Characterization and Experimental Host Range of a Brazilian Tomato Isolate of Tomato severe rugose virus

We report the complete molecular characterization of the DNA-A and DNA-B of a Brazilian tomato isolate of Tomato severe rugose virus (ToSRV) and the experimental host range of the virus determined using white-fly transmission tests. Genome analysis showed that ToSRV has a close evolutionary relationship with Tomato rugose mosaic virus. Of 33 plants species inoculated with viruliferous Bemisia tabaci biotype B, 13 species were susceptible to ToSRV, nine asymptomatically. Therefore, ToSRV disease management strategy should include the control of infected weeds close to tomato fields.

Experimental and theoretical investigations of adsorption hysteresis and criticality in MCM-41: Studies with O-2, Ar, and CO2

MCM-41 materials of six different pore diameters were prepared and characterized using X-ray diffraction, transmission electron microscopy, helium pycnometry, small-angle neutron scattering, and gas adsorption (argon at 77.4 and 87.4 K, nitrogen and oxygen at 77.4 K, and carbon dioxide at 194.6 K). A recent molecular continuum model of the authors, previously used for adsorption of nitrogen at 77.4 K, was applied here for adsorption of argon, oxygen, and carbon dioxide. While model predictions of single-pore adsorption isotherms for argon and oxygen are in satisfactory agreement with experimental data, significant deviation was found for carbon dioxide, most likely due to its high quadrupole moment. Predictions of critical pore diameter, below which reversible condensation occurs: were possible by the model and found to be consistent with experimental estimates, for the adsorption of the various gases. On the other hand, existing models such as the Barrett-Joyner-Halenda (BJH), Saito-Foley, and Dubinin-Astakhov models were found to be inadequate, either predicting an incorrect pore diameter or not correlating the isotherms adequately. The wall structure of MCM-41 appears to be close to that of amorphous silica, as inferred from our skeletal density measurements.

Transmission electron microscopy of a transformation toughened white cast iron

Transmission electron microscopy has been used to study the microstructure of an experimental white cast iran, in which a combination of modified alloy composition and unconventional heat treatment has resulted in a fracture toughness of 40 MPa m(-1/2). Microstructural features of the alloy that contribute to the toughness improvement and hence distinguish it from conventional white irons have been investigated. In the as-cast condition the dendrites are fully austenitic and the eutectic consists of M7C3 carbides and martensite. During heat treatment at 1130 degrees C the austenite is partially destabilized by precipitation of chromium-rich M7C3 carbides. This results in a dendritic microconstituent consisting of bulk retained austenite and secondary carbides which are sheathed with martensite. The martensite sheaths, which contain interlath films of retained austenite, are irregular in shape with some laths extending into the bulk retained austenite. Emphasis has been placed on the morphology, distribution, and stability of the retained austenite and its transformation products in the dendrites. The implications of these findings on the transformation toughening mechanism in this alloy are discussed.

Experimental infection of the rabbit tick, Haemaphysalis leporispalustris, with the bacterium Rickettsia rickettsii, and comparative biology of infected and uninfected tick lineages

The present study consisted of two experiments that evaluated experimental infections of Haemaphysalis leporispalustris ticks by a Brazilian strain of Rickettsia rickettsii, and their effect on tick biology. In experiment I, ticks were exposed to R. rickettsii during the larval, nymphal or adult stages by feeding on rabbits (Oryctolagus cuniculus) needle-inoculated with R. rickettsii, and thereafter reared on uninfected rabbits for the entire next tick generation. Regardless of the tick stage that acquired the infection, all subsequent tick stages were shown to be infected by PCR (infection rates varying from 1.3 to 41.7%), and were able to transmit R. rickettsii to uninfected rabbits, as demonstrated by rabbit seroconversion, guinea pig inoculation with rabbit blood, and PCR on rabbit blood. In Experiment II, ticks were exposed to R. rickettsii during the larval stage by feeding on rabbits co-infested with R. rickettsii-infected adult ticks, and thereafter reared on uninfected rabbits until the next generation of larvae. Again, all subsequent tick stages were shown to be infected by PCR (infection rates varying from 3.0 to 40.0%), and were able to transmit R. rickettsii to uninfected rabbits. Thus, it was demonstrated that larvae, nymphs, and adults of H. leporispalustris were able to acquire and maintain the R. rickettsii infection by transstadial and transovarial transmissions within the tick population, with active transmission of the bacterium to susceptible rabbits by all parasitic stages. Analyses of biological parameters of uninfected and R. rickettsii-infected tick lineages were performed in order to evaluate possible deleterious effects of R. rickettsii to the infected tick lineages. Surprisingly, all but one of the four R. rickettsii-experimental groups of the present study showed overall better biological performance than their sibling uninfected control ticks. Results of the present study showed that H. leporispalustris could support infection by a high virulent strain of R. rickettsii for at least two generations, in which infected tick lineages tended to have better performance than uninfected ticks. Our results support a possible role of H. leporispalustris in the enzootic maintenance of R. rickettsii in Latin America, as previously suggested by earlier works.

Experimental infection of dogs (Canis familiaris) with sporulated oocysts of Neospora caninum

Neospora caninum is widely distributed in the world and this parasite is one of the major causes of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum and several species of domestic and wild animals are intermediate hosts. Dogs can become infected by the ingestion of tissues containing cysts and then excrete oocysts. It is not yet known whether sporulated oocysts are able to induce a patent infection in dogs, i.e. a shedding of N. caninum oocysts in feces. The objective of this study was to experimentally examine the infection of dogs by sporulated oocysts. The oocysts used in the experiment were obtained by feeding dogs with brain of buffaloes (Bubalus bubalis) positive for anti-N. caninum antibodies by indirect fluorescent antibody test (IFAT >= 200). Oocysts shed by these dogs were confirmed to be N. caninum by molecular methods and by bioassay in gerbils, and sporulated N. caninum oocysts were used for the oral infection of four dogs. The dogs were 8 weeks old and negative for antibodies to N. caninum and Toxoplasma gondii. Dogs 1 and 4 received an inoculum of 10,000 sporulated oocysts each; dog 2 an inoculum of 5000 sporulated oocysts and dog 3 received 1000 sporulated oocysts of N. caninum. The total feces excreted by these dogs were collected and examined daily for a period of 30 days. No oocysts were found in their feces. The dogs were monitored monthly for a 6-month period to observe a possible seroconversion and when this occurred the animals were eliminated from the experiment. Dogs 1 and 4 seroconverted 1 month after the infection with titer, in the IFAT, of 1600 and 800, respectively; the other two dogs presented no seroconvertion during the 6-month period. Dogs 1 and 2 were euthanized 180 days after infection and were examined for the detection of N. caninum in tissues (brain, muscle, lymph node, liver, lung, heart and bone marrow) by immunohistochemistry and PCR with negative results in both techniques. Bioassay in gerbils with brain of these dogs was also performed and again the results were negative. In conclusion, dogs infected with sporulated oocysts of N. caninum were not able to shed oocysts in feces. However, a higher dose of infection stimulated the production of antibodies against N. caninum in the dogs. (C) 2010 Elsevier B.V. All rights reserved.

Experimental infection of pregnant queens with two major Brazilian clonal lineages of Toxoplasma gondii

Toxoplasma gondii isolates from Brazil are biologically and genetically different from European and North America isolates. Recently, four genotypes were considered the common clonal lineages in Brazil and were designated as types BrI, BrII, BrIII, and BrIV. The pathogenicity of two major Brazilian lineages was investigated after oral inoculation of queens in the middle third of their pregnancies with T. gondii cysts. Twelve pregnant queens without T. gondii antibodies were distributed in group A (infected with a type BrI isolate); group 2 (infected with type BrIII isolate), and group 3 (non-infected control). Infection with type BrI isolate caused toxoplasmosis manifestations and abortion from one litter. Toxoplasmosis manifestations besides premature stillbirth of one litter were observed in queens infected with type BrIII isolate. Indirect fluorescence antibody test showed T. gondii antibodies in all eight infected queens at 30 days after inoculation. In two 10-day-old kittens of the same litter (group 1), titers of 16 and 64 were detected. At the same time, titers of 16, 32, and 32 were detected in three kittens from the same litter (group 2). Experimental infection with tissue cysts from a type BrI and type BrIII isolates of T. gondii developed similar reproductive disturbance in primary infected pregnant queens.

Microstructural and Ultrastructural Assessment of Inferior Alveolar Nerve Damage Following Nerve Lateralization and Implant Placement: An Experimental Study in Rabbits

Purpose: The present study assessed damage to the inferior alveolar nerve (IAN) following nerve lateralization and implant placement surgery through optical and transmission electron microscopy (TEM). Materials and Methods: IAN lateralization was performed in 16 adult female rabbits (Oryctolagus cuniculus). During the nerve lateralization procedure, one implant was placed through the mandibular canal, and the IAN was replaced in direct contact with the implant The implant was placed in the right mandible, and the left side was used as a control (no surgical procedure) After 8 weeks, the animals were sacrificed and samples were prepared for optical and TEM analysis of IAN structural damage Histomorphometric analysis was performed to determine the number and cross-sectional dimensions of nerve fascicles and myelin sheath thickness between experimental and control grouos. The different parameters were compared by one-way analysis of variance at the 95% significance level Results: Alterations in the perineural and endoneural regions of the IAN, with higher degrees of vascularization., were observed in the experimental group TEM showed that the majority of the myelinated nerve fibers were not affected in the experimental samples. No significant variation in the number of fascicles was observed, significantly larger fascicle height and width were observed in the control group, and significantly thicker myelin sheaths were observed in the experimental samples Conclusion: IAN lateralization resulted in substantial degrees of tissue disorganization at the microstructural level because of the presence of edema However, at the ultrastructural level, small amounts of fiber degeneration were observed. INT J ORAL MAXILLOFAC IMPLANTS 2009,24-859-865

Experimental human infection with the dog hookworm, Ancylostoma caninum

Objective: To investigate possible routes for human infection by the dog hookworm (Ancylostoma caninum). Design, setting and participant. Relatively small numbers of infective larvae were administered orally and percutaneously to an informed healthy volunteer (J K L) under medical supervision, at intervals between May 1998 and May 1999. Main outcome measures: Symptoms; weekly blood eosinophil counts; faecal microscopy. Results: A marked blood eosinophilia followed a single oral exposure to 100 infective larvae, while faecal examination remained negative. Eosinophil counts then declined gradually, although a rapid, spontaneous rise several months later, at the beginning of spring, possibly indicated reactivation of dormant larvae. Blood eosinophil numbers did not rise significantly after percutaneous infection with 200 larvae. A subsequent, smaller, oral inoculum of 20 larvae provoked an eosinophil response similar to that of the first experiment. Conclusions: Our findings suggest that, following ingestion, some infective larvae of A. caninum develop directly into adult worms in the human gut (as they do in dogs). While the percutaneous route might be the most common means of human exposure to canine hookworm larvae, leading generally to subclinical infection, oral infection may be more likely to provoke symptomatic eosinophilic enteritis.

The Australian paralysis may be the missing link tick in the transmission of Hendra virus from bats to horses to humans

Hendra virus is a new virus of the family Paramyxoviridae. This virus was first detected in Queensland, Australia, in 1994; although, it seems that the virus has infected fruit-eating bats (flying-foxes) for a very long time. At least 2 humans and 15 horses have been killed by this virus since it first emerged as a virus that may infect mammals other than flying-foxes. Hendra virus is thought to have moved from flying-foxes to horses, and then from horses to people. There is a reasonably strong hypothesis for horse-to-human transmission: transmission of virus via nasal discharge, saliva and/or urine. In contrast, there is no strong hypothesis for flying-fox-to-human transmission. I present evidence that the Australian paralysis tick, Ixodes holocyclus, which has apparently only recently become a parasite of flying-foxes, may transmit Hendra virus and perhaps related viruses from flying-foxes to horses and other mammals. (C) 2003 Elsevier Science Ltd. All rights reserved.

Caracterização experimental e numérica do desempenho térmico de um edifício "gaioleiro": identificação de oportunidades de melhoria

O sector dos edifícios é responsável por uma percentagem significativa dos consumos de energia primária e de energia eléctrica em Portugal, associada principalmente ao conforto térmico dos seus ocupantes. A União Europeia pretende uma redução de 20%, até 2020, dos consumos de energia e consequentes emissões de CO2 através da melhoria da eficiência energética dos edifícios públicos e residenciais. Em Portugal, o Plano Nacional para a Eficiência Energética (PNAEE) tem por objectivo obter uma poupança anual de energia de pelo menos 1% até ao ano de 2016, tomando como base a média de consumos de energia final, registados entre 2001 e 2005. Neste contexto, os edifícios anteriores a 1990 (primeira versão do RCCTE) podem apresentar um potencial significativo de melhoria da sua eficiência energética com base na sua reabilitação. Os edifícios “Gaioleiros” (1880 – 1930) constituindo uma parte importante do património histórico da cidade de Lisboa, para os quais a informação sobre o seu desempenho térmico é limitada, considerou-se pertinente efectuar um estudo destinado à sua caracterização experimental e numérica, face à especificidade do comportamento térmico das suas paredes caracterizadas pela elevada espessura. No presente trabalho, apresenta-se a metodologia e os resultados experimentais da medição da resistência térmica das paredes e da medição das necessidades térmicas de aquecimento da habitação. Estes resultados experimentais foram utilizados na validação do modelo de simulação térmica da habitação, que posteriormente serviu para avaliar as suas necessidades térmicas de aquecimento (Nic) e de arrefecimento (Nvc), identificar oportunidades de melhoria e avaliar o respectivo potencial de reabilitação. Neste trabalho, como contributos para uma reabilitação sustentável, apresentam-se avaliações de oportunidades de melhoria com base em estratégias de reforço do isolamento térmico. Dos resultados obtidos concluiu-se que melhorando o isolamento térmico das paredes e vãos envidraçados é possível baixar consideravelmente os consumos de energia associados à habitação, cumprindo assim as exigências estabelecidas no RCCTE ao nível dos requisitos de qualidade térmica da envolvente e consumos energéticos para edifícios novos e grandes reabilitações.

A traffic differentiation add-on to the IEEE 802.15.4 protocol: implementation and experimental validation over a real-time operating system

The IEEE 802.15.4 is the most widespread used protocol for Wireless Sensor Networks (WSNs) and it is being used as a baseline for several higher layer protocols such as ZigBee, 6LoWPAN or WirelessHART. Its MAC (Medium Access Control) supports both contention-free (CFP, based on the reservation of guaranteed time-slots GTS) and contention based (CAP, ruled by CSMA/CA) access, when operating in beacon-enabled mode. Thus, it enables the differentiation between real-time and best-effort traffic. However, some WSN applications and higher layer protocols may strongly benefit from the possibility of supporting more traffic classes. This happens, for instance, for dense WSNs used in time-sensitive industrial applications. In this context, we propose to differentiate traffic classes within the CAP, enabling lower transmission delays and higher success probability to timecritical messages, such as for event detection, GTS reservation and network management. Building upon a previously proposed methodology (TRADIF), in this paper we outline its implementation and experimental validation over a real-time operating system. Importantly, TRADIF is fully backward compatible with the IEEE 802.15.4 standard, enabling to create different traffic classes just by tuning some MAC parameters.

Low-loss austempered ductile iron gears: experimental evaluation comparing materials and lubricants

An experimental study to evaluate the power dissipation of gears was performed. Three low-loss gear models were manufactured using standard 20° pressure angle tools. Austempered ductile iron (ADI) and 20MnCr5 carburized steel gears were tested in an FZG gear test machine using mineral, ester and polyalphaolephine (PAO)-based oils. The results compare power dissipation, the influence of different tooth flank geometries, materials and lubricants. This work concludes that conventional power-transmission gears can be replaced by these improved and more efficient low–loss models, which can be produced using common tools and that steel gears can be successfully replaced by austempered ductile iron gears.

Experimental life cycle of Lagochilascaris minor Leiper, 1909

The life cycle of Lagochilascaris minor was studied using material collected from human lesion and applying the experimental model: rodents (mice, hamsters), and carnivorae (cats, dogs). In mice given infective eggs, orally, hatch of the third stage larvae was noted in the gut wall, with migration to liver, lungs, skeletal musculature and subcutaneous tissue becoming, soon after, encysted. In cats infected with skinned carcasses of mice (60 to 235 days of infection) it was observed: hatch of third stage larvae from the nodules (cysts) in the stomach, migration through the oesophagus, pharynx, trachea, related tissues (rhino-oropharynx), and cervical lymphonodes developing to the mature stage in any of these sites on days 9-20 post inoculation (P.I.). There was no parasite development up to the mature stage in cats inoculated orally with infective eggs, which indicates that the life cycle of this parasite includes an obligatory intermediate host. In one of the cats (fed carcass of infected mice) necropsied on day 43 P.I., it was observed the occurence of the self-infective cycle of L. minor in the lung tissues and in the cervical region which was characterized by the finding of eggs in different stages of development, third stage larvae and mature worms. It's believed that some component of the carnivorae gastrointestinal tracts may preclude the development of third stage larvae from L. minor eggs what explains the interruption of the life cycle in animals fed infective eggs. It's also pointed out the role of the intermediate host in the first stages of the life cycle of this helminth.

Ultrastructural study on experimental infection of rotavirus in a murine heterologous model

Viral replication, histopathological and ultrastructural changes were observed for a period of nine days in the small intestine of suckling mice infected with a simian rotavirus (SA11). Samples taken from duodenum, jejunun and ileum were prepared for light microscopy, transmission and scanning electron microscopy analysis. Histopathologic effect could be detected within 8 hr post-infection, when only a few altered cells were observed. Damage was extensive after 16 hr post-infection, showing swollen enterocytes and reduced and irregularly oriented microvilli at intestinal villi tips. Virus particles were detected at 16 and 48 hr post-infection, budding from the viroplasm into the rough endoplasmic reticulum cisternae in ileum enterocytes. Clear evidence of viral replication, observed by electron microscopy was not described before in heterologous murine models. Regeneration of the intestinal villi began at the third day post-infection. Despite some differences observed in clinical symptoms and microscopic analysis of homologous and heterologous rotavirus infections, we concluded that mechanisms of heterologous rotavirus infection in mice follow similar patterns to those observed in the homologous models.

Interaction between Didelphis albiventris and Triatoma infestans in relation to Trypanosoma cruzi transmission

This paper attempts to prove if a high Trypanosoma cruzi prevalence of opossums might be reached with few potential infective contacts. One non-infected Didelphis albiventris to T. cruzi and 10 infected nymphs of Triatoma infestans were left together during 23 hr in a device that simulated a natural opossum burrow. Twenty-six replicates were perfomed using marsupials and triatomines only once. Potentially infective contacts occurred in all the trials. From the 26 opossums used in trials, 54% did not eat any bug. Of the 260 bugs used, 21% were predated. In the 25 trials involving 205 surving bugs, 36 % of them did not feed. In 15/25 cases, maior ou igual a 60% of the triatomines were able to feed. The parasitological follow-up of 24 opossums showed that among 10 that had eaten bugs, 4 turned out infected and among the 14 that had not predate, 3 (21%) became positive. In sum, 7/24 (29%) of the marsupials acquired the infection after the experiment. This infection rate was similar to the prevalences found for the opossum population of Santiago del Estero, Argentina, suggesting that the prevalences observed in the field might be reached if each marsupial would encounter infected bugs just once in its lifetime.