Occurrence of 15 Haplotypes of Linepithema micans (Hymenoptera: Formicidae) in Southern Brazil.

The ant genus Linepithema is widely known, thanks to the pest species Linepithema humile (Mayr), which is easily mistaken for Linepithema micans (Forel) due to their morphological similarity. Like L. humile, L. micans is associated to the main grapevine pest in Brazil, Eurhizococcus brasiliensis (Wille), also known as ground pearl. Therefore, the present study uses mtDNA fragments to expand the knowledge of haplotype diversity and distribution of L. micans in the state of Rio Grande do Sul (Brazil), to understand the genetic differences of the populations identified in this study. We identified 15 haplotypes of L. micans spread across different localities. Twelve of these haplotypes were new for the species. The high haplotype diversity uncovered in Rio Grande do Sul (Brazil) for this species was predictable, as L. micans is in its native environment. Additional studies that take gene flow into account may reveal interesting aspects of diversity in these populations.

Comparison Between Ground Ant (Hymenoptera: Formicidae) Communities Foraging in the Straw Mulch of Sugarcane Crops and in the Leaf Litter of Neighboring Forests.

In many sugarcane plantations in Brazil, the straw is left on the soil after harvesting, and vinasse, a by-product of the production of sugar and ethanol, is used for fertigation. Our goal was to compare ant community composition and species richness in the straw mulch of sugarcane crops with the leaf litter of neighboring forests. We tested the hypothesis that ant communities in the straw mulch of vinasse-irrigated sugarcane crops and in the forest leaf litter were similar, because the combination of straw mulching and vinasse irrigation has a positive effect on soil fauna. Straw mulch and leaf litter were collected from 21 sites and placed in Berlese funnels. In total, 61 species were found in the forest leaf litter, whereas 34 and 28 species were found in the straw mulch of sugarcane fields with and without vinasse, respectively. Ant communities differed between forest and crop fields, but the species in the sugarcane straw mulch were a subset of the species found in the forest leaf litter. Although vinasse is rich in organic matter, it did not increase ant diversity. Seven feeding and/or foraging types were identified and, among the different types, surface-foraging omnivorous ants were the most prevalent in all habitats. Vinasse-irrigated sugarcane straw mulch had more predatory species than mulch from vinasse-free fields, but fewer than forest leaf litter. However, this positive effect of vinasse irrigation should be carefully evaluated because vinasse has negative effects on the environment.

Vinasse and Its Influence on Ant (Hymenoptera: Formicidae) Communities in Sugarcane Crops.

Sugarcane is an important crop within the Brazilian socioeconomic landscape. There is a constant need for approaches to increase sustainability at all steps of the production chain. Irrigating sugarcane crops with vinasse is one of these approaches, because vinasse is a residue of sugarcane processing that can be used to fertilize these same crops. However, due to its chemical properties, vinasse may be harmful to soil fauna. Analyzing the structure and functional organization of ant communities is a fast and practical way to monitor sites affected by the addition of chemicals. This study compared the structure of soil ant communities in vinasse-irrigated sugarcane crops to those in secondary forests adjacent to the crops. In total, 32 genera and 107 species of ants were observed; of these, 30 species foraged in crop fields and 102 foraged in forests. Twenty-five percent of the species were present in both crops and forests. Ant communities in crop soil had poorer taxonomic composition and lower richness in each functional group compared to communities in forest remnants. However, regardless of vegetation type, epigeic ants were more diverse, and Dorymyrmex brunneus (crop) and Pachycondyla striata (forest) were very frequent. Vinasse did not increase the diversity of epigeic and hypogeic ants, but it may affect the community composition.

Molecular Profile of the Brazilian Weaver Ant Camponotus textor Forel (Hymenoptera, Formicidae).

Camponotus textor Forel is, to date, the only weaver ant recorded from Brazil, and all existing studies on the species are restricted to describing its weaving and nesting behaviors. The aim of this work is to establish the molecular profile of the species. We sampled eight different colonies by sequencing mitochondrial genes (COI, transfer DNA (tRNA), and an intergenic spacer) and the nuclear gene 28S ribosomal DNA (rDNA). We then assessed haplotype diversity and also analyzed distribution patterns of this species based on the correlation between genetic and geographic distances. Our results provide an additional tool for species identification by identifying new regions that can be used as molecular markers for barcoding (such as the intergenic spacer (IGS) and tRNA-Leu). In addition, the phylogenetic analysis revealed that C. textor has features that could be associated with deep population divergences. We identified a wide range of mitotypes and three distinct groups, suggesting a possible reduction of gene flow between colonies.

Seasonal Life Cycle of Linepithema micans (Forel) (Hymenoptera: Formicidae) in Vineyards.

Linepithema micans (Forel) is considered to be the main ant species responsible for the spread of Eurhizococcus brasiliensis (Wille), a soil scale that damages vine plants in southern Brazil. In this work, the seasonal life cycle of L. micans was evaluated in vineyards to define the most appropriate time for the application of toxic baits for control. Ant nests were transported to the laboratory in plastic bags, and individual ants were separated into groups of eggs, larvae, pupae, workers, males, and queens, recording the number of individuals per colony. The seasonal life cycle of L. micans in the vineyards began in spring with the deposition of eggs by queens. During the warm months of the year (spring and summer), it was possible to observe nests consisting primarily of eggs and pupae. During the autumn and winter months, the colonies were composed predominantly of larvae and a reduction in eggs was observed. Because the main control strategy targets the larval stage of the insect, the application of toxic baits for the control of L. micans must be performed during the winter.

Occurrence of three haplotypes of Linepithema micans (Forel) (Hymenoptera: Formicidae) in Southern Brazil.

Linepithema micans (Forel) (Hymenoptera: Formicidae) is reported to occur from eastern Brazil to central Argentina in pasture or grassland, forest and second growth riparian forest, nesting under stones, rotting wood, and sandy soil. However, information on this species is poor and its ecological interactions and role as pests are unknown. Linepithema humile (Mayr), a closely related species to L. micans, known as the Argentine ant, is native to South America, and was accidentally introduced to several regions of the world. Recent studies have shown that other related species, such as L. micans, could become as pestiferous as L. humile because of its phylogenetic proximity. Samples of L. micans from different habitats in Southern Brazil were characterized by sequencing of the mitochondrial DNA. Sequences were compared to previously obtained sequences from samples of L. humile and the genetic distance and differences in the tRNALeu structure were investigated. Our data identified three haplotypes of L. micans, two of which were observed in ant populations closely associated with the Brazilian ground pearl Eurhizococcus brasiliensis (Hempel) (Hemiptera: Margarodidae), a soil scale that is a serious pest of vineyards. The third haplotype was identified in ants from populations invading residences in urban habitats.

Coptotermes gestroi (Isoptera: Rhinotermitidae) in Brazil: possible origins inferred by mitochondrial cytochrome oxidase II gene sequences.

The Asian subterranean termite, Coptotermes gestroi, originally from northeast India through Burma, Thailand, Malaysia, and the Indonesian archipelago, is a major termite pest introduced in several countries around the world, including Brazil. We sequenced the mitochondrial COII gene from individuals representing 23 populations. Phylogenetic analysis of COII gene sequences from this and other studies resulted in two main groups: (1) populations of Cleveland (USA) and four populations of Malaysia and (2) populations of Brazil, four populations of Malaysia, and one population from each of Thailand, Puerto Rico, and Key West (USA). Three new localities are reported here, considerably enlarging the distribution of C. gestroi in Brazil: Campo Grande (state of Mato Grosso do Sul), Itajaí (state of Santa Catarina), and Porto Alegre (state of Rio Grande do Sul).

Relationship among Apis mellifera L. stings, swarming and climate conditions in the city of Rio Claro, SP, Brazil

The presence of bees (Apis mellifera L.) in urban areas has increased in recent years due to environmental disturbances caused by humans. Bee migration to cities may provoke serious accidents, since some people present allergic reactions to their venoms. In Rio Claro city, São Paulo state, Brazil, the number of calls to the fire brigade for removal of bee swarms, and the number admissions in local hospitals due to bee stings were investigated during 2002 and 2003, and a correlation between these data and the average temperature, rainfall and relative humidity was found. The study period was divided into three phases according to the number of times that the fire brigade was called to remove swarms (263 times): January to July 2002 - 51 calls (19.39 percent); August 2002 to July 2003 - 140 calls (53.23 percent); and August to December 2003 - 72 calls (27.38 percent). A significant correlation among the number of calls, the local temperature and rainfall was detected. The number of accidents was not associated with environmental variables. Based on the current results, public activities for prevention of bee attacks may be developed to avoid unwanted contact between humans and these insects, and/or provide the appropriate management of the colonies.

Ant communities (Hymenoptera: Formicidae) in an urban ecosystem near the Atlantic Rainforest

The relationships between an urban ecosystem located near the Atlantic Rainforest in southeastern Brazil and ant communities were studied with the objective of quantifying the ant richness and abundance in the household environment and its surroundings. Eighty residences were sampled, where 58 species and 28 genera pertaining to 7 sub-families were found to be present. Inside the residences, the species richness was found to be lower (26), although the abundance was greater (10,670), with the wash area and kitchen being the locales that contributed with the greatest number of hits. The opposite was true in the areas outside the residences, where 54 species and 3,747 ants were observed. Inside houses, the species known as Tramp ants were found, in the following order of importance: Solenopsis -saevissima, Tapinoma melanocephalum, Linepithema humile, Paratrechina fulva, Wasmannia -auropunctata, P. -longicornis, Pheidole megacephala, Monomorium pharaonis and M. floricola. Externally, mainly in the yards and gardens, species such as Octostruma rugifera, Heteroponera dolo, Hypoponera sp.1 and sp.6, Gnamptogenys sp. 4, G. striatula, Odontomachus meinerti, Pachycondyla constricta and P. striata were found. In general, a greater number of species and lower abundance of individuals were observed in the neighborhoods nearer the mountains than in those closer to the urban center.

A relação entre um ecossistema urbano localizado próximo à Mata Atlântica na região sudeste do Brasil e as comunidades de formigas foi estudada com o objetivo de quantificar a riqueza e a abundância de formigas no interior e no entorno das residências. Oitenta casas foram amostradas, tendo sido encontradas 58 espécies e 28 gêneros pertencentes a 7 subfamílias. No interior das residências foi encontrada uma baixa riqueza de espécies (26) e um alto número de indivíduos (10.670), sendo a área de serviço e a cozinha os locais que mais contribuíram para esse resultado. Já no entorno das residências foram encontradas 54 espécies e 3.747 indivíduos. No interior das casas, as espécies conhecidas como 'Tramp - ants" foram encontradas na seguinte ordem de importância: Solenopsis -saevissima, Tapinoma -melanocephalum, Linepithema humile, Paratrechina fulva, Wasmannia auropunctata, P. longicornis, Pheidole -megacephala, Monomorium pharaonis e M. floricola. Externamente, principalmente no jardim e quintal, foram encontradas espécies como: Octostruma rugifera, Heteroponera dolo, Hypoponera sp.1 e sp.6, Gnamptogenys sp.4, G. -striatula, Odontomachus meinerti, Pachycondyla constricta e P. striata. Em geral, o maior número de espécies e baixa abundância de indivíduos foram observados em bairros próximos às áreas de mata.

Ant communities (Hymenoptera: Formicidae) in an urban ecosystem near the Atlantic Rainforest.

The relationships between an urban ecosystem located near the Atlantic Rainforest in southeastern Brazil and ant communities were studied with the objective of quantifying the ant richness and abundance in the household environment and its surroundings. Eighty residences were sampled, where 58 species and 28 genera pertaining to 7 sub-families were found to be present. Inside the residences, the species richness was found to be lower (26), although the abundance was greater (10,670), with the wash area and kitchen being the locales that contributed with the greatest number of hits. The opposite was true in the areas outside the residences, where 54 species and 3,747 ants were observed. Inside houses, the species known as Tramp ants were found, in the following order of importance: Solenopsis saevissima, Tapinoma melanocephalum, Linepithema humile, Paratrechina fulva, Wasmannia auropunctata, P. longicornis, Pheidole megacephala, Monomorium pharaonis and M. floricola. Externally, mainly in the yards and gardens, species such as Octostruma rugifera, Heteroponera dolo, Hypoponera sp.1 and sp.6, Gnamptogenys sp. 4, G. striatula, Odontomachus meinerti, Pachycondyla constricta and P. striata were found. In general, a greater number of species and lower abundance of individuals were observed in the neighborhoods nearer the mountains than in those closer to the urban center.

Susceptibility of the ant-cultivated fungus Leucoagaricus gongylophorus (Agaricales: Basidiomycota) towards microfungi.

The aim of this study was to select virulent strains of microfungi against Leucoagaricus gongylophorus, a symbiotic fungus cultivated by leaf-cutting ants. The results from in vitro assays showed that microfungal strains had a variable and significant impact on the colony development of L. gongylophorus. Specifically, Trichoderma harzianum, Escovopsis weberi CBS 810.71 and E. weberi A088 were more effective, inhibiting the L. gongylophorus colonies by 75, 68 and 67%, respectively (P < 0.05) after 15 days. Strain E. weberi A086 and Acremonium kiliense were less effective: 43 and 26%, respectively (P < 0.05). In spite of the current negative perspective of a microbiological control approach for these ants, the present work discusses the possibility of using mycopathogenic fungi for the control of these insects, and points out the importance of encouraging more studies in this area.

Starch metabolism in Leucoagaricus gongylophorus, the symbiotic fungus of leaf-cutting ants.

Leucoagaricus gongylophorus, the symbiotic fungus of the leaf-cutting ants, degrades starch, this degradation being supposed to occur in the plant material which leafcutters forage to the nests, generating most of the glucose which the ants utilize for food. In the present investigation, we show that laboratory cultures of L. gongylophorus produce extracellular alpha-amylase and maltase which degrade starch to glucose, reinforcing that the ants can obtain glucose from starch through the symbiotic fungus. Glucose was found to repress alpha-amylase and, more severely, maltase activity, thus repressing starch degradation by L. gongylophorus, so that we hypothesize that: (1) glucose down-regulation of starch degradation also occurs in the Atta sexdens fungus garden; (2) glucose consumption from the fungus garden by A. sexdens stimulates degradation of starch from plant material by L. gongylophorus, which may represent a mechanism by which leafcutters can control enzyme production by the symbiotic fungus. Since glucose is found in the fungus garden inside the nests, down-regulation of starch degradation by glucose is supposed to occur in the nest and play a part in the control of fungal enzyme production by leafcutters.

Variability of non-mutualistic filamentous fungi associated with Atta sexdens rubropilosa nests.

A survey of the filamentous fungi other than the symbiotic one found in association with Atta sexdens rubropilosa colonies was carried out. Different fungal species (27 taxa) were isolated a few days after treating the workers with toxic baits (sulfluramid; Mirex-S), from 40 laboratory and 20 field nests. Syncephalastrum racemosum (54%) and Escovopsis weberi (21%), Trichoderma harzianum (38%) and Fusarium oxysporum (23%) were the prevalent species in laboratory and field nests, respectively. Acremonium kiliense, Acremonium strictum, E. weberi, F. oxysporum, Fusarium solani, Moniliella suaveolens and T. harzianum were found in both nests' groups. We revealed that many filamentous fungi can co-exist in a dormant state inside the nests of these insects and some of them appear to be tightly associated with this environment.

Low variation in ribosomal DNA and internal transcribed spacers of the symbiotic fungi of leaf-cutting ants (Attini: Formicidae)

Leaf-cutting ants of the genera Atta and Acromyrmex (tribe Attini) are symbiotic with basidiomycete fungi of the genus Leucoagaricus (tribe Leucocoprineae), which they cultivate on vegetable matter inside their nests. We determined the variation of the 28S, 18S, and 5.8S ribosomal DNA (rDNA) gene loci and the rapidly evolving internal transcribed spacers 1 and 2 (ITS1 and ITS2) of 15 sympatric and allopatric fungi associated with colonies of 11 species of leafcutter ants living up to 2,600 km apart in Brazil. We found that the fungal rDNA and ITS sequences from different species of ants were identical (or nearly identical) to each other, whereas 10 GenBank Leucoagaricus species showed higher ITS variation. Our findings suggest that Atta and Acromyrmex leafcutters living in geographic sites that are very distant from each other cultivate a single fungal species made up of closely related lineages of Leucoagaricus gongylophorus. We discuss the strikingly high similarity in the ITS1 and ITS2 regions of the Atta and Acromyrmex symbiotic L. gongylophorus studied by us, in contrast to the lower similarity displayed by their non-symbiotic counterparts. We suggest that the similarity of our L. gongylophorus isolates is an indication of the recent association of the fungus with these ants, and propose that both the intense lateral transmission of fungal material within leafcutter nests and the selection of more adapted fungal strains are involved in the homogenization of the symbiotic fungal stock.

Low variation in ribosomal DNA and internal transcribed spacers of the symbiotic fungi of leaf-cutting ants (Attini: Formicidae).

Leaf-cutting ants of the genera Atta and Acromyrmex (tribe Attini) are symbiotic with basidiomycete fungi of the genus Leucoagaricus (tribe Leucocoprineae), which they cultivate on vegetable matter inside their nests. We determined the variation of the 28S, 18S, and 5.8S ribosomal DNA (rDNA) gene loci and the rapidly evolving internal transcribed spacers 1 and 2 (ITS1 and ITS2) of 15 sympatric and allopatric fungi associated with colonies of 11 species of leafcutter ants living up to 2,600 km apart in Brazil. We found that the fungal rDNA and ITS sequences from different species of ants were identical (or nearly identical) to each other, whereas 10 GenBank Leucoagaricus species showed higher ITS variation. Our findings suggest that Atta and Acromyrmex leafcutters living in geographic sites that are very distant from each other cultivate a single fungal species made up of closely related lineages of Leucoagaricus gongylophorus. We discuss the strikingly high similarity in the ITS1 and ITS2 regions of the Atta and Acromyrmex symbiotic L. gongylophorus studied by us, in contrast to the lower similarity displayed by their non-symbiotic counterparts. We suggest that the similarity of our L. gongylophorus isolates is an indication of the recent association of the fungus with these ants, and propose that both the intense lateral transmission of fungal material within leafcutter nests and the selection of more adapted fungal strains are involved in the homogenization of the symbiotic fungal stock.

Occurrence of killer yeasts in leaf-cutting ant nests.

Killer activity was screened in 99 yeast strains isolated from the nests of the leaf-cutting ant Atta sexdens against 6 standard sensitive strains, as well as against each other. Among this yeast community killer activity was widespread since 77 strains (78%) were able to kill or inhibit the growth of at least one standard strain or nest strain. Toxin production was observed in representatives of all the studied genera including Aureobasidium, Rhodotorula, Tremella and Trichosporon, whose killer activity has not yet been described.

Toxicity of synthetic piperonyl compounds to leaf-cutting ants and their symbiotic fungus.

The development of Leucoagaricus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens was inhibited in vitro by synthetic compounds containing the piperonyl group. In addition, worker ants that were fed daily on an artificial diet to which these compounds were added had a higher mortality rate than the controls. The inhibition of the fungal growth increased with the size of the carbon side chain ranging from C1 through C8 and decreasing thereafter. 1-(3,4-Methylenedioxybenzyloxy)octane (compound 5) was the most active compound and inhibited the fungal development by 80% at a concentration of 15 micrograms ml-1. With worker ants the toxic effects started with compound 5 and increased with the number of carbons in the side chain. Thus, for the same concentration (100 micrograms ml-1) the mortality rates observed after 8 days of diet ingestion were 82%, 66% and 42%, for 1-(3,4-methylenedioxybenzyloxy)decane, 1-(3,4-methylenedioxybenzyloxy)dodecane and compound 5, respectively, whereas with commercial piperonyl butoxide the mortality was 68%. The latter compound, which is known as a synergist insecticide, was as inhibitory to the symbiotic fungus as the synthetic compound 5. The possibility of controlling these insects in the future using compounds that can target simultaneously both organisms is discussed.

Anthrone and oxanthrone C,O-diglycosides from Picramnia teapensis.

Two C,O-diglycosylated compounds, the anthrone picramnioside F, and the oxanthrone mayoside C, were isolated from the stem bark of Picramnia teapensis, along with the previously reported anthraquinones, 1-O-beta-D- and 8-O-beta-D-glucopyranosyl emodin. The compounds were separated by recycling-HPLC, and their structures were determined on the basis of spectroscopic analysis. CD measurements were used to establish the absolute configuration of the anthrone and oxanthrone. The antifungal activity of 1-O-beta-D- and 8-O-beta-D-glucopyranosyl emodin against Leucoagaricus gongilophorus was shown to be similar to that of the lignan sesamin.

Yeasts associated with nests of the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908.

A total of 137 yeasts associated with the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 were characterized, being selected 93 for analysis. Twenty four species belonging to seven genera (Candida, Cryptococcus, Rhodotorula, Sporobolomyces, Tremella, Trichosporon, Pichia) were isolated from the different analysed material. The genus Candida was widely distributed, with C. homilentoma, C. colliculosa-like, C. famata and C. colliculosa being the most prevalent. A few isolates did not fit the standard descriptions and probably some of them could be new biotypes or even new species. Three strains of black yeasts were also isolated, and four other were identified as being Candida spp. The effective number of yeast species was higher in newer sponge. The origin, distribution and relative importance of these microorganisms for the ants are discussed.

Toxicity of lignans to symbiotic fungus of leaf-cutting ants.

Lignans fromVirola sebifera Aubl.,Virola sp., andOtoba parvifolia (Mkfg.) A. Gentry (Myristicaceae) inhibited the in vitro growth of the fungus cultivated by leaf-cutting ants of the speciesAtta sexdens rubropilosa Forel (Hymenoptera: Formicidae). A comparison of activity among the lignans was obtained.