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Established interspecific metabolic allometric relationships do not adequately describe the complexity and variable physiological states of developing animals. Consequently, intraspecific allometric relationships of oxygen consumption and carbon dioxide production as a function of body mass; the respiratory quotient; the function of the silk cocoon; and body composition were investigated for each distinct developmental stage of the silkworm, Bombyx mori. Whole animal O2 consumption in Bombyx ranged from 0.00064 + 0.000047 ml O2 .hr-1 at larval instar I to 0.77 + 0.06 ml O2 .hr-1 in pre-pupal, falling to 0.21+ 0.01 ml O2 .hr-1 in the pupae. Those instars having a significant relationship between O2 consumption as a function of body mass, the slope of the line relating O2 consumption to body mass varied between 0.99 and 1.02, while across all instars the slope was 0.82. Developmental allometry should be presented for individual developmental stages because the individual allometric exponents of the stages can be significantly different from the overall allometric exponent throughout development and in some cases, the overall allometric exponent can be a statistical artifact. The first larval instar of Bombyx mori has the lowest cross sectional area of high metabolic tissue of the midgut (27%) and had one of the highest percentages of some metabolically inert tissues (i.e. lipid, 7.5%). Body composition of the first instar does not support the idea that smaller mass animals having the highest O2 consumption are composed of a greater percentage of metabolically active organs when compared to larger animals. However, this developmental stage has the highest percentage of the mitochondrial marker cytochrome oxidase, which correlates well with the high O2 consumption rate of the smaller mass. Therefore, established interspecific principles should not be assumed to function as valid models for intraspecific developmental relationships of metabolism as a function of body mass. Developmental allometry should include an analysis of individual stages of development as well as an analysis of development as a whole to gain a comprehensive understanding of the complexity of allometry of the developing animal such as the silkworm.

Orientador: Daniela Carvalho dos Santos / Resumo: O bicho-da-seda, Bombyx mori (Lepidoptera: Bombycidae), é o inseto de maior importância econômica na produção de seda. A lagarta se alimenta de folhas de amoreira e é altamente sensível a agrotóxicos, assim o uso destes em culturas agrícolas circunvizinhas às plantações de amoreira pode afetar o desenvolvimento de B. mori, acarretando em desequilíbrio nas suas funções metabólicas e, consequentemente, comprometendo a produção de casulos. Entre estes agrotóxicos, destaca-se o Novaluron, inseticida inibidor da síntese de quitina nos insetos e empregado no controle de insetos pragas de culturas agrícolas próximas as plantações de amoreira. Assim, investigamos os efeitos tóxicos de Novaluron no desenvolvimento de B. mori. Lagartas de B. mori, foram separadas em dois grupos experimentais: grupo controle (GC) e grupo tratamento (GT: tratado com 0, 15 mL/L de Novaluron). Após ecdise do 2° para o 3° instar, lagartas do GT foram alimentadas por 24 horas com folhas de amoreira tratadas com o inseticida. Paralelamente foi realizada uma nova exposição, porém em lagartas que realizavam a ecdise do 4° para o 5° instar. Lagartas, pupas e adultos de B. mori foram anestesiadas e segmentos do intestino médio, glândula da seda e órgãos reprodutores retirados e processados convencionalmente para técnicas de microscopias de luz, eletrônica e imunohistoquímica. Além disso, os efeitos no desenvolvimento, reprodução e qualidade do casulo também foram avaliados. O Novaluron provocou efeitos citotóxico... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The silkworm, Bombyx mori (Lepidoptera: Bombycidae is the insect of major economic importance in the production of silk. The larvae feeds on mulberry leaves and is highly sensitive to agrochemicals, thus the use of these in agricultural crops surrounding the mulberry plantations can affect the development of B. mori, causing an imbalance in its metabolic functions and, consequently, compromising the production of cocoons. Among these agrochemicals, stands out the Novaluron, an insecticide inhibitor of the synthesis of chitin in insects and used in the control of insect pests of crops near mulberry plantations. Thus, we investigated the lethal and sublethal effects of Novaluron on the development of B. mori. Larvae were selected into two experimental groups: control group (CG) and treatment group (TG: treated with 0, 15 mL/L Novaluron). After ecdysis from the 2nd to the 3rd instar, the TG larvae were fed for 24 hours with mulberry leaves treated with the insecticide. In parallel, a new exposition was carried out, however in larvae that carry out the ecdysis from the 4th to the 5th instar. B. mori larvae, pupae and adults were anesthetized and segments of the midgut, silk gland and reproductive organs were removed and processed conventionally for light microscopy, electron microscopy and immunohistochemistry. In addition, the effects on the development, reproduction and quality of the cocoon were also evaluated. Novaluron caused cytotoxic effects on the midgut and the silk glan... (Complete abstract click electronic access below) / Doutor

Made available in DSpace on 2017-07-10T14:17:10Z (GMT). No. of bitstreams: 1 AMPOLA DOS _ MALPIGHI DE Bombyx mori PELO ALPHABACULOVIRUS.pdf: 2176663 bytes, checksum: 866566cf54573cb621db0597bbb30516 (MD5) Previous issue date: 2015-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Bombyx mori nucleopolyhedrovirus (BmNPV) is an entomopathogenic virus Baculoviridae family, Alphabaculovirus genre, which infects the silkworm, Bombyx mori and causes nuclear polyhedrosis disease. A viral geographic isolated from BmNPV was identified in the state of Paraná, Brazil, and a number of organs and target tissues were identified, but there is no information about the infection of Malpighian tubules (MT). MT make up the excretory system of B. mori, acting in the elimination of toxic substances and electrolyte homeostasis. This study aimed to analyze the behavior on the susceptibility and cytopathology of MT B. mori to BmNPV. Therefore, hybrid B.mori caterpillars at 5º instar, were inoculated experimentally with a viral suspension of BmNPV. On different days post-inoculation (dpi), the 4° to 9° dpi, MT segments were collected, divided in regions of the ampullae, proximal, medial and distal; following the processing for light and electron microscopy transmission. Material control followed the same process that the inoculated. The results showed that B. mori MT showed variations in their morphology and susceptibility to BmNPV according to the analyzed region. Thus, the region of the ampullae in its transition area to the proximal, appeared susceptible to BmNPV, from 6° dpi, while other regions showed no evidence of infection by the pathogen. The transition area is a small region which is located at the end of the ampullae and the beginning of the proximal region of MT. The cellular morphology of simple lining epithelium showed continuous variation of pavement, characteristic of the ampullae, the cylindrical, in the transition area; already continuous with the proximal region of MT, this morphological change was abruptly. The cytopathology revealed in hypertrophic nucleus, viroplasm containing the nucleocapsid and viral polyhedra. At the end of the infectious cycle was observed cytolysis of infected cells, releasing polyhedra in the extracellular medium, both in hemocele as in MT lumen, a factor that may favor the transmission of the disease. Thus, infection of the ampullae in its transition area to the proximal region, will contribute to greater scientific knowledge of cytopathology aspects and targets of infection from this virus isolated geographical, for the establishment of its infectious cycle of the insect's body BmNPV / Bombyx mori nucleopolyhedrovirus (BmNPV) é um vírus entomopatogênico da família Baculoviridae, gênero Alphabaculovirus, que infecta o bicho-da-seda, Bombyx mori (Lepidoptera: Bombycidae) e causa a doença poliedrose nuclear. Um isolado geográfico viral do BmNPV foi identificado no estado do Paraná, Brasil, e uma série de órgãos e tecidos alvos foram identificados, porém não há informações sobre a infecção dos túbulos de Malpighi (TM). Os TM compõem o sistema excretor de B. mori, atuando na eliminação de substâncias tóxicas e na homeostase hidroeletrolítica. Assim, este estudo objetivou analisar o comportamento referente à susceptibilidade e citopatologia dos TM de B. mori ao BmNPV. Para tanto, lagartas híbridas de B. mori, 5° instar, foram inoculadas experimentalmente com uma suspensão viral de BmNPV. Em diferentes dias pós-inoculação (dpi), do 4° ao 9° dpi, segmentos dos TM foram coletados, subdivididos nas regiões da ampola, proximal, média e distal; seguindo o processamento para as microscopias de luz e eletrônica de transmissão. Material controle seguiu o mesmo processamento que o inoculado. Os resultados demonstraram que os TM de B. mori, apresentaram variação na sua morfologia e susceptibilidade ao BmNPV de acordo com a região analisada. Assim, a região da ampola, na sua área de transição com a proximal, se apresentou susceptível ao BmNPV, a partir do 6° dpi, enquanto as demais regiões não evidenciaram indícios de infecção pelo patógeno. A área de transição é uma pequena região que se localiza ao término da ampola e inicio da região proximal do TM. A morfologia celular do epitélio de revestimento simples apresentou variação continua de pavimentoso, característico da ampola, a cilíndrico, na sua área de transição; já na continuidade com a região proximal do TM, esta variação morfológica foi abrupta. A citopatologia revelou no núcleo hipertrófico, o viroplasma, contendo os nucleocapsídeos e os poliedros virais. Ao final do ciclo infeccioso foi verificado a citólise de células infectadas, liberando poliedros no meio extracelular, tanto na hemocele como no lúmen do TM, fator que pode favorecer a transmissão da doença. Assim, a infecção da ampola, na sua área de transição com a região proximal, irá contribuir com um maior conhecimento científico dos aspectos citopatológicos e dos alvos de infecção deste isolado geográfico viral, visando o estabelecimento do seu ciclo infeccioso do BmNPV no corpo do inseto.

The goal of the thesis is to incorporate materials with either fluorescent or conductive properties in silk fibers, by feeding silkworms with a diet containing these materials. To achieve this, one would have to breed (rear) silkworm from eggs into larvaes, then to feed the silkworms with this special diet containing fluorescent or conductive materials. Samples of silk were then collected either from spun cocoons or via removing the silk producing organs (silk glands) from the silkworms via dissection. The samples were then analyzed with absorbance spectrometer, spectrofluorometer or via photoluminesecent measurement to determine if any materials had been incorporated into the silk fibers.   Silkworms were successfully reared from eggs up to moths, once the silkworm larvae had grown enough in size their diet were switches from their regular food (silkworm chow) to food containing conjugated molecules or polymers with fluorescent or conductive properties. A total of 14 materials were tested. One material gave a clear positive result and that was from the fluorescent compound Rhodamine B. Other fluorescent materials, Nile red and POWT yielded some results indicating their presence in the silk but the results were not conclusive. The rest of the materials all failed with being incorporated within the silk fibers; this was due to their lethality, size, lack of zwitterionic properties and such.  The properties of the materials are of great importance for the uptake process, where a small zwitterionic molecule has a great change of being taken up and incorporated in the silk fibers. Whereas a big materials such as a polymer without any zwitterionic will in most cases just follow through the food in the digestive track without any uptake.

In our study, we examined the polymorphism of gene Ser2 from domesticated silkmoth Bombyx mori and its closest wild relative B. mandarina. As a starting material for our work, we used the restriction map of allele C isolated from hybrid lineages 200 and 300 of European silkmoth B. mori (Michaille et al. 1990a). We also used the published sequence of allele D which was isolated from {\clq}qDaizo`` p50 strain of B. mori (Kludkiewicz et al 2009). Based on the published sequence, we designed PCR primers and performed sequence analysis of 4 different alleles of gene Ser2, including the almost complete genomic sequence of allele C (except for two short regions containing repetitive DNA). We then compared the sequences and found that the alleles differ significantly not only in intron lengths and the sizes of exon 9, but also in exon arrangements. While the allele C contains 12 exons (1, 2, 3, 4, 5, 6, 7, 8a, 9, 10, 11, 8b), the alelle D and other alleles analysed have 13 exons (1, 2, 3, 4, 5, 6, 7, 8, 9a, 10a, 9b, 10b, 11). Allele C showed very unique arrangement of exons, which suggested that it may represent an ancestral form of Ser2 gene. We analysed the arrangment of exons located at the 3´end of the Ser2 gene by PCR and electrophoresis in 70 available strains of B. mori and B. mandarina. We also tried to find a similar allele to the allele C and we amplified and sequenced 2 kb region from 55 Bombyx strains. Phyllogenetic analysis of gene Ser2 gene suggested that the C allele of now probably extinct European silkmoth hybrids 200 and 300 does not seem to be an ancestral form of Ser2 gene, but it rather seems to be the result of complex secondary rearrangements.

Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(工学) / 甲第17556号 / 工博第3715号 / 新制||工||1566(附属図書館) / 30322 / 京都大学大学院工学研究科機械理工学専攻 / (主査)教授 富田 直秀, 教授 井手 亜里, 教授 安達 泰治 / 学位規則第4条第1項該当

This diploma thesis is focused on the silk production of invertebrates. The aim of the work is to systematically introduce important species and describe the purpose of their silk production. The ability to produce silk is widespread in many insect families. The production of proteinaceous fibers also occurs in arachnids and (in the form of byssus threads) also in mollusks. The Domestic Silk Moth (Bombyx mori) is an economically important insect, being a primary producer of silk. Hence, an integral part of the thesis focuses on Domestic Silk Moth. In this thesis all developmental stages of Silk Moth (from an egg to an adult) are described. In the past, efforts to breed silkworms also appeared in the Czech lands. Therefore, a short chapter of the thesis is devoted to the history of sericulture. Furthermore, this thesis describes methods of rearing silkworm larvae. For students of Natural Sciences and Biology, it is crucial to be in touch with live objects. Because the silkworm rearing is quite simple, it is possible to incorporate this topic to the school projects about holometabolous insects. The practical part of this thesis contains schedule of practical exercise about silk moths. And in final section of practical part, worksheets for lower-secondary students are presented. Keywords: silkworm...