An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus: goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa

The aim of this study was to determine the relative cryopreservation success of koala and wombat spermatozoa and to investigate reasons for their respective post-thaw survival by examining the sperm's response to a range of osmotic media and determining the presence and distribution of F-actin. An hypothesis was proposed that F-actin may be imparting a degree of structural inflexibility to the koala sperm plasma membrane; hence, exposure of spermatozoa to cytochalasin D (5 mu M), a F-actin depolymerisation agent, should result in increased plasticisation of the membrane and greater tolerance of cell volume changes that typically occur during cryopreservation. In experiment 1, koala (n = 4) and wombat (n = 4) spermatozoa packaged in 0.25 mL straws were cryopreserved using two freezing rates (fast-3 cm above liquid N2 interface; slow-6 degrees C/min in a freezing chamber) and two glycerol concentrations (8 and 14% v/v) in a tris-citrate glucose buffer with 15% (v/v) egg yolk. Wombat spermatozoa showed better (P < 0.01) post-thaw survival (% motile, % intact plasma membranes, % decondensed sperm heads) than koala spermatozoa. When exposed to media of varying osmolality, koala spermatozoa were less tolerant (% intact plasma membrane) of hyper-osmotic conditions (920 and 1410mOsmol/kg) than wombat spermatozoa. F-actin was localised using a monoclonal antibody but only found in the wombat sperm head. When koala and wombat spermatozoa were exposed to media of varying osmolality, cytochalasin D had no beneficial effect on sperm survival (% intact plasma membranes). This study has demonstrated that wombat spermatozoa are highly tolerant of cryopreservation when compared to koala sperm but that spermatozoa from both species show greatest post-thaw survival when frozen slowly in 14% glycerol. Koala sperm are also particularly susceptible to hyper-osmotic environments but lack of detectable F-actin in the koala spermatozoan suggests that poor cryopreservation success in this species is unlikely to be associated with F-actin induced plasma membrane inflexibility. (c) 2006 Elsevier Inc. All rights reserved.

Cloning and sequence analysis of pituitary prolactin cDNA from the northern brown bandicoot (Isoodon macrourus)

The nuclectide sequence for pituitary prolactin cDNA from the marsupial bandicoot (Isoodon macrourus) was determined by reverse transcription-polymerase chain reaction and 5'/3' rapid amplification of cDNA ends. The deduced amino acid sequence showed high sequence identity with brushtail possum prolactin (95%) and all of the expected structural features of a quadruped prolactin. A prolactin gene tree was constructed and rates of evolution calculated for bandicoot, possum, opossum and several mammalian and non-mammalian prolactins. Bootstrap analysis provided strong support for marsupials as a sister group with eutherian mammals and weak support for opossum and bandicoot as an independent grouping from the brushtail possum. The rates of molecular evolution for marsupial prolactins were comparable to the slow rate seen in the majority of quadruped prolactins that have been sequenced. (c) 2005 Elsevier Inc. All rights reserved.

Male reproductive tactics and female choice in the solitary, promiscuous bridled nailtail wallaby (Onychogalea fraenata)

Several behavioral studies of large, gregarious, and sexually dimorphic macropods have shown that males form dominance hierarchies and large males have the highest reproductive success. The bridled nailtail wallaby (Onychogalea fraenata) is a smaller and strongly sexually dimorphic macropod, but is also highly solitary and males do not form dominance hierarchies that are maintained temporally or spatially. Genetic studies of paternity have shown that large males are the most reproductively successful and only one-quarter of males sire offspring at any one time. The aim of this study was to investigate the tactics that males adopt to secure access to females at the time of estrus and to investigate whether females can influence which males have access to them. This study was conducted using 2 wild, free-ranging populations of bridled nailtail wallabies. Females in estrus were located and observed. and the total number of males present, the relative weight rank of each mate, and interactions between individuals were recorded. Females showed a preference for large males and incited male-male competition when the group of males present was large. Unlike other dimorphic macropods, fights among males were rare and were restricted to males of similar size. Large males gained access to females by guarding and following them closely and threatening other males who attempted to gain access. Smaller males spent less time with females, suggesting that small males may leave multimale groups in an attempt to locate unguarded females. Given the solitary nature of this species and the lack of a stable dominance hierarchy to influence male reproductive success. mate searching and mate guarding may be important male reproductive tactics in this species.

Transposon-free regions in mammalian genomes

Despite the presence of over 3 million transposons separated on average by similar to 500 bp, the human and mouse genomes each contain almost 1000 transposon-free regions (TFRs) over 10 kb in length. The majority of human TFRs correlate with orthologous TFRs in the mouse, despite the fact that most transposons are lineage specific. Many human TFRs also overlap with orthologous TFRs in the marsupial opossum, indicating that these regions have remained refractory to transposon insertion for long evolutionary periods. Over 90% of the bases covered by TFRs are noncoding, much of which is not highly conserved. Most TFRs are not associated with unusual nucleotide composition, but are significantly associated with genes encoding developmental regulators, suggesting that they represent extended regions of regulatory information that are largely unable to tolerate insertions, a conclusion difficult to reconcile with current conceptions of gene regulation.

Use of a GnRH agonist and hCG to obtain an index of testosterone secretory capacity in the koala (Phascolarctos cinereus)

Testosterone secretion in mammals typically occurs in random pulses such that a single blood sample provides limited information on reproductive endocrine status. However, it has been shown in several species that an index of the prevailing testosterone biosynthetic capacity of the testes can be obtained by measuring the increase in circulating testosterone after injection of a GnRH agonist or human chorionic gonadotrophin (hCG). Hence, the aims of the present study were to examine fluctuations in testosterone secretion in the koala (n = 6) over a 24-hour period and then characterise testosterone secretion after injection of the GnRH agonist buserelin (4 mu g) or hCG (1000 IU). The latter was used to establish an index of the prevailing testosterone biosynthetic capacity of the koala testis. Individual koalas showed major changes in blood testosterone concentrations over 24 hours, but there was no apparent diurnal pattern of testosterone secretion (P >.05). Injection of buserelin and hCG resulted in an increase (P

An extracellular transglutaminase is required for apple pollen tube growth

An extracellular form of the calcium-dependent protein-cross-linking enzyme TGase (transglutaminase) was demonstrated to be involved in the apical growth of Malus domestica pollen tube. Apple pollen TGase and its substrates were co-localized within aggregates on the pollen tube surface, as determined by indirect immunofluorescence staining and the in situ cross-linking of fluorescently labelled substrates. TGase-specific inhibitors and an anti-TGase monoclonal antibody blocked pollen tube growth, whereas incorporation of a recombinant fluorescent mammalian TGase substrate (histidine-tagged green fluorescent protein: His6-Xpr-GFP) into the growing tube wall enhanced tube length and germination, consistent with a role of TGase as a modulator of cell wall building and strengthening. The secreted pollen TGase catalysed the cross-linking of both PAs (polyamines) into proteins (released by the pollen tube) and His6-Xpr-GFP into endogenous or exogenously added substrates. A similar distribution of TGase activity was observed in planta on pollen tubes germinating inside the style, consistent with a possible additional role for TGase in the interaction between the pollen tube and the style during fertilization.

Isolation and characterisation of bacteria associated with flying insects in hospitals, with particualr emphasis on Clostridum difficile

Clostridium difficile is a bacterial healthcare-associated infection, which houseflies Musca domestica may transfer due to their synanthropic nature. The aims of this thesis were to determine the ability of M. domestica to transfer C. difficile mechanically and to collect and identify flying insects in UK hospitals and classify any associated bacteria. M. domestica exposed to independent suspensions of vegetative cells and spores of C. difficile were able to mechanically transfer the bacteria on to agar for up to 4 hours following exposure. C. difficile could be recovered from fly excreta for 96hrs and was isolated from the M. domestica alimentary canal. Also confirmed was the carriage of C. difficile by M. domestica larvae, although it was not retained in the pupae or in the adults that subsequently developed. Flying insects were collected from ultra-violet light flytraps in hospitals. Flies (order Diptera) were the most commonly identified. Chironomidae were the most common flies, Calliphora vicina were the most common synanthropic fly and ‘drain flies’ were surprisingly numerous and represent an emerging problem in hospitals. External washings and macerates of flying insects were prepared and inoculated onto a variety of agars and following incubation bacterial colonies identified by biochemical tests. A variety of flying insects, including synanthropic flies (e.g. M. domestica and C. vicina) collected from UK hospitals harboured pathogenic bacteria of different species. Enterobacteriaceae were the group of bacteria most commonly isolated, followed by Bacillus spp, Staphylococci, Clostridia, Streptococci and Micrococcus spp. This study highlights the potential for M. domestica to contribute to environmental persistence and spread of C. difficile in hospitals. Also illustrated is the potential for flying insects to contribute to environmental persistence and spread of other pathogenic bacteria in hospitals and therefore the need to implement pest control as part of infection control strategies.

Farmacocinética de oxitetraciclina en dosificación oral múltiple en cerdos. Análisis PK-PD

La oxitetraciclina, antibiótico de amplio uso en medicina veterinaria, pertenece al grupo de las tetraciclinas. Inhibe la síntesis de proteínas en la bacteria a nivel ribosomal. La oxitetraciclina presenta principalmente una acción bacteriostática frente a bacterias Gram-positivas y Gram-negativas, así como también frente a otros microorganismos tales como micoplasmas, espiroquetas, clamidias y rickettsias. Se clasifica en la actualidad de acuerdo con la cinética de muerte bacteriana como co-dependiente. En la bibliografía científica existen trabajos publicados de farmacocinética de oxitetraciclina en distintas especies animales, sin embargo hay escasos trabajos realizados en el cerdo. Dado que es necesario conocer la disposición de un fármaco en la especie animal estudiada para diseñar un adecuado régimen de dosificación, los objetivos del presente trabajo han sido: (i) describir el perfil farmacocinético de la oxitetraciclina tras administración oral única y múltiple en cerdos (Sus scrofa domestica) y (ii) realizar el análisis PK-PD para predecir la eficacia terapeútica del régimen de dosificación...

Acoustic signalling reflects personality in a social mammal

Social interactions among individuals are often mediated through acoustic signals. If acoustic signals are consistent and related to an individual's personality, these consistent individual differences in signalling may be an important driver in social interactions. However, few studies in non-human mammals have investigated the relationship between acoustic signalling and personality. Here we show that acoustic signalling rate is repeatable and strongly related to personality in a highly social mammal, the domestic pig (Sus scrofa domestica). Furthermore, acoustic signalling varied between environments of differing quality, with males from a poor-quality environment having a reduced vocalization rate compared with females and males from an enriched environment. Such differences may be mediated by personality with pigs from a poor-quality environment having more reactive and more extreme personality scores compared with pigs from an enriched environment. Our results add to the evidence that acoustic signalling reflects personality in a non-human mammal. Signals reflecting personalities may have far reaching consequences in shaping the evolution of social behaviours as acoustic communication forms an integral part of animal societies.

Ottimizzazione e progettazione di un sistema di cogenerazione statica di energia elettrica tramite celle ad effetto Seebeck

La presente trattazione descrive l’ottimizzazione del prototipo di un generatore termoelettrico da integrare in caldaie a biomassa. La tecnologia termoelettrica consente di generare potenza elettrica a partire da un flusso di calore. Sebbene tale tecnologia presenti caratteristiche di elevata affidabilità e silenziosità, essa richiede ulteriori sforzi tecnologici nella ricerca dell’integrazione ottimale con un impianto termico, ed è inoltre necessaria una progettazione accurata dell’elettronica di controllo. Nella prima sezione del testo vengono descritti i principi fisici alla base di questa tecnologia e le tecniche per la conversione della potenza generata dai moduli termoelettrici (o ad effetto Seebeck) al fine di renderla fruibile da una tipica utenza domestica. Nel secondo capitolo si riporta lo stato dell’arte dell’applicazione della tecnologia termoelettrica in caldaie a biomassa e alcuni casi di implementazione recenti, sottolineando le configurazioni utilizzate, i principali problemi riscontrati nello sviluppo e nella sperimentazione, e le modifiche conseguentemente adottate per superarli. Il terzo capitolo analizza il prototipo sviluppato presso il Dipartimento di Ingegneria Industriale dell’Università di Bologna e descrive il processo di dimensionamento dei componenti meccanici e le proposte di ottimizzazione a seguito dei risultati dei primi collaudi. Viene infine sviluppato un modello matematico per confrontare, in funzione dei costi di produzione e di esercizio, le possibilità di sostituzione dei metodi di generazione di energia elettrica convenzionali con l’impianto di cogenerazione termoelettrica realizzato, considerando anche aspetti di impatto ambientale. Tali analisi e ottimizzazioni consistono in un primo miglioramento necessario a rendere la tecnologia termoelettrica applicabile alle caldaie a biomassa stand-alone e a permettere una generazione continua di energia elettrica nelle regioni in cui le infrastrutture sono poco affidabili.

Utilizzo di composti d'aroma per aumentare la sicurezza igienico-sanitaria di germogli di ravanello

Questo elaborato ha come obiettivo valutare l’efficacia di alcuni composti d’aroma, ossia componenti di oli essenziali la cui attività antimicrobica è nota in letteratura, nell’aumentare la sicurezza igienico-sanitaria di germogli di ravanello Daikon. In particolare sono stati utilizzati timolo e carvacrolo, costituenti di oli di piante tra cui timo e origano. Vista la grande espansione nel mercato alimentare di semi germogliati pronti all’uso, matrice veicolante numerosi casi di tossinfezione alimentare, sono state effettuate prove per valutare il rischio di sviluppo di Listeria. Si è lavorato simulando un processo casalingo, facendo germinare i semi in acqua addizionata o meno di diverse concentrazioni dei due terpeni fenolici. I semi utilizzati venivano precedentemente inoculati con Listeria innocua, utilizzata come surrogato del patogeno Listeria monocytogenes. Il ricambio dell’acqua di ammollo avveniva ogni 24 ore, e dopo 6/24/32/48 ore venivano prelevate quantità note di semi e acqua, al fine di valutarne il contenuto in L. innocua, carica microbica totale ed enterobatteri. La prima prova, che ha previsto l’uso di timolo come unico composto antimicrobico, ha mostrato una riduzione di un ciclo logaritmico di L. innocua ed enterobatteri nei semi, mentre nell’acqua essi erano presenti solo dopo 24 e 48 ore. Risultati analoghi sono stati ottenuti utilizzando solo carvacrolo. Infine è stato testato l’eventuale effetto sinergico di timolo e carvacrolo, che però non ha dato differenze particolari rispetto a campioni trattati coi singoli composti. I risultati ottenuti hanno evidenziato l’elevato rischio microbiologico associato ai semi germogliati con produzione domestica, poiché le condizioni adottate favoriscono lo sviluppo di specie patogene. L’utilizzo dei composti di aroma, se pure in grado di ridurre sia le cinetiche di crescita che la quantità di Listeria, non è tuttavia in grado di controllare in maniera significativa il rischio associato a tali prodotti.

Produção textual de adolescentes acerca da saúde sexual e reprodutiva: subsídios para atuação de enfermagem

Trata-se de um estudo exploratório-descritivo, com abordagem qualitativa, que teve por objetivo analisar as mensagens, acerca da promoção da saúde sexual e reprodutiva, produzidas por adolescentes de escolas públicas e particulares da cidade do Rio Grande, num concurso de redação e música promovido pelo Grupo Gestor Municipal (GGM) do Projeto Saúde e Prevenção nas Escolas (SPE), nos anos de 2007 e 2008. Após autorização pelo GGM para realização deste estudo, foram disponibilizadas para reprodução, via xérox, as 29 redações e as três letras de músicas inscritas nos concursos. Para o tratamento dos dados utilizou-se a técnica de análise de conteúdo na modalidade temática. Participaram 35 adolescentes, sendo 25 moças e dez rapazes, com idades entre onze e dezessete anos. Quanto à escolaridade, dois frequentavam a quinta série; doze a sexta, doze a sétima e nove a oitava. Apreendeu-se que, em sua produção textual, os(as) adolescentes revelaram as vulnerabilidades e fortalezas referentes à saúde sexual e reprodutiva. Entre os inúmeros fatores que aumentam a vulnerabilidade individual, social e programática, discorreram sobre a carência de informações, a dificuldade para transformar o conhecimento em prática, a sensação de imunidade, a violência familiar, a conduta repressora de pais e mães, as mensagens de cunho sexual veiculadas pela mídia, a necessidade de serem aceitos(as) pelo grupo, preconceitos, e falta de ações governamentais direcionadas a adolescentes. No que se refere às fortalezas, sabem que a informação é uma importante aliada para a promoção da saúde sexual e reprodutiva citando, entre as fontes acessíveis, os serviços públicos de saúde, a família e a escola. Demonstraram conhecimento acerca da alarmante propagação da epidemia da AIDS entre jovens, conhecendo os sinais e sintomas das DSTs mais comuns e as formas de prevenção. As moças enfatizaram a necessidade de compartilhar a responsabilidade preventiva com os rapazes, bem como de amor próprio e respeito mútuo. O acesso aos serviços de saúde também foi apresentado como indispensável ao adolescer saudável. Os(as) jovens demonstraram conhecimento sobre drogas seus efeitos e consequências. Referem-se à adolescência como um período gostoso, repleto de dúvidas, mas também cheio de potencialidades. Assim, os mesmos componentes apresentados como desencadeadores de vulnerabilidade podem torná-los(as) fortes e capazes de superar os desafios comuns a essa etapa da vida. Para que tal superação ocorra, é necessário que tenham acesso à informação e a problematizem; que sejam capazes de incorporá-las ao cotidiano, adotando práticas protegidas e protetoras; que haja diálogo, despido de tabus, censuras e preconceitos no ambiente familiar; que as escolas adotem de forma transversalizada temáticas referentes à saúde sexual e reprodutiva; que os serviços de saúde tenham infraestrutura para assegurar os direitos contidos no Estatuto da Criança e do Adolescente; entre outras estratégias fortalecedoras.

Alphaherpesvirus-associated disease in greater bilbies (Macrotis lagotis)

Case report: A captive breeding colony of 9 greater bilbies (Macrotis lagotis) exhibited mild upper respiratory signs and sudden deaths with 100% mortality over a 2-week period. Histologically, acute necrotising and erosive epithelial lesions throughout the upper respiratory system and bronchi were associated with eosinophilic intranuclear inclusion bodies. Inclusions were also present in hepatocytes and adrenocortical cells, but were not always associated with necrosis. Transmission electron microscopy of lung sections revealed nucleocapsids forming arrays within some nuclei. A pan-herpesvirus PCR yielded a 440-bp product, with sequencing confirming homology with the alphaherpesviruses. Viral culture in a marsupial cell line resulted in cytopathic effect consistent with an alphaherpesvirus. Conclusion: This is the first report of a herpesvirus-associated disease in greater bilbies. © 2016 Australian Veterinary Association.

Effect of time and diet change on the bacterial community structure throughout the gastrointestinal tract and in faeces of the northern brown bandicoot, Isoodon macrourus

A significant gap, in not only peramelid nutritional physiology but marsupial nutrition as a whole, is the lack of information relating to microorganisms of the gastrointestinal tract. This research is a preliminary investigation that will provide a baseline for comparisons among peramelids. The high degree of 16S rRNA gene clones identified in this research that are closely related to culturable bacteria suggests that additional research will enable a more complete description of the gastrointestinal bacteria of I. macrourus. Most identifiable clones belonged to Clostridium and Ruminococcus. This research has confirmed that the hindgut of I. macrourus, the caecum, proximal colon and distal colon, are the main sites for microbial activity.

Validação de metodologia analítica por CLAE-DAD para determinação de compostos fenólicos e atividade biológica em coprodutos agroindustriais

The southern region of Brazil, especially the states of Parana and Santa Catarina stand out for growing grapes and apples for fresh consumption and in order to add value to these products, process the material for the production of wine, juices and jellies . As a result large quantities of by-products, such as peels, seeds and pulp are produced becoming environmental problems. Studies reuse of these by-products have attracted interest because they have shown a high biological potential, due to the presence of high levels of phenolic compounds, which are associated with a lower incidence of disease caused by oxidative stress, due to its antioxidant, antiinflammatory and antibacterial properties. Currently, few studies are presented on the phenolic composition and biological potential of waste grape variety Bordô (Vitis labrusca) and apple (Malus domestica) Gala variety, cultivated in southern Brazil. Within this context, the objectives of this study were: compare the efficiency of solidliquid and liquid-liquid extraction, perform the optimization and validation of analytical methodology by HPLC-DAD for the separation, identification and quantification of multiclass phenolic compounds, evaluate the activity antioxidant by sequestering methods of free radical 2,2-diphenyl-1 picrilhidrazina (DPPH) and 2,2-azino-bis (3- ethyl-benzthiazoline-6-sulphonic acid) (ABTS) solution, reduction of Fe3+ in Fe2+ method (FRAP), ORAC, RP-HPLC-ABTS online, Rancimat and determination of total phenolics three agro-industrial byproducts, pomace and stems grape Bordô produced in Paraná Southwest region and Gala apple pomace coming from the Santa Catarina West. Optimization and validation of chromatographic method showed satisfactory quality parameters for the compounds of interest and the solidliquid extraction was more efficient in extracting phenolic evaluated. The three byproducts evaluated showed significant levels of phenolic compounds when analyzed by HPLC, especially flavonoids, catechin and epicatechin besides that showed significant antioxidant capacity. The grape stems extract had the highest sequestration capacity of DPPH and ABTS radical and reduced iron, and high content of phenolic compounds. The apple pomace extract showed the best response to the Rancimat method, which indicates a high potential to protect the oil from lipid oxidation, was no significant difference when compared to synthetic antioxidant TBHQ. The results of this study showed that the agro-industrial coproducts analyzed are rich in phenolic compounds of high antioxidant capacity and therefore must be better explored by the food and pharmaceutical industries.