Estudio microtomográfico de la anatomía funcional de dos especies de insectos plaga, el psílido asiático de los cítricos, diaphorina citri (hemiptera liviidae) y la broca del café, hypothenemus hampei (coleoptera, curculionidae)

Abstract

ABSTRACT Traditionally, the study of the internal anatomy of insects has been based on dissection techniques with their own limitations. The computerized microtomography (micro-CT), is a relatively new technique that is based on obtaining X-ray images that after being processed allows to reconstruct the external and internal anatomy and the specimen, as well as to separate with different color the structures. Being able to observe them in their anatomical position from any perspective and without the need for dissections and therefore without the deformations and destruction that these imply. All this has opened up a range of possibilities previously unimaginable, and impossible, with the classic methods of dissection and/or microscopy techniques. The main objective of this doctoral thesis has been to carry out a detailed microtomographic study (with high resolution 3D images and videos,) of the external and internal anatomy of two species of insect pests: the Asian citrus psyllid (Diaphorina citri Kuwayama, 1908) and the coffee berry borer (Hypothenemus hampei Ferrari, 1867), of great importance worldwide due to the enormous damage and economic losses they cause. Additionally, during the study of H. hampei's activity within the coffee cherry, we found specimens of the coffee bean weevil (Araecerus fasciculatus De Geer in 1775) emerging from coffee cherries. This discovery motivated a study of the bean weevil and its activity within the coffee berry, which was the object of an additional publication on the subject and which is included as additional material. The results obtained here, together with those of our previous work (in which we carried out a microtomographic study of the honey bee and compared the results with the classic publication of Snodgrass on the anatomy of the species), confirm that the micro-CT techniques constitute an excellent alternative to the traditional methods based on dissection. As it is a non-destructive technique that allows to observe the structures (external and internal) in their real anatomical position, from any perspective, as many times as required, without the risk of deforming or breaking them. And without the inconvenience of the deformations produced during the manipulations required for dissection. Likewise, during the microtomographic study of both species it has been possible to corroborate structures already described by other authors and at the same time others have been discovered, never described before, which if it were not for the micro-CT could hardly have been made evident. The results obtained, for the most part already published, in a summarized form, have been the following: The Asian Citrus Psyllid (Diaphorina citri): External anatomy (the head, with detailed description of the structures, including the oral system, eyes and antennae; the thorax, morphology of the different segments, legs, sclerites, distribution of the spiracles, and the wings; the abdomen -segments, distribution of the spiracles, and terminalia of both sexes-). Internal anatomy (digestive, pharynx, esophagus, filter chamber, midgut, and hindgut and anus; excretory system, the tubular structures -appendices- of the midgut, classically considered as Malpigium tubules; internal structures of the cibarial chamber (precibarial -salivary - pump, cibarial pump, maxillary and mandibular cones and associated muscles; stylets bundle, glands -salivary, coxal and antennal-, coxal and antennal glands are described and cited for the first time in Psylloidea, discussing their possible function as producers of sexual pheromones; tracheal system -spiracles, and disposition of the abdominal tracheal tubes-; circulatory system -dorsal vessel, aorta and aliform muscles-; Nervous system -brain, ventral ganglion nerve chain and nerves-, the existence of sexual differences in the conformation of the brain and ventral ganglion nerve chain are described for the first time in Psylloidea; male reproductive system -testicles, seminal vesicles, accessory glands, spermatic pump, connection ducts and edeagus-; female reproductive system -ovaries, accessory glands, spermatheca, colleterial/cement gland, connecting ducts and the ovipositor-, the functional anatomy of the spermatheca is described and analyzed for the first time in Psylloidea; bacteriome -situation, external and internal organization-; being the first three-dimensional reconstruction of it; different details of the musculature). Anatomical-morphological and functional aspects derived from the chopper-sucker feeding (structures involved, perforation point, the stylets inside the plant vessels and the salivary sheaths). The coffee berry borer (Hypothenemus hampei): Study of the vital activity of H. hampei inside the coffee berry (a behavioral pattern was observed in which the female is placed with the abdomen directed to the outside, blocking the entrance of the tunnels. Likewise, the construction of tunnels presents a zig-zag pattern that makes the penetration of possible parasites difficult. On the other hand, the use of the food resources of the coffee cherry is maximized thanks to the construction sequence of the tunnels and oviposition chambers. That in a sequential way begins in the seed periphery and moves towards the interior). External anatomy (the head - structures and sexual differences in the number and arrangement of ommatidia; the thorax - different segments, respiratory spiracles and first observation on sexual differences in the morphology of the pronotum; the abdomen - respiratory spiracles, and sexual differences in the number of segments. Internal anatomy (digestive system -esophagus, crop, proventricle, midgut, hindgut, gastric cecum, rectal ampulla and anus-, sexual differences in the conformation and number of circumvolutions of the midgut are described for the first time; excretory system -number and arrangement of Malpigio's tubules-; male reproductive system -testes, accessory glands, seminal vesicles, spermatic pump, ejaculatory duct and first description of the edeagus-; female reproductive system - ovaries, lateral oviducts, accessory gland, spermatheca, spermatic gland, common oviduct and gonopore; respiratory system - with a detailed study of the respiratory spiracles; the filtering systems they present are described for the first time in detail, and the whole system of tracheal tubes is reconstructed for the first time, naming the different branches, some of them never described before. Having calculated the total length of the tubes and studied the distribution of internal light by size, the circulatory system -dorsal and aortic vessels-; the nervous system -brain, ventral nerve chain nodes and nerves, sexual differences in the size and shape of the brain are described for the first time; the musculature -showing for the first time a reduction in the muscles involved in flight in males-. The Coffee Bean Weevil (Araecerus fasciculatus): The species was described in 1775 as Curculio fasciculatus, and this is the first time in the two and a half centuries since then that thanks to modern microtomography techniques it has been possible to "freeze" the insect's activity inside a coffee berry and study it in detail. This study has revealed interesting aspects of the biology of the bark beetle inside the coffee berry. Having reconstructed 5th instar larvae and females inside the coffee berry. 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