Illthrift in Suckling Lambs Secondary to Umbilical Infections and Possible Implication of Cryptosporidiosis as a Risk Factor

Illthrift was observed in 20/60 lambs aged 40–45 days in a dairy sheep flock in Greece. Cryptosporidiosis had been diagnosed and successfully treated with halofuginone lactate a month earlier. Parasitological examinations were negative for endoparasites while hematology and biochemistry were unremarkable. Necropsy of 5 lambs revealed lung and liver abscessation, presumably secondary to umbilical infections due to poor farm hygiene, though umbilical lesions were not observed. No new cases were observed following treatment of the umbilicus of newborn lambs with chlorexidine. Although umbilical infections are common, this is the first reported case of illthrift in lambs attributed to umbilical infection; illthrift may be the only clinical manifestation of such infections. The prior presence of cryptosporidiosis may have contributed to the severity of the infection through the reduction of local immunity. Recognition of this possibly underdiagnosed or underappreciated condition may improve medical, production, and welfare standards in the sheep industry

Gut bacteriome analysis of Anastrepha fraterculus sp. 1 during the early steps of laboratory colonization

Microbial communities associated to insect species are involved in essential biological functions such as host nutrition, reproduction and survivability. Main factors have been described as modulators of gut bacterial community, such as diet, habit, developmental stage and taxonomy of the host. The present work focuses on the complex changes that gut microbial communities go through when wild insects are introduced to artificial rearing conditions. Specifically, we analyzed the effect of the laboratory colonization on the richness and diversity of the gut bacteriome hosted by the fruit fly pest Anastrepha fraterculus sp. 1. Bacterial profiles were studied by amplicon sequencing of the 16S rRNA V3–V4 hypervariable region in gut samples of males and females, in teneral (1-day-old, unfed) and post-teneral (15-day-old, fed) flies. A total of 3,147,665 sequence reads were obtained and 32 bacterial operational taxonomic units (OTUs) were identified. Proteobacteria was the most abundant phylum (93.3% of the total reads) and, Wolbachia and Enterobacter were the most represented taxa at the genus level (29.9% and 27.7%, respectively, of the total read counts). Wild and laboratory flies showed highly significant differences in the relative abundances of bacteria. The analysis of the core bacteriome showed the presence of five OTUs in all samples grouped by origin, while nine and five OTUs were exclusively detected in laboratory and wild flies, respectively. Irrespective of fly origin or sex, a dominant presence of Wolbachia was observed in teneral flies, whereas Enterobacter was highly abundant in post-teneral individuals. We evidenced significant differences in bacterial richness and diversity among generations under laboratory colonization (F0, F1, F3 and F6) and compared to laboratory and wild flies, displaying also differential patterns between teneral and post-teneral flies. Laboratory and wild A. fraterculus sp. 1 harbor different gut bacterial communities. Laboratory colonization has an important effect on the microbiota, most likely associated to the combined effects of insect physiology and environmental conditions (e.g., diet and colony management).Instituto de GenéticaFil: Salgueiro, Julieta. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pimper, Lidia Elena. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Segura, Diego Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Milla, Fabian Horacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Russo, Romina María. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Asimakis, Elias D. University of Patras. Department of Environmental Engineering; GreciaFil: Stathopoulou, Panagiota. University of Patras. Department of Environmental Engineering; GreciaFil: Bourtzis, Kostas. Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture. Insect Pest Control Laboratory; AustriaFil: Cladera, Jorge Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Tsiamis, George. University of Patras. Department of Environmental Engineering; GreciaFil: Lanzavecchia, Silvia Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret". Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Wolbachia pipientis associated with tephritid fruit fly pests: from basic research to applications

Members of the true fruit flies (family Tephritidae) are among the most serious agricultural pests worldwide, whose control and management demands large and costly international efforts. The need for cost-effective and environmentally friendly integrated pest management (IPM) has led to the development and implementation of autocidal control strategies. These approaches include the widely used sterile insect technique and the incompatible insect technique (IIT). IIT relies on maternally transmitted bacteria (namely Wolbachia) to cause a conditional sterility in crosses between released mass-reared Wolbachia-infected males and wild females, which are either uninfected or infected with a different Wolbachia strain (i.e., cytoplasmic incompatibility; CI). Herein, we review the current state of knowledge on Wolbachia-tephritid interactions including infection prevalence in wild populations, phenotypic consequences, and their impact on life history traits. Numerous pest tephritid species are reported to harbor Wolbachia infections, with a subset exhibiting high prevalence. The phenotypic effects of Wolbachia have been assessed in very few tephritid species, due in part to the difficulty of manipulating Wolbachia infection (removal or transinfection). Based on recent methodological advances (high-throughput DNA sequencing) and breakthroughs concerning the mechanistic basis of CI, we suggest research avenues that could accelerate generation of necessary knowledge for the potential use of Wolbachia-based IIT in area-wide integrated pest management (AW-IPM) strategies for the population control of tephritid pests.Instituto de GenéticaFil: Mateos, Mariana. Texas A&M University. Departments of Ecology and Conservation Biology, and Wildlife and Fisheries Sciences; Estados UnidosFil: Martinez Montoya, Humberto. Universidad Autónoma de Tamaulipas. Unidad Académica Multidisciplinaria Reynosa Aztlan. Laboratorio de Genética y Genómica Comparativa; MéxicoFil: Lanzavecchia, Silvia Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Conte, Claudia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Guillén, Karina. El Colegio de la Frontera Sur; MéxicoFil: Morán-Aceves, Brenda M. El Colegio de la Frontera Sur; MéxicoFil: Toledo, Jorge. El Colegio de la Frontera Sur; MéxicoFil: Liedo, Pablo. El Colegio de la Frontera Sur; MéxicoFil: Asimakis, Elias D. University of Patras. Department of Environmental Engineering; GreciaFil: Doudoumis, Vangelis. University of Patras. Department of Environmental Engineering; GreciaFil: Kyritsis, Georgios A. University of Thessaly. Department of Agriculture Crop Production and Rural Environment. Laboratory of Entomology and Agricultural Zoology; GreciaFil: Papadopoulos, Nikos T. University of Thessaly. Department of Agriculture Crop Production and Rural Environment. Laboratory of Entomology and Agricultural Zoology; GreciaFil: Augustinos, Antonios A. Hellenic Agricultural Organization. Institute of Industrial and Forage Crops. Department of Plant Protection; GreciaFil: Segura, Diego Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética. Laboratorio de Genética de Insectos de Importancia Económica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Tsiamis, George. University of Patras. Department of Environmental Engineering; Greci

Study of symbiotic bacterial communities in insects of agricultural and environmental importance

In recent years, our knowledge of the role of endosymbiotic bacteria in their eukaryotic host organisms has expanded to such an extent that we now know a multitude of functions in which they participate, greatly influencing important aspects of biology, ecology and evolution of their hosts. Symbiotic bacteria affect growth and nutrition, reproduction, speciation, communication and the role of insects as carriers of infectious diseases, they confer protection from natural enemies and pathogenic microorganisms and induce resistance to extreme environmental conditions as well as pesticides or other chemical agents. These properties could be used to improve the application of innovative techniques, with limited environmental impact, with the aim of controlling insects that act as agricultural pests or carriers of infectious diseases. The flies of the genera Bactrocera, Dacus and Zeugodacus are important pests of cultivated plant species, capable of affecting a large number of different fruits and vegetables, which they use mainly to fulfill their biological cycle, during larval development. Although many species originate from regions of Southeast Asia and Africa, they have greatly expanded their distribution into many tropical, subtropical and temperate regions of the planet, displacing indigenous fly populations in many cases. Moreover, with the continued rise in temperatures, new opportunities for expansion are constantly emerging for these highly invasive species. Chemical insecticides are widely used to deal with these pests, even though they exert multiple negative effects on humans, animals and the environment, and their application becomes less effective over time due to the development of resistance from the pests. The purpose of this thesis is to exploit the symbiotic bacteria of these insects in order to develop and/or improve environmentally friendly methods aimed at controlling their populations. Techniques such as the Sterile Insect Technique (SIT), which uses ionizing radiation to sterilize and release mass reared insects, and the Incompatible Insect Technique (IIT), which employs symbiotic bacteria that cause reproductive alterations to their insect-hosts, are very promising technologies in this direction. The application and improvement of innovative technologies depends on the constant production of new knowledge that will reduce defects and imperfections. For example, ionizing radiation affects host fitness, resulting in reduced competitiveness of mass-reared insects compared to wild populations. In this case, symbiotic bacteria can be provided in order to improve the fitness of mass-reared strains and thus the efficiency of the technique. On the other hand, for the design and better implementation of the Incompatible Insect Technique, it is essential to identify the numerous interactions between different bacterial genera that are present in a host and cause reproductive disorders (reproductive parasites), between the different strains of a reproductive parasite that might be present, as well as the reproductive parasites and the insect-host. More specifically, the presence of four symbiotic bacteria, Wolbachia, Spiroplasma, Cardinium and Arsenophonus, which contain strains that cause reproductive alterations, was examined in 14 species of flies, belonging to the genera Bactrocera, Dacus and Zeugodacus and collected from 30 natural populations originating from three countries in southeast Asia, as well as 19 laboratory populations. A total of 801 adults from natural populations and 380 from laboratory colonies were analyzed. In samples from natural populations, occurrence of symbiotic bacteria varied greatly between different species, populations and countries. Overall, infections with Wolbachia were the most prevalent, followed by the three Entomoplasmatales genera and Cardinium species, while no infections with Arsenophonus were observed. In contrast, laboratory populations did not harbor infections with the symbiotic bacteria. The molecular characterization of the symbiotic bacteria showed that most of them were classified into common phylogenetic groups, indicating possible horizontal transfer of symbiotic bacteria between different species of fruit flies. In three species (B. correcta, B. dorsalis and B. zonata), multiple strains of Wolbachia were observed. Finally, Wolbachia pseudogenes were identified, some of which were similar to previously identified pseudogenes, which had been incorporated into the insect-host genome. Moreover, the effect of ionizing radiation on the composition of bacterial communities of adult male and female Zeugodacus cucurbitae flies, that were mass-reared under laboratory conditions was investigated. The effect of radiation was studied in relation to the diet applied at the larval stage as well as the sex of the flies. The effect of two different diets was studied: an artificial diet based on bran and one based on a natural host, sweet gourd. Significant differences were observed in the bacterial communities between samples reared on different diets and different sexes before treatment with radiation. After treatment with irradiation, bacterial communities showed significant differentiation in flies reared on the artificial diet, showing also a significant decrease in species richness and diversity, whereas the samples grown on the natural host remained nearly unchanged. Gender specific changes in the composition of the bacterial communities, but on a smaller degree, were also observed among samples, with males showing greater diversity and females more richness. Both males and females exhibited less or the same diversity and richness after radiation treatment, except for females that were reared on sweet gourd, that showed an interesting increase in both traits. Similarly, we investigated the effect of ionizing radiation on the structure of bacterial communities of adult male and female Bactrocera dorsalis flies from laboratory populations. Differences in the composition of bacterial communities based on the developmental stage of the flies (larvae and adults) as well as in adults based on their irradiation status were studied. No differences in diversity and richness indices were observed between adult flies and larvae, whereas the two developmental stages exhibited different bacterial profiles. Adult individuals exhibited higher levels of diversity and richness after irradiation than non-irradiated adult samples, whereas the two categories showed significant differences in their bacterial profile composition. Finally, the future perspectives of the results of this dissertation are discussed with regard to improving or developing applications based on the Sterile and Incompatible Insect Techniques for the effective control of these parasitic insects. The ecological and evolutionary implications of symbiotic bacterial infections in natural populations from southeast Asia are also commented.Τα τελευταία χρόνια οι γνώσεις μας σχετικά με τον ρόλο των ενδοσυμβιωτικών βακτηρίων στους ευκαρυωτικούς οργανισμούς-ξενιστές τους, έχουν αυξηθεί σε τέτοιο βαθμό ώστε να γνωρίζουμε πλέον ένα πλήθος από λειτουργίες στις οποίες συμμετέχουν, επηρεάζοντας σε μεγάλο βαθμό σημαντικές πτυχές της βιολογίας, οικολογίας και εξέλιξης των ξενιστών τους. Τα ενδοσυμβιωτικά βακτήρια επιδρούν στην ανάπτυξη και τη διατροφή, στην αναπαραγωγή και την ειδογένεση, στην επικοινωνία, στην προστασία ενάντια σε φυσικούς εχθρούς, παθογόνους μικροοργανισμούς και παράσιτα, στην αντοχή σε ακραίες περιβαλλοντικές συνθήκες και στον ρόλο των εντόμων σαν φορείς μολυσματικών ασθενειών για τον άνθρωπο και τα ζώα. Οι λειτουργίες αυτές μας επιτρέπουν να βελτιώσουμε την εφαρμογή καινοτόμων τεχνικών, με μικρό περιβαλλοντικό αντίκτυπο, για την αντιμετώπιση εντόμων που δρουν σαν εχθροί καλλιεργειών ή φορείς μολυσματικών ασθενειών. Οι μύγες των γενών Bactrocera, Dacus και Zeugodacus αποτελούν σημαντικούς εχθρούς καλλιεργούμενων φυτικών ειδών, με ικανότητα να προσβάλλουν μεγάλο αριθμό διαφορετικών φρούτων και λαχανικών, τα οποία χρησιμοποιούν κυρίως για την εκπλήρωση του βιολογικού τους κύκλου, κατά την ανάπτυξη των προνυμφών τους. Αν και πολλά είδη κατάγονται από περιοχές της νοτιοανατολικής Ασίας και της Αφρικής, έχουν επεκτείνει σε μεγάλο βαθμό την κατανομή τους σε πολλές τροπικές, υποτροπικές και εύκρατες περιοχές του πλανήτη, εκτοπίζοντας γηγενείς πληθυσμούς μυγών, ενώ με την συνεχή αύξηση της θερμοκρασίας, τους παρουσιάζονται συνεχώς νέες ευκαιρίες εξάπλωσης. Για την αντιμετώπισή τους χρησιμοποιούνται ευρέως χημικά εντομοκτόνα, τα οποία εμφανίζουν πολύπλευρες αρνητικές συνέπειες στο περιβάλλον, τον άνθρωπο και τα ζώα, ενώ με το πέρασμα του χρόνου η εφαρμογή τους καθίσταται αναποτελεσματική λόγω ανάπτυξης αντίστασης από τα έντομα-στόχους. Σκοπός της παρούσας Διατριβής είναι η εκμετάλλευση του συμβιωτικού βακτηριακού φορτίου των συγκεκριμένων εντόμων με σκοπό την ανάπτυξη ή και βελτίωση περιβαλλοντικά φιλικότερων τεχνικών που αποσκοπούν στον έλεγχο των πληθυσμών τους. Τέτοιες τεχνικές, όπως η τεχνική στείρου εντόμου (Sterile Insect Technique, SIT), που χρησιμοποιεί ιονίζουσα ακτινοβολία για την στείρωση και μαζική απελευθέρωση εντόμων, και η τεχνική ασύμβατου εντόμου (Incompatible Insect Technique, IIT), που χρησιμοποιεί συμβιωτικά βακτήρια που προκαλούν αναπαραγωγικές αλλοιώσεις στα έντομα-ξενιστές τους, αποτελούν πολλά υποσχόμενες τεχνολογίες προς αυτή την κατεύθυνση. Η εφαρμογή καινοτόμων τεχνολογιών απαιτεί συνεχώς παραγωγή νέας γνώσης στην κατεύθυνση της βελτίωσής τους, της μείωσης των ατελειών και των μειονεκτημάτων τους. Συγκεκριμένα, η ιονίζουσα ακτινοβολία επιδρά στην αρμοστικότητα των εντόμων, με συνέπεια την μειωμένη ανταγωνιστικότητα των μαζικά εκτρεφόμενων εντόμων σε σύγκριση με τα άτομα των άγριων πληθυσμών. Σε αυτή την περίπτωση, συμβιωτικά βακτήρια μπορούν να χρησιμοποιηθούν στην παρεχόμενη τροφή για την βελτίωση της αρμοστικότητας των μαζικά εκτρεφόμενων εργαστηριακών στελεχών και κατά συνέπεια της αποδοτικότητας της τεχνικής. Από την άλλη μεριά, για τον σχεδιασμό αλλά και την καλύτερη εφαρμογή της τεχνικής ασύμβατου εντόμου, είναι σημαντική η γνώση των πολυποίκιλων αλληλεπιδράσεων μεταξύ των διαφορετικών ειδών βακτηρίων που προκαλούν αναπαραγωγικές αλλοιώσεις (αναπαραγωγικά παράσιτα), μεταξύ των διαφορετικών στελεχών ενός αναπαραγωγικού παρασίτου, καθώς και των αναπαραγωγικών παρασίτων με το έντομο-ξενιστή. Συγκεκριμένα, στην παρούσα Διατριβή, ελέγχθηκε η παρουσία τεσσάρων συμβιωτικών βακτηρίων, των Wolbachia, Spiroplasma, Cardinium και Arsenophonus τα οποία περιέχουν στελέχη που προκαλούν αναπαραγωγικές αλλοιώσεις, σε 14 είδη μυγών που ανήκουν στα γένη Bactrocera, Dacus και Zeugodacus που συλλέχθηκαν από 30 φυσικούς πληθυσμούς, με καταγωγή από τρεις χώρες της νοτιοανατολικής Ασίας, καθώς και από 19 εργαστηριακούς πληθυσμούς. Συνολικά αναλύθηκαν 801 ενήλικα άτομα από φυσικούς πληθυσμούς και 380 άτομα από εργαστηριακές αποικίες. Τα αποτελέσματα στους φυσικούς πληθυσμούς έδειξαν πολύ μεγάλη απόκλιση στην εμφάνιση και τη σχετική συχνότητα των συμβιωτικών βακτηρίων ανά είδος, πληθυσμό και χώρα. Συνολικά οι μολύνσεις με Wolbachia ήταν οι επικρατέστερες, ακολουθούμενες από γένη των Entomoplasmatales και είδη Cardinium, ενώ δεν παρατηρήθηκαν μολύνσεις με είδη Arsenophonus. Αντίθετα, οι εργαστηριακοί πληθυσμοί δεν περιείχαν μολύνσεις με τα συμβιωτικά βακτήρια. Ο μοριακός χαρακτηρισμός των συμβιωτικών βακτηρίων απέδειξε ότι τα περισσότερα κατατάσσονταν σε κοινή φυλογενετική ομάδα, γεγονός που υποδηλώνει οριζόντια μεταφορά συμβιωτικών βακτηρίων μεταξύ διαφορετικών ειδών. Σε τρία είδη (B. correcta, Β. dorsalis και B. zonata) παρατηρήθηκαν μολύνσεις με πολλαπλά στελέχη του βακτηρίου Wolbachia. Τέλος, αναγνωρίστηκαν περιπτώσεις ψευδογονιδίων της Wolbachia, ορισμένα από τα οποία ήταν όμοια με ψευδογονίδια που εντοπίστηκαν προηγουμένως σε άλλα έντομα-ξενιστές, στα οποία μάλιστα είχαν βρεθεί ενσωματωμένα στο γονιδίωμά τους. Επίσης, πραγματοποιήθηκε έλεγχος της επίδρασης ιονίζουσας ακτινοβολίας στη σύσταση των βακτηριακών κοινοτήτων ενήλικων αρσενικών και θηλυκών μυγών του είδους Zeugodacus cucurbitae που αναπτύσσονται μαζικά στο εργαστήριο. Η επίδραση της ακτινοβολίας μελετήθηκε σε συνάρτηση με την δίαιτα που εφαρμόστηκε στο προνυμφικό στάδιο, καθώς και το φύλο των μυγών. Μελετήθηκε η επίδραση δύο διαφορετικών διατροφών: μιας συνθετικής με βάση άλευρα πιτύρου και μιας φυσικής από κολοκύθα. Αρχικά, παρατηρήθηκαν διαφορές στις βακτηριακές κοινότητες των εντόμων πριν την εφαρμογή ακτινοβολίας, τόσο μεταξύ μυγών που είχαν αναπτυχθεί σε διαφορετικές δίαιτες, καθώς και μεταξύ ατόμων διαφορετικού φύλου. Μετά την επίδραση ακτινοβολίας, οι βακτηριακές κοινότητες σημείωσαν στατιστικά σημαντική διαφοροποίηση στις μύγες που αναπτύχθηκαν με την συνθετική δίαιτα, εμφανίζοντας επίσης στατιστικά σημαντική μείωση στην αφθονία και την ποικιλότητα, ενώ το βακτηρίωμα των δειγμάτων που αναπτύχθηκαν με τον φυσικό ξενιστή παρέμεινε σχεδόν αναλλοίωτο. Αλλαγές στη σύνθεση των βακτηριακών κοινοτήτων, αλλά μικρότερης κλίμακας, παρατηρήθηκαν και μεταξύ ατόμων διαφορετικού φύλου, με τα αρσενικά άτομα να εμφανίζουν επίσης στην πλειοψηφία τους μεγαλύτερη ποικιλότητα ενώ τα θηλυκά μεγαλύτερη αφθονία. Τόσο τα αρσενικά όσο και τα θηλυκά άτομα εμφάνισαν μικρότερη ή ίδια ποικιλότητα και αφθονία μετά την επίδραση ακτινοβολίας, με εξαίρεση τα θηλυκά άτομα που αναπτύχθηκαν με κολοκύθα, στα οποία παρατηρήθηκε αύξηση και στα δύο χαρακτηριστικά. Ομοίως, έγινε έλεγχος της επίδρασης ιονίζουσας ακτινοβολίας στην δομή των βακτηριακών κοινοτήτων ενήλικων αρσενικών και θηλυκών ατόμων τους είδους Bactrocera dorsalis από εργαστηριακούς πληθυσμούς. Μελετήθηκαν διαφορές στη σύσταση των βακτηριακών κοινοτήτων ανάλογα με το αναπτυξιακό στάδιο στο οποίο βρίσκονται οι μύγες (προνυμφικό στάδιο και ενήλικες) καθώς και στις ενήλικες μετά την επίδραση ιονίζουσας ακτινοβολίας. Αρχικά, δεν παρατηρήθηκαν διαφορές στους δείκτες ποικιλότητας και αφθονίας, μεταξύ ενηλίκων μυγών και προνυμφών, ενώ τα δύο αναπτυξιακά στάδια που μελετήθηκαν ανέπτυξαν διαφορετικά βακτηριακά προφίλ. Τα ενήλικα άτομα μετά την ακτινοβόληση εμφάνισαν υψηλότερους δείκτες ποικιλότητας και αφθονίας σε σχέση με τα ενήλικα δείγματα που δεν ακτινοβολήθηκαν, ενώ οι δύο κατηγορίες εμφάνισαν και στατιστικά σημαντική διαφοροποίηση στη σύσταση του βακτηριακού τους προφίλ. Τέλος, γίνεται σχολιασμός της προοπτικής χρησιμοποίησης των αποτελεσμάτων της παρούσας Διατριβής με σκοπό τη βελτίωση ή ανάπτυξη τεχνικών στείρου και ασύμβατου εντόμου για την αντιμετώπιση των συγκεκριμένων ειδών εντόμων. Επίσης σχολιάζονται οι οικολογικές και εξελικτικές προεκτάσεις των μολύνσεων με συμβιωτικά βακτήρια στους φυσικούς πληθυσμούς από την νοτιοανατολική Ασία

Ovine and Caprine Toxoplasmosis: Experimental Study

Eighteen mature ewes of the Mytilene breed and 18 mature Local- Damascus crossbred goats, seronegative for Toxoplasma gondii (T. gondii) by ELISA were used. All animals were mated after synchronization of estrus. On day 90 of pregnancy, animals were randomly assigned to 3 experimental groups; 6 ewes (S1) and 6 goats (G1) were orally inoculated by stomach tube with 1000 oocysts; 6 ewes (S2) and 6 goats (G2) were orally inoculated with a non-infected control inoculum. On day 140+2 of pregnancy, the remaining 6 ewes (S3) and 6 goats (G3) were inoculated by stomach tube with 3000 oocysts. Positive T. godii DNA was detected in 94% of fetal and maternal blood, 95% fetal tissue, 89% pre-colostral udder secretions and 12.5% milk samples using Polymerase Chain Reaction (PCR). Infected animals and their live newborns was seropositive (ELISA) until the end of the study. PCR was able to detect T. gondii DNA in maternal blood of infected animals 3-5 days before abortion occurred. This time period may be used to implement preventive and therapeutic measure to reduce abortion rate and associated economic losses. Since milk and milk products are important food sources in rural areas and in many cases it is used unpasteurized before consumption. The T. gondii DNA, detected by PCR in milk samples of infected animals, increases the possibility that the parasite is transmitted through consumption of unpasteurized milk which is a highly relevant result for public health considerations and providing valuable information for future research

Mediterranean Plants as Potential Source of Biopesticides: An Overview of Current Research and Future Trends

The development and implementation of safe natural alternatives to synthetic pesticides are urgent needs that will provide ecological solutions for the control of plant diseases, bacteria, viruses, nematodes, pests, and weeds to ensure the economic stability of farmers and food security, as well as protection of the environment and human health. Unambiguously, production of botanical pesticides will allow for the sustainable and efficient use of natural resources and finally decrease the use of chemical inputs and burden. This is further underlined by the strict regulations on pesticide residues in agricultural products and is in harmony with the Farm to Fork strategy, which aims to reduce pesticide use by 50% by 2030. Thus, the present work aims to compile the scientific knowledge of the last 5 years (2017–February 2023) regarding the Mediterranean plants that present biopesticidal effects. The literature review revealed 40 families of Mediterranean plants with at least one species that have been investigated as potential biopesticides. However, only six families had the highest number of species, and they were reviewed comprehensively in this study. Following a systematic approach, the extraction methods, chemical composition, biopesticidal activity, and commonly used assays for evaluating the antimicrobial, pesticidal, repellant, and herbicidal activity of plant extracts, as well as the toxicological and safety aspects of biopesticide formulation, are discussed in detail. Finally, the aspects that have not yet been investigated or are under-investigated and future perspectives are highlighted

Detection of Wolbachia Infections in Natural and Laboratory Populations of the Moroccan Hessian Fly, Mayetiola destructor (Say)

Mayetiola destructor (Hessian fly) is a destructive pest of wheat in several parts of the world. Here, we investigated the presence of reproductive symbionts and the effect of the geographical location on the bacterial community associated to adult Hessian flies derived from four major wheat producing areas in Morocco. Using specific 16S rDNA PCR assay, Wolbachia infection was observed in 3% of the natural populations and 10% of the laboratory population. High throughput sequencing of V3-V4 region of the bacterial 16S rRNA gene revealed that the microbiota of adult Hessian flies was significantly influenced by their native regions. A total of 6 phyla, 10 classes and 79 genera were obtained from all the samples. Confirming the screening results, Wolbachia was identified as well in the natural Hessian flies. Phylogenetic analysis using the sequences obtained in this study indicated that there is one Wolbachia strain belonging to supergroup A. To our knowledge, this is the first report of Wolbachia in Hessian fly populations. The observed low abundance of Wolbachia most likely does not indicate induction of reproductive incompatibility. Yet, this infection may give a new insight into the use of Wolbachia for the fight against Hessian fly populations

Cytogenetic and symbiont analysis of five members of the B. dorsalis complex (Diptera, Tephritidae): no evidence of chromosomal or symbiont-based speciation events

The Bactrocera dorsalis species complex, currently comprising about 90 entities has received much attention. During the last decades, considerable effort has been devoted to delimiting the species of the complex. This information is of great importance for agriculture and world trade, since the complex harbours several pest species of major economic importance and other species that could evolve into global threats. Speciation in Diptera is usually accompanied by chromosomal rearrangements, particularly inversions that are assumed to reduce/eliminate gene flow. Other candidates currently receiving much attention regarding their possible involvement in speciation are reproductive symbionts, such as Wolbachia, Spiroplasma, Arsenophonus, Rickettsia and Cardinium. Such symbionts tend to spread quickly through natural populations and can cause a variety of phenotypes that promote pre-mating and/or post-mating isolation and, in addition, can affect the biology, physiology, ecology and evolution of their insect hosts in various ways. Considering all these aspects, we present: (a) a summary of the recently gained knowledge on the cytogenetics of five members of the B. dorsalis complex, namely B. dorsaliss.s., B. invadens, B. philippinensis, B. papayae and B. carambolae, supplemented by additional data from a B. dorsaliss.s. colony from China, as well as by a cytogenetic comparison between the dorsalis complex and the genetically close species, B. tryoni, and, (b) a reproductive symbiont screening of 18 different colonized populations of these five taxa. Our analysis did not reveal any chromosomal rearrangements that could differentiate among them. Moreover, screening for reproductive symbionts was negative for all colonies derived from different geographic origins and/or hosts. There are many different factors that can lead to speciation, and our data do not support chromosomal and/or symbiotic-based speciation phenomena in the taxa under study

Impact of the Post-Transplant Period and Lifestyle Diseases on Human Gut Microbiota in Kidney Graft Recipients

Gaining long-term graft function and patient life quality remain critical challenges following kidney transplantation. Advances in immunology, gnotobiotics, and culture-independent molecular techniques have provided growing insights into the complex relationship of the microbiome and the host. However, little is known about the over time-shift of the gut microbiota in the context of kidney transplantation and its impact on both graft and health stability. Here we aimed to characterize the structure of gut microbiota within stable kidney graft recipients. We enrolled forty kidney transplant patients after at least three months of transplantation and compared them to eighteen healthy controls. The overall microbial community structure of the kidney transplanted group was clearly different from control subjects. We found lower relative abundances of Actinobacteria, Bacteroidetes, and Verrucomicrobia within the patient group and a higher abundance of Proteobacteria compared to the control group. Both richness and Shannon diversity indexes were significantly lower in the kidney graft recipients than in healthy controls. Post-graft period was positively correlated with the relative abundance of the Proteobacteria phylum, especially Escherichia.Shigella genus. Interestingly, only Parabacteroides was found to significantly differentiate patients that were not suffering from lifestyle diseases and those who suffer from post-graft complications. Furthermore, network analysis showed that the occurrence of lifestyle diseases was significantly linked with a higher number of negative interactions of Sutterella and Succinivibrio genera within patients. This study characterizes gut microbiome fluctuation in stable kidney transplant patients after a long post-allograft period. Analysis of fecal microbiota could be useful for nephrologists as a new clinical tool that can improve kidney allograft monitoring and outcomes

High Throughput Analysis Reveals Changes in Gut Microbiota and Specific Fecal Metabolomic Signature in Hematopoietic Stem Cell Transplant Patients

There is mounting evidence for the emerging role of gut microbiota (GM) and its metabolites in profoundly impacting allogenic hematopoietic stem cell transplantation (allo-HSCT) and its subsequent complications, mainly infections and graft versus host-disease (GvHD). The present study was performed in order to investigate changes in GM composition and fecal metabolic signature between transplant patients (n = 15) and healthy controls (n = 18). The intestinal microbiota was characterized by NGS and gas chromatography–mass spectrometry was employed to perform untargeted analysis of fecal metabolites. We found lower relative abundances of Actinobacteria, Firmicutes, and Bacteroidetes and a higher abundance of Proteobacteria phylum after allo-HSCT. Particularly, the GvHD microbiota was characterized by a lower relative abundance of the short-chain fatty acid-producing bacteria, namely, the Feacalibacterium, Akkermansia, and Veillonella genera and the Lachnospiraceae family, and an enrichment in multidrug-resistant bacteria belonging to Escherichia, Shigella, and Bacteroides. Moreover, network analysis showed that GvHD was linked to a higher number of positive interactions of Blautia and a significant mutual-exclusion rate of Citrobacter. The fecal metabolome was dominated by lipids in the transplant group when compared with the healthy individuals (p < 0.05). Overall, 76 metabolites were significantly altered within transplant recipients, of which 24 were selected as potential biomarkers. Furthermore, the most notable altered metabolic pathways included the TCA cycle; butanoate, propanoate, and pyruvate metabolisms; steroid biosynthesis; and glycolysis/gluconeogenesis. Specific biomarkers and altered metabolic pathways were correlated to GvHD onset. Our results showed significant shifts in gut microbiota structure and fecal metabolites characterizing allo-HSCT