The effects of physiological levels of intermittent pressure on the development of articular cartilage in vitro

A semi-continuous perfusion system has been designed and constructed to deliver physiological levels of hydrostatic pressure to regenerating cell/polymer constructs of articular cartilage over the long term (weeks to months). Prior to being transferred to the pressurized system, equine articular chondrocytes were dynamically seeded onto nonwoven meshes of polyglycolic acid which provide the cells with a three-dimensional growth environment similar to that found in vivo. When applied in an intermittent manner, physiological pressurization at 500 psi was found to stimulate articular chondrocytes to produce regenerated constructs with greater concentrations of sulfated glycosaminoglycan compared to control (unpressurized) constructs, while the concentration of collagen was not significantly different between pressurized and control samples. Foal articular chondrocytes were found to produce significantly greater amounts of the extracellular matrix (sulfated glycosaminoglycan and collagen) than chondrocytes isolated from adult horses (greater than two years of age) at similar pressure levels. Increasing the level of physiological intermittent pressure from 500 psi to 1000 psi was found to further increase the concentration of sulfated glycosaminoglycan in regenerated constructs and, for the first time, was shown to significantly increase collagen concentrations above control samples, suggesting that a minimum level of dynamic force may be needed to stimulate collagen production. By combining pressurized and stirred culture environments, tissue constructs were developed which had greater concentrations of the extracellular matrix than constructs regenerated in single culture environments and masses greater than those previously developed in stagnant (minimum medium perfusion) pressurized cultures. A correlation was noted between the compressive modulus, which is a measure of the strength of a regenerated construct, and the concentration of sulfated glycosaminoglycan in constructs cultured mostly in a pressurized environment. The fact that this correlation has not been shown in control samples or in samples cultured mostly in mixed cultures, suggests that intermittent pressurization may influence the structural arrangement of the extracellular matrix. This form of dynamic loading creates regenerated tissue which is more mechanically stable than constructs cultured in the absence of forces similar to those experienced in the native environment

Communications infrastructure requirements for telemedicine/telehealth in the context of planning for and responding to natural disasters: Considering the need for shared regional networks

During the course of recent years the frequency and magnitude of major disasters - of natural, technological, or ecological origin - have made the world community dramatically aware of the immense losses of human life and economic resources that are caused regularly by such calamities. Particularly hard hit are developing countries, for whom the magnitude of disasters frequently outstrips the ability of the society to cope with them. In many cases this situation can be prevented, and the recent trend in disaster management has been to emphasize the importance of preparedness and mitigation as a means of prevention. In cases of disaster, a system is needed to respond to relief requirements, particularly the delivery of medical care. There is no generic telecommunications infrastructure appropriate for the variety of applications in medical care and disaster management. The need to integrate telemedicine/telehealth into shared regional disaster management telecommunications networks is discussed. Focus is on the development of infrastructure designed to serve the needs of disaster prone regions of the developing world

Effect of Rock Cover on Small Mammal Abundance in a Montana Grassland

We examined the influence of rock cover, as an indicator of presumable retreat site availability on the abundance of deer mice (Peromyscus maniculatus) and prevalence of Sin Nombre virus (SNV) using long-term live trapping and habitat data from three live trapping grids and a shortterm (three month), spatially replicated study across three slopes in Cascade County, Montana. In our long-term study, we found that deer mice were more abundant at a live-trapping grid with greater rock cover, than two grids with less rock cover. There was a non-significant trend (P = 0.053) for deer mice to be more abundant in rocky sites in the short term study. In the long-term study, average SNV antibody prevalence among deer mice was slightly greater (5.0 vs. 3.5 % on average) at the live trapping grid with more rock cover, than the grid with less rock cover. We were unable to demonstrate differences in SNV antibody prevalence among treatments in the short-term study. Further studies are needed to elucidate the multiple determinants of deer mouse abundance and SNV prevalence in grassland ecosystem and other habitat types

Environmental drivers of Ross River virus in south-east Tasmania, Australia: Towards strengthening public health interventions

In Australia, Ross River virus (RRV) is predominantly identified and managed through passive health surveillance. Here, the proactive use of environmental datasets to improve community-scale public health interventions in southeastern Tasmania is explored. Known environmental drivers (temperature, rainfall, tide) of the RRV vector Aedes camptorhynchus are analysed against cumulative case records for five adjacent local government areas (LGAs) from 1993 to 2009. Allowing for a 0- to 3-month lag period, temperature was the most significant driver of RRV cases at 1-month lag, contributing to a 23. 2% increase in cases above the long-term case average. The potential for RRV to become an emerging public health issue in Tasmania due to projected climate changes is discussed. Moreover, practical outputs from this research are proposed including the development of an early warning system for local councils to implement preventative measures, such as public outreach and mosquito spray programmes

Prior Virus Exposure Alters the Long-Term Landscape of Viral Replication during Feline Lentiviral Infection

We developed a feline model of lentiviral cross-species transmission using a puma lentivirus (PLV or FIVPco) which infects domestic cats but does not cause disease. Infection with PLV protects cats from CD4+ T-cell decline caused by subsequent infection with virulent feline immunodeficiency virus (FIV). Previous studies implicate innate immune and/or cellular restriction mechanisms for FIV disease attenuation in PLV-infected cats. In this study, we evaluated viral infection and cytokine mRNA transcription in 12 different tissue reservoirs approximately five months post infection. We quantitated tissue proviral load, viral mRNA load and relative transcription of IL-10, IL-12p40 and IFNγ from tissues of cats exposed to FIV, PLV or both viruses and analyzed these parameters using a multivariate statistical approach. The distribution and intensity of FIV infection and IFNγ transcription differed between single and co-infected cats, characterized by higher FIV proviral loads and IFNγ expression in co-infected cat tissues. Variability in FIV mRNA load and IFNγ was significantly more constrained in co-infected versus singly infected cat tissues. Single-infected:co-infected ratios of FIV mRNA load compared to FIV proviral load indicated that active viral transcription was apparently inhibited during co-infection. These results indicate that previous PLV infection increases activation of tissue innate immunity and constrains the ability of FIV to productively infect tissue reservoirs of infection for months, independent of FIV proviral load, supporting a model in which innate immunity and/or modulation of target cell susceptibility play a key role in PLV-induced protection from FIV disease

Closing the gap on causal processes of infection risk from cross-sectional data:structural equation models to understand infection and co-infection

BACKGROUND: Epidemiological studies of disease exposure risk are frequently based on observational, cross-sectional data, and use statistical approaches as crucial tools for formalising causal processes and making predictions of exposure risks. However, an acknowledged limitation of traditional models is that the inferred relationships are correlational, cannot easily distinguish direct from indirect determinants of disease risk, and are often considerable simplifications of complex interrelationships. This may be particularly important when attempting to infer causality in patterns of co-infection through pathogen-facilitation. METHODS: We describe analyses of cross-sectional data using structural equation models (SEMs), a contemporary advancement on traditional regression approaches, based on our study system of feline gammaherpesvirus (FcaGHV1) in domestic cats. RESULTS: SEMs strongly supported a latent (host phenotype) variable associated with FcaGHV1 exposure and co-infection risk, suggesting these individuals are simply more likely to become infected with multiple pathogens. However, indications of pathogen-covariance (potential facilitation) were also variably detected: potentially among FcaGHV1, Bartonella spp and Mycoplasma spp. CONCLUSIONS: Our models suggest multiple exposures are primarily driven by host phenotypic traits, such as aggressive male phenotypes, and secondarily by pathogen-pathogen interactions. The results of this study demonstrate the application of SEMs to understanding epidemiological processes using observational data, and could be used more widely as a complementary tool to understand complex cross-sectional information in a wide variety of disciplines

A Sarcoptes scabiei specific isothermal amplification assay for detection of this important ectoparasite of wombats and other animals

Background The globally distributed epidermal ectoparasite, Sarcoptes scabiei, is a serious health and welfare burden to at-risk human and animal populations. Rapid and sensitive detection of S. scabiei infestation is critical for intervention strategies. While direct microscopy of skin scrapings is a widely utilised diagnostic method, it has low sensitivity. PCR, alternatively, has been shown to readily detect mite DNA even in microscopy-negative skin scrapings. However, a limitation to the latter method is the requirements for specialised equipment and reagents. Such resources may not be readily available in regional or remote clinical settings and are an important consideration in diagnosis of this parasitic disease. Methodology A Loop Mediated Isothermal Amplification (LAMP) assay targeting the ITS-2 gene for S. scabiei was developed and evaluated on clinical samples from various hosts, previously screened with conventional S. scabies-specific PCR. Species specificity of the newly developed LAMP assay was tested against a range of DNA samples from other arthropods. The LAMP assays were performed on a real-time fluorometer as well as thermal cycler to evaluate an end-point of detection. Using skin scrapings, a rapid sample processing method was assessed to eliminate extensive processing times involved with DNA extractions prior to diagnostic assays, including LAMP. Results The S. scabiei LAMP assay was demonstrated to be species-specific and able to detect DNA extracted from a single mite within a skin scraping in under 30 minutes. Application of this assay to DNA extracts from skin scrapings taken from a range of hosts revealed 92.3% congruence (with 92.50% specificity and 100% sensitivity) to the conventional PCR detection of S. scabiei. Preliminary results have indicated that diagnostic outcome from rapidly processed dry skin scrapings using our newly developed LAMP is possible in approximately 40 minutes. Discussion We have developed a novel, rapid and robust molecular assay for detecting S. scabiei infesting humans and animals. Based on these findings, we anticipate that this assay will serve an important role as an ancillary diagnostic tool at the point-of-care, complementing existing diagnostic protocols for S. scabiei

Association between canine leishmaniosis and Ehrlichia canis co-infection: a prospective case-control study

Abstract Background In the Mediterranean basin, Leishmania infantum is a major cause of disease in dogs, which are frequently co-infected with other vector-borne pathogens (VBP). However, the associations between dogs with clinical leishmaniosis (ClinL) and VBP co-infections have not been studied. We assessed the risk of VBP infections in dogs with ClinL and healthy controls. Methods We conducted a prospective case-control study of dogs with ClinL (positive qPCR and ELISA antibody for L. infantum on peripheral blood) and clinically healthy, ideally breed-, sex- and age-matched, control dogs (negative qPCR and ELISA antibody for L. infantum on peripheral blood) from Paphos, Cyprus. We obtained demographic data and all dogs underwent PCR on EDTA-blood extracted DNA for haemoplasma species, Ehrlichia/Anaplasma spp., Babesia spp., and Hepatozoon spp., with DNA sequencing to identify infecting species. We used logistic regression analysis and structural equation modelling (SEM) to evaluate the risk of VBP infections between ClinL cases and controls. Results From the 50 enrolled dogs with ClinL, DNA was detected in 24 (48%) for Hepatozoon spp., 14 (28%) for Mycoplasma haemocanis, 6 (12%) for Ehrlichia canis and 2 (4%) for Anaplasma platys. In the 92 enrolled control dogs, DNA was detected in 41 (45%) for Hepatozoon spp., 18 (20%) for M. haemocanis, 1 (1%) for E. canis and 3 (3%) for A. platys. No Babesia spp. or “Candidatus Mycoplasma haematoparvum” DNA was detected in any dog. No statistical differences were found between the ClinL and controls regarding age, sex, breed, lifestyle and use of ectoparasitic prevention. A significant association between ClinL and E. canis infection (OR = 12.4, 95% CI: 1.5–106.0, P = 0.022) was found compared to controls by multivariate logistic regression. This association was confirmed using SEM, which further identified that younger dogs were more likely to be infected with each of Hepatozoon spp. and M. haemocanis, and dogs with Hepatozoon spp. were more likely to be co-infected with M. haemocanis. Conclusions Dogs with ClinL are at a higher risk of co-infection with E. canis than clinically healthy dogs. We recommend that dogs diagnosed with ClinL should be tested for E. canis co-infection using PCR

Three Pathogens in Sympatric Populations of Pumas, Bobcats, and Domestic Cats: Implications for Infectious Disease Transmission

Anthropogenic landscape change can lead to increased opportunities for pathogen transmission between domestic and non-domestic animals. Pumas, bobcats, and domestic cats are sympatric in many areas of North America and share many of the same pathogens, some of which are zoonotic. We analyzed bobcat, puma, and feral domestic cat samples collected from targeted geographic areas. We examined exposure to three pathogens that are taxonomically diverse (bacterial, protozoal, viral), that incorporate multiple transmission strategies (vector-borne, environmental exposure/ ingestion, and direct contact), and that vary in species-specificity. Bartonella spp., Feline Immunodeficiency Virus (FIV), and Toxoplasma gondii IgG were detected in all three species with mean respective prevalence as follows: puma 16%, 41% and 75%; bobcat 31%, 22% and 43%; domestic cat 45%, 10% and 1%. Bartonella spp. were highly prevalent among domestic cats in Southern California compared to other cohort groups. Feline Immunodeficiency Virus exposure was primarily associated with species and age, and was not influenced by geographic location. Pumas were more likely to be infected with FIV than bobcats, with domestic cats having the lowest infection rate. Toxoplasma gondii seroprevalence was high in both pumas and bobcats across all sites; in contrast, few domestic cats were seropositive, despite the fact that feral, free ranging domestic cats were targeted in this study. Interestingly, a directly transmitted species-specific disease (FIV) was not associated with geographic location, while exposure to indirectly transmitted diseases – vectorborne for Bartonella spp. and ingestion of oocysts via infected prey or environmental exposure for T. gondii – varied significantly by site. Pathogens transmitted by direct contact may be more dependent upon individual behaviors and intra-specific encounters. Future studies will integrate host density, as well as landscape features, to better understand the mechanisms driving disease exposure and to predict zones of cross-species pathogen transmission among wild and domestic felids