COVID-19 pandemic - Inequities and inequalities to exercise and their consequences on the physical and mental health of women with cardiovascular disease: recommendations on how to address the needs of women.

The coronavirus disease (COVID-19) pandemic disproportionately affects those with pre-existing conditions and has exacerbated gender inequalities. Cardiovascular disease (CVD) is the leading cause of death among Canadian women. Exercise improves physical and mental health and CVD management. Amid the pandemic, women are experiencing an increase in caregiving responsibilities, job insecurities, and domestic violence creating competing demands for prioritizing their health. Recommendations on how to meet the unique needs of Canadian women with CVD through exercise are provided. Novelty: Exercise recommendations amid the pandemic for women with CVD need to be flexible, feasible, and fun.

Profiling of human burned bones: oxidising versus reducing conditions.

Complementary optical and neutron-based vibrational spectroscopy techniques (Infrared, Raman and inelastic neutron scattering) were applied to the study of human bones (femur and humerus) burned simultaneously under either aerobic or anaerobic conditions, in a wide range of temperatures (400 to 1000 °C). This is the first INS study of human skeletal remains heated in an oxygen-deprived atmosphere. Clear differences were observed between both types of samples, namely the absence of hydroxyapatite's OH vibrational bands in bone burned anaerobically (in unsealed containers), coupled to the presence of cyanamide (NCNH2) and portlandite (Ca(OH)2) in these reductive conditions. These results are expected to allow a better understanding of the heat effect on bone´s constituents in distinct environmental settings, thus contributing for an accurate characterisation of both forensic and archaeological human skeletal remains found in distinct scenarios regarding oxygen availability.

New and Interesting Fungi. 2.

One order, seven families, 28 new genera, 72 new species, 13 new combinations, four epitypes, and interesting new host and / or geographical records are introduced in this study. Pseudorobillardaceae is introduced for Pseudorobillarda (based on P. phragmitis). New genera include: Jeremyomyces (based on J. labinae) on twigs of Salix alba (Germany); Neodothidotthia (based on N. negundinicola) on Acer negundo (Ukraine); Neomedicopsis (based on N. prunicola) on fallen twigs of Prunus padus (Ukraine); Neophaeoappendicospora (based on N. leucaenae) on Leucaena leucocephala (France) (incl. Phaeoappendicosporaceae); Paradevriesia (incl. Paradevriesiaceae) (based on P. americana) from air (USA); Phaeoseptoriella (based on P. zeae) on leaves of Zea mays (South Africa); Piniphoma (based on P. wesendahlina) on wood debris of Pinus sylvestris (Germany); Pseudoconiothyrium (based on P. broussonetiae) on branch of Broussonetia papyrifera (Italy); Sodiomyces (based on S. alkalinus) from soil (Mongolia), and Turquoiseomyces (incl. Turquoiseomycetales and Turquoiseomycetaceae) (based on T. eucalypti) on leaves of Eucalyptus leptophylla (Australia); Typhicola (based on T. typharum) on leaves of Typha sp. (Germany); Xenodevriesia (incl. Xenodevriesiaceae) (based on X. strelitziicola) on leaves of Strelitzia sp. (South Africa). New species include: Bacillicladium clematidis on branch of Clematis vitalbae (Austria); Cercospora gomphrenigena on leaves of Gomphrena globosa (South Africa); Cyphellophora clematidis on Clematis vitalba (Austria); Exophiala abietophila on bark of Abies alba (Norway); Exophiala lignicola on fallen decorticated trunk of Quercus sp. (Ukraine); Fuscostagonospora banksiae on Banksia sp. (Australia); Gaeumannomycella caricicola on dead leaf of Carex remota (Germany); Hansfordia pruni on Prunus persica twig (Italy) (incl. Hansfordiaceae); Microdochium rhopalostylidis on Rhopalostylis sapida (New Zealand); Neocordana malayensis on leaves of Musa sp. (Malaysia); Neocucurbitaria prunicola on fallen twigs of Prunus padus (Ukraine); Neocucurbitaria salicis-albae on Salix alba twig (Ukraine); Neohelicomyces deschampsiae on culm base of dead leaf sheath of Deschampsia cespitosa (Germany); Pararoussoella juglandicola on twig of Juglans regia (Germany); Pezicula eucalyptigena on leaves of Eucalyptus sp. (South Africa); Phlogicylindrium dunnii on leaves of Eucalyptus dunnii (Australia); Phyllosticta hagahagaensis on leaf litter of Carissa bispinosa (South Africa); Phyllosticta austroafricana on leaf spots of unidentified deciduous tree host (South Africa); Pseudosigmoidea alnicola on Alnus glutinosa leaf litter (Germany); Pseudoteratosphaeria africana on leaf spot on unidentified host (Angola); Porodiplodia vitis on canes of Vitis vinifera (USA); Sodiomyces alkalinus from soil (Mongolia), Sodiomyces magadiensis and Sodiomyces tronii from soil (Kenya), Sympodiella quercina on fallen leaf of Quercus robur (Germany) and Zasmidium hakeicola on leaves of Hakea corymbosa (Australia). Epitypes are designated for: Cryptostictis falcata on leaves of E. alligatrix (Australia), Hendersonia phormii on leaves of Phormium tenax (New Zealand), Sympodiella acicola on needles of Pinus sylvestris (Netherlands), and Sphaeria scirpicola var. typharum on leaf of Typha sp. (Germany). Several taxa originally described from rocks are validated in this study. New taxa include: Extremaceae fam. nov., and new genera, Arthrocatena, Catenulomyces, Constantinomyces, Extremus, Hyphoconis, Incertomyces, Lapidomyces, Lithophila, Monticola, Meristemomyces, Oleoguttula, Perusta, Petrophila, Ramimonilia, Saxophila and Vermiconidia. New species include: Arthrocatena tenebrosa, Catenulomyces convolutus, Constantinomyces virgultus, C. macerans, C. minimus, C. nebulosus, C. virgultus, Exophiala bonariae, Extremus adstrictus, E. antarcticus, Hyphoconis sterilis, Incertomyces perditus, Knufia karalitana, K. marmoricola, K. mediterranea, Lapidomyces hispanicus, Lithophila guttulata, Monticola elongata, Meristemomyces frigidus, M. arctostaphyli, Neodevriesia bulbillosa, N. modesta, N. sardiniae, N. simplex, Oleoguttula mirabilis, Paradevriesia compacta, Perusta inaequalis, Petrophila incerta, Rachicladosporium alpinum, R. inconspicuum, R. mcmurdoi, R. monterosanum, R. paucitum, Ramimonilia apicalis, Saxophila tyrrhenica, Vermiconidia antarctica, V. calcicola, V. foris, and V. flagrans.

Qualitative analysis of the vaginal microbiota of healthy cattle and cattle with genital-tract disease.

The microbial community of the reproductive appara-tus, when known, can provide information about the health of the host. Metagenomics has been used to characterize and obtain genetic infor-mation about microbial communities in various environments and can relate certain diseases with changes in this community composition. In this study, samples of vaginal surface mucosal secretions were col-lected from five healthy cows and five cows that showed symptoms of reproductive disorders. Following high-throughput sequencing of the isolated microbial DNA, data were processed using the Mothur soft-ware to remove low-quality sequences and chimeras, and released to the Ribosomal Database Project for classification of operational taxo-nomic units (OTUs). Local BLASTn was performed and results were loaded into the MEGAN program for viewing profiles and taxonomic microbial attributes. The control profile comprised a total of 15 taxa, with Bacteroides, Enterobacteriaceae, and Victivallis comprising the highest representation of OTUs; the reproductive disorder-positive profile comprised 68 taxa, with Bacteroides, Enterobacteriaceae, His-tophilus, Victivallis, Alistipes, and Coriobacteriaceae being the taxa with the most OTU representation. A change was observed in both the community composition as well as in the microbial attributes of the profiles, suggesting that a relationship might exist between the patho-gen and representative taxa, reflecting the production of metabolites to disease progression.

Uso popular de espécies medicinais da família Verbenaceae no Brasil / Popular use of medicinal species of the Verbenaceae family in Brazil

RESUMO Com o presente trabalho objetivou-se realizar um levantamento bibliográfico acerca das espécies da família Verbenaceae utilizadas na medicina popular nas diversas regiões do Brasil. Os trabalhos acadêmicos utilizados para elaboração desta pesquisa foram extraídos de revistas indexadas junto ao Scielo publicadas no período de 2000 a 2012, sendo também realizada busca no Google acadêmico. Os dados foram compilados e organizados em ordem alfabética por espécies, seguidas do nome popular, forma de uso, parte usada, indicações e área de estudo. Foram encontrados um total de 85 publicações indicando 55 espécies distribuídas em oito gêneros. A espécie com maior número de citações foi Lippia alba (Mill.) N.E.Br., com 56 registros. Das plantas descritas na literatura, a folha foi o órgão mais citado como parte usada e o chá, o modo de preparo mais utilizado. Dentre as categorias de indicações medicinais, as plantas que mereceram destaque foram às relacionadas ao tratamento das doenças do sistema respiratório e digestório. Constatou-se que o Rio de Janeiro foi o estado que contou com o maior número de publicações na área. O uso de plantas medicinais está cada vez mais presente entre as diversas comunidades. Com isso, as informações contidas neste trabalho servirão para auxiliar na elaboração de banco de dados sobre o modo de uso das plantas pertencentes à família Verbenaceae com potencial medicinal encontradas nos Estados brasileiros.

ABSTRACT The current work aimed to conduct a literature review about the Verbenaceae family species used in popular medicine in different regions of Brazil. The academic papers employed for the preparation of this research were taken from journals indexed by the Scielo published from 2000 to 2012, and a research in Google scholar tool was also performed. The data were compiled and organized alphabetically by species, followed by the popular name, manner of use, employed part, indications and study area. A total of 85 publications indicating 55 species were found and distributed into eight genera. The species with the highest number of citations was the Lippia alba (Mill.) N.E.Br., with 56 records. From the plants described in the literature, the leaf was the most cited part of the vegetables, and the tea was the most widely used preparation method. Among the categories of medicinal recommendations, the highlighted plants were the ones related to the treatment of diseases of the respiratory and the digestive systems. It was observed that Rio de Janeiro was the state in which the largest number of publications in the area was concentrated. The use of medicinal plants is increasingly present among several communities. Thus, the information in this work will be applied to assist in database development on how to use the plants belonging to the family Verbenaceae with medicinal potential found in Brazilian states.

An in vivo comparison of radiographic and clinical examination with separation for assessment of approximal caries in primary teeth.

AIM: To compare in vivo, clinical examination (CE) and bitewing radiographs (BW) for the detection of cavitations and initial approximal lesions in deciduous molars with validation after temporary separation (TS) and direct visual examination (DE) of the approximal surfaces. MATERIALS AND METHODS: Thirty children aged 4 to 8 years without visual cavitations or restorations and with at least one approximal contact. Two calibrated examiners evaluated 355 surfaces of primary molars, which were then validated through TS and DE. RESULTS: Of the 280 surfaces considered sound by CE, 24.6% were cavitated after separation. Comparing BW and DE, it was observed that, when the radiolucency involved the inner half of dentin, 68.4% of lesions were cavitated. The sensitivity and specificity of CE and BW were 0.14 and 0.43, and 0.80 and 0.75, respectively. CONCLUSION: DE identified a greater number of pre-cavitated lesions and cavities. BW were useful for detection of decayed surfaces, but with low specificity for diagnosis of sound surfaces, while detection of sound or decayed approximal areas by means of CE alone was not possible. The combination of methods was effective in obtaining an accurate diagnosis of caries in the primary dentition.

Comparison of microleakage of different margin types around Class V resin restorations in primary teeth.

AIM: This in vitro study compared the effect of a concave with a straight-bevelled cavity margin on the microleakage of Class V composite resin restorations in primary teeth. MATERIALS AND METHODS: Standardised Class V cavity preparations were made in vitro on the buccal (all margins placed in enamel) and on the lingual (margins placed in enamel and cementum) surfaces of 20 sound primary molars. The teeth were randomly assigned to two groups of 10 each: in Group 1, a concave bevel was made with a high-speed No. 04 tungsten carbide bur and in Group 2, a straight bevel was made with a high-speed No. 556 tungsten carbide bur. The teeth were restored incrementally with Adper Single Bond 2 (3M) adhesive and Filtek Z 350 (3M) composite resin. All specimens were subsequently thermocycled and immersed in 50% silver nitrate solution. Microleakage of the restorations was then assessed by silver penetration. A grading scale of 0 to 4 was used as the scoring criterion. RESULTS: At the enamel margins no statistically significant differences were observed between the two groups (p>>0.05). Occlusal walls in enamel, in both groups, exhibited less leakage than the cervical walls in cementum (p<0.01) and Group 1 showed better results than Group 2 in decreasing microleakage at the cementum margins (p <0.05). CONCLUSION: Based on the results, it was concluded that concave-beveled cavity preparations may reduce but did not totally eliminate microleakage at the cementum margins of Class V composite resin restorations in primary teeth.

Phylogenetic analysis of substrate-binding subunits in an osmoprotectant system.

Substrate-binding subunits are important components of the solute importation system, known as the osmoprotectant system, which consists of a membrane protein belonging to the ABC superfamily. These molecules recognize specific substrates that have different physiological roles in prokaryotes, i.e., roles that contribute to the survival of these organisms in environments with high concentrations of salt. Using the MEGA software, this study performed a phylogenetic analysis of 431 nucleotide sequences of these subunits, orthologous to each other, collected from the http://www.genome.jp/kegg/ database. This analysis allowed phylogenetic trees to be generated, clearly demonstrating that there was horizontal transfer of some genes through sharing by different organisms. Furthermore, two probable ancestral sequences were generated that showed homology with permeases that transport choline, glycine betaine, and carnitine, which are trimethylamines currently present in various prokaryotes. Therefore, this system probably arose in prokaryotic organisms with the basic function of capturing nutrients, and by performing this basal function and being shared with other organisms, it was fixed in the genome. However, because of prokaryote habitat diversification, this system contributed decisively to the adaptation of these organisms to different environments, especially environments that had a high salt concentration, thus acting as an osmoprotection system, which is how they are currently categorized.

Retrospective study on the occurrence of primary incisor trauma in preschool children of a low-income area in Brazil.

AIM: The objective of this study was to investigate the occurrence of trauma in primary incisors in a sample of preschool Brazilian children living in a low-income area of Rio de Janeiro, Brazil. METHODS: For the study 527 dental records of a Municipal Children's Hospital were evaluated, and the following related factors were recorded: gender age, tooth affected, type and cause of trauma. Data were tabulated and subjected to statistical analysis (chi-square test). RESULTS: There were no significant differences in the prevalence of injuries among boys and girls. Children 10 to 24 months-old were the most affected (p < 0.05). The maxillary central incisor was the most affected tooth (p < 0.05), with predominance of lateral luxation and concussions (p > 0.05). The most common cause of trauma was associated with child's own-height fall (p < 0.05). CONCLUSION: Based on the results, it was concluded that the high incidence of incisor trauma in deciduous teeth should be viewed with concern, requiring the implementation of health policies aimed primarily at the prevention of such accidents.

Duplication of a permanent maxillary incisor root caused by trauma to the predecessor primary tooth: clinical case report.

AIM: This report describes a permanent maxillary incisor tooth with two roots possibly caused by trauma to the predecessor primary tooth. SUMMARY: Diagnosis, aetiology, clinical implications and endodontic and aesthetic treatment of an incisor tooth with two roots are presented. Diagnosis, early intervention by a multidisciplinary team and clinical and radiographic examination of patients who suffer trauma to primary teeth are of importance to minimize or avoid damage to successor teeth. KEY LEARNING POINTS: • A possible sequelae of trauma to a primary tooth is root duplication in the permanent tooth. • Root duplication is a rare anomaly that has its diagnosis based on the radiographic examination. • Careful diagnosis and follow-up are necessary to prevent future complications related to the successor tooth.

First Report of Soft Rot of Potatoes Caused by Dickeya dadantii in Zimbabwe.

A survey was carried out in the potato- (Solanum tuberosum L.) growing regions of Zimbabwe in April 2009 to assess the prevalence of bacterial soft rot. A total of 125 tubers with soft rot symptoms were collected. The disease caused severe economic losses ranging from 20 to 60% on tubers in the field and in storage. Affected tubers had symptoms that ranged from light vascular discoloration to complete seed piece decay. Infected tuber tissue was often cream colored and soft to the touch. In the field, plants showed severe wilting, often accompanied by a slimy, brown necrosis of the lower stems. Seventy-five of 125 isolations from diseased tubers yielded pectolytic bacteria on crystal violet pectate (CVP) medium and colonies were characterized after purification on King's B medium. All 75 isolates were gram-negative rods, oxidase negative, facultatively anaerobic, able to degrade pectate, and rot potato slices. They grew at 37°C, were sensitive to erythromycin, positive for phosphatase, indole production, cis-aconitate, lactose, d-arabinose, meso-tartrate, casein, d-melibiose, myo-inositol, and malonate utilization, while negative for acid production from trehalose, inuline, and α-methyl glucose. Dickeya dadantii (Erwinia chrysanthemi 3937 from the Scottish Research Institute) was included in all biochemical and pathogenicity tests. These characteristics are typical for two species, D. zeae and D. dadantii (2). Thus, the 75 isolates were further identified by PCR amplification with BOX and REP primers (3) and five isolates by gyrB sequence analysis (1). These analyses give support for the isolates being D. dadantii. Partial gyrB sequence analysis showed that the analyzed isolates had 96% sequence identity with the D. dadantii type strain Ech 586T (GenBank Accession No. CP001836.1). One-microliter suspensions (108 CFU per ml) of 20 samples were injected into the stolon end of potato tubers (S. tuberosum L.) cv. BP1. Each isolate was inoculated into three tubers, which were maintained at 25°C. Three control tubers were inoculated with sterile distilled water. Soft rot symptoms identical to those observed in the field and in storage appeared on all inoculated tubers 1 to 2 days after inoculation but not on the control tubers. A bacterium with identical characteristics to those described above was consistently reisolated from the rotted tissue of inoculated tubers. To our knowledge, this is the first report of soft rot on potato in Zimbabwe caused by D. dadantii, formerly referred to as E. chrysanthemi. This finding has implications for import and export of potato material into and out of Zimbabwe. Zimbabwe imports seed from various countries because of the current seed shortage and exports table potatoes to other African states. References: (1) C. Brady et al. Int. J. Syst. Evol. Micobiol. 59:2339, 2009. (2) R. Samson et al. Int. J. Syst. Evol. Microbiol. 55:1415, 2005. (3) J. Versalovic et al. 1991. Nucleic Acids Res. 19:6823, 1991.

Isolation of Enterobacter cowanii from Eucalyptus showing symptoms of bacterial blight and dieback in Uruguay.

AIMS: This study was performed to identify bacterial strains isolated simultaneously with Pantoea species from Eucalyptus trees showing symptoms of bacterial blight and dieback in Uruguay. METHODS AND RESULTS: Several molecular techniques including 16S rRNA and rpoB gene sequencing and DNA-DNA hybridization were used to characterize the gram-negative, facultatively anaerobic, slime-producing bacterial strains isolated along with Pantoea species from Eucalyptus. Hypersensitivity reactions (HR) and pathogenicity tests were performed on tobacco and Eucalyptus seedlings, respectively. The isolates clustered closely with the type strain of Enterobacter cowanii in both phylogenetic trees constructed. The DNA-DNA similarity between the isolates and the type strain of Ent. cowanii ranged from 88% to 92%. A positive HR was observed on the tobacco seedlings, but no disease symptoms were visible on the inoculated Eucalyptus seedlings. CONCLUSIONS: Enterobacter cowanii was isolated from trees with symptoms of bacterial blight although strains of this bacterial species do not appear to be the causal agent of the disease. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides the first report of Ent. cowanii isolated from Eucalyptus. Its presence in Eucalyptus tissue suggests that it is an endophyte in trees showing symptoms of blight.

Plant-pathogenic bacteria as biological weapons - real threats?

At present, much attention is being given to the potential of plant pathogens, including plant-pathogenic bacteria, as biological weapons/bioterror weapons. These two terms are sometimes used interchangeably and there is need for care in their application. It has been claimed that clandestine introduction of certain plant-pathogenic bacteria could cause such crop losses as to impact so significantly on a national economy and thus constitute a threat to national security. As a separate outcome, it is suggested that they could cause serious public alarm, perhaps constituting a source of terror. Legislation is now in place to regulate selected plant-pathogenic bacteria as potential weapons. However, we consider it highly doubtful that any plant-pathogenic bacterium has the requisite capabilities to justify such a classification. Even if they were so capable, the differentiation of pathogens into a special category with regulations that are even more restrictive than those currently applied in quarantine legislation of most jurisdictions offers no obvious benefit. Moreover, we believe that such regulations are disadvantageous insofar as they limit research on precisely those pathogens most in need of study. Whereas some human and animal pathogens may have potential as biological or bioterror weapons, we conclude that it is unlikely that any plant-pathogenic bacterium realistically falls into this category.

Serum uric acid is associated with microvascular function in hypertensive individuals.

We investigated the relationship of serum uric acid (UA) with resting forearm blood flow (FBF), reactive hyperaemia (RH) and flow-mediated dilation (FMD) of the brachial artery in hypertensive adults (n=506, mean age 62 years, 59% women). UA was measured by a colorimetric method. FBF, RH and FMD were measured by brachial artery ultrasound. Regression analyses were used to assess whether UA was associated with FBF, RH and FMD before and after adjustment for age, sex, systolic BP, diabetes, total and high-density lipoprotein cholesterol, smoking, body mass index (BMI), C-reactive protein (CRP), serum creatinine, alcohol intake, statin and diuretic use and brachial artery diameter (BAD). UA was significantly associated with FBF (P<0.0001) and RH (P=0.0001) but not with FMD (P=0.43). After adjustment for the covariates listed above, higher UA level remained independently associated with a higher FBF (P=0.012) and lower RH (P=0.004). The independent predictors were as follows: (a) higher FBF: lower age, higher BMI, history of smoking, statin use, higher CRP, higher BAD and higher UA levels; (b) lower RH: higher BMI, diabetes and higher UA levels; (c) lower FMD: greater age, male sex, higher BMI, history of smoking, statin use and higher BAD. We conclude that in hypertensive individuals, higher UA levels are associated with higher resting FBF and lower RH, markers of microvascular function, but not with brachial artery FMD.

Isolation and Identification of the Causal Agent of Brown Stalk Rot, A New Disease of Maize in South Africa.

During 2004 to 2005, an unreported disease of maize (Zea mays) was observed on commercial fields in the Northwest and Mpumalanga Provinces of South Africa. Infected plants were stunted, with a vertical crack at the first internode. Inside the stem, a dark-brown, narrow lesion was present along the crack. Internal browning inside the stem extended upward, reaching the top internode in some plants. Seed cobs were underdeveloped. Diseased plants were scattered in the fields and 10 to 70% of the crop was affected. Gram-negative, facultatively anaerobic bacteria were consistently isolated from diseased tissues. Pathogenicity tests established that representative strains induced disease symptoms similar to those observed on maize plants in the field. Physiological and biochemical characterization using the API 20E and API 50CHE systems and 16S rRNA gene sequence analyses showed that the strains belonged to the genus Pantoea. The results of these tests also separated the strains into two groups. The first group, giving a positive reaction in the indole test, was similar to Pantoea ananatis. The second group of strains was indole negative and resembled P. agglomerans. The fluorescent amplified fragment length polymorphism (F-AFLP) genomic fingerprints generated by the indole-positive strains and P. ananatis reference strains were similar and clustered together in the dendrogram, confirming that the indole-positive bacteria causing brown stalk rot on maize were P. ananatis. The F-AFLP fingerprints produced by the indole-negative strains were distinctly different from those generated by P. ananatis, P. agglomerans, P. dispersa, P. citrea, P. stewartii subsp. stewartii, and P. stewartii subsp. indologenes. The results indicated that indole-negative bacteria causing brown stalk rot on maize might belong to a previously undescribed species of the genus Pantoea. This is the first report of a new disease on maize, brown stalk rot, caused by two bacterial species, P. ananatis and an undescribed Pantoea sp.

DNA sequence and RFLP data reflect geographical spread and relationships of Amylostereum areolatum and its insect vectors.

The white rot fungus, Amylostereum areolatum (Basidiomycetes), is best known for its symbiotic relationship with various siricid wood wasp species. In this study, the relationship between isolates of A. areolatum associated with two wood wasp species, Sirex noctilio and S. juvencus, are considered to identify possible intraspecific groups. Isolates from the northern (native) and southern (exotic) hemispheres are included to determine patterns of geographical spread and origin of introductions into the southern hemisphere. The phylogenetic relationships of these isolates to authentic isolates of A. chailletii, A. laevigatum and A. ferreum were also investigated. Sequence and restriction fragment length polymorphism (RFLP) analyses of the variable nuc-IGS-rDNA region provided markers to distinguish intraspecific groups within A. areolatum. Isolates of A. areolatum associated with S. noctilio and S. juvencus contained four heterogenic sequences in the DNA region analysed. These sequences occurred in one of five combinations in each isolate. Some of these sequences were unique to isolates of A. areolatum from either wasp species, while others were present in both groups. This shows the ancient and specialized evolutionary relationship that exists between these insects and fungi. Isolates from the southern hemisphere all share the same sequence group. This supports previous hypotheses that S. noctilio has spread between countries and continents of this region. At the interspecific level, the IGS-rDNA sequence analysis showed that A. ferreum and A. laevigatum are closely related to each other, and they in turn are related to A. chailletii. Amylostereum areolatum was the most distinctly defined species in the genus. This can be attributed to the obligate relationship between A. areolatum and its insect vectors. Polymerase chain reaction-RFLP analysis was also shown to be an effective tool to distinguish between the different species of Amylostereum.

Bacterial Blight and Dieback of Eucalyptus Species, Hybrids, and Clones in South Africa.

During 1998, a new disease appeared on trees representing a Eucalyptus grandis × E. nitens (GN) hybrid in a nursery in KwaZulu/Natal. The disease has subsequently spread to other Eucalyptus species, hybrids, and clones in nurseries and plantations throughout South Africa. Typical symptoms of the disease include dieback of young shoots and leaf blight. This ultimately leads to stunting of trees. The objective of this study was to isolate and identify the causal agent of the disease. A bacterium was consistently isolated from infected tissue. Pathogenicity tests were undertaken with a range of bacterial strains. Four pathogenic strains were selected from different geographical regions and Eucalyptus hosts for further study. The bacterium causing Eucalyptus leaf and shoot blight is gram negative and rod-shaped, varying in size from 0.5 to 0.75 µm wide and 1.0 to 2.0 µm long. Colonies of this bacterium have a yellow pigment. The results from the Biolog tests identified the bacterium as Pantoea agglomerans with a similarity index of 0.315. The 16S rDNA sequences of the purported Pantoea sp. were compared with those of other related Enterobacteriaceae from GenBank/EMBL. Phylogenetic analysis using PAUP revealed that the isolates group together with P. agglomerans, P. ananatis, and P. stewartii subsp. stewartii. The fatty acid profiles and phenotypic characteristics of the new pathogen are similar to P. ananatis, and % G + C is within the range of this species. DNA:DNA hybridization between the four strains and the type strain of P. ananatis conclusively showed that the bacterium causing blight and dieback of Eucalyptus in South Africa belongs to this species. This is the first report in which P. ananatis has been found as a causal agent of a disease on Eucalyptus.

The root rot fungus Armillaria mellea introduced into South Africa by early Dutch settlers.

Dead and dying oak (Quercus) and numerous other woody ornamental trees and shrubs showing signs and symptoms of Armillaria root rot were identified in the Company Gardens, Cape Town, South Africa, which were established in the mid-1600s by the Dutch East Indies Trading Company. Nineteen isolates from dying trees or from mushrooms were collected and analysed to identify and characterize the Armillaria sp. responsible for the disease. The AluI digestion of the amplified product of the first intergenic spacer region (IGS-1) of the rRNA operon of 19 isolates from the Company Gardens was identical to that of some of the European isolates of A. mellea s. s. The IGS-1 region and the internal transcribed spacers (ITS) were sequenced for some of the Cape Town isolates. Phylogenetic analyses placed the Cape Town isolates in the European clade of A. mellea, which is distinct from the Asian and North American clades of this species. Identification based on sexual compatibility was conducted using A. mellea tester strains in diploid-haploid pairings, which showed some compatibility between the Cape Town isolates and testers from Europe. Somatic compatibility tests (diploid-diploid pairings) and DNA fingerprinting with multilocus, microsatellite probes indicated that the Cape Town isolates were genetically identical and may have resulted from vegetative (clonal) spread from a single focus in the centre of the original Company Gardens (c. 1652). The colonized area is at least 345 m in diameter. Assuming a linear spread rate underground of 0.3 m/year to 1.6 m/year, the genet (clone) was estimated to be between 108 and 575 years old. These data suggest that A. mellea was introduced into Cape Town from Europe, perhaps on potted plants, such as grapes or citrus, planted in the Company Gardens more than 300 years ago.

Gibberella fujikuroi mating population E is associated with maize and teosinte.

Summary Isolates of Fusarium subglutinans mating population E are usually found on maize. This fungus forms part of the so-called Gibberella fujikuroi species complex. Previously, F. subglutinans has been associated with two additional mating populations (B and H) and a variety of plant hosts. This was mainly due to a lack of diagnostic morphological characters, but the use of DNA sequence information showed that the strains making up mating populations B, E and H, as well as those associated with the different plant hosts, represent separate species. Recently, another putative mating population has been reported on the wild teosinte relatives of maize. Based on sexual compatibility studies, these isolates were apparently closely related to the pitch canker fungus, F. subglutinans f. sp. pini (= F. circinatum;G. fujikuroi mating population H). The aim of the current study was to determine whether the population of F. subglutinans from teosinte constitutes a new or an existing lineage within the G. fujikuroi complex. For this purpose, portions of the mitochondrial small subunit ribosomal DNA, calmodulin and beta-tubulin genes from the fungi were sequenced. Phylogenetic analyses and comparison with sequences from public domain databases indicated that the F. subglutinans isolates from teosinte are most closely related to strains of G. fujikuroi mating population E. These results were confirmed using sexual compatibility studies. The putative mating population from the wild relatives of maize therefore forms part of the existing E-mating population and does not constitute a new lineage in the G. fujikuroi species complex.

PCR-based identification of MAT-1 and MAT-2 in the Gibberella fujikuroi species complex.

All sexually fertile strains in the Gibberella fujikuroi species complex are heterothallic, with individual mating types conferred by the broadly conserved ascomycete idiomorphs MAT-1 and MAT-2. We sequenced both alleles from all eight mating populations, developed a multiplex PCR technique to distinguish these idiomorphs, and tested it with representative strains from all eight biological species and 22 additional species or phylogenetic lineages from this species complex. In most cases, either an approximately 800-bp fragment from MAT-2 or an approximately 200-bp fragment from MAT-1 is amplified. The amplified fragments cosegregate with mating type, as defined by sexual cross-fertility, in a cross of Fusarium moniliforme (Fusarium verticillioides). Neither of the primer pairs amplify fragments from Fusarium species such as Fusarium graminearum, Fusarium pseudograminearum, and Fusarium culmorum, which have, or are expected to have, Gibberella sexual stages but are thought to be relatively distant from the species in the G. fujikuroi species complex. Our results suggest that MAT allele sequences are useful indicators of phylogenetic relatedness in these and other Fusarium species.