Thesis (M.Sc. (Hons.)) -- University of Western Sydney, 2003. / "A thesis presented in partial fulfilment of the requirements for the degree of Master of Science (Honours), College of Science, Technology and the Environment, University of Western Sydney ... January, 2003" Bibliography : leaves 142-150.

Bark beetles are major pests of pine forests in North America that can inflict considerable damage and cause severe economic loss. The checkered beetle Thanasimus dubius Fabricius (Coleoptera: Cleridae), an abundant predator in the Eastern USA and Canada, has been suggested to influence the dynamics of several bark beetles, including the economically important southern pine beetle (SPB), Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae). Because of the importance of T. dubius in pine forests, I studied several aspects of T. dubius biology including both its chemical and dispersal ecology. I first present an experiment that was conducted to improve the rearing system of T. dubius so that sufficient numbers of predators could be produced more efficiently for both behavioral and ecological studies. Improvements to the rearing method potentially could be achieved by adding a preservative to increase the shelf-life of the diet used to feed larval T. dubius thereby allowing feeding intervals to be extended. To evaluate this, I added sorbic acid, a preservative, to the larval diet at three different concentrations (0, 0.1 and 0.2%) and for two different intervals between feedings (2-3 vs. 5 days). Additionally, I measured potential deleterious effects of this preservative on predator performance (i.e., female fecundity). I then assessed the effect of this predator on SPB survival by releasing newly hatched T. dubius larvae at several densities (0, 50 100, and 200) on pine logs infested by SPB. I also studied the chemotactic response of this predator toward various volatiles such as prey pheromones or tree volatiles in a wind tunnel. I developed a method that associated upwind flight behavior and antennal behavior to quantify the relative attractiveness of each tested source of chemical. I then conducted three experiments to investigate the ability of individual T. dubius to respond to different prey olfactory signals (bark beetle pheromones) and assess their potential for learning. First, I examined individual predator preferences toward three major prey pheromones (ipsenol, ipsdienol, and frontalin) in a study that combined a mark-release-recapture experiment with a field choice test. I also measured the responses of individual wild T. dubius in a wind tunnel, which allowed the testing of a wider range of semiochemicals (frontalin, ipsenol, ipsdienol, sulcatol, and α-pinene). A final wind tunnel experiment tested the ability of naive predators to learn two prey pheromones, frontalin and ipsenol, that were presented alone or associated with a reward. In the last chapter, I quantified the dispersal of this predator and its behavior along an edge separating a pine forest from a non-suitable habitat. I analyzed T. dubius distribution and movement in the field relative to its bark beetle prey Ips grandicollis Eichhoff (Coleoptera: Curculionidae) and to the root feeder Dendroctonus valens LeConte (Coleoptera: Curculionidae). These three insects are associated with the phenomenon of red pine decline in the Great Lakes area. Thus, understanding the relative pattern of movements of these three beetles could provide considerable knowledge on the spatial and temporal progression of red pine decline by, for instance, assessing the connection between existing pockets. I estimated the dispersal quantiles for this predator relative to its bark beetle prey and determined whether dispersal behavior was relatively homogeneous (one kind of disperser) or heterogeneous (two kinds). In a second part, I sampled the abundance of T. dubius, I. grandicollis, and D. valens along transects set between a matrix area constituted by a clearing and a habitat zone composed of red pine forest. I further developed a diffusion model, including a constant k (corresponds to the ratio of densities along the edge), that permits characterization of beetle behavior around the boundary. Studies developed in the present dissertation, therefore, investigated several aspects of T. dubius ecology: predation on SPB, ability to respond to an olfactory signal, and dispersal traits. I first showed that augmentation with larval T. dubius can successfully reduce the SPB ratio of increase in infested pine logs. I also developed a system of rearing T. dubius that was more efficient and could allow larger numbers of predators to be produced. Sorbic acid did not reduce adult lifetime or size but did affect female fecundity (20 - 40% reduction). Increasing feeding time interval to 5 days (instead of 2-3 days) improved the efficiency of the system, even when sorbic acid was not added in the diet. Wind tunnel experiments present a methodology that could enable future quantification of the relative attractiveness of various semiochemical treatments. I identified a particular antennal behavior, which I called "stand up antennal" that was typically produced when the insect was attracted to a semiochemical. The choice tests in the field and the wind tunnel experiment using wild adults both showed that T. dubius individuals are generalists that are flexible in their response to a broad range of semiochemical signals (frontalin, ipsenol, ipsdienol, sulcatol, and α-pinene). I demonstrate that naive predators are more attracted toward ipsenol after having experienced conditioning with this kairomone via a reward. Finally, experiments on dispersal and edge behavior revealed that T. dubius and one of its common prey, the bark beetle I. grandicollis, exhibit a different pattern of movement within red pine stands and surrounding habitats. First, adult T. dubius can disperse substantially further (50% dispersed beyond 1.54 km) than I. grandicollis adults (50% dispersed beyond 0.13 km). Second, T. dubius was highly affected by the presence of an edge, showing a step-like pattern with very predators caught in unsuitable areas. Conversely, the behavior of I. grandicollis was less limited by such a boundary. As with T. dubius, the root colonizer D. valens showed a strong edge behavior that was characterized by avoiding matrix zones.

Cairomônios são semioquímicos envolvidos nas interações interespecíficas dos insetos, que beneficiam o receptor em detrimento do emissor. Uma destas interações envolve a busca hospedeira por parasitoides, que detectam estes compostos como estratégia para localizar seus hospedeiros. No caso do ectoparasitoide larval, Habrobracon hebetor Say (Hymenoptera: Braconidae), o \'frass\' de Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) libera um cairomônio que atrai as fêmeas do parasitoide. Considerando que A. kuehniella é muito utilizada para produção de ovos, visando à criação de inimigos naturais, este cairomônio é indesejável, pois acaba favorecendo altas infestações por H. hebetor, podendo causar prejuízos expressivos ao longo do processo. Uma das soluções para o controle de tais infestações poderia ser o emprego de armadilhas com o semioquímico sintético. Entretanto, como o cairomônio ainda não foi identificado, este trabalho teve como objetivo o seu isolamento - a etapa inicial para sua identificação. O cairomônio foi extraído por meio da lavagem com solvente e aeração do \'frass\' de A. kuehniella. Os extratos foram, então, testados em olfatometria de quatro vias com fêmeas do parasitoide, registrando-se o tempo de permanência em cada tratamento. O extrato de lavagem ativo foi fracionado em coluna de sílica em cinco frações, cuja atratividade também foi averiguada para proceder com o isolamento do cairomônio. Os extratos e frações foram então analisados por cromatografia gasosa por ionização em chama (GC-FID) e espectrometria de massas (GC-MS), a fim de localizar compostos exclusivos nas amostras ativas. Além disso, as frações provenientes do extrato de lavagem foram aeradas para a extração de seus compostos voláteis e também analisadas por GC-MS. O cairomônio foi isolado na fração 75% éter, indicando ser um composto polar. Por comparação dos cromatogramas, apenas um pico na coluna HP-5ms foi selecionado como pertencente ao cairomônio. Em coluna Rtx-1ms, o pico bifurcou-se, indicando que, pelo menos, um dos compostos faz parte do cairomônio. As combinações metodológicas deste trabalho conduziram ao isolamento do cairomônio presente no \'frass\' de A. kuehniella e à seleção de pelo menos um composto que compõe o semioquímico responsável pela atração do parasitoide H. hebetor, resultando nas primeiras informações de sua estrutura química para sua futura identificação. / Kairomones are semiochemicals involved in interspecific interactions of insects that benefit the receiver to the detriment of the emitter. One of these interactions is host searching by parasitoids which detect these compounds as a strategy to locate their hosts. In the case of the larval ectoparasitoid, Habrobracon hebetor Say (Hymenoptera: Braconidae), the \'frass\' of Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) releases a kairomone that attracts females parasitoids. Considering that A. kuehniella has been used for egg production in order to keep natural enemy rearing, this kairomone is undesirable, because it may lead to high H. hebetor infestation and causes important losses during the rearing process. One of the attempts to control these infestations could be the use semiochemical-baited traps. However, as the kairomone is still unidentified, the aim of this work was to isolate it - the first step for its identification. The kairomone was extracted by solvent washing and aeration of A. kuehniella \'frass\'. Then the extracts were tested in a four-way olfactometer with female parasitoids, registering the time taken in each treatment. The active washing extract was fractionated in five fractions in a silica column, and its attractivity was evaluated to proceed to isolation. The extracts and fractions were then analysed by gas chromatography with flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC-MS), in order to locate exclusive compounds in active samples. Furthermore, the fractions from the washed extract were aerated to extract their volatile compounds that were also analysed by GC-MS. The kairomone was isolated in the 75% ether fraction, indicating that it is a polar compound. Comparing the chromatograms, only one peak on the HP-5ms column was selected as belonging to the kairomone. On the Rtx-1ms column, the peak bifurcated, indicating that at least one of the compounds is part of the kairomone. The methodological combinations of this work provided the isolation of the kairomone present in A. kuehniella \'frass\' and the selection of at least one compound that comprises the semiochemical that attracts H. hebetor parasitoid, resulting in the first source of information about its chemical structure for its future identification.

Among several early-season insect pests threatening apple production in eastern North America, plum curculio (PC), Conotrachelus nenuphar Herbst (Coleoptera: Curculionidae), is one of the most devastating pests of pome and stone fruits. Historically, several methods have been used to monitor and control plum curculio in apple orchards. The odor-baited trap tree approach has proven to serve as an effective monitoring and attract-and-kill system for plum curculio. The synergistic lure consisting of benzaldehyde (a plant volatile) and grandisoic acid (the PC aggregation pheromone) is used in this approach. However, the high cost of the lure and the degradation of benzaldehyde by UV light and heat have resulted in limited grower adoption. The major goals of this study were (1) to evaluate under field conditions, the attractiveness of methyl salicylate (MES) to PC, and to determine if it can replace the benzaldehyde (BEN), and (2) to test whether we can manipulate insect pest behavior through host plant preference by grafting selected perimeter- row trees with multiple cultivars. My results indicated that (1) methyl salicylate in combination with grandisoic acid was as attractive to PC as the standard synergistic lure composed of benzaldehyde and grandisoic acid, therefore MES can be used as a replacement for BEN, (2) trees grafted with six cultivars were more attractive to PC and concentrated more PC fruit injuries, compared to non-grafted trees, and (3) ‘Wickson’ and ‘Red Astrachan’ received numerically more PC injuries while ‘Dabinett’ received the lowest level of injury compared with other cultivars. These results highlight that a semiochemical-based strategy is a low-cost and grower-friendly approach to monitoring and controlling the PC population.

Thesis (PhD (Chemistry and Polymer Science))--University of Stellenbosch, 2010. / Video 1: Twin-bearing Döhne Merino ewe butting only one of her lambs in typical rejection behavior. Rejection of one twin is not a common phenomenon. (Video by Dr Marlize Viviers) / Video 2: Twin-bearing ewe # 22 accepting alien lamb # 179. The alien lamb wears a jacket her other lamb has been wearing for the preceding 24 hours. The excessive sniffing at the jacket, bleating and looking around show that the ewe is not totally satisfied with the situation. (Video by Dr Marlize Viviers) / Video 3: Ewe # 22 rejecting alien lamb # 179 after removal of the jacket it has worn in the previous video. (Video by Dr Marlize Viviers) / ENGLISH ABSTRACT: Sheep husbandry substantially contributes to South Africa’s economy. Lamb mortality is regarded as a major constraint to sustainable sheep production, and an important factor in lamb survival is the strong ewe–lamb bonding. This is a well-known phenomenon in all sheep races and strong evidence exists that the selective ewe–lamb bond formation has a pheromonal basis, in that the ewe can distinguish her lamb by its unique, familiar smell. Sheep farmers and scientists have examined various methods of making alien lambs acceptable to foster ewes. The survival rate of offspring could be substantially increased if a reliable method could be devised to prevent ewes from rejecting their own lambs or if ewes could be induced to accept alien lambs. A better understanding of the semiochemicals involved in this communication between ewe and offspring could facilitate the development of a method by which ewes can be coaxed into adopting foster lambs. This would positively contribute to efficient sheep production in South Africa. The objective of this study was to characterise the volatile constituents present in the wool and the amniotic fluid collected from the Döhne Merino lambs to achieve a better understanding of the ewe–lamb semiochemical communication. The volatile organic constituents associated with the wool and amniotic fluid were characterised and a total of 133 volatile organic compounds were identified. Several compound classes are represented in these samples and the majority of these compounds have previously been identified in the excretions and secretions of numerous mammalian species. A number of different aspects of ewe–lamb kin recognition were investigated. First, the odour profiles of twin lambs were compared to those of other twins. The results of these analyses revealed that the wool volatiles of twin lambs are qualitatively as well as quantitatively practically identical, but different from those of other lambs in the flock. Second, the change in the odour profiles of lambs was studied over a seven-day period. It was found that the odour profiles of lambs are not fixed, and for as long as a ewe makes use of olfactory recognition as the main mode of identification of her lamb she has to continue learning the odour of her lamb. Third, the amniotic fluid was investigated as a possible source of maternal labels, i.e. as chemical codes consisting of mixtures of compounds in specific quantitative ratios that are imprinted on lambs by their mothers at birth. It was found that the amniotic fluids of first- and second-born lambs are qualitatively and quantitatively different. It is possible that the duration of the lambing birth process could contribute to this change in amniotic fluid composition. Fourth, the role of residual proteins in the dissemination of the semiochemicals involved in ewe– lamb communication was investigated and six proteins were identified as amniotic fluid and wool-associated proteins. Bioassays carried out during the lambing season of 2009 confirmed the role of the odour of the lamb in ewe-lamb recognition, but bioassays carried out with synthetic pheromone formulations gave inconclusive results. / AFRIKAANSE OPSOMMING: Skaapboerdery lewer aansienlike bydrae tot die ekonomie van Suid Afrika, maar lamsterftes is een van die kwellende probleme waarmee kleinveeboere te kampe het. Die eksklusiewe band wat binne die eerste paar uur na 'n lam se geboorte tussen die ooi en haar lam ontwikkel, is die hooffaktor waarvan lammers se oorlewing afhang. Hierdie sterk binding tussen lam en ooi is 'n bekende verskynsel in alle skaap-rasse en dit is bekend dat 'n feromoon wat deur die lam vrygestel word, dit vir die ooi moontlik maak om haar lam aan sy unieke reuk uit te ken. Lamsterftes sou aansienlik verminder kon word as hierdie feromoon geïdentifiseer en gebruik kan word om ooie te verhoed om hulle eie lammers te verwerp en om die aanvaarding van vreemde lammers deur ooie moontlik te maak. Die doel van hierdie studie was die chemiese karakterisering van die vlugtige organiese verbindings aanwesig in die amniotiese vloeistof en wol van Döhne Merino lammers ten einde 'n beter begrip van die semiochemiese kommunikasie tussen ooie en lammers te verkry. In die amniotiese vloeistof is 70 vlugtige organiese verbindings geïdentifiseer. Dieselfde verbindings asook 'n verdere 63 verbindings is in lammerwol geïdentifiseer. Die meerderheid van hierdie verbindings is reeds voorheen in verskeie soogdierafskeidings gevind. Die studie het verskeie interessante aspekte van hierdie semiochemiese kommunikasie aan die lig gebring. Eerstens is daar gevind dat die samestelling van die vlugtige organiese materiaal in die wol van tweelinglammers, oftewel hul reukprofiele, feitlik identies is. Verder verskil die reukprofiele van 'n tweeling van dié van ander tweelinge en enkelinge in die trop. Tweedens is gevind dat die reukprofiel van 'n lam nie by geboorte vasgelê word en so behoue bly nie, maar dat dit geleidelik verander oor ten minste die periode van sewe dae waaroor hierdie verskynsel bestudeer is. Dit noodsaak die ooi om haar geheue met betrekking tot die lam se reuk voortdurend te verfris gedurende die tydperk wat sy hoofsaaklik van haar reuksin gebruik maak om haar lam te herken. Derdens is die amniotiese vloeistof ondersoek as moontlike bron van ‘n sogenaamde moedermerk, waardeur chemiese mengsel van verbindings wat in ‘n spesifieke kwantitatiewe samestelling by geboorte deur die ooi op haar lam agter gelaat word. Ten opsigte van kwalitatiewe en kwantitatiewe samestelling, verskil die vlugtige organiese fraksie van die amniotiese vloeistof van die tweelinglam wat eerste gebore word van die amniotiese vloeistof van die lam wat tweede gebore word. Dit is onwaarskynlik dat amniotiese vloeistowwe wat ten opsigte van hul vlugtige organiese fraksies van mekaar verskil, nagenoeg identiese moedermerke op tweeling lammers sou kon laat. Vierdens is vasgestel dat spore van ses proteïene uit die amniotiese vloeistof op die wol van lammers agtergelaat word. Daar kon ongelukkig nie vasgestel word of hierdie proteïene 'n rol in die vrystelling van semiochemiese inligting speel nie. Gedragstoetse wat tydens die lamseisoen van 2009 uitgevoer is, het bevestig dat die lam se reukprofiel deurslaggewende rol in ooi-lam herkenning speel, maar gedragstoetse wat met sintetiese feromoonmengsels uitgevoer is, het onbesliste resultate opgelewer.

Thesis (MSc)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: Bontebok, Damaliscus pygargus pygargus, formerly classified as D. dorcas dorcas, are territorial animals with interdigital glands between the toes of the forelegs. Males regularly defecate on dung heaps, on which they often lie, to communicate with other members of their species. They also communicate by means of visual displays, scent marking and occasionally with scraping or pawing of dung heaps. It is assumed that scent marking with the interdigital secretion serves to define territories frequented by these antelope. These glands secrete a complex mixture of volatile and non-volatile compounds and the volatile compounds in the secretion serve as a chemical signal for other bontebok. It has been suggested that the interdigital secretion is not produced in its final composition by the interdigital gland alone, but that microbial activity is responsible for many of the compounds present in the secretion. In general, many compounds can be attributed to the by-products of microbial hydrolysis of triglycerides, a common characteristic of sebum. It is well documented that micro-organisms inhabit the deep recesses of sebaceous glands and the presence of micro-organisms has been found to be consistent in all antelope exocrine glandular areas. This study involved the chemical characterisation of the volatile metabolites produced in vitro by micro-organisms from the interdigital cavity of the bontebok. Various comparative studies were made, one of which was comparison of the metabolites produced by the individual microbial species as well as the total community of bacteria incubated in different media. A comparison of the compounds identified in the interdigital secretion and the metabolites produced by the micro-organisms in the different media was also made. The volatile metabolite extracts of the individual bacterial species and of the total community were chemically characterised by low-resolution gas chromatography-mass spectroscopy. Classes of compounds identified from the volatile metabolite extracts include: • Acids - Aliphatic (saturated and unsaturated) • Alcohols - Aliphatic (saturated and unsaturated) • Aldehydes - Aliphatic (saturated and unsaturated) • Aromatic compounds • Ketones - Aliphatic (saturated and unsaturated) • Pyrazines • Dimethyldisulphide • Squalene and cholesterol Several qualitative differences were found between the compounds identified in the volatile metabolite extracts of the micro-organisms when incubated in tryptic soy broth (TSB) and minimal salt medium (MSM). In particular, when the microbes were incubated in TSB medium a number of pyrazines were found that were not present when utilising MSM as a medium. Additional qualitative differences were found between the compounds identified in the metabolite extracts of the individual bacterial species and the total community of bacteria, when incubated in both TSB and MSM media. A comparison of the interdigital secretion and the metabolite extracts of the microbial communities incubated in TSB and MSM revealed that many compounds produced in MSM corresponded to the compounds identified in the interdigital secretion. These corresponding compounds were found to be saturated and unsaturated acids, aldehydes and squalene. Furthermore, there was only one corresponding compound in the case of TSB as medium. / AFRIKAANSE OPSOMMING: Die bontebok, Damaliscus pygargus pygargus, voorheen geklassifiseer as D. dorcas dorcas, is 'n territoriale dier met interdigitale kliere tussen die kloutjies van die voorpote. Ramme ontlas gereeld op mishope, waarop hulle dikwels lê, om met ander lede van die spesie te kommunikeer. Hulle kommunikeer ook deur middel van visuele seine, reukmerking en soms deur mishope met die voorpote te kap of te skraap. Reukmerking met die interdigitale afskeiding dien klaarblyklik om gebiede wat deur hierdie diere bewoon word, af te baken. Die interdigitale kliere skei 'n komplekse mengsel van vlugtige en nie-vlugtige verbindings af en die vlugtige verbindings dien as chemiese sein vir ander bontebokke. Die vermoede bestaan dat die interdigitale klier nie alleen verantwoordelik is vir die finale samestelling van die interdigitale afskeiding nie, maar dat mikrobiese aktiwiteit bydra tot die produksie van baie van die verbindings wat in die afskeiding aanwesig is. Sekere verbindings kan in die algemeen toegeskryf word aan die vorming van die neweprodukte van mikrobiese hidrolise van trigliseriede, 'n algemene eienskap van sebum. Dit is bekend dat die diep holtes van vetkliere 'n goeie teelaarde is vir mikroorganismes en daar is gevind dat mikroorganismes feitlik deurgaans voorkom in alle anteloop eksokriene klierareas. Hierdie studie behels die chemiese karakterisering van die vlugtige metaboliete wat in vitro deur mikroorganismes van die interdigitale klierholte van die bontebok geproduseer word. Verskeie vergelykende studies is uitgevoer waarvan een die vergelyking was van die metaboliete wat deur die individuele mikrobiese spesies sowel as die totale gemeenskap van bakterieë geproduseer word tydens inkubasie in verskillende media. Vergelyking van die verbindings wat in die interdigitale afskeiding geïdentifiseer is met die metaboliete wat in verskillende media geproduseer is, het ook deel van die studie uitgemaak. Die vlugtige metaboliet ekstrakte van die individuele bakteriese spesies en van die totale gemeenskap is chemies gekarakteriseer deur middel van laeresolusie gaschromatografie-massaspektrometrie. Die volgende groepe verbindings is onder andere in die vlugtige metaboliet ekstrakte geïdentifiseer: • Sure - Alifaties (versadig en onversadig) • Alkohole - Alifaties (versadig en onversadig) • Aldehiede - Alifaties (versadig en onversadig) • Aromatiese verbindings • Ketone - Alifaties (versadig en onversadig) • Pirasiene • Dimetieldisulfied • Skwaleen en cholesterol Verskeie kwalitatiewe verskille is gevind tussen die verbindings wat geïdentifiseer is in die vlugtige metaboliet ekstrakte van die mikroorganismes onderskeidelik in TSB medium en MSM geïnkubeer. Opvallend was byvoorbeeld die voorkoms van pirasiene in gevalle waar mikroorganismes in TSB medium geïnkubeer is, terwyl hierdie groep verbindings afwesig was wanneer MSM gebruik is. Onderlinge kwalitatiewe verskille is ook gevind tussen die verbindings wat geïdentifiseer is in die metaboliet ekstrakte van die individuele bakteriese spesies en die totale gemeenskap van bakterieë, wanneer in TSB medium sowel as in MSM geïnkubeer is. Vergelyking van die verbindings in die interdigitale afskeiding en in die metaboliet ekstrakte van die mikrobiese gemeenskappe, het getoon dat 'n aantal verbindings wat in MSM geproduseer is, ooreenstem met verbindings wat in die interdigitale afskeiding geïdentifiseer is. Daar is gevind dat hierdie verbindings versadigde en onversadigde sure en aldehiede en skwaleen is. Met TSB as medium was daar slegs een ooreenstemmende verbinding.

Rodent commensal species produce great damage in agriculture and urban areas. As invasive species they can endanger local species and are carriers and vectors of several important zoonoses. Control methods rely mainly on the use of warfarins, which can be inadvertently be taken up by untargeted species. Warfarins have also lost their efficacy in rodents due to the development of genetic resistance. In addition, these methods are considered inhumane as they cause a slow and painful death due to haemorrhages. Olfaction is a main source for environmental risk assessment by rodents, and it can be used to modify their use of space. My aim in this thesis was to identify behavioural reactions of the house mouse (Mus musculus), using laboratory strains as models of wild animals, to ecologically meaningful chemical messages, including predator and plant chemical olfactory cues. My results showed that mice avoided complex ferret olfactory cues and ethanol which is a ubiquitous chemical related to fruit rotting and ripening. The feline protein Fel d 1, which belongs to the secretoglobin family and is a major cat allergen in humans, did not elicit significant avoidance or alter foraging behaviour in mice. However, Trimethylthiazoline purified from fox faeces, elicited clear avoidance behaviour and stress responses. I carried out a bibliographic review to evaluate and discuss rodent pest control methods from an ethical standpoint. This literature showed that many of the current methods of pest control are considered inhumane, and do not tally with current society concerns and welfare standards in other domains such as farms or laboratory animals. These results raise new research questions to identify ferret and plant chemical compounds that can induce rodent avoidance, and to carry out next stage of research with wild animals both under laboratory and field conditions.

As respostas dos nematoides entomopatogênicos, Steinernema carpocapsae (Weiser) e Heterorhabditis indica (Poinar, Jackson e Klein), aos voláteis emitidos por raízes de cana-de-açúcar (Saccharum officinarum L.) danificadas ou não danificadas por ninfas da cigarrinha-da-raiz, Mahanarva fimbriolata (Stål), foram estudadas em laboratório. Plantas de cana-de-açúcar foram transplantadas para câmaras laterais do olfatômetro de seis vias e submetidas ou não a herbivoria por M. fimbriolata. Cerca de 10000 nematoides entomopatogênicos foram liberados na câmara central do olfatômetro e após 24 horas os nematoides foram contabilizados em cada câmara lateral do dispositivo. Raízes de plantas previamente danificadas ou não danificadas por M. fimbriolata foram destacadas das plantas, congeladas com nitrogênio liquido e maceradas. O material foi então aerado por 8 horas para a coleta dos voláteis. O perfil dos voláteis dos extratos naturais foi analisado em um cromatógrafo gasoso acoplado a um espectrômetro de massas (GC-MS). Os nematoides de ambas as espécies foram atraídos pelos voláteis radiculares de plantas danificadas pela cigarrinha-das-raízes, em contraste com plantas não danificadas e areia umedecida (controle). As análises em GC-MS revelaram que não houve diferença qualitativa no perfil de voláteis radiculares entre os tratamentos. Por outro lado, dos 11 compostos presentes nas raízes não danificadas, di-hidro mircenol e Beta isso metioneno reduziram, significativamente, sua concentração após o ataque de M. fimbriolata. Estes resultados sugerem que o recrutamento de nematoides entomopatogênicos neste sistema pode ter sido influenciado pela diminuição da concentração destes dois compostos radiculares nas plantas sob herbivoria. / The responses of entomopathogenic nematodes Steinernema carpocapsae (Weiser) and Heterorhabditis indica (Poinar, Jackson e Klein), to volatiles emitted by sugar cane roots damaged or undamaged by Mahanarva fimbriolata (Stål) nymphs, were investigated under laboratory conditions. Thus, sugarcane plants damaged or undamaged by M. fimbriolata were transplanted to lateral arms of six arm olfactometer. Approximately 10000 entomopathogenic nematodes were released in the central chamber of the olfactometer and after 24 hours, the nematodes were counted in each arm. Roots of previously damaged or undamaged plants were detached, frozen with liquid nitrogen and macerated. The material was then aerated for 8 hours to collect volatiles. The volatiles profile was analyzed by a gas chromatograph-mass spectrometer. Most nematodes of both species moved to plants that had been previously damaged over undamaged plants, and there was no significant difference in the choice of undamaged plant and only moisted sand. The GC-MS analyzes showed no qualitative difference between the root treatments volatiles profile. In both treatments, the same 11 compounds were found. Although, two compounds, dihydro myrcenol and Beta-iso-methionene, showed quantitative difference between treatments, observing greater production in undamaged plants, i.e. herbivory caused reduction in tested sugar cane roots volatiles releasing. These data provide new information about the tritrophic relationships occurring in the soil which, in the future, can be used in order to develop field management for M. fimbriolata.

O estudo do comportamento sexual dos insetos abrange todos os eventos ocorridos desde a formação dos casais por meio do cortejo, até a separação dos mesmos após a cópula. Esses estudos são importantes e representam o primeiro passo em pesquisas que visam à elucidação de feromônios da espécie alvo. No caso da broca-do-café, Hypothenemus hampei (Ferrari), que é considerada mundialmente a principal praga do café, existe uma lacuna de informações sobre o comportamento sexual dessa espécie. Além disso, as pesquisas têm sido direcionadas para o uso de armadilhas contendo alcoóis (ex: etanol e metanol) visando o manejo dessa praga, mas com resultados conflitantes. Em razão disso, o objetivo do presente trabalho foi estudar o comportamento sexual de H. hampei, como suporte para estudos subseqüentes visando a elucidação do feromônio da espécie. As cópulas de H. hampei ocorreram em qualquer horário dentro de um ciclo de 24 horas. Embora em menor freqüência, 60% dos casais realizaram pelo menos uma cópula na idade de 0-24 horas, chegando a 100% dos casais nas demais idades. A maior atividade sexual foi observada nas idades de 48-72 e 72-96 horas, onde, nessa última, ocorreram 6,9 ± 0,47 cópulas por casal com duração de 2,04 ± 0,13 minutos cada. Os padrões comportamentais exibidos foram semelhantes a outros grupos de curculionídeos, com pré-cópula, cópula e pós-cópula. Os resultados indicaram ainda a presença de um feromônio produzido pelos machos mediando o acasalamento. A possibilidade de as fêmeas de H. hampei realizarem a recópula após sua saída dos frutos abre novas perspectivas sobre a viabilidade deste feromônio no manejo desta praga. / The sexual behavior study in insects includes all the events occurred since of pair encounter by means of courtship until their separation after mating. These studies are important and represent the first step of researches that aim to elucidate pheromones of target species. Regarding the coffee berry borer, Hypothenemus hampei (Ferrari), which is the most important coffee pest around the world, little is known about its sexual behavior. Furthermore, the researchers have been focused on the use of traps based on alcohols (e.g. ethanol and methanol) with aims of managing this pest, but its results are conflicting. For that reason, the aim of the present work was to study the sexual behavior of H. hampei, whose it will give support to subsequent studies about evaluation of pheromone this species. Daily mating rhythm of H. hampei occurred on any time of 24- hour cycle. Although at low rates, 60% of paired adults mated at least once when they were 0-24 hours, reaching 100% in older pairs. The highest sexual activity was observed in the ages of 48-72 and 72-96 hours, being in this last one the occurrence of 6,9 ± 0,47 matings/pair with a mean duration of 2,04 ± 0,13 minutes each. The pattern behavior displayed was similar to other groups of Curculionidae, with premating, mating and postmating. The results also indicated the presence of a pheromone produced by the males that mediates mating. The possibility of the H. hampei females recopulates after they leave the fruits opens new perspectives to investigate the viability of using this pheromone in management of this pest.

Sphenophorus levis (Coleoptera: Curculionidae) é uma das principais pragas de importância econômica na cana-de-açúcar. Seu ataque pode causar a morte da planta, promovendo falhas na rebrota e dependendo do nível de infestação, inviabilizar novos cortes. Além disso, é considerado um inseto de difícil manejo. Dentre as razões, destacam-se o hábito críptico de suas larvas, que se alimentam no interior do colmo da cana, e dos adultos, que permanecem boa parte de seu tempo abaixo da palhada sobre o solo. Por esta razão, os métodos tradicionais de controle, como os agroquímicos têm sido pouco eficientes. Neste sentido, a busca por novos conhecimentos e novas estratégias para o manejo desta praga tem se tornado frequente. Assim, o objetivo principal deste trabalho foi o de reinvestigar o feromônio de S. levis, visando a possível existência de novos compostos químicos na sua composição. Além disso, foram estudados ainda a morfologia e morfometria de S. levis, visando uma ágil e fácil separação por sexo nesta espécie, e uma amostragem populacional das formas biológicas de S. levis em campo, ao longo de uma safra, tendo em vista o manejo desta praga. As fêmeas apresentaram maior tamanho corporal que os machos, porém, houve sobreposição entre o tamanho mínimo da fêmea e máximo do macho, comprometendo uma distinção pelo tamanho. A separação dos sexos em S. levis pode ser feita observando-se a parte ventral do abdome, com os machos apresentando toda a região pilosa e as fêmeas somente com pelos na região apical. Uma maior concavidade presente no final do abdome dos machos, também pode ser utilizada como um parâmetro auxiliar nesta separação. Observando-se a flutuação populacional das diferentes fases biológicas de S. levis, por meio de trincheiras, verificou-se que as larvas ocorrem praticamente durante todos os meses do ano, enquanto que pupas e adultos recém emergidos, em menos da metade das amostras. O pico populacional de larvas, pupas e adultos recém emergidos ocorreu em outubro. Para os adultos, a amostragem por meio de iscas indicou que o pico de maior intensidade foi em dezembro. Na etapa de reinvestigação do feromônio de agregação de S. levis, foram obtidas respostas positivas nas antenas de machos e fêmeas para três compostos químicos. Um destes compostos foi o álcool 2-metil-4-octanol, já identificado anteriormente por Zarbin et al. (2003). Os outros dois, são novos compostos denominados 2,3-butanodiol diacetil (2,3 diacetilbutano) e meso 2,3 butanodiol diacetil (meso 2,3 diacetilbutano). Os três compostos sintéticos foram testados em GC-EAG, porém ainda sem uma resposta conclusiva. Novos testes necessitam ainda ser conduzidos para elucidar a composição química do feromônio de agregação de S. levis. / Sphenophorus levis (Coleoptera: Curculionidae) is one of the main pests of economic importance in sugarcane in Brazil. The attack can result plant death promoting the regrowth and failures depending on the level of infestation, derail further cuts. S. levis is a hard management pest. Among the reasons, it highlights the cryptic habits of larva, which feed inside the sugarcane stalk, and adults live in the soil surface, above of straw. Due to this, the traditional control methods such as agrochemicals have been inefficient. The search for new knowledge and new strategies for the management of this pest has become frequent. Thus, the main objective of this work was to reinvestigate the S. levis pheromone, aiming the possible existence of new chemical compounds in its composition. Beside that, this study aimed to morphometric and morphological comparison between males and females, seeking sex separation in a fast and effective manner. Moreover, population sample of S. levis in field was performed, aiming at an improvement in planning of the management of this pest. Females of S. levis have a greater body size than males; however this parameter cannot be used as a sex indicator, because there was overlap between the minimum and maximum body sizes of females and males respectively. S. levis sex separation can be carried observing abdomen ventral side, where males present hairy venter and females just a tuft in the apical region, and by a greater concavity of the end of the abdomen in males. The fluctuation of S. levis biological stages through trenches, it was found that the larvae occurs during almost all months of the year, while pupae and recently emerged adults in less than half of the samples. The population peak for larvae, pupae and recently emerged adults was in October. For adults, the sampling through baits indicated that the peak intensity was higher in December. Then, in the reinvestigate the S. levis pheromone, natural extracts from males showed responses of antennas of males and females of the species for three chemical compounds. One of these compounds was the alcohol 2-methyl-4-octanol, previously identified by Zarbin et al. (2003). The other two are new compounds, called 2.3-butanediol diacetil (2.3 diacetilbutano) and meso 2.3 butanediol diacetyl (meso 2.3 diacetilbutano). The three synthetic compounds were tested in GC-EAG, but there is still no conclusive answer. New tests still need to be conducted to elucidate the chemical composition of S. levis aggregation pheromone.