Effect of larval handling on black soldier fly life history traits and bioconversion efficiency.

Introduction: The black soldier fly is considered the most promising insect species for mass production; however, information on the effects of handling, which is unavoidable during experimental trials and rearing practices, is still limited. Materials and methods: To address this gap, three different manipulation intensities were tested on 100 6-day-old larvae per replica (6 replicates/treatments) fed on Gainesville diet: (1) hard-handled (HH), larvae underwent continuous manipulation until the end of larval stage, (2) soft-handled (SH), larvae were manipulated after the appearance of the first prepupa, (3) no-handled (NH), larvae remained untouched. Every 4 days from the beginning to the end of the larval stage, the manipulations lasted 30 min and occurred under laboratory conditions (20°C). During the sampling operations, at least 30 larvae were randomly extracted, washed, dried, and weight-mimicked. At the end of larval stage, all the boxes remained untouched until the adult fly stage, and the emergency rate and sex ratio were evaluated on dead flies. Data were statistically analyzed using IBM SPSS V20.0.0 software and the considered significance level was p < 0.05. Results: The larval stage lasted 8.2 days for both HH and SH (p > 0.05). Despite the HH larvae being the most manipulated, no difference was also observed in final weight (HH, 160 mg; SH, 150 mg; p > 0.05) and survival rate (HH, 96.2%; SH, 94.5%; p > 0.05). The manipulation did not influence the bioconversion capacity of the larvae (bioconversion efficiency corrected for the residue: HH, 14.3%; SH, 12.91%; reduction rate: HH, 58.4%; SH, 55.9%; waste reduction index: HH, 7.28%/day; SH, 7.25%/day; p > 0.05). Finally, the development time from larva to fly (about 20.7; p > 0.05), the emergency rate (NH: 92.8%; SH: 89.5%; HH: 82.7%) and sex ratio (~1.2% to male flies) were not affected by the handling (p > 0.05). Discussion: In conclusion, the handling procedures used in the current study did not influence the life history traits of the black soldier fly. However, further studies are needed to evaluate if different experimental protocols on various scales, the colony strain or other handling procedures may suggest a different scenario or confirm the results.

Developmental Biology and Identification of a Garden Pest, Otiorhynchus (Podoropelmus) smreczynskii Cmoluch, 1968 (Coleoptera, Curculionidae, Entiminae), with Comments on Its Origin and Distribution.

The mature larva and pupa of Otiorhynchus smreczynskii are described and illustrated with complete chaetotaxy for the first time. Five larval instars and the factors for larval growth are determined, and the larval development of this species is comprehensively described. In order to confirm species affiliation, selected larvae were subjected to genetic analysis (mtCOI). Host plants and unique feeding signs of some Entiminae species are presented, and all available data on development are documented and interpreted. Additionally, the morphometry of 78 specimens (48 of O. smreczynskii and 30 of O. rotundus) was examined in order to verify the usefulness of morphological features in distinguishing both species. The female genitalia of both species are illustrated, described and compared with each other for the first time. Finally, the updated distribution of O. smreczynskii is given, and a possible origin of O. smreczynskii and O. rotundus is proposed.

RNA-Sequencing of Heterorhabditis nematodes to identify factors involved in symbiosis with Photorhabdus bacteria.

BACKGROUND: Nematodes are a major group of soil inhabiting organisms. Heterorhabditis nematodes are insect-pathogenic nematodes and live in a close symbiotic association with Photorhabdus bacteria. Heterorhabditis-Photorhabdus pair offers a powerful and genetically tractable model to study animal-microbe symbiosis. It is possible to generate symbiont bacteria free (axenic) stages in Heterorhabditis. Here, we compared the transcriptome of symbiotic early-adult stage Heterorhabditis nematodes with axenic early-adult nematodes to determine the nematode genes and pathways involved in symbiosis with Photorhabdus bacteria. RESULTS: A de-novo reference transcriptome assembly of 95.7 Mb was created for H. bacteriophora by using all the reads. The assembly contained 46,599 transcripts with N50 value of 2,681 bp and the average transcript length was 2,054 bp. The differentially expressed transcripts were identified by mapping reads from symbiotic and axenic nematodes to the reference assembly. A total of 754 differentially expressed transcripts were identified in symbiotic nematodes as compared to the axenic nematodes. The ribosomal pathway was identified as the most affected among the differentially expressed transcripts. Additionally, 12,151 transcripts were unique to symbiotic nematodes. Endocytosis, cAMP signalling and focal adhesion were the top three enriched pathways in symbiotic nematodes, while a large number of transcripts coding for various responses against bacteria, such as bacterial recognition, canonical immune signalling pathways, and antimicrobial effectors could also be identified. CONCLUSIONS: The symbiotic Heterorhabditis nematodes respond to the presence of symbiotic bacteria by expressing various transcripts involved in a multi-layered immune response which might represent non-systemic and evolved localized responses to maintain mutualistic bacteria at non-threatening levels. Subject to further functional validation of the identified transcripts, our findings suggest that Heterorhabditis nematode immune system plays a critical role in maintenance of symbiosis with Photorhabdus bacteria.

Integrative Transcriptomics and Proteomics Analyses to Reveal the Developmental Regulation of Metorchis orientalis: A Neglected Trematode With Potential Carcinogenic Implications.

Metorchis orientalis is a neglected zoonotic parasite of the gallbladder and bile duct of poultry, mammals, and humans. It has been widely reported in Asian, including China, Japanese, and Korea, where it is a potential threat to public health. Despite its significance as an animal and human pathogen, there are few published transcriptomic and proteomics data available. Transcriptome Illumina RNA sequencing and label-free protein quantification were performed to compare the gene and protein expression of adult and metacercariae-stage M. orientalis, resulting in 100,234 unigenes and 3,530 proteins. Of these, 13,823 differentially expressed genes and 1,445 differentially expressed proteins were identified in adult versus metacercariae. In total, 570 genes were differentially expressed consistent with the mRNA and protein level in the adult versus metacercariae stage. Differential gene transcription analyses revealed 34,228 genes to be expressed in both stages, whereas 66,006 genes showed stage-specific expression. Compared with adults, the metacercariae stage was highly transcriptional. GO and KEGG analyses based on transcriptome and proteome revealed numerous up-regulated genes in adult M. orientalis related to microtubule-based processes, microtubule motor activity, and nucleocytoplasmic transport. The up-regulated genes in metacercariae M. orientalis were mainly related to transmembrane receptor protein serine/threonine kinase activity, transmembrane receptor protein serine/threonine kinase signaling pathway. Transcriptome and proteome comparative analyses showed numerous up-regulated genes in adult stage were mainly enriched in actin filament capping, spectrin, and glucose metabolic process, while up-regulated genes in metacercariae stage were mainly related to cilium assembly, cilium movement, and motile cilium. These results highlight changes in protein and gene functions during the development of metacercariae into adults, and provided evidence for the mechanisms involved in morphological and metabolic changes at both the protein and gene levels. Interestingly, many genes had been proved associated with liver fibrosis and carcinogenic factors were identified highly expressed in adult M. orientalis, which suggests that M. orientalis is a neglected trematode with potential carcinogenic implications. These data provide attractive targets for the development of therapeutic or diagnostic interventions for controlling M. orientalis.

Mechanism of the Passage of Angiostrongylus cantonensis across the Final Host Blood-Brain Barrier Using the Next-Generation Sequencing.

BACKGROUND: Multicellular parasites Angiostrogylus cantonensis larvae develop in the final host rat brain at the fourth stage (L4) and migrate to the lungs by the adult stage. The potential mechanism of its blood-brain barrier (BBB) passage remains unclear. METHODS: By using Illumina Hiseq/Miseq sequencing, we obtained the transcriptomes of 3 groups of adult males and 3 groups of female of A. cantonensis to generate similarly expressed genes (SEGs) between 2 genders at the adult stage. Next 2 groups of L4 expressed genes were used to compared with SEGs to create differentially expressed genes (DEGs) between 2 life stages to unlock potential mechanism of BBB passage. RESULTS: In total, we obtained 381 581 802 clean reads and 56 990 699 010 clean bases. Of these, 331 803 unigenes and 482 056 transcripts were successfully annotated. A total of 3 166 DEGs between L4 and adults SEGs were detected. Annotation of these DEGs showed 167 were down-regulated and 181 were up-regulated. Pathway analysis exhibited that calcium signaling pathway, the ECM-receptor interaction, focal adhesion, and cysteine and methionine metabolism were highly associated with DEGs. The function of these pathways might be related to BBB traversal, as well as neuro-regulation, interactions between parasite and host, environmental adaption. CONCLUSION: This study expanded the regulatory characteristics of the two important life stages of A. cantonensis. This information may provide a better appreciation of the biological features of the stages of the parasitic A. cantonensis.

Main Pathways and Ion Channels Differentially Expressed in the Transcriptome of Male and Female Adult Angiostrongylus cantonensis Using a Deep Sequencing Approach.

BACKGROUND: The adult stage is an important period in the life cycle of Angiostrongylus cantonensis, as it is at this stage that male and female worms produce thousands of fertilized eggs daily. METHODS: To explore the transcriptional details of adult male and female A. cantonensis, three groups of male and female adult worms were collected, and their transcriptome profiles were analyzed using an Illumina next-generation sequencing platform. A total of 283,910,174 clean reads were obtained, and 137,626 unigenes and 237,059 transcripts were then generated. Unigenes were successfully annotated by querying the Gene Ontology (GO), the Kyoto Encyclopedia of Genes and Genomes (KEGG), NCBI non-redundant protein sequences (NR), PFAM, STRING, and SWISS-PROT databases. Then, differentially expressed genes (DEGs) between the 2 genders were identified. The GO and KEGG databases were used for DEG annotation, and a number of DEG annotations were enriched. RESULTS: The results obtained from querying DEGs using the GO and KEGG databases revealed that male and female adult worms exhibited differences in metabolism and production. Protein phosphorylation, ion transport, and calcium transport were all significantly enriched according to GO annotation. A number of other pathways were also enriched according to KEGG enrichment annotation, including the pentose phosphate pathway, nitrogen metabolism, oocyte meiosis pathway, neuroactive ligand-receptor interaction, calcium signaling pathway, transforming growth factor ß (TGF-ß) signaling pathway etc. CONCLUSION: We hypothesized that the nervous system of the worm plays a key role in the physiological regulation of adult A. cantonensis, and based on this, the function of the calcium-signaling pathway should be investigated.

Integrated management of blood-feeding arthropods in veterinary teaching facilities - Part 2: overview of control methods against adults and immature stages.

Numerous arthropod species are involved in the vector-borne transmission of pathogens either to animals and/or to humans. Part 1 of this paper was dedicated to a review of these species, and their role in the transmission of pathogens in North-western Europe. This part will discuss the different anti-arthropod control methods, which are either general, used as good management practices, or arthropod-specific. The majority of these measures are efficient against several arthropod groups. Management of the environment is crucial for controlling the immature stages of winged arthropods, fleas and lice, but also ticks. Spraying pesticides should be considered carefully, because of the risk of emergence of resistance and the negative impact on the environment and non-targeted insects. Monitoring of haematophagous arthropods is useful when considering its use in the validation of control measures, the follow-up of endemic populations, vigilance for emergence of new species, and the detection of pathogens and, indirectly, resistance to chemicals. Monitoring also helps to determine the most appropriate timing and location for implementing control measures. It is strongly advised to combine control methods targeting adults and others addressing immature stages. Even if challenging, their combination under an integrated pest management programme should be preferred. Indeed, integrated vector management aims at making vector control more efficient, cost-effective, ecologically sound and sustainable.

Nombre d'espèces d'arthropodes participent à la transmission vectorielle d'agents pathogènes aux animaux et/ou aux humains. Dans une première partie de leur étude, les auteurs ont donné une vue d'ensemble de ces espèces et expliqué le rôle qu'elles jouent dans la transmission d'agents pathogènes dans le nord de l'Europe occidentale. Cette deuxième partie est consacrée aux différentes méthodes de contrôle des arthropodes ; elles se répartissent entre d'une part, des méthodes générales qui correspondent aux bonnes pratiques de gestion, et d'autre part, des méthodes ciblant spécifiquement les arthropodes. La plupart de ces mesures sont efficaces contre plusieurs groupes d'arthropodes. Il est essentiel de respecter les principes de bonne gestion de l'environnement afin de parvenir à contrôler les stades immatures des arthropodes ailés tels que les puces et les poux, mais aussi les tiques. L'utilisation de pesticides par aérosols devrait être envisagée avec prudence en raison du risque d'émergence de résistances et de l'impact négatif de ces produits sur l'environnement et sur les insectes non-cibles. La surveillance des arthropodes hématophages présente un intérêt lorsqu'on l'applique pour la validation des mesures de contrôle, le suivi des populations endémiques, la veille face à l'émergence d'espèces nouvelles et la détection d'agents pathogènes, ainsi qu'indirectement pour détecter les résistances aux agents chimiques. La surveillance permet également de déterminer quand et où mettre en oeuvre les mesures de contrôle pour plus de résultats. Il est conseillé d'associer les méthodes ciblant les adultes et celles ciblant les stades immatures. Malgré les difficultés d'une telle association, il conviendra de la privilégier dans le cadre d'un programme intégré de gestion des nuisibles. En effet, la gestion intégrée des vecteurs vise à rendre la lutte anti-vectorielle à la fois plus efficace, plus rentable, plus rationnelle au plan écologique et plus durable.

Numerosas especies de artrópodo participan en la transmisión vectorial de patógenos a animales o personas. En la primera parte de esta serie de artículos se hace un repaso de estas especies y se explica el papel que cumplen en la transmisión de patógenos en el noroeste de Europa. En esta segunda parte se examinan los diferentes métodos de lucha contra los artrópodos, que pueden ser genéricos, esto es, utilizados como buena práctica de gestión, o ir dirigidos selectivamente contra artrópodos. La mayoría de estas medidas son eficaces contra varios grupos de artrópodos. La gestión de las condiciones del entorno es crucial para combatir a las fases inmaduras de artrópodos alados, pulgas y piojos, así como a las garrapatas. El rociado con plaguicidas es un recurso que conviene estudiar con detenimiento, dado el riesgo de que surjan resistencias y su negativo impacto en el medio ambiente y en otros insectos que no se pretende dañar. La vigilancia de las poblaciones de artrópodos hematófagos resulta útil cuando se contempla su uso para la validación de medidas de lucha, el seguimiento de poblaciones endémicas, la vigilancia de la aparición de nuevas especies o la detección de patógenos e, indirectamente, la resistencia a productos químicos. La vigilancia también ayuda a determinar el momento y lugar idóneos para aplicar medidas de lucha. Se aconseja vivamente combinar métodos de lucha dirigidos contra los adultos con otros que combatan a las fases inmaduras. La aplicación de este tipo de planteamiento combinado, por dificultosa que resulte, será siempre la opción preferible para formar parte de un programa de gestión integrada de plagas. La lucha integrada contra los vectores, en efecto, tiene por objetivo hacer más eficaz, rentable, ecológicamente sensata y sostenible la lucha contra los vectores.

Molecular Characterization of Copepod Photoreception.

Copepod crustaceans are an abundant and ecologically significant group whose basic biology is guided by numerous visually guided behaviors. These behaviors are driven by copepod eyes, including naupliar eyes and Gicklhorn's organs, which vary widely in structure and function among species. Yet little is known about the molecular aspects of copepod vision. In this study we present a general overview of the molecular aspects of copepod vision by identifying phototransduction genes from newly generated and publicly available RNA-sequencing data and assemblies from 12 taxonomically diverse copepod species. We identify a set of 10 expressed transcripts that serve as a set of target genes for future studies of copepod phototransduction. Our more detailed evolutionary analyses of the opsin gene responsible for forming visual pigments found that all of the copepod species investigated express two main groups of opsins: middle-wavelength-sensitive (MWS) opsins and pteropsins. Additionally, there is evidence from a few species (e.g., Calanus finmarchicus, Eurytemora affinis, Paracyclopina nana, and Lernaea cyprinacea) for the expression of two additional groups of opsins-the peropsins and rhodopsin 7 (Rh7) opsins-at low levels or distinct developmental stages. An ontogenetic analysis of opsin expression in Calanus finmarchicus found the expression of a single dominant MWS opsin, as well as evidence for differences in expression across development in some MWS, pteropsin, and Rh7 opsins, with expression peaking in early naupliar through early copepodite stages.

Conservation and diversification of the transcriptomes of adult Paragonimus westermani and P. skrjabini.

BACKGROUND: Paragonimiasis is an important and widespread neglected tropical disease. Fifteen Paragonimus species are human pathogens, but two of these, Paragonimus westermani and P. skrjabini, are responsible for the bulk of human disease. Despite their medical and economic significance, there is limited information on the gene content and expression of Paragonimus lung flukes. RESULTS: The transcriptomes of adult P. westermani and P. skrjabini were studied with deep sequencing technology. Approximately 30 million reads per species were assembled into 21,586 and 25,825 unigenes for P. westermani and P. skrjabini, respectively. Many unigenes showed homology with sequences from other food-borne trematodes, but 1,217 high-confidence Paragonimus-specific unigenes were identified. Analyses indicated that both species have the potential for aerobic and anaerobic metabolism but not de novo fatty acid biosynthesis and that they may interact with host signaling pathways. Some 12,432 P. westermani and P. skrjabini unigenes showed a clear correspondence in bi-directional sequence similarity matches. The expression of shared unigenes was mostly well correlated, but differentially expressed unigenes were identified and shown to be enriched for functions related to proteolysis for P. westermani and microtubule based motility for P. skrjabini. CONCLUSIONS: The assembled transcriptomes of P. westermani and P. skrjabini, inferred proteins, and extensive functional annotations generated for this project (including identified primary sequence similarities to various species, protein domains, biological pathways, predicted proteases, molecular mimics and secreted proteins, etc.) represent a valuable resource for hypothesis driven research on these medically and economically important species.

Checklist of Ephemeroptera (Insecta) from São Mateus River Basin, Espírito Santo, Brazil

ABSTRACTIn order to expand the knowledge on the composition of Ephemeroptera from large rivers, we present, herein, the first survey of mayflies from the São Mateus River Basin, Espírito Santo State. Adults were collected biannually in 2012 and 2013 with the aid of Pennsylvania light trap in eleven points distributed in the main river of the river basin, São Mateus River and its two main tributaries, Cotaxé River (Braço Norte) and Cricaré River (Braço Sul). Thirty-three species were identified (22 nominal and 11 morphospecies) in 24 genera and five families. One genus and one species are reported for the first time from Brazil, three species from Southeastern Region of Brazil, and two species from the state. Five species and one genus not previously described were also found. Moreover, around 20% of the known species of mayfly registered from the state were found. This work reinforces the need to give more attention to research on large rivers due to the high potential for diversity, not only for Ephemeroptera, but also for other aquatic insects.

La proliferación de los miocitos ventriculares del corazón de mamífero adulto: un fenómeno esporádico pero factible / Proliferation of adult mammalian ventricular cardiomyocytes: A sporadic but feasible phenomenon

La proliferación de los miocitos que forman parte de los ventrículos cardíacos del mamífero adulto ha sido descartada por algunos investigadores con el argumento de que estas células están diferenciadas en forma terminal; sin embargo, este dogma ha sido puesto en duda a partir de los hallazgos de otros investigadores quienes han observado que estos miocitos pueden presentar los procesos necesarios para la proliferación, es decir síntesis de ADN, mitosis y citocinesis, cuando el miocardio se daña en forma experimental con estrategias de tipo farmacológico o quirúrgico, o debido a condiciones patológicas relacionadas con el sistema cardiovascular. Esta revisión integra algunos de los trabajos disponibles en la literatura que han evaluado la síntesis del ADN, mitosis y citocinesis en estas células, en el miocardio dañado, para saber si su proliferación puede ser considerada como un fenómeno factible. La revisión concluye con una reflexión sobre las perspectivas del conocimiento generado en esta área de estudio.

Proliferation of adult mammalian ventricular cardiomyocytes has been ruled out by some researchers, who have argued that these cells are terminally differentiated; however, this dogma has been rejected because other researchers have reported that these cells can present the processes necessary to proliferate, that is, DNA synthesis, mitosis and cytokinesis when the heart is damaged experimentally through pharmacological and surgical strategies or due to pathological conditions concerning the cardiovascular system. This review integrates some of the available works in the literature evaluating the DNA synthesis, mitosis and cytokinesis in these myocytes, when the myocardium is damaged, with the purpose of knowing if their proliferation can be considered as a feasible phenomenon. The review is concluded with a reflection about the perspectives of the knowledge generated in this area.

[Proliferation of adult mammalian ventricular cardiomyocytes: a sporadic but feasible phenomenon]. / La proliferación de los miocitos ventriculares del corazón de mamífero adulto: un fenómeno esporádico pero factible.

Proliferation of adult mammalian ventricular cardiomyocytes has been ruled out by some researchers, who have argued that these cells are terminally differentiated; however, this dogma has been rejected because other researchers have reported that these cells can present the processes necessary to proliferate, that is, DNA synthesis, mitosis and cytokinesis when the heart is damaged experimentally through pharmacological and surgical strategies or due to pathological conditions concerning the cardiovascular system. This review integrates some of the available works in the literature evaluating the DNA synthesis, mitosis and cytokinesis in these myocytes, when the myocardium is damaged, with the purpose of knowing if their proliferation can be considered as a feasible phenomenon. The review is concluded with a reflection about the perspectives of the knowledge generated in this area.

Ciclo biológico de Argas miniatus em laboratório: estágio adulto / Life cycle of Argas (Persicargas) miniatus in the laboratory: adult stage

Os parâmetros do ciclo biológico de Argas (Persicargas) miniatus, fase adulta, foram estudados em condições controladas (27±1°C e 80±10 por cento UR) e em condições ambientais não controladas (ambiente de laboratório nas estações chuvosa e seca). Nessas condições, os machos se originaram preferencialmente de N2 e as fêmeas de N3. As fêmeas realizaram até 18 posturas (em condições controladas), enquanto que as fêmeas mantidas nas estações chuvosa e seca realizaram até 9 e 12 posturas, respectivamente. O número total de ovos foi 1350 (em condições controladas), 443 ovos na estação chuvosa e 894 ovos na estação seca. Em jejum, os carrapatos machos sobreviveram por 165 dias (condições controladas) e 135 dias em ambiente de laboratório; as fêmeas sobreviveram por 300 dias em condições controladas e 240 dias no ambiente de laboratório. Nas condições ambientais não controladas, os parâmetros biológicos do estágio adulto de Argas (Persicargas) miniatus sofreram alterações negativas, principalmente durante a estação chuvosa.

The life cycle of Argas (Persicargas) miniatus in adult stage was studied in controlled conditions (27±1°C and 80±10 percent RH) and uncontrolled temperature and humidity conditions (room conditions during wet and dry season). Males originated preferentially from N2 and females from N3. Females in controlled conditions laid eggs up to 18 times, while females in uncontrolled room conditions, during wet and dry seasons, laid eggs 9 to 12 times, respectively. In controlled conditions, females laid 1350 eggs, whereas in uncontrolled conditions females laid 443 eggs during the wet season and 894 eggs during the dry season. Male ticks survived without feeding for 165 days in controlled conditions and 135 days in uncontrolled conditions, while females survived for 300 days in controlled conditions and 240 days in uncontrolled conditions. In ambient conditions, biological parameters of Argas (Persicargas) miniatus adult stage were negatively affected especially during the wet season when compared with adult stage kept in controlled condition.

Efficacy of Mebendazole Composite Cream in Treatment of Trichiniasis in Mice by Skin Administration / 第二军医大学学报

Male mice weighting 25-32g were infected with larvae of Trichinella spiralis. Skin administrations of mebendazole composite cream were carried out on the mice in 3 different experiment groups. It was found that there were the reduction rate of worms at 97.3%, 100% and 99. 7% in the adult worm group (50 mg/kg x 1 d), in the invasive larvae group (25 mg/kg x 7 d) and in the encysted larvae group (25 mg/kg x 7 d), respectively. These results indicate that mebendazole composite cream is very effective against both intestinal and muscular phases of T. spiralis.