Molecular analysis of the mitochondrial markers COI, 12S rDNA and 16S rDNA for six species of Iranian scorpions

Objectives Annually, 1.2 million humans are stung by scorpions and severely affected by their venom. Some of the scorpion species of medical importance have a similar morphology to species with low toxicity. To establish diagnostic tools for surveying scorpions, the current study was conducted to generate three mitochondrial markers, Cytochrome Oxidase I (COI gene), 12S rDNA and 16S rDNA for six species of medically important Iranian scorpions: Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus, M. eupeus, Odontobuthus doriae, and Scorpio maurus. Results Phylogenetic analyses of the obtained sequences corroborated the morphological identification. For the first time, 12S rDNA sequences are reported from Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus and M. eupeus and also the 16S rDNA sequence from Hottentotta saulcyi. We conclude that the mitochondrial markers are useful for species determination among these medically important species of scorpions

Virus Diversity and Loads in Crickets Reared for Feed: Implications for Husbandry

Insects generally have high reproductive rates leading to rapid population growth and high local densities; ideal conditions for disease epidemics. The parasites and diseases that naturally regulate wild insect populations can also impact when these insects are produced commercially, on farms. While insects produced for human or animal consumption are often reared under high density conditions, very little is known about the microbes associated with these insects, particularly those with pathogenic potential. In this study we used both target-free and targeted screening approaches to explore the virome of two cricket species commonly reared for feed and food, Acheta domesticus and Gryllus bimaculatus. The target-free screening of DNA and RNA from a single A. domesticus frass sample revealed that only 1% of the nucleic acid reads belonged to viruses, including known cricket, insect, bacterial and plant pathogens, as well as a diverse selection of novel viruses. The targeted screening revealed relatively high levels of Acheta domesticus densovirus, invertebrate iridovirus 6 and a novel iflavirus, as well as low levels of Acheta domesticus volvovirus, in insect and frass samples from several retailers. Our findings highlight the value of multiple screening approaches for a comprehensive and robust cricket disease monitoring and management strategy. This will become particularly relevant as-and-when cricket rearing facilities scale up and transform from producing insects for animal feed to producing insects for human consumption

Pan-european assessment, monitoring, and mitigation of stressors on the health of bees

As part of the activities foreseen by the POSHBEE project, we performed experiments to verify the toxicokinetics of three pesticides (the insecticide sulfoxaflor, the fungicide azoxystrobin and the herbicide glyphosate) in the bodies of social bees (Apis mellifera and Bombus terrestris) and solitary bees (Osmia bicornis). For each species all castes and sexes were studied. Based on the results from dose-response assays, sublethal doses were used to treat individuals, which were exposed to the chosen pesticides orally and by contact. Bees were then sampled at multiple time points post- exposure, to capture the breakdown of the active ingredients in the bodies of the organisms. Results of the chemical analyses on bee samples were used for the evaluation of the dynamics of the oral and contact acute exposure in the three species. Overall, the active ingredients sulfoxaflor, azoxystrobin and glyphosate degrade in all species, sex or caste. Nevertheless, there were some exceptions: in honey bee workers, glyphosate administered topically and azoxystrobin administered via the oral route seemed to remain stable in the bees’ bodies even 10 days after exposure (11% and 13% of degradation rate, respectively). We also observed a low degradation of sulfoxaflor following topical exposure in bumble bee queens (26% of degradation rate) and in Osmia bee females (22% of degradation rate). In bumble bees exposed topically to glyphosate degradation was lower than 50% for all sexes and castes. These results deserve further attention by researchers to understand the destiny of these molecules in the bee body and their effects therein.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation actio

Culturable bacteria associated with Anopheles darlingi and their paratransgenesis potential

Background Malaria remains a major public health problem in South America, mostly in the Amazon region. Among newly proposed ways of controlling malaria transmission to humans, paratransgenesis is a promising alternative. Paratransgenesis aims to inhibit the development of parasites within the vector through the action of genetically modified bacteria. The first step towards successful paratransgenesis in the Amazon is the identification of Anopheles darlingi symbiotic bacteria, which are transmitted vertically among mosquitoes, and are not pathogenic to humans. Methods Culturable bacteria associated with An. darlingi and their breeding sites were isolated by conventional microbiological techniques. Isolated strains were transformed with a GFP expressing plasmid, pSPT-1-GFP, and reintroduced in mosquitoes by feeding. Their survival and persistence in the next generation was assessed by the isolation of fluorescent bacteria from eggs, larvae, pupae and adult homogenates. Results A total of 179 bacterial strains were isolated from samples from two locations, Coari and Manaus. The predominant genera identified in this study were Acinetobacter, Enterobacter, Klebsiella, Serratia, Bacillus, Elizabethkingia, Stenotrophomonas and Pantoea. Two isolated strains, Serratia-Adu40 and Pantoea-Ovo3, were successfully transformed with the pSPT-1-GFP plasmid and expressed GFP. The fluorescent bacteria fed to adult females were transferred to their eggs, which persisted in larvae and throughout metamorphosis, and were detected in adult mosquitoes of the next generation. Conclusion Serratia-Adu40 and Pantoea-Ovo3 are promising candidates for paratransgenesis in An. darlingi. Further research is needed to determine if these bacteria are vertically transferred in nature

Pan-european assessment, monitoring, and mitigation of stressors on the health of bees

Within the PoshBee Project, we have worked out precise protocols for testing the toxicological endpoints and metabolisation rate of agrochemicals on and in bees. Three agrochemical classes (insecticides, fungicides and herbicides) were tested on three bee groups (honey bees, bumble bees and solitary mason bees). The specific protocols, different for each combination of the pesticide class and bee group, were designed based on the existing toxicity testing methods and modified according to the specificities of the different bee groups, their castes and the different exotoxicological features of the compounds. Sulfoxaflor, Azoxystrobin, and Glyphosate were used respectively as examples of the three pesticide classes. Apis mellifera, Bombus terrestris and Osmia bicornis were chosen as the examples of the three bee groups. The protocols for honey bees and bumble bees were designed based on the official existing guidelines edited by OECD. Nevertheless, given that the OECD guidelines provide recommendations to fit with most agrochemicals, many of the parameters are defined with low precision in order to be flexible and adapt to different categories of molecules. While OECD has no guidelines for testing solitary bees, we used information from the unpublished ring test protocols for Osmia bicornis by the ICPPR non- Apis working group.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action

Virus Prospecting in Crickets-Discovery and Strain Divergence of a Novel Iflavirus in Wild and Cultivated Acheta domesticus

Orthopteran insects have high reproductive rates leading to boom-bust population dynamics with high local densities that are ideal for short, episodic disease epidemics. Viruses are particularly well suited for such host population dynamics, due to their supreme ability to adapt to changing transmission criteria. However, very little is known about the viruses of Orthopteran insects. Since Orthopterans are increasingly reared commercially, for animal feed and human consumption, there is a risk that viruses naturally associated with these insects can adapt to commercial rearing conditions, and cause disease. We therefore explored the virome of the house cricket Acheta domesticus, which is both part of the natural Swedish landscape and reared commercially for the pet feed market. Only 1% of the faecal RNA and DNA from wild-caught A. domesticus consisted of viruses. These included both known and novel viruses associated with crickets/insects, their bacterial-fungal microbiome, or their plant food. Relatively abundant among these viral Operational Taxonomic Units (OTUs) was a novel Iflavirus, tentatively named Acheta domesticus Iflavirus (AdIV). Quantitative analyses showed that AdIV was also abundant in frass and insect samples from commercially reared crickets. Interestingly, the wild and commercial AdIV strains had short, extremely divergent variation hotspots throughout the genome, which may indicate specific adaptation to their hosts' distinct rearing environments

The environment and species affect gut bacteria composition in laboratory co-cultured Anopheles gambiae and Aedes albopictus mosquitoes

The midgut microbiota of disease vectors plays a critical role in the successful transmission of human pathogens. The environment influences the microbiota composition; however, the relative mosquito-species contribution has not been rigorously disentangled from the environmental contribution to the microbiota structure. Also, the extent to which the microbiota of the adult sugar food source and larval water can predict that of the adult midgut and vice versa is not fully understood. To address these relationships, larvae and adults of Anopheles gambiae and Aedes albopictus were either reared separately or in a co-rearing system, whereby aquatic and adult stages of both species shared the larval water and sugar food source, respectively. Despite being reared under identical conditions, clear intra- and interspecies differences in midgut microbiota-composition were observed across seven cohorts, collected at different time points over a period of eight months. Fitting a linear model separately for each OTU in the mosquito midgut showed that two OTUs significantly differed between the midguts of the two mosquito species. We also show an effect for the sugar food source and larval water on the adult midgut microbiota. Our findings suggest that the mosquito midgut microbiota is highly dynamic and controlled by multiple factors

Global similarity, and some key differences, in the metagenomes of Swedish varroa-surviving and varroa-susceptible honeybees

There is increasing evidence that honeybees (Apis mellifera L.) can adapt naturally to survive Varroa destructor, the primary cause of colony mortality world-wide. Most of the adaptive traits of naturally varroa-surviving honeybees concern varroa reproduction. Here we investigate whether factors in the honeybee metagenome also contribute to this survival. The quantitative and qualitative composition of the bacterial and viral metagenome fluctuated greatly during the active season, but with little overall difference between varroa-surviving and varroa-susceptible colonies. The main exceptions were Bartonella apis and sacbrood virus, particularly during early spring and autumn. Bombella apis was also strongly associated with early and late season, though equally for all colonies. All three affect colony protein management and metabolism. Lake Sinai virus was more abundant in varroa-surviving colonies during the summer. Lake Sinai virus and deformed wing virus also showed a tendency towards seasonal genetic change, but without any distinction between varroa-surviving and varroa-susceptible colonies. Whether the changes in these taxa contribute to survival or reflect demographic differences between the colonies (or both) remains unclear

Traditional and new prognosticators in breast cancer. Nottingham index, Mib-1 and estrogen receptor signaling relmain the best predictors of relapse and survival in a series of 289 cases.

7Histopathological and immunohistochemical findings on tissue microarrays, overall survival (OS), disease-free survival (DFS) and incidence of relapses (R) were recorded and statistically analyzed in 289 breast cancers. A higher R and a shorter DFS were significantly related to larger tumors, lymph node invasion, higher tumor grade, absence of estrogen receptors (ER), triple negative tumors, and presence of lymphovascular invasion (LVI). Longer OS was observed to be significantly associated with smaller tumor size (T), lymph node negativity, lower tumor grade, absence of LVI, lower Mib-1 expression and with the presence of ER. At multivariate analysis, only T for DFS and lymph node status and triple negativity either for DFS or OS had independent prognostic value. In the 194 lymph node-negative women DFS and OS were inversely related to tumor grade, absence of ER, Mib-1 expression in more than 15% of neoplastic cells and, only for DFS, presence of LVI. In the 95 lymph node-positive the number of involved nodes was the most discriminating parameter either for DFS or OS; T, Her-2 status and presence of LVI were significantly related to DFS. ER negativity was related to higher grade, progesterone receptors (PR) negativity, Her-2 negativity, hence to triple negativity, to basal-like type, Mib-1expression over 15% of neoplastic cells. Her-2 positivity was related to higher grade, ER positivity and PR positivity. Basal-like type was not an independent prognosticator, while triple negative type has a significant relation to shorter OS. The Nottingham prognostic index accurately identifies prognostic groupings and Mib-1 expression and ER signaling are the key biological predictors even in single cases.reservedmixedMEGHA T; NERI A; MALAGNINO V; CARUSO S; ONORATI M; F. ROVIELLO; TOSI PMegha, T; Neri, Alessandro; Malagnino, Valeria; Caruso, Stefano; Onorati, M; Roviello, Franco; Tosi, Pier

Educatio siglo XXI : revista de la Facultad de Educación

Investigación realizada en un colegio de Educación Infantil y Primaria de Murcia cuyo objetivo es conocer la estructura de valores transmitida por la familia así como la posible influencia de dichos valores en los resultados de aprendizaje del alumno. Para ello, se utiliza un cuestionario dirigido a los padres y madres con ochenta ítems repartidos en diez dimensiones y una entrevista semiestructurada dirigida a los profesores tutores. Todas las dimensiones del cuestionario fueron valoradas altamente, lo que pone de manifiesto que el tipo de valores contemplado en el instrumento aplicado es transmitido correctamente por las familias. Los autores insisten en la importancia de una educación en valores que no puede llevarse a cabo de forma aislada en el ámbito familiar, del mismo modo que resultaría poco fructífera si solo se trabajase en la escuela.MurciaUniversidad de Murcia, Facultad de Educación; Campus Universitario de Espinardo; 30100 Murcia; +34968364019; +34968364146; [email protected]