Vectorial Competence of Larvae and Adults of Alphitobius diaperinus in the Transmission of Salmonella Enteritidis in Poultry

Introduction: The ingestion of food products originating from poultry infected with Salmonella spp. is one of the major causes of food poisoning in humans. The control of poultry salmonellosis is particularly difficult since birds are asymptomatic and numerous factors may expedite the maintenance of bacteria in poultry production facilities. Objective: The aim of the study was to determine the vectorial capacity of adults and larvae of Alphitobius diaperinus (Coleoptera: Tenebrionidae) in the experimental transmission of Salmonella Enteritidis phage type 4 to 1-day-old specific pathogen-free White Leghorn chicks. Methods: Adult insects and larvae were starved for 1 day, fed for 24 h or 7 days on sterile ration that had been treated with Salmonella Enteritidis phage type 4, and the levels of bacterial infection were determined. Infected adult insects and larvae were fed to groups of day-old chicks, after which bacteria were recovered from cecum, liver, and spleen samples over a 7-day period. Results: Infected larvae were more efficient than adult insects in transmitting Salmonella Enteritidis to chicks. Higher concentrations of bacteria could be reisolated from the cecum, liver, and spleen of chicks that had ingested infected larvae compared with those that had ingested infected adults. Conclusions: The control of A. diaperinus, and particularly of the larvae, represents a critical factor in the reduction of Salmonella spp. in poultry farms.

Notes on Population Dynamics of Amblyomma Ticks (Acari: Ixodidae) in Brazil

Previous population dynamics data, generated for Amblyomma parvum Aragao and Aniblyomma cajennense (Fabricius) in Argentina and southeastern Brazil, have indicated that these ticks complete I generation per year, with larvae predominating in autumn, nymphs in winter, and mostly adults during spring and summer. The present study reports population dynamics data for free-living Amblyomma spp. ticks in northern Brazil (Amazon forest, latitude 10 degrees S, 63 degrees W), and for Amblyomma spp. ticks collected oil birds in Southeastern Brazil (latitude 23 degrees S, 45 degrees W). In northern Brazil, adult ticks predominated from mid-spring to mid-autumn, larvae predominated in early winter, and nymphs from mid-winter to mid-spring. Seven Amblyomma spp. were identified, although A. cajannense predominated in I of the 2 sites sampled. In southeastern Brazil, larval infestations on birds peaked in autumn, followed by a nymphal infestation peak in late winter. At least 32% and 75% of these larvae and nymphs, respectively, were identified as Amblyomma longirostre (Koch). Similar to previous work, the present study showed that Amblyomma spp. larvae and nymphs predominated during autumn-winter months, and mostly adults during spring-summer months, a pattern compatible with 1 genration/yr, even at latitude 10 degrees S in northern Brazil.

DESCRIPTION OF ADULTS AND NYMPH, AND REDESCRIPTION OF THE LARVA, OF ORNITHODOROS MARINKELLEI (ACARI: ARGASIDAE), WITH DATA ON ITS PHYLOGENETIC POSITION

The argasid tick Ornithodoros marinkellei Kohls, Clifford, and Jones, 1969 was described 4 decades ago based on larval specimens collected from bats (Pteronotus spp.) in Colombia and Panama. Thereafter, larval O. marinkellei parasitizing bats were reported from Venezuela, Guyana, and Brazil. Herein, we describe the adults and nymph, and redescribe the larva of O. marinkellei based on specimens recently collected in the western Brazilian Amazon region. In contrast to all other known adult argasids, the idiosoma of both males and females of O. marinkellei is covered with sclerotized plaques. The idiosoma of the nymph of O. marinkellei is entirely micromamillated, and differs from the adults by the absence of plaques. The larva of O. marinkellei is morphologically similar to the larvae of the 2 other species belonging to the subgenus Subparmatus, i.e., Ornithodoros viguerasi Cooley and Kohls, 1941 and Ornithodoros mormoops Kohls, Clifford, and Jones, 1969. Because of the long and narrow dorsal plate, the larva of O. marinkellei is readily distinguished from O. viguerasi and O. mormoops. Comparison of our larvae from Brazil with O. marinkellei paratype specimens from Colombia confirmed their taxonomic identification. However, a few morphological differences, particularly in the size of the gnathosoma, were observed. Further studies are necessary to clarify whether O. marinkellei is a complex of different species, or a single species represented by morphologically polymorphic, and geographically distinct populations. Partial mitochondrial 16S rDNA gene sequences were generated for O. marinkellei specimens from Brazil, and compared with available homologous sequences in GenBank. Phylogenetic analyses revealed O. marinkellei to be distinct from the remaining argasid species available in GenBank, including other bat-associated tick species that are found in sympatry with O. marinkellei in the Neotropical region.

Experimental Infection of Rhipicephalus sanguineus Ticks with the Bacterium Rickettsia rickettsii, Using Experimentally Infected Dogs

We evaluated if Rickettsia rickettsii-experimentally infected dogs could serve as amplifier hosts for Rhipicephalus sanguineus ticks. In addition, we checked if Rh. sanguineus ticks that acquired Ri. rickettsii from dogs could transmit the bacterium to susceptible hosts (vector competence), and if these ticks could maintain the bacterium by transstadial and transovarial transmissions. Uninfected larvae, nymphs, and adults of Rh. sanguineus were allowed to feed upon three groups of dogs: groups 1 (G1) and 2 (G2) composed of Ri. rickettsii-infected dogs, infected intraperitoneally and via tick bites, respectively, and group 3 composed of uninfected dogs. After larval and nymphal feeding on rickettsemic dogs, 7.1-15.2% and 35.8-37.9% of the molted nymphs and adults, respectively, were shown by polymerase chain reaction (PCR) to be infected by Ri. rickettsii, confirming that both G1 and G2 dogs were efficient sources of rickettsial infection (amplifier host), resulting in transstadial transmission of the agent. These infected nymphs and adults successfully transmitted Ri. rickettsii to guinea pigs, confirming vector competence after acquisition of the infection from rickettsemic dogs. Transovarial transmission of Ri. rickettsii was observed in engorged females that had been infected as nymphs by feeding on both G1 and G2 dogs, but not in engorged females that acquired the infection during adult feeding on these same dogs. In the first case, filial infection rates were generally <50%. No tick exposed to G3 dogs was infected by rickettsiae in this study. No substantial mortality difference was observed between Ri. rickettsii-infected tick groups (G1 and G2) and uninfected tick group (G3). Our results indicate that dogs can be amplifier hosts of Ri. rickettsii for Rh. sanguineus, although only a minority of immature ticks (<45%) should become infected. It appears that Rh. sanguineus, in the absence of horizontal transmission, would not maintain Ri. rickettsii through successive generations, possibly because of low filial infection rates.

Deep sequencing of New World screw-worm transcripts to discover genes involved in insecticide resistance

Background: The New World screw-worm (NWS), Cochliomyia hominivorax, is one of the most important myiasis-causing flies, causing severe losses to the livestock industry. In its current geographical distribution, this species has been controlled by the application of insecticides, mainly organophosphate (OP) compounds, but a number of lineages have been identified that are resistant to such chemicals. Despite its economic importance, only limited genetic information is available for the NWS. Here, as a part of an effort to characterize the C. hominivorax genome and identify putative genes involved in insecticide resistance, we sampled its transcriptome by deep sequencing of polyadenylated transcripts using the 454 sequencing technology. Results: Deep sequencing on the 454 platform of three normalized libraries (larval, adult male and adult female) generated a total of 548,940 reads. Eighteen candidate genes coding for three metabolic detoxification enzyme families, cytochrome P450 monooxygenases, glutathione S transferases and carboxyl/cholinesterases were selected and gene expression levels were measured using quantitative real-time polymerase chain reaction (qRT-PCR). Of the investigated candidates, only one gene was expressed differently between control and resistant larvae with, at least, a 10-fold down-regulation in the resistant larvae. The presence of mutations in the acetylcholinesterase (target site) and carboxylesterase E3 genes was investigated and all of the resistant flies presented E3 mutations previously associated with insecticide resistance. Conclusions: Here, we provided the largest database of NWS expressed sequence tags that is an important resource, not only for further studies on the molecular basis of the OP resistance in NWS fly, but also for functional and comparative studies among Calliphoridae flies. Among our candidates, only one gene was found differentially expressed in resistant individuals, and its role on insecticide resistance should be further investigated. Furthermore, the absence of mutations in the OP target site and the high frequency of mutant carboxylesterase E3 indicate that metabolic resistance mechanisms have evolved predominantly in this species.

A sampling device to quantify offspring release of sessile marine invertebrates

Quantifying the rate of propagule release is of most importance to estimate reproductive output of natural populations, but simple methods to obtain such data are seldom reported. We designed and tested an inexpensive apparatus capable of reliably measure the release of gametes, eggs or larvae of sessile marine invertebrates in vertical walls. A population of the acom barnacle Chthamalus bisinuatus was sampled with this trap over 68d to obtain a time series of naupliar release. An apparent semilunar trend is shown, indicating the effectiveness of this sampling method.

Retention-favorable timing of propagule release in barnacles and periwinkles

Recent studies on the ecology of marine larvae suggest that retention near coastal areas and self-recruitment are probably much more common than previously thought. In light of this, the advective potential of pelagic stages can be partially determined by the timing of propagule release. We sampled the upper-shore levels of a subtropical coastline in southeastern Brazil to examine the temporal patterns of propagule release for the common barnacle Chthamalus bisinuatus and the periwinkle Nodilittorina lineolata. The release timing in both species was very consistent between sites a few kilometers apart. Barnacles released nauplii in a rhythmic pattern, mostly coinciding with neap tides, when the speed of tidal currents was lowest. There was no variation in propagule release in relation to diel or flood-ebb tidal cycles. Periwinkles released propagules in a very irregular pattern, which remarkably matched a time series of wave heights. Egg capsules were released during periods of rough seas, when onshore surface currents were expected to prevail. Eggs were released mostly during ebb tides, and there was no diel variation. Propagule release rhythms are usually viewed as a means to ensure fast offshore advection, thus avoiding presumed hazardous conditions for larvae. However, propagule release strategies in these upper-shore species would more likely contribute to the retention of offspring close to parental stocks.

Shelf and estuarine transport mechanisms affecting the supply of competent larvae in a suite of brachyuran crabs with different life histories

Supply of competent larvae to the benthic habitat is a major determinant of population dynamics in coastal and estuarine invertebrates with an indirect life cycle. Larval delivery may depend not only on physical transport mechanisms, but also on larval behavior and physiological progress to the competent stage. Yet, the combined analysis of such factors has seldom been attempted. We used time-series analyses to examine tide- and wind-driven mechanisms responsible for the supply of crab megalopae to an estuarine river under a major marine influence in SW Spain, and monitored the vertical distribution of upstream moving megalopae, their net flux and competent state. The species Panopeus africanus (estuarine), Brachynotus sexdentatus (euryhaline) and Nepinnotheres pinnotheres (coastal) comprised 80% of the whole sample, and responded in a similar way to tide and wind forcing. Tidal range was positively correlated to supply, with maxima 0 to 1 d after spring tides, suggesting selective tidal stream transport. Despite being extensively subjected to upwelling, downwind drift under the effect of westerlies, not Ekman transport, explained residual supply variation at our sampling area. Once in the estuary, net flux and competence state matched the expected trends. Net upstream flux increased from B. sexdentatus to P. africanus, favoring transport to a sheltered coastal habitat (N. pinnotheres), or to the upper estuary (P. africanus). Competence state was highest in N. pinnotheres, intermediate in B. sexdentatus and lowest in P. africanus, as expected if larvae respond to cues from adequate benthic habitat. P. africanus megalopae were found close to the bottom, not above, rendering slower upstream transport than anticipated.

Morphological Studies on the Mature Worker Larvae of Paratrechina fulva (Hymenoptera, Formicidae)

The ant Paratrechina fulva is native to Brazil, but has spread to other countries where it has become a serious pest. Their larvae are known from a preliminary description made without the aid of scanning electron microscopy. The present investigation aimed at describing the mature worker larvae of this species in deeper detail, while identifying useful morphological characters for taxonomy and cladistics. General morphological traits typical of Paratrechina larvae described so far were also confirmed with this species, particularly regarding body shape, types of hairs, and distribution pattern of head sensilla, but P fulva presented unusually shaped mandibles.

Description of the immatures of Linepithema humile Mayr (Hymenoptera: Formicidae)

Linepithema humile Mayr is an ant species originally native to South America that has been spread accidently throughout the globe through international trade. It is a serious urban and crop pest. Despite its economic importance, little is known about the larvae of this species apart from a brief description based on a few specimens. The present investigation is aimed at describing every immature stage of L. humile. Three larval instars were determined through the frequency distribution of the maximum width of head capsules from a sample of 525 larvae. The morphological descriptions were based on 150 eggs, 70 larvae, and 90 pupae examined by light and scanning electron microscopy. Some morphological characteristics reported to be typical of Linepithema Mayr larvae were confirmed - dolichoderoid body shape, presence of dorsal protuberance, sparse simple body hairs, presence of nine pairs of spiracles and dolichoderoid mandibles. We concluded that an earlier published description was based on queen larvae, and that the protuberance is only present in worker larvae. The information provided in this study may aid ant systematics and phylogenetics, as well provide a better understanding of the biology of this species.

Description of the Immatures of Workers of the Weaver Ant, Camponotus textor (Hymenoptera: Formicidae)

The weaver ants Camponotus textor (Forel) (Hymenoptera: Formicidae) are native to Central and South America, where they use their larvae to build silken nests by sewing tree leaves together. Few Studies have been conducted with this species, and little is known about the morphology of their larvae. Tie present paper estimated the number of larval instars of C. textor and presents a detailed morphological description of each immature stage based on light and electron microscopic observations. The number of larval in stars was estimated as four based oil the frequency distribution of larval head widths. Tic larvae of this species presented some typical characteristics of Camponotus (Mayr) larvae: body shape `pogonomyrmecoid`, ten pairs of spiracles, antennae with three sensilla, conspicuous `chiloscleres` on the labrum, and mature larvae with pronounced labial pseudopalps. Unprecedented characteristics would include: great diversity of body hair types, and `camponotoid` mandibles but with four medial denticles. This information can aid biological and taxonomic studies with these ants, and may be useful for ant systematics.

Description of the Immatures of Workers of the Ant Linepithema micans Forel (Hymenoptera: Formicidae)

Linepithema micans Forel is a poorly studied species that is now considered a pest in vineyards in South Brazil. Larval characters have been used in a few phylogenetic studies, and their importance greatly depends in the amount of available information on different species. This study presents a complete panorama on the external morphology of the immatures of L. micans based on observations by light and scanning electron microscopy. The number of larval instars was estimated as three based on the frequency distribution of head widths of 965 larvae. Larvae of L. micans were similar to other Linepithema Mayr in the general format of body and mandibles, presenting a dorsal abdominal protuberance, nine pairs of spiracle, and unbranched hairs. On the other hand, L. micans was unique for having shorter hairs, predominantly denticulate, intraspecific variation in the number of antennal sensilla and in the types of sensilla on the labial palps were reported. Microsc. Res. Tech. 74:337-342, 2011. (C) 2010 Wiley-Liss, Inc.

Saxitoxins accumulation by freshwater tilapia (Oreochromis niloticus) for human consumption

The accumulation of saxitoxins (STXs) in fish from freshwater aquaculture was investigated for the first time in the present study. Cyanotoxins have been monitored in liver and muscle samples of Oreochromis miloticus by chromatographic methods, both before and after the deputation process. The results show that tilapia can accumulate STXs. Our findings suggest that deputation with clean water is an alternative process to eliminate STXs from fish and, therefore, improve the safety of tilapia for consumers. (C) 2009 Elsevier Ltd. All rights reserved.

Pupal Development of Ceratitis capitata (Diptera: Tephritidae) and Diachasmimorpha longicaudata (Hymenoptera: Braconidae) at Different Moisture Values in Four Soil Types

This study aimed to evaluate adult emergence and duration of the pupal stage of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and emergence of the fruit fly parasitoid, Diachasmimorpha longicaudata (Ashmead), under different moisture conditions in four soil types, using soil water matric potential Pupal stage duration in C capitata was influenced differently for males and females In females, only soil type affected pupal stage duration, which was longer in a clay soil In males, pupal stage duration was individually influenced by moisture and soil type, with a reduction in pupal stage duration in a heavy clay soil and in a sandy clay, with longer duration in the clay soil As allude potential decreased, duration of the pupal stage of C capitata males increased, regardless of soil type C capitata emergence was affected by moisture, regardless of soil type, and was higher in drier soils The emergence of D longicaudata adults was individually influenced by soil type and moisture factors, and the number of emerged D longicaudata adults was three times higher in sandy loam and lower in a heavy clay soil Always, the number of emerged adults was higher at higher moisture conditions C capitata and D longicaudata pupal development was affected by moisture and soil type, which may facilitate pest sampling and allow release areas for the parasitoid to be defined under field conditions.

Breeding of Ornidia obesa (Diptera: Syrphidae: Eristalinae) on Pig Carcasses in Brazil

On Mho obesa F. (Diptera: Syrphidae) is usually neglected in forensic entomology, although adults are rather frequent on vertebrate carrion. In this study, conducted in southeastern Brazil in 2008, we used two pig carcasses, one killed by cocaine overdose and the other by shooting, to evaluate mainly the possible influences of the type of death on the larval development of O. obesa in the pig remains. We recorded the breeding of 218 adult specimens of this syrphid fly from the carcass killed by shooting, and none from the carcass killed by cocaine. These observations may open a new perspective for the use of O. obesa in forensic studies, considering its breeding preferences and its complete development on vertebrate carrion.