 1994). Specific rabbit polyclonal antisera were used to test for Chickpea chlorotic dwarf virus (CpCDV, genus Mastrevirus , family Geminiviridae ; provided by H. J. Vetten, BBA, Braunschweig, Germany); for Broad bean stain virus (BBSV, genus Comovirus , family Comoviridae ); for Bean yellow mosaic virus (BYMV, genus Potyvirus , family Potyviridae ) and for Pea seed borne mosaic virus (PSbMV, genus Potyvirus , family Potyviridae ). In addition, three specific monoclonal antibodies were used to detect Faba bean necrotic yellows virus (FBNYV, genus Nanovirus , family Nanoviridae ) (Franz et al ., 1996); Bean leaf roll virus (BLRV, family Luteoviridae ) (4B10; Katul, 1992) and Beet western yellows virus (BWYV, genus Polerovirus , family Luteoviridae ) (ATCC PVAS-647; American Type Culture Collection, Manassas, VA, USA). Serological tests showed that BYMV, an aphid-transmitted and seedborne virus that was identified in 89% of samples tested, was the most common virus. In most of the fields surveyed, BYMV symptoms were noted to occur at high levels (80–100% infection). PSbMV was detected in nine samples (one from Fayoum and eight from Beni Suef); FBNYV was detected in 11 samples (all from Beni Suef). CpCDV, a leafhopper-transmitted virus, was, by contrast, detected in only two samples collected from the El-Nobareia governorate showing leaf rolling, yellowing and stunting. BWYV, BLRV and BBSV were not detected in any of the samples tested. In Egypt, FBNYV, BYMV and PSbMV have previously been reported in faba bean (Makkouk et al ., 1994), but this is the first record of CpCDV affecting faba bean in Egypt. CpCDV is the only member of the Geminivirus that is reported to naturally infect faba bean.

Human nasal epithelial cells derived from multiple subjects exhibit differential responses to H3N2 influenza virus infection in vitro

Ptaeroxylon obliquum (Thunb.) Radlk. (Rutaceae) is traditionally used to treat human and animal diseases in South Africa. In this study, the activity of leaf extracts, fractions, and isolated compounds was determined against nonpathogenic mycobacterial species and nosocomial bacterial pathogens. An acetone leaf extract was partitioned by liquid-liquid fractionation, and obliquumol, a mixture of lupeol and β-amyrin, and eranthin were isolated. Antimicrobial activity was determined using a serial microdilution assay against Mycobacterium smegmatis (American Type Culture Collection [ATCC] 1441), M. bovis (BCG P1172), M. aurum (NCTC 10437), M. fortuitum (ATCC 6841), Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Pseudomonas aeruginosa (ATCC 25922), and Escherichia coli (ATCC 27853). The n-hexane fraction had minimal inhibitory concentration (MIC) values as low as 20 and 40 µg/mL against M. fortuitum and S. aureus, respectively. The chloroform fraction also had promising activity with an MIC value of 80 µg/mL against both P. aeruginosa and M. fortuitum. Obliquumol had excellent activity (MIC 8 µg/mL) against M. fortuitum. Fractionation of the crude extract potentiated the antimicrobial activity of the nonpolar fractions. The isolated compound, obliquumol, had good antimicrobial and excellent antimycobacterial activities. The antimicrobial activity provides some scientific rationale for the use of P. obliquum against infectious diseases and related symptoms. This is the first report on the antibacterial activity of obliquumol.

<i>Phoma tracheiphila</i>

In this paper the Subcommittee on the Taxonomy of Mollicutes proposes minimum standards for descriptions of new cultivable species of the class Mollicutes (trivial term, mollicutes) to replace the proposals published in 1972 and 1979. The major class characteristics of these organisms are the lack of a cell wall, the tendency to form fried-egg-type colonies on solid media, the passage of cells through 450- and 220-nm-pore-size membrane filters, the presence of small A-T-rich genomes, and the failure of the wall-less strains to revert to walled bacteria under appropriate conditions. Placement in orders, families, and genera is based on morphology, host origin, optimum growth temperature, and cultural and biochemical properties. Demonstration that an organism differs from previously described species requires a detailed serological analysis and further definition of some cultural and biochemical characteristics. The precautions that need to be taken in the application of these tests are defined. The subcommittee recommends the following basic requirements, most of which are derived from the International Code of Nomenclature of Bacteria, for naming a new species: (i) designation of a type strain; (ii) assignment to an order, a family, and a genus in the class, with selection of an appropriate specific epithet; (iii) demonstration that the type strain and related strains differ significantly from members of all previously named species; and (iv) deposition of the type strain in a recognized culture collection, such as the American Type Culture Collection or the National Collection of Type Cultures. The publication of the description should appear in a journal having wide circulation. If the journal is not the International Journal of Systematic Bacteriology, a reprint must be submitted to that journal so that the name may be considered for inclusion in a validation list as required by the International Code of Nomenclature of Bacteria.

Mycoplasmas serologically different from the known Mycoplasma species were found to be important causes of mastitis in California. We describe the cultural, morphological, biological, and serological characteristics of one of these isolates, strain ST-6. In growth inhibition and immunofluorescence tests, we found that strain ST-6 differed from all 75 currently accepted Mycoplasma species and serogroups. Therefore, we regard this strain as a member of a new species for which we propose the name Mycoplasma californicum. Strain ST-6, the type strain, has been deposited in the Collaborating Center for Animal Mycoplasmas, Institute of Medical Microbiology, University of Aarhus, Denmark, under the accession number AMRC-C 1077 and in the American Type Culture Collection under the number ATCC 33461.

Pisum sativum L. subsp. elatius (Steven ex M. Bieb.) Asch. & Graebn. is a wild pea species that is native to Bulgaria. It readily crosses to the cultivated pea species P. sativum subsp. sativum. Field pea is an important component in the crop rotation system of the northeast region of Bulgaria. Little is known or published on the diseases of wild Pisum subspecies. In June 1997, brown to reddish brown, irregularly shaped lesions 5 to 10 mm in diameter were found on the leaves and stems of P. sativum subsp. elatius growing under native conditions in the low growing vegetation in a mixed forest habitat on the Black Sea coast at Albena, Bulgaria (43°22'26″N; 28°05'02″E) at an elevation of about 50 m. Black pycnidia were observed within lesions and contained hyaline, primarily two-celled conidia that measured 7 to 17 × 3 to 5 μm. On artificially inoculated pea stem pieces incubated on 2% water agar (WA) at 22 to 24°C for 28 days, pseudothecia developed with hyaline, two-celled ascospores constricted at the septum and measuring 12 to 17 × 4 to 7 μm. Black chlamydospores produced singly or in chains also formed in infected foliar tissues and on potato dextrose agar (PDA) and WA. Isolations were made from the lesions on pea tissue onto WA and PDA after disinfesting in 0.25% NaOCl for 5 min. Koch's postulates were fulfilled by inoculating the foliage of P. sativum subsp. sativum cvs. Dark Skin Perfection and Sounder and P. sativum subsp. elatius (W6-20047), and reisolating the fungus from lesions that developed on the inoculated leaves and stems. The wild Pisum fungus was identified as Mycosphaerella pinodes (Berk. & Blox.) Vestergr. based on cultural and morphological characteristics (2), pathogenicity tests, and by comparing random amplified polymorphic DNA (RAPD) markers with those of American Type Culture Collection (ATCC) isolates 201628 to 201633 of M. pinodes. The fungus was identified as a pathogen of cultivated peas in Bulgaria by Kovachevsky and Hristov (1) in 1949. This is the first report of M. pinodes infecting P. sativum subsp. elatius in Bulgaria and other countries where P. sativum subsp. elatius is a native plant species.

During the late stages of infection, Salmonella secretes numerous effectors through a type III secretion system that is encoded within Salmonella pathogenicity island 2 (SPI2). Despite the importance of SPI2 as a major virulence factor leading to the systemic spread of the bacteria and diseases, a global view of its effects on host responses is still lacking. Here, we measured global impacts of SPI2 effectors on the host phosphorylation and protein expression levels in RAW264.7 and in HeLa cells, as macrophage and nonphagocytic models of infection. We observe that SPI2 effectors differentially modulate the host phosphoproteome and cellular processes (e.g. protein trafficking, cytoskeletal regulation, and immune signaling) in a host cell-dependent manner. Our unbiased approach reveals the involvement of many previously unrecognized proteins, including E3 ligases (HERC4, RanBP2, and RAD18), kinases (CDK, SIK3, and WNK1), and histones (H2B1F, H4, and H15), in late stages of Salmonella infection. Furthermore, from this phosphoproteome analysis and other quantitative screens, we identified HSP27 as a direct in vitro and in vivo molecular target of the only type III secreted kinase, SteC. Using biochemical and cell biological assays, we demonstrate that SteC phosphorylates multiple sites in HSP27 and induces actin rearrangement through this protein. Together, these results provide a broader landscape of host players contributing to specific processes/pathways mediated by SPI2 effectors than was previously appreciated.

Previous studies have demonstrated that topoisomerase I is cleaved late during apoptosis, but have not identified the proteases responsible or examined the functional consequences of this cleavage. Here, we have shown that treatment of purified topoisomerase I with caspase-3 resulted in cleavage at DDVD146 downward arrowY and EEED170 downward arrowG, whereas treatment with caspase-6 resulted in cleavage at PEDD123 downward arrowG and EEED170 downward arrowG. After treatment of Jurkat T lymphocytic leukemia cells with anti-Fas antibody or A549 lung cancer cells with topotecan, etoposide, or paclitaxel, the topoisomerase I fragment comigrated with the product that resulted from caspase-3 cleavage at DDVD146 downward arrowY. In contrast, two discrete topoisomerase I fragments that appeared to result from cleavage at DDVD146 downward arrowY and EEED170 downward arrowG were observed after treatment of MDA-MB-468 breast cancer cells with paclitaxel. Topoisomerase I cleavage did not occur in apoptotic MCF-7 cells, which lack caspase-3. Cell fractionation and band depletion studies with the topoisomerase I poison topotecan revealed that the topoisomerase I fragment remains in proximity to the chromatin and retains the ability to bind to and cleave DNA. These observations indicate that topoisomerase I is a substrate of caspase-3 and possibly caspase-6, but is cleaved at sequences that differ from those ordinarily preferred by these enzymes, thereby providing a potential explanation why topoisomerase I cleavage lags behind that of classical caspase substrates such as poly(ADP-ribose) polymerase and lamin B1.

The first non-radioactive digoxigenin labeled cDNA probes were prepared from part of coat protein gene of the American (No.45058, American Type Culture Collection), Czech (A52/E6), German (Chandler/E6) and Norwegian (Mimek/Alpine) isolates of strawberry vein banding virus (SVBV). All five American different air-dried SVBV leaf samples originating from National Clonal Germplasm Repository (N.C.G.R., Corvallis, U.S.A) and six from twelve Czech strawberry isolates exhibiting SVBV-like infection symptoms reacted positively in dot blot hybridization with American probe. Twenty six Czech strawberry SVBV samples were used in dot blot with probes prepared against the Czech, American, German and Norwegian isolates. Our results showed that the samples reacted identically, independently on the origin of the probe. They confirm the spread of SVBV in Central Europe and introduce the first reliable screening method for this virus.

To assess the effect of Lactobacillus acidophilus (American Type Culture Collection (ATCC) 700396) on enterotoxigenic Escherichia coli (ETEC) infection, in the present study, a parallel, double-blind, placebo-controlled 4-week intervention was performed in healthy males. The subjects largely consumed their habitual diet, but had to abstain from consuming dairy foods generally high in Ca. The subjects were randomised into the L. acidophilus (dose 10⁹ colony-forming units twice daily; n 20) or the placebo (n 19) group. After an adaptation period of 2 weeks, the subjects were orally infected with a live, but attenuated, ETEC vaccine, able to induce mild, short-lived symptoms. Before and after the challenge, the subjects recorded stool consistency, bowel habits, and frequency and severity of gastrointestinal complaints. The ETEC challenge led to a significant increase in faecal output on the 2nd day and a concomitant increase in Bristol stool scale scores. Likewise, abdominal pain, bloating, flatulence, fever, headache and nausea peaked 1 d after the oral challenge. The concentrations of faecal calprotectin and IgA peaked 2 d after and that of serum IgM peaked 9 and 15 d after the oral challenge. The concentrations of serum IgA and IgG were unaffected. The ETEC challenge led to a reduction in the number of Bacteroides-Prevotella, Bifidobacterium, Clostridium cluster XIVab and total faecal bacteria. Probiotic treatment was associated with a larger increase in Bristol stool scale scores and more fever, headache and nausea after the ETEC challenge compared with the placebo treatment. These differences were, however, small and with substantial variation within the groups. Oral application of an attenuated live ETEC vaccine provides a useful model for food-borne infections. Supplementation with L. acidophilus ATCC 700396, however, was ineffective in reducing ETEC infection symptoms in healthy men.

Febrile or Exanthematous Illness Associated with Zika, Dengue, and Chikungunya Viruses, Panama.