Plants harbor diverse communities of fungi and other microorganisms. Fungi are known to occur both on plant surfaces (epiphytes) and inside plant tissues (endophytes), but the two communities have rarely been compared. We compared epiphytic and endophytic fungal communities associated with leaves of coffee (Coffea arabica) in Puerto Rico. We asked whether the dominant fungi are the same in both communities, whether endophyte and epiphyte communities are equally diverse, and whether epiphytes and endophytes exhibit similar patterns of spatial heterogeneity among sites. Leaves of naturalized coffee plants were collected from six sites in Puerto Rico. Epiphytic and endophytic fungi were isolated by placing leaf pieces on potato dextrose agar without and with surface sterilization, respectively. A total of 821 colonies were isolated and grouped into 131 morphospecies. The taxonomic affinities of the four most common nonsporulating fungi were determined by sequencing the nuclear ribosomal internal transcribed spacer (ITS) region: two grouped with Xylaria and one each with Botryosphaeria and Guignardia. Of the most common genera, Pestalotia and Botryosphaeria were significantly more common as epiphytes; Colletotrichum, Xylaria, and Guignardia were significantly more common as endophytes. Suprisingly, more morphospecies occurred as endophytes than as epiphytes. Differences among sites in number of fungi per plant were significant. Thus epiphytic and endophytic communities differed greatly on a single leaf, despite living only millimeters apart, and both communities differed from site to site. Significant correlations between occurrence of fungal morphospecies suggested that fungi may have positive or negative effects on their neighbors. This is the first quantitative comparison of epiphytic and endophytic fungal floras in any plant, and the first to examine endophytic fungi or epiphytic fungi in leaves of coffee, one of the world's most valuable crops.

It is known that soil is a recipient of solid wastes able to contain enteric pathogens in high concentrations. Although the role of soil as a reservoir of certain bacterial pathogens is not in question, recent findings show that soil may have a larger role in the transmission of enteric diseases than previously thought. Many of the diseases caused by agents from soil have been well characterized, although enteric diseases and their link to soil have not been so well studied. Gastrointestinal infections are the most common diseases caused by enteric bacteria. Some examples are salmonellosis (Salmonella sp.), cholera (Vibrio cholerae), dysentery (Shigella sp.) and other infections caused by Campylobacter jejuni, Yersinia sp. and Escherichia coli O157:H7 and many other strains. Viruses are the most hazardous and have some of the lowest infectious doses of any of the enteric pathogens. Hepatitis A, hepatitis E, enteric adenoviruses, poliovirus types 1 and 2, multiple strains of echoviruses and coxsackievirus are enteric viruses associated with human wastewater. Among the most commonly detected protozoa in sewage are Entamoeba histolytica, Giardia intestinalis and Cryptosporidium parvum. This article reviews the existing literature of more than two decades on waste disposal practices that favor the entry of enteric pathogens to soil and the possible consequent role of the soil as a vector and reservoir of enteric pathogens.

Grassland-based production systems use ∼26% of land surface on earth. However, there are no evaluations of these systems as a source of antibiotic pollution. This study was conducted to evaluate the presence, diversity, and distribution of tetracycline resistance genes in the grasslands of the Colombian Andes, where administration of antibiotics to animals is limited to treat disease and growth promoters are not included in animals’ diet. Animal (ruminal fluid and feces) and environmental (soil and water) samples were collected from different dairy cattle farms and evaluated by PCR for the genes tet(M), tet(O), tetB(P), tet(Q), tet(W), tet(S), tet(T), otr(A), which encode ribosomal protection proteins (RPPs), and the genes tet(A), tet(B), tet(D), tet(H), tet(J), and tet(Z), encoding efflux pumps. A wide distribution and high frequency for genes tet(W) and tet(Q) were found in both sample types. Genes tet(O) and tetB(P), detected in high frequencies in feces, were detected in low frequencies or not detected at all in the environment. Other genes encoding RPPs, such as tet(M), tet(S), and tet(T), were detected at very low frequencies and restricted distributions. Genes encoding efflux pumps were not common in this region, and only two of them, tet(B) and tet(Z), were detected. DGGE–PCR followed by comparative sequence analysis of tet(W) and tet(Q) showed that the sequences detected in animals did not differ from those coming from soil and water. Finally, the farms sampled in this study showed more than 50% similarity in relation to the tet genes detected. In conclusion, there was a remarkable presence of tet genes in these production systems and, although not all genes detected in animal reservoirs were detected in the environment, there is a predominant distribution of tet(W) and tet(Q) in both animal and environmental reservoirs. Sequence similarity analysis suggests the transmission of these genes from animals to the environment.

(Accepted 31st July 1999)In the ‘elfin’ cloud forest of Serrania de Macuira, exchangeable ammonium and nitrate, and the rates of soil nitrogen mineralization and nitrification were measured in soil samples under different water, temperature and mineral nutrient additions. The effects of nitrogen, phosphorus and nitrogen plus phosphorus fertilization on radial trunk growth were measured in three tree species. In the cloud forest soils, concentrations of ammonium were much higher than those of nitrate. Nitrate was higher in samples collected during the afternoon than during the morning, probably as a result of leaching during the night or nitrification during the day. When samples were incubated under different water and temperature treatments, rates of nitrogen mineralization and nitrification increased more with changes in soil water content than with changes in temperature. Nitrification was significantly increased in soils amended with ammonium or with ammonium plus phosphorus, suggesting that nitrification is substrate-limited. Fertilization with nitrogen and phosphorus resulted in significantly increased girth increments in Guapira fragrans (Dum. -Cours.) Little and Rapanea guianensis Aublet. Myrcianthes fragrans (Sw.) D.C. did not respond to the fertilization. The results of this study support the hypothesis that the characteristics of montane rain forest in small and large tropical mountains (the ‘Massenerhebung’ effect) are greatly controlled by soil water conditions and related soil nitrogen availability.

Isotopic analysis and molecular-based bioassay methods were used in conjunction with geochemical data to assess intrinsic reductive dechlorination processes for a chlorinated-solvent contaminated site in Tucson, Arizona. Groundwater samples were obtained from monitoring wells within a contaminant plume comprising tetrachloroethene and its metabolites trichloroethene, cis-1,2-dichloroethene, vinyl chloride, and ethene, as well as compounds associated with free-phase diesel present at the site. Compound specific isotope (CSI) analysis was performed to characterize biotransformation processes influencing the transport and fate of the chlorinated contaminants. PCR analysis was used to assess the presence of indigenous reductive dechlorinators. The target regions employed were the 16s rRNA gene sequences of Dehalococcoides sp. and Desulfuromonas sp., and DNA sequences of genes pceA, tceA, bvcA, and vcrA, which encode reductive dehalogenases. The results of the analyses indicate that relevant microbial populations are present and that reductive dechlorination is presently occurring at the site. The results further show that potential degrader populations as well as biotransformation activity is non-uniformly distributed within the site. The results of laboratory microcosm studies conducted using groundwater collected from the field site confirmed the reductive dechlorination of tetrachloroethene to dichloroethene. This study illustrates the use of an integrated, multiple-method approach for assessing natural attenuation at a complex chlorinated-solvent contaminated site.