Molecular phylogenetics and evolution of host plant use in the Neotropical rolled leaf 'hispine' beetle genus Cephaloleia (Chevrolat) (Chrysomelidae: Cassidinae)

Here, we report the results of a species level phylogenetic study of Cephaloleia beetles designed to clarify relationships and patterns of host plant taxon and tissue use among species. Our study is based on up to 2088bp of mtDNA sequence data. Maximum parsimony, maximum likelihood, and Bayesian methods of phylogenetic inference consistently recover a monophyletic Cephaloleia outside of a basal clade of primarily palm feeding species (the 'Arecaceae-feeding clade'), and C. irregularis. In all three analyses, the 'Arecaceae-feeding clade' includes Cephaloleia spp. with unusual morphological features, and a few species currently placed in other cassidine genera and tribes. All three analyses also recover a clade that includes all Zingiberales feeding Cephaloleia and most Cephaloleia species (the 'Zingiberales-feeding clade'). Two notable clades are found within the 'Zingiberales-feeding clade.' One is comprised of beetles that normally feed only on the young rolled leaves of plants in the families Heliconiaceae and Marantaceae (the 'Heliconiaceae & Marantaceae-feeding clade'). The other is comprised of relative host tissue generalist, primarily Zingiberales feeding species (the 'generalist-feeding clade'). A few species in the 'generalist-feeding clade' utilize Cyperaceae or Poaceae as hosts. Overall, relatively basal Cephaloleia (e.g., the 'Arecaceae clade') feed on relatively basal monocots (e.g., Cyclanthaceae and Arecaceae), and relatively derived Cephaloleia (e.g., the 'Zingiberales-feeding clade') feed on relatively derived monocots (mostly in the order Zingiberales). Zingiberales feeding and specialization on young rolled Zingiberales leaves have each apparently evolved just once in Cephaloleia.     

A new species of bromeliad-feeding Cephaloleia Chevrolat (Coleoptera,Chrysomelidae, Cassidinae) from Costa Rica: evidence from DNA barcodes,larval and adult morphology and insect diets

The Neotropical genus Cephaloleia Chevrolat (Coleoptera: Chrysomelidae: Cassidinae) includes 214 species distributed from the south of Mexico to Argentina. Cephaloleia beetles feed mostly on plants from the order Zingiberales. The interactions between Cephaloleia beetles and their Zingiberales host plants is proposed as one of the oldest and most conservative associations. Here we describe a new species of Cephaloleia (Cephaloleia kuprewiczae sp. n.) that feeds on two species of bromeliads (Pitcairnia arcuata and Pitcairnia brittoniana, Bromeliaceae: Pitcairnioideae). Cephaloleia kuprewiczae was previously described as Cephaloleia histrionica. This study includes evidence from DNA barcodes (COI), larval and adult morphology and insect diets that separates Cephaloleia kuprewiczae from Cephaloleia histrionica as a new species.     

Horizontal Transmission of Bursaphelenchus xylophilus between Sexes of Monochamus alternatus

Four experiments were conducted using nematode-infested and nematode-free adults of the cerambycid beetle, Monochamus alternatus, to determine horizontal transmission pathways of Bursaphelenchus xylophilus. When nematode-infested beetles of one sex and nematode-free beetles of the opposite sex were paired in containers for 48 or 72 hours, the number of nematodes carried by nematode-free beetles tended to increase with increased number of nematodes carried by nematode-infested beetles. The nematodes acquired by "nematode-free" beetles could be transmitted to pine. A female beetle that received 13 nematodes from a male transmitted one nematode to a Pinus densiflora bolt via an oviposition wound. When the nematode-infested and nematode-free beetles were observed continuously, it was observed that the number of nematodes carried by nematode-free beetles at the end of the first sexual mounting increased as the number of nematodes carried by nematode-infested beetles just before mounting increased. The number of nematodes transferred to nematode-free beetles was positively related to duration time of mounting. There was no difference in transmission efficacy between male-to-female transmission and female-to-male transmission. The horizontal transmission pathways are discussed relative to the persistence of B. xylophilus in resistant pine forests and the control of pine wilt disease.     

Inhibitory Effect of Bursaphelenchus mucronatus (Nematoda: Aphelenchoididae) on B. xylophilus Boarding Adult Monochamus alternatus (Coleoptera: Cerambycidae)

Inhibitory effects of Bursaphelenchus mucronatus on the number of B. xylophilus carried by an adult Monochamus alternatus were investigated using artificial pupal chambers. When pupal chambers were infested with either B. xylophilus or B. mucronatus, the load of B. xylophilus onto the beetle was greater (P < 0.001) than that of B. mucronatus. However, within the pupal chamber there was no difference in the abundance of the third-stage dispersal juveniles, which would molt to the fourth-stage dispersal juveniles to board beetles. The nematode load on beetles that emerged from pupal chambers infested with both Bursaphelenchus species was smaller (P = 0.015) than that of beetles with B. xylophilus alone but greater (P < 0.001) than that of beetles with B. mucronatus alone, suggesting an inhibitory effect of B. mucronatus. As a result of this study, the rate of inhibition of B. mucronatus on molting of third-stage dispersal juveniles of B. xylophilus to fourth-stage dispersal juveniles was 0.65, which resulted in great inhibition on boarding beetles at a rate of 0.7.     

Decomposition and nutrient release in halophytes of a Mediterranean salt marsh

This study dealt with the decomposition and nutrient release from the halophytes Atriplex portulacoides, Arthrocnemum macrostachyum, Limoniastrum monopetalum, and Spartina densiflora, the dominant species in the Castro Marim salt marsh, Portugal. Environmental effects on decomposition were also assessed. The study was carried out for one year using the in situ litterbag technique. S. densiflora showed a lower decomposition rate (k = 0.003 day⁻¹) than the other study species (k = 0.005-0.009). Study species showed similar decomposition patterns, that is, the weight loss mostly occurred during the autumn-winter period (study beginning in November). This indicates that temperature in this period did not hamper the decomposition process. The decomposition rate was positively affected by the initial N concentration (r² = 0.87, P < 0.05) and negatively by the C:N ratio (r² = 0.86, P < 0.05) in decomposing materials. At the end of the study, S. densiflora and L. monopetalum, the species with lower initial N concentrations, retained much higher proportion of initial N (89-109%) than the others (5-14%). Also, S. densiflora with the lowest P concentration retained higher proportion of initial P (48%) than the others (5-20%). Release of K and Mg were also slower from S. densiflora and was associated with their initial low concentration in this species. The lowest Mn release was observed from A. macrostachyum and also in relation to the lowest initial concentration. Our study supports the hypothesis that decomposition patterns of marsh species are mostly associated with differences regarding their morphology and chemical composition. Given the higher resistance of S. densiflora to decomposition, its progressive spreading may result in accumulation of organic detritus overtime in invaded salt marshes.     

Ecological Study of Nematode Parasitism in Ips Beetles from California and Idaho

Nematodes found in Ips paraconfusus from ponderosa pine in California were an undescribed species of Parasitaphelenchus, Contortylenchus elongatus, C. reversus, and C. brevicomi. C. elongatus, the most commonly found contortylenchid, was present in 98.2% of the contortylenchid-parasitized beetles. Only one nematode parasite of the gut, a Parasitorhabditis sp., was isolated. Although significant differences in parasitism were observed, they were by collection sites, rather than by elevation or bole sources (slash or standing). Significant changes in parasitism between fall and spring collections were observed but not at every site. Nematode parasitism in the F₁ generation of I. paraconfusus by Parasitaphelenchus, Contortylenchus, or Parasitorhabditis increased or decreased from the parent generation depending upon the experiment.Nematode parasites from I. pini included an undescribed Parasitaphelenchus sp., two undescribed Contortylenchus spp., C. reversus and Parasitylenchus (= Neoparasitylenchus) ovarius from the hemocel, and Parasitorhabditis ipini from the gut. Parasitaphelenchus sp. was found in 99% and 45.3% of the beetles from Idaho and California, respectively. Of the 1,000 I. pini from Idaho and California, 157 were parasitized by the contortylenchid species or P. ovarius.     

Fishborne Trematode Metacercariae Detected in Freshwater Fish from Vientiane Municipality and Savannakhet Province, Lao PDR

Freshwater fish from Vientiane Municipality and Savannakhet Province, Lao PDR were examined by the muscle compression and artificial digestion methods to know the infection status with trematode metacercariae. In the fish from Savannakhet, 2 species of metacercariae, Opisthorchis viverrini and Haplorchis taichui, were detected. O. viverrini metacercariae were found in 6 species of fish, Puntius brevis, Hampala dispar, Esomus metallicus, Mystacoleucus marginatus, Puntioplites falcifer, and Cyclocheilichthys armatus. H. taichui metacercariae were detected in 3 species of fish, P. brevis, P. falcifer, and M. marginatus. In the fish from Vientiane, 4 species of metacercariae, O. viverrini, H. taichui, Haplorchis yokogawai, and Centrocestus formosanus, were detected. Among them, O. viverrini metacercariae were found in 7 species of fish, Onychostoma elongatum, C. armatus, H. dispar, P. brevis, Cyclocheilichthys repasson, Osteochilus hasseltii, and Hypsibarbus lagleri. The metacercariae of H. taichui were detected in 6 species of fish, C. repasson, O. elongatum, C. armatus, H. dispar, Labiobarbus leptocheila, and Cirrhinus molitorella. The metacercariae of H. yokogawai were found in 9 species of fish, C. repasson, O. elongatum, C. armatus, H. dispar, Labiobarbus leptocheila, O. hasseltii. C. molitorella, Hypsibarbus wetmorei, and H. lagleri. The metacercariae of C. formosanus were detected in 4 species of fish, C. repasson, P. brevis, O. hasseltii, and C. molitorella. From these results, it is confirmed that fishborne trematode metacercariae, i.e. O. viverrini, H. taichui, H. yokogawai and C. formosanus, are prevalent in various species of freshwater fish from Savannakhet Province and Vientiane Municipality, Lao PDR.     

Heterodera achilleae n. sp. (Nematoda: Heteroderidae) from Yarrow in Yugoslavia

Heterodera achilleae n. sp., a member of the H. rostochiensis group, is described and illustrated from roots of yarrow, Achillea millefolium L. in Sarajevo, Yugoslavia. This new, round-cyst species differs from closely related species especially as follows: (1) from H. leptonepia, by having stouter larvae (a = 21), with longer styler (25 μ), and with outlet of dorsal esophageal gland averaging 5.7 μ from base o f styler; (2) from H. millefolii, in having excretory pore at base of neck and small, straight vulval slit of 5 μ; (3) from H. rostochiensis, in having a B/A ratio (Granek''s ratio) of 1.6 ; (4) from H. tabacum, by longer female stylet, two annules on female head, and males with outlet of dorsal esophageal gland further back (5.7 μ). In addition, H. achilleae n. sp. differs from the latter three species in having prominent longitudinal striae on the anterior half, or more, of cysts and females.     

Checklist of tortoise beetles (Coleoptera,Chrysomelidae, Cassidinae) from Colombia with new data and description of a new species

A new tortoise beetle species, Cyrtonota abrili, is described from the Antioquia and Caldas departments in Colombia. New faunistic data are provided for 87 species, including 16 new additions to the country’s fauna. A checklist of the known 238 species of tortoise beetles recorded from Colombia is given.     

Susceptibility of the Mediterranean fruit fly (Ceratitis capitata) and the Natal fruit fly (Ceratitis rosa) to entomopathogenic nematodes

The potential of entomopathogenic nematodes, Heterorhabditis bacteriophora, Heterorhabditis zealandica and Steinernema khoisanae, to infect pupariating larvae, pupae and adults of Ceratitis capitata and Ceratitis rosa was investigated in laboratory bioassays. Pupariating larvae and adult flies were susceptible to nematode infection, with no infection recorded for the pupae. Pupariating larvae of C. capitata were generally more susceptible to infection than those of C. rosa. Significantly more larvae of C. capitata were infected by H. bacteriophora. For C. rosa, highest infectivity of larvae was obtained with H. zealandica. In contrast, adults of both species were highly infected by S. khoisanae.     

The ant genus Carebara Westwood (Hymenoptera,Formicidae): synonymisation of Pheidologeton Mayr under Carebara,establishment and revision of the C. polita species group

In this paper the genus Pheidologeton Mayr, 1862 is synonymized under Carebara Westwood, 1840 and the Carebara polita group is established and revised. This species group currently includes six species from the Afrotropical region (C. madibai, C. nicotianae, C. perpusilla, C. polita, C. silvestrii, and C. villiersi) and two species from the Neotropical region (C. brevipilosa and C. urichi). The polita group clearly links Carebara and Pheidologeton, and, due to a lack of autapomorphic characters for the latter, a separation of the two genera is no longer justified. As a result Carebara is presented as a monophyletic and better defined genus that can be separated from other genera with more confidence. We present an overview of the distribution and biology of Carebara as well as images from the various genera currently in synonymy under Carebara, and discuss the characters they share. The polymorphism present in Afrotropical and Malagasy Carebara is discussed and one new species from Africa, C. madibai sp. n., is described. The subspecies Carebara perpusilla arnoldiana syn. n., Carebara perpusilla concedens syn. n., and Carebara perpusilla spinosa syn. n. are new synonyms of Carebara perpusilla. Oligomyrmex politus nicotianae is re-elevated to species level and transferred into Carebara, C. nicotianae comb. n., stat. rev.; C. punctata is a new synonym of C. silvestrii comb. n. and C. pygmaea albipes comb. n., syn n., C. pygmaea bugnioni comb. n., syn. n., and C. simularensis syn. n. are new synonyms of C. pygmaea comb. n.. The following names are transferred from Pheidologeton to Carebara as new combinations (with the species epithets adjusted to female endings where necessary): aberrans, affinis, affinis javana, affinis minor, affinis spinosior, affinis sumatrensis, ceylonensis, dentiviris, diversa, diversa draco, diversa ficta, diversa laotina, diversa macgregori, diversa philippina, diversa standfussi, diversa taprobanae, diversa tenuirugosa, diversa williamsi, hammoniae, hostilis, kunensis, latinoda, maccus, mayri, melanocephala, melasolena, nana, nanningensis, obscura, petulens, pullata, pungens, pygmaea, rubra, rugiceps, rugosa, schossnicensis, silena, silvestrii, solitaria, transversalis, trechideros, varia, vespilla, volsellata, yanoi, and zengchengensis. Three new combinations are creating secondary junior homonyms and are here replaced with new names: C. mayri (Santschi, 1928) = C. gustavmayri nom. n., C. rugosa (Karavaiev, 1935) = C. rugoflabella nom. n., and C. silvestrii (Wheeler, 1929b) = C. luzonensis nom. n. Two new combinations are creating secondary junior homonyms among species already in Carebara: C. taprobanae (Forel, 1911a) = C. sinhala nom. n., and C. nana Santschi, 1919 = C. pumilia nom. n.     

Pathogenicity Tests on Nine Mosquito Species and Several Non-target Organisms with Strelkovimermis spiculatus (Nemata Mermithidae)

Nine species of mosquitoes and several species of non-target aquatic organisms were tested for susceptibility to the mernaithid nematode, Strelkovimermis spiculatus. All species of Anopheles, Aedes, Culex, and Toxorhynchites exposed to S. spiculatus were susceptible. Of the nine mosquito species tested, C. pipiens quinquefasciatus had the greatest tolerance to initial invasion and the highest percent infection of those that survived. High levels of infection were also achieved with Aedes taeniorhynchus and A. albopictus, but these mosquitoes were significantly less tolerant to parasitism than C. pipiens quinquefasciatus. Strelkovimermis spiculatus did not infect or develop in any of the non-target hosts tested.     

FMRFamide-like Immunoactivity in Heterodera glycines (Nemata: Tylenchida)

Material antigenically related to the neuromodulatory peptide FMRFamide was detected and examined in preparations of the soybean cyst nematode, Heterodera glycines, and in the free-living nematodes Caenorhabditis elegans and Panagrellus redivivus. FMRFamide-related peptides were quantified by an enzyme-linked immunosorbent assay. Specific activities were remarkably similar among all of the vermiform members of the three species. FMRFamide-related peptide immunoactivity was present in both sexes and all stages of H. glycines examined. The highest specific activity was present in second-stage juveniles and in males, and the lowest in white and yellow females. Total FMRFamide-related peptide level per individual was highest in brown females, with 90% of the activity associated with the eggs. Peptide levels in these eggs and in second-stage juveniles were comparable and increased in adults, especially in females. Chromatographic analysis of FMRFamide-related peptide preparations from H. glycines juveniles, C. elegans, and P. redivivus revealed distinct qualitative differences between the infective plant parasite and the free-living nematodes.     

Systematics of the Neotropical genus Catharylla Zeller (Lepidoptera,Pyralidae s. l., Crambinae)

The Neotropical genus Catharylla Zeller, 1863 (type species: Crambus tenellus Zeller, 1839) is redescribed. Catharylla contiguella Zeller, 1872, C. interrupta Zeller, 1866 and Myelois sericina Zeller, 1881, included by Munroe (1995) in Catharylla, are moved to Argyria Hübner. Catharylla paulella Schaus, 1922 and C. tenellus (Zeller, 1839) are redescribed. Six new species are described by Léger and Landry: C. bijuga, C. chelicerata, C. coronata, C. gigantea, C. mayrabonillae and C. serrabonita. The phylogenetic relationships were investigated using morphological as well as molecular data (COI, wingless, EF-1α genes). The median and subterminal transverse lines of the forewing as well as the short anterior and posterior apophyses of the female genitalia are characteristic of the genus. The monophyly of Catharylla was recovered in all phylogenetic analyses of the molecular and the combined datasets, with three morphological apomorphies highlighted. Phylogenetic analyses of the morphology of the two sexes recovered three separate species groups within Catharylla: the chelicerata, the mayrabonillae, and the tenellus species groups. The possible position of Micrelephas Schaus, 1922 as sister to Catharylla, based on both morphological and molecular data, and the status of tribe Argyriini are discussed. The biogeographical data indicate that the chelicerata species group is restricted to the Guyanas and the Amazonian regions whereas the tenellus group is restricted to the Atlantic Forest in the South-Eastern part of Brazil. The mayrabonillae group is widespread from Costa Rica to South Bolivia with an allopatric distribution of the two species. COI barcode sequences indicate relatively strong divergence within C. bijuga, C. mayrabonillae, C. serrabonita and C. tenellus.     

Fishborne Trematode Metacercariae in Freshwater Fish from Guangxi Zhuang Autonomous Region, China

A survey was performed to investigate the infection status of fishborne trematode (FBT) metacercariae in freshwater fish from Guangxi Zhuang Autonomous Region, China. A total of 307 freshwater fish of 31 species were collected from 5 administrative regions of Guangxi Zhuang Autonomous Region. They were examined by artificial digestion method from July 2003 to August 2004. No metacercariae were detected in fish from Fusui-xian. In fish from Mashan-xian and a market in Nanning, 3 species of metacercariae, Haplorchis taichui, Haplorchis pumilio, and Centrocestus formosanus, were mainly detected. Metacercariae (8 in number) of Clonorchis sinensis were found in 1 Chanodichthys dabryi purchased from a market in Nanning. In fish from Yangshuo, Metagonimus yokogawai metacercariae were detected from all 18 fish species examined. Total 13 C. sinensis metacercariae were found in 3 out of 10 Hemibarbus maculatus from Yangshuo. All 7 Zacco platypus from Yangshuo were infected with 8-112 Echinochasmus perfoliatus metacercariae. In fish from Binyang-xian, H. pumilo metacercariae were mainly detected in all 5 fish species examined, and only 1 metacercaria of C. sinensis was found in a Hemiculter leucisculus. From the above results, it was confirmed that some species of freshwater fish play a role of second intermediate hosts for FBT in Guangxi Zhuang Autonomous Region, China. In particular, 4 species of intestinal flukes, M. yokogawai, H. taichui, H. pumilio, and C. formosanus, were prevalent in fish hosts, whereas C. sinensis metacercariae were detected only in 3 fish species.     

Temperature Effects on Heterorhabditis megidis and Steinernema carpocapsae Infectivity to Galleria mellonella

The effect of temperature on the infection of larvae of the greater wax moth, Galleria mellonella, by Heterorhabditis megidis H90 and Steinernema carpocapsae strain All, was determined. For both species, infection, reproduction, and development were fastest at 20 to 24 °C. Infection by both H. megidis and S. carpocapsae occurred between 8 and 16 °C; however, neither species reproduced at 8 °C. Among the nematodes used in experiments at 8 °C, no H. megidis and very few S. carpocapsae developed beyond the infective juvenile stage. Compared with H. megidis, S. carpocapsae invaded and killed G. mellonella larvae faster at 8 to 16 °C. By comparing invasion rates, differences in infectivity between the two nematode species were detected that could not be detected in conventional petri dish bioassays where mortality was measured after a specified period. Invasion of G. mellonella larvae by H. megidis was faster at 24 than at 16 °C.     

Horizontal transmission of a microsporidium from the convergent lady beetle, Hippodamia convergens Guérin-Méneville (Coleoptera: Coccinellidae), to three coccinellid species of Nova Scotia

Convergent lady beetles, Hippodamia convergens Guérin-Méneville, are a popular choice for aphid control in North America. An unidentified microsporidium was found in H. convergens adults that were purchased from a commercial insectary in 2004. This study examined egg cannibalism and egg predation as a means of horizontal transmission of the unidentified microsporidium among H. convergens larvae and three coccinellid species found in Nova Scotia: Coccinella septempunctata (seven-spotted lady beetle), C. trifasciata perplexa (three-banded lady beetle), and Harmonia axyridis (multicolored Asian lady beetle). The microsporidium was transmitted with 100% efficiency when first instars fed on microsporidia-infected eggs. Mean spore count data from smear preparations of infected beetles suggest that the infection was as heavy in C. trifasciata perplexa (a native coccinellid) (11.2 ± 0.96 spores/100 μm²) as it was in H. convergens (the natural host) (12.8 ± 1.16) but lighter in the introduced species C. septempunctata (7.5 ± 0.65) and H. axyridis (0.8 ± 0.11). For all of the beetle species examined, larval development was significantly longer for microsporidia-infected individuals than for their uninfected cohorts. The microsporidium had no effect on larval mortality. Based on the results of this study, field-collected H. convergens should be examined for microsporidia and uninfected individuals should be used to rear individuals for release in biological control programs. However, this is unlikely to happen because H. convergens are relatively easy and inexpensive to collect from their overwintering sites for redistribution.