From species descriptions to diversity patterns: the validation of taxonomic data as a keystone for ant diversity studies reproducibility and accuracy.

Research findings in natural sciences need to be comparable and reproducible to effectively improve our understanding of ecological and behavioural patterns. In this sense, knowledge frontiers in biodiversity studies are directly tied to taxonomic research, especially in species-rich tropical regions. Here we analysed the taxonomic information available in 470 studies on Brazilian ant diversity published in the last 50 years. We aimed to quantify the proportion of studies that provide enough data to validate taxonomic identification, explore the frequency of studies that properly acknowledge their taxonomic background, and investigate the primary resources for ant identification in Brazil. We found that most studies on Brazilian ant diversity (73.6%) explicitly stated the methods used to identify their specimens. However, the proportion of papers that provide complete data for the repository institutions and vouchered specimens is vanishingly small (5.8%). Additionally, only 40.0% of the studies consistently presented taxon authorities and years of description, rarely referencing taxonomic publications correctly. In turn, the number of specialists and institutions consulted for ant identification in Brazil has increased in the last years, along with the number of studies that explicitly provide their taxonomic procedures for ant identification. Our findings highlight a shift between generations regarding the recognition of taxonomy as fundamental science, deepening our understanding of biodiversity.

A compilation of host plants and their gall-inducing insects for the Caatinga Biome / Uma compilação de plantas hospedeiras e seus insetos galhadores para o bioma Caatinga

Caatinga is a seasonally dry tropical forest, one of the richest in plant species. Unfortunately, many groups of herbivorous insects associated with these plants are poorly known. This study aimed to investigate the diversity of gall-inducing insects (GII) and host plants (HP) in the Caatinga. For this, we compiled the information available in the literature of inventories on GII and their HP communities, and the described gall midge species. We found 100 species, 72 genera, and 32 families of HP hosting a total of 156 morphospecies of GII and 12 species of described cecidomyiids. Plant species with only one GII species represented 74% of hosts, but in super HP (i.e., HP with a high number of GII), despite the small number of HP species, there were many GII species. Fabaceae was also the most specious family, with 30% of HP species and 40% of GII. Furthermore, our results showed a low number of species of HP and GII for the Brazilian Caatinga, that we discussed this pattern with the following arguments, first, it is likely that the number of galling insect inventories for the Caatinga is under-sampled, second the Caatinga has a relatively smaller number of plant species when compared to other biomes, and finally, we argue that the Caatinga is a seasonally dry tropical forest where the deciduousness represents a relevant factor in the colonization and performance rates of GII.

A Caatinga é uma floresta tropical sazonalmente seca, uma das mais ricas em espécies vegetais. Infelizmente, muitos grupos de insetos herbívoros associados a essas plantas são pouco conhecidos. Este estudo teve como objetivo investigar a diversidade de insetos galhadores (IG) e plantas hospedeiras (PH) na Caatinga. Para isso, nós compilamos as informações disponíveis na literatura de inventários sobre a comunidade de IG e suas PH, e as espécies descritas de cecidomiídeos. Nós encontramos 100 espécies, 72 gêneros e 32 famílias de PH abrigando um total de 156 morfoespécies de IG e 12 espécies descritas de cecidomiídeos. Espécies de plantas com apenas uma espécie de IG representaram 74% das hospedeiras, mas para as super PH (ou seja, PH com alto número de IG), apesar do pequeno número de espécies de PH, havia muitas espécies de IG. Fabaceae foi a família mais rica, com 30% das espécies de PH e 40% dos IG. Além disso, nossos resultados mostraram um baixo número de espécies de PH e IG para a Caatinga brasileira, e discutimos esse padrão com os seguintes argumentos, primeiro, é provável que o número de inventários de insetos galhadores para a Caatinga seja subamostrado, segundo a Caatinga possui um número relativamente menor de espécies vegetais quando comparada a outros biomas e, por fim, argumentamos que a Caatinga é uma floresta tropical sazonalmente seca onde a deciduidade representa um fator relevante para as taxas de colonização e desempenho dos IG.

Plant-galling insect interactions: a data set of host plants and their gall-inducing insects for the Cerrado.

Recent decades have seen increased research interest in the processes and mechanisms related to insect gall richness and host plants. The data set provided here includes 968 records of interactions between galling insects and host plants for the Cerrado biome. The data set comprises 505 species of 222 genera and 67 families of host plants. The botanical families most represented in the data set are Asteraceae, Fabaceae, Myrtaceae, Malpighiaceae, and Melastomataceae, which account for ~48.5% of all records and 52% of the total number of species. The gall-inducing insects listed in the data set include 48 species of Cecidomyiidae and one species of Tephritidae. This data set is the first to compile inventories of plant-galling insect communities and information about the diversity and distribution of insect galls and their host plants in the Cerrado. The data set reveals knowledge gaps and opportunities for future research on patterns of diversity and distribution, and provides a basis for generating and testing new ecological hypotheses. Please cite this data paper when using the current data in publications and let us know how the data are used in the publications. There are no copyright restrictions.

An overview of inventories of gall-inducing insects in Brazil: looking for patterns and identifying knowledge gaps.

We compiled published Brazilian gall-inducing insect inventories aiming to understand trends and biases in this field research and to investigate the factors that potentially explain the diversity of gall-inducing insects among different sampling sites. A total of 51 studies with gall-inducing insect inventories were compiled for Brazil, which sampled 151 sites in 88 municipalities, 13 states and five regions. The number of papers published on gall-inducing insects per year has increased over the last 30 years, being Cecidomyiidae (Diptera) the main galling taxon, Fabaceae the main host-plant family and Protium heptaphyllum (Burseraceae) the most important super-host species in these inventories. We found a great bias in the geographical distribution of Brazilian inventories, with the majority of studies in the Southeast region, and Atlantic Forest and Cerrado biomes. The total richness of gall-inducing insects differed significantly among regions and biomes, with higher gall richnesses being recorded in the North region and Amazon biome. However, Brazilian regions and biomes did not vary in richness of gall-inducing insect morphotypes per plant species. According our results, sampling by cecidologists in less studied regions of Brazil is needed, particularly in the North and South regions and subsampled biomes such as the Amazon, Pampas and Pantanal.

Shifts in Plant Assemblages Reduce the Richness of Galling Insects Across Edge-Affected Habitats in the Atlantic Forest.

Impacts of habitat loss and fragmentation on specialist herbivores have been rarely addressed. Here we examine the structure of plant and galling insect assemblages in a fragmented landscape of the Atlantic forest to verify a potential impoverishment of these assemblages mediated by edge effects. Saplings and galling insects were recorded once within a 0.1-ha area at habitat level, covering forest interior stands, forest edges, and small fragments. A total of 1,769 saplings from 219 tree species were recorded across all three habitats, with differences in terms of sapling abundance and species richness. Additionally, edge-affected habitats exhibited reduced richness of both host-plant and galling insects at plot and habitat spatial scale. Attack levels also differed among forest types at habitat spatial scale (21.1% of attacked stems in forest interior, 12.4% in small fragments but only 8.5% in forest edges). Plot ordination resulted in three clearly segregated clusters: one formed by forest interior, one by small fragments, and another formed by edge plots. Finally, the indicator species analysis identified seven and one indicator plant species in forest interior and edge-affected habitats, respectively. Consequently, edge effects lead to formation of distinct taxonomic groups and also an impoverished assemblage of plants and galling insects at multiple spatial scales. The results of the present study indicate that fragmentation-related changes in plant assemblages can have a cascade effects on specialist herbivores. Accordingly, hyperfragmented landscapes may not be able to retain an expressive portion of tropical biodiversity.

Evidence of Allopolyploidy in Urochloa humidicola Based on Cytological Analysis and Genetic Linkage Mapping.

The African species Urochloa humidicola (Rendle) Morrone & Zuloaga (syn. Brachiaria humidicola (Rendle) Schweick.) is an important perennial forage grass found throughout the tropics. This species is polyploid, ranging from tetra to nonaploid, and apomictic, which makes genetic studies challenging; therefore, the number of currently available genetic resources is limited. The genomic architecture and evolution of U. humidicola and the molecular markers linked to apomixis were investigated in a full-sib F1 population obtained by crossing the sexual accession H031 and the apomictic cultivar U. humidicola cv. BRS Tupi, both of which are hexaploid. A simple sequence repeat (SSR)-based linkage map was constructed for the species from 102 polymorphic and specific SSR markers based on simplex and double-simplex markers. The map consisted of 49 linkage groups (LGs) and had a total length of 1702.82 cM, with 89 microsatellite loci and an average map density of 10.6 cM. Eight homology groups (HGs) were formed, comprising 22 LGs, and the other LGs remained ungrouped. The locus that controls apospory (apo-locus) was mapped in LG02 and was located 19.4 cM from the locus Bh027.c.D2. In the cytological analyses of some hybrids, bi- to hexavalents at diakinesis were observed, as well as two nucleoli in some meiocytes, smaller chromosomes with preferential allocation within the first metaphase plate and asynchronous chromosome migration to the poles during anaphase. The linkage map and the meiocyte analyses confirm previous reports of hybridization and suggest an allopolyploid origin of the hexaploid U. humidicola. This is the first linkage map of an Urochloa species, and it will be useful for future quantitative trait locus (QTL) analysis after saturation of the map and for genome assembly and evolutionary studies in Urochloa spp. Moreover, the results of the apomixis mapping are consistent with previous reports and confirm the need for additional studies to search for a co-segregating marker.

Microsatellite markers for Urochloa humidicola (Poaceae) and their transferability to other Urochloa species.

BACKGROUND: Urochloa humidicola is a warm-season grass commonly used as forage in the tropics and is recognized for its tolerance to seasonal flooding. This grass is an important forage species for the Cerrado and Amazon regions of Brazil. U. humidicola is a polyploid species with variable ploidy (6X-9X) and facultative apomixis with high phenotypic plasticity. However, this apomixis and ploidy, as well as the limited knowledge of the genetic basis of the germplasm collection, have constrained genetic breeding activities, yet microsatellite markers may enable a better understanding of the species' genetic composition. This study aimed to develop and characterize new polymorphic microsatellite molecular markers in U. humidicola and to evaluate their transferability to other Urochloa species. FINDINGS: A set of microsatellite markers for U. humidicola was identified from two new enriched genomic DNA libraries: the first library was constructed from a single sexual genotype and the second from a pool of eight apomictic genotypes selected on the basis of previous results. Of the 114 loci developed, 72 primer pairs presented a good amplification product, and 64 were polymorphic among the 34 genotypes tested. The number of bands per simple sequence repeat (SSR) locus ranged from 1 to 29, with a mean of 9.6 bands per locus. The mean polymorphism information content (PIC) of all loci was 0.77, and the mean discrimination power (DP) was 0.87. STRUCTURE analysis revealed differences among U. humidicola accessions, hybrids, and other Urochloa accessions. The transferability of these microsatellites was evaluated in four species of the genus, U. brizantha, U. decumbens, U. ruziziensis, and U. dictyoneura, and the percentage of transferability ranged from 58.33% to 69.44% depending on the species. CONCLUSIONS: This work reports new polymorphic microsatellite markers for U. humidicola that can be used for breeding programs of this and other Urochloa species, including genetic linkage mapping, quantitative trait loci identification, and marker-assisted selection.

Pattern of attack of a galling insect reveals an unexpected preference-performance linkage on medium-sized resources / Padrão de ataque de um inseto galhador revela uma inesperada ligação entre preferência e performance sobre recursos de tamanho médio

Pattern of attack of a galling insect reveals an unexpected preference-performance linkage on medium-sized resources. The Plant Vigor Hypothesis (PVH) predicts oviposition preference and higher offspring performance on longer and fast-growing shoots, and although several studies have tested its predictions, long-term studies concerning the patterns of host selection by galling species are still lacking. The PVH was tested in this study using Bauhinia brevipes (Fabaceae) as the host of a leaf gall midge, Asphondylia microcapillata (Diptera, Cecidomyiidae) during three consecutive years. Shoots were collected from the same 80 plants between 2001 and 2003 and shoot length, number of healthy and galled leaves, gall number, and mortality factors were recorded. Nearly 600 galls were found on the 5,800 shoots collected. Medium-sized shoots supported from 46 to 70 percent of all galls, with greater gall survival rate in 2002 and 2003. A decrease in parasitism rate coupled with an increase in gall predation lead to a constant similar gall survivorship rate in all years (x = 22.7 percent). Although gall abundance varied among years (122 in 2001, 114 in 2002 and 359 in 2003) preference for longer shoots was not observed because the percentage of galled shoots and galled leaves were higher on medium shoot length classes in all years. The observed distribution of gall abundance and galled shoots were always greater than the expected distribution on medium shoot length classes. These findings do not support the PVH, and show that A. microcapillata can maximize the female preference and larval performance on medium-sized shoots of B. brevipes.

A Hipótese do Vigor de Plantas (HVP) prevê uma oviposição preferencial e alta performance da prole em ramos longos e de crescimento rápido da planta hospedeira. Embora diversos estudos tenham testado suas predições, estudos de longa duração focados no padrão de seleção de planta hospedeira por insetos galhadores ainda são escassos. A HVP foi testada neste estudo usando Bauhinia brevipes (Fabaceae) como planta hospedeira de uma galha foliar induzida por Asphondylia microcapillata (Diptera: Cecidomyiidae) por três anos consecutivos. Ramos foram coletados das mesmas 80 plantas entre 2001 e 2003 e o tamanho do ramo, número de ramos galhados e sadios, número de galhas e fatores de mortalidade foram registrados. Aproximadamente 600 galhas de A. microcapillata foram coletadas em 5.800 ramos de B. brevipes. Ramos médios suportaram entre 46 por cento e 70 por cento do total de galhas, e a taxa de sobrevivência das galhas foi maior em 2002 e 2003. Uma diminuição da taxa de parasitismo associada com um aumento da taxa de predação de galhas resultou em uma sobrevivência similar em todos os anos (x = 22,7 por cento). A abundância de galhas variou entre os anos (122 em 2001, 114 em 2002 e 359 em 2003). A preferência por ramos longos não foi observada porque a porcentagem de ramos galhados e folhas galhadas foram maiores em ramos de classes de tamanho intermediário por dois anos. A distribuição observada da abundância de galhas e ramos galhados foi sempre maior do que a distribuição esperada em classes de ramos médios. Estes resultados não suportam a HVP, e mostram que A. microcapillata pode maximizar a preferência da fêmea e a performance larval em ramos de tamanho médio de B. brevipes.

A 15-year post evaluation of the fire effects on ant community in an area of Amazonian forest

Fire represents an important disturbance to ant communities in areas of fire regime. Otherwise, little is known about the effects of fire on ant communities in areas of non-fire regimes, such as in the Amazonian region. We evaluated the long-term effect of fire on ant species richness in a rain forest (Bacaba Plateau) burned 15-years ago and compare our data with the data of primary unburned forest. A total of 85 ant species distributed in 21 genera and 14 tribes were collected; among them, 72 and 44 species were found on the litter and vegetation, respectively. The fire damaged forest studied supports an intermediate richness of ants when compared to a primary unburned rain forest in the same region. A comparative analysis of ant species richness showed that the Bacaba Plateau presented a different ant fauna when compared with the primary unburned forests, suggesting that fire can alter ant species composition. Although, our results cannot be conclusive on the effects of fire on ant community, they represent a pioneer data on human induced fire in tropical rain forests.

O fogo representa uma importante perturbação para a comunidade de formigas em áreas de regime de fogo. No entanto, pouco se conhece sobre os efeitos do fogo na comunidade de formiga em áreas de não-regime, tal como a região da Amazônia. Nós analisamos o efeito de longo prazo do fogo sobre a riqueza de formiga numa floresta tropical queimada 15 anos atrás e comparamos nossos dados, com os de uma floresta primária não-queimada. Foram coletadas um total de 85 espécies de formigas distribuídas em 21 gêneros e 14 tribos, dentre eles 72 e 44 espécies foram encontradas na liteira e vegetação, respectivamente. Esta área de floresta queimada, com 85 espécies, pode suportar uma riqueza intermédia de formigas quando comparadas com uma floresta tropical primária não-queimada, com 29, 22 e 98 espécies na mesma região. Uma análise comparativa da riqueza de espécies de formigas mostrou que o platô Bacaba apresentou uma diferente fauna de formigas quando comparado com as florestas primárias não queimadas, sugerindo que o fogo pode alterar a composição das espécies. Embora, nossos resultados sobre os efeitos do fogo não sejam conclusivos, eles representam dados pioneiros sobre incêndios induzidos por humanos em floresta tropical.

Distribution and frequency of galls induced by Anisodiplosis waltheriae Maia (Diptera: Cecidomyiidae) on the invasive plant Waltheria indica L. (Sterculiaceae).

The frequency of galls induced by Anisodiplosis waltheriae Maia, a recently described species, on Waltheria indica L. was studied. W indica is an invasive weed in regeneration areas of Atlantic Forest in southeastern Brazil. Plants were collected in May 2004 and above-ground biomass, main stem length, number of leaves, number of galls per leaf and leaf area of each individual were recorded. Nearly 90% of all plants and 25% of all leaves were attacked by the gall midge, with an average of 0.67 galls/leaf. Leaf area had a weak effect on gall abundance while the number of leaves had no effect on gall abundance. Only 31% of the variation in gall abundance was explained by plant biomass. Natural enemies killed one third of the sampled galls. Predation accounted for 22.9% of gall mortality, unknown factors killed 7.6%, microhymenopteran parasitoids killed 2.5% and fungi only 1%. Mortality factors were not influenced by leaf area or gall density.

Influence of Apion sp. (Brentidae, Apioninae) stem-galls on induced resistance and leaf area of Diospyros hispida (Ebenaceae)

We addressed the influence of the stem galls induced by an unidentified species of Apion sensu lato (Brentidae, Apioninae) on the host plant, Diospyros hispida (Ebenaceae) leaf area and induced resistance against a Cecidomyiidae (Diptera) leaf galls. The study was performed in a cerrado vegetation in Serra do Cipó, southeastern Brazil. Although the number of leaves produced on galled and ungalled shoots did not differ statically (p>0.05), the presence of the apionid galls influenced the area of the leaves on the attacked shoots of D. hispida. Leaves on galled stems were approximately 50 percent smaller compared to leaves in healthy stems. The average of the cecidomyiid leaf galls successfully induced on healthy shoots was higher compared to galls successfully induced on shoots galled by the apionid. The same pattern was found for the abundance of hypersensitive reactions against the cedidomyiid gall induction. Therefore, the ability of the cecidomyiid to successfully induce galls was not influenced by the apionid galler.

Neste estudo verificamos a influência de galhas de ramos induzida por uma espécie não identificada de Apion sp. (Brentidae, Apioninae) sobre a área foliar da planta hospedeira Diospyros hispida (Ebenaceae) e na resistência induzida contra galhas de folhas de um Cecidomyiidae (Diptera). Este estudo foi conduzido em uma vegetação de cerrado na Serra do Cipó, sudeste do Brasil. Embora o número de folhas produzidas nos ramos galhados e não galhados não tenha diferido estaticamente (p>0,05), a presença das galhas de Apion sp. influenciou a área foliar nos ramos atacados de D. hispida. Folhas de ramos com galhas foram aproximadamente 50 por cento menores quando comparadas a folhas em ramos saudáveis. A média de galhas foliares do Cecidomyiideae que se formaram com sucesso em folhas de ramos saudáveis foi maior quando comparado à média de galhas bem sucedidas em folhas de ramos galhados pelo Apion sp.. O mesmo padrão foi encontrado para a abundância de reações hipersensibilidade contra a formação de galhas do cecidomyiídeo. Portanto, a habilidade do cecidomyiídeo de induzir galhas não foi influenciada pelas galhas de Apion sp..

Distribution and frequency of galls induced by Anisodiplosis waltheriae Maia (Diptera: Cecidomyiidae) on the invasive plant Waltheria indica L. (Sterculiaceae)

The frequency of galls induced by Anisodiplosis waltheriae Maia, a recently described species, on Waltheria indica L. was studied. W. indica is an invasive weed in regeneration areas of Atlantic Forest in southeastern Brazil. Plants were collected in May 2004 and above-ground biomass, main stem length, number of leaves, number of galls per leaf and leaf area of each individual were recorded. Nearly 90 percent of all plants and 25 percent of all leaves were attacked by the gall midge, with an average of 0.67 galls/leaf. Leaf area had a weak effect on gall abundance while the number of leaves had no effect on gall abundance. Only 31 percent of the variation in gall abundance was explained by plant biomass. Natural enemies killed one third of the sampled galls. Predation accounted for 22.9 percent of gall mortality, unknown factors killed 7.6 percent, microhymenopteran parasitoids killed 2.5 percent and fungi only 1 percent. Mortality factors were not influenced by leaf area or gall density.

Estudou-se a freqüência de galhas induzidas por Anisodiplosis waltheriae Maia, espécie recentemente descrita, em Waltheria indica L. W. indica é uma erva invasora em áreas de regeneração de Floresta Atlântica no Sudeste do Brasil. As plantas foram coletadas em maio de 2004 e a biomassa aérea, comprimento do caule, número de folhas, número de galhas por folha e área foliar de cada indivíduo foram registrados. Quase 90 por cento das plantas e 25 por cento das folhas foram atacadas, com a média de 0,67 galhas/folha. A área foliar pouco influenciou o número de galhas, porém o número de folhas não afetou a abundância de galhas (P > 0,05). Apenas 31 por cento da variação no número de galhas foi explicada pela biomassa da planta. Um terço das galhas foi morta por inimigos naturais. A predação explicou 22,9 por cento da mortalidade, fatores desconhecidos mataram 7,6 por cento, microhimenópteros parasitóides mataram 2,5 por cento e fungos apenas 1 por cento. Os fatores de mortalidade não foram influenciados pela área foliar nem densidade de galhas.

Foraging patterns of the leaf-cutter ant Atta laevigata (Smith) (Myrmicinae: Attini) in an area of cerrado vegetation / Padrões de forrageamento da formiga cortadeira Atta laevigata (Smith) (Myrmicinae: Attini) em uma área de cerrado

A taxa de forrageamento pode interferir diretamente no impacto que Atta laevigata (Smith) exerce no ambiente. Os fatores que mais afetam a taxa de forrageamento foram avaliados utilizando-se dados de A. laevigata coletados por três meses no Parque Nacional da Serra do Cipó, MG. Um padrão bimodal de forrageamento foi observado em dias ensolarados. Amostras do peso seco coletadas nasentradas dos ninhos foram em média maiores nos dia ensolarados (0.9713 ± 0.1358 g), quando comparado aos dias chuvosos (0.4485 ± 0.1120g). Ninhos grandes (0.9713 ± 0.1358 g) também possuíam maiores coletas de peso seco comparadas aos ninhos pequenos (0.2752 ± 0.0463 g). Indivíduos de A. laevigata trocam de entradas de ninhos para entradas com melhores condições microhabitacionais tanto em diaschuvosos como ensolarados. A combinação da temperatura, umidade, tamanho de ninho e especializaçãode microhabitats parecem ser os fatores reguladores mais importantes dos padrões de forrageamento de A. laevigata no Parque Nacional da Serra do Cipó.

Foraging rate can directly affect the impact that Atta laevigata (Smith) has on the environment. We attempted to assess the factors which most affect foraging rate of A. laevigata using data collected over a three month period in the Parque Nacional da Serra do Cipó in Minas Gerais, Brazil. A bimodal foraging pattern was observed on clear sunny days, based on collected dry weights ofvegetative material being carried towards nest entrances. Sampled entrances on clear sunny days yielded a daily average dry weight twice as high (0.9713 ± 0.1358 g) as on rainy days (0.4485 ± 0.1120 g). Larger nests (0.9713 ± 0.1358 g) yielded higher dry weights than smaller nests (0.2752 ± 0.0463 g). Field observations showed Ants switched to nest entrances with more suitable microhabitat conditions inboth rainy and sunny days. A combination of temperature, humidity, nest size, and microhabitat specialization were the most important factors influencing A. laevigata foraging pattern.

Ovipositing behaviour of Compsobracon mirabilis (Szépligeti) (Hymenoptera, Braconidae) in a cerrado habitat, southeastern Brazil

O comportamento de oviposição do braconídeo parasitóide, Compsobracon mirabilis (Szépligeti, 1901) é descrito. As observações foram conduzidas em uma região de cerrado localizada em Três Marias, Minas Gerais, Brasil. A oviposição foi feita em um ramo de Alibertia concolor (Cham.) K. Schum.1889 (Rubiaceae), no interior do qual havia treze larvas de uma espécie de Lepidoptera não identificada.

The oviposition behaviour of the braconid parasitoid, Compsobracon mirabilis ( Szépligeti, 1901) is described. Observations were conducted in a cerrado region located in Três Marias, Minas Gerais, Brazil. The oviposition occurred in a branch of Alibertia concolor (Cham.) K. Schum. 1889 (Rubiaceae), inside of which there were thirteen larvae of an unidentified species of Lepidoptera.