Lesion development and immunohistochemical changes in granulomas from cattle experimentally infected with Mycobacterium bovis

Mycobacterium bovis, the causative agent of bovine tuberculosis, persists within granulomas. Formation of granulomas involves a complex array of immune activation and cellular migration. To examine temporal changes in granuloma development, we inoculated 32 cattle with M. bovis of deer origin. Tissues from 4 calves each were examined at 15, 28, 42, 60, 90, 180, 270, and 370 days after inoculation. Granulomas in the medial retropharyngeal lymph node were staged (I-IV) on the basis of cellular composition and the presence or absence of necrosis and peripheral fibrosis. Immunohistochemistry for inducible nitric oxide synthase (iNOS), CD68, CD4, CD8, and gamma/delta T cells was performed. Fifteen days after inoculation only stage I granulomas were seen, while between 28 and 60 days, there was a steady progression through granuloma stages such that by day 60, granulomas of all 4 stages were seen. Acid-fast bacilli were present in moderate-to-large numbers in stage I granulomas 15-60 days after inoculation. Stage IV granulomas contained large numbers of acid-fast bacteria. Abundant iNOS immunoreactivity was associated with granulomas from day 15 through day 60 but was minimal from day 90 to the termination of the experiment. The relative number of CD4+ and CD68+ cells remained constant throughout the study. In contrast, at time points >60 days, numbers of CD8+ and gamma/delta T cells diminished. Tuberculous granulomas are dynamic lesions that follow an orderly progression through disease stages. Diminished expression of iNOS and reduced numbers of CD8+ and gamma/delta T cells late in the progression of tuberculous granulomas may represent a failure of the host response to control infection.     

In vivo endothelial cell infection by Anaplasma marginale

Anaplasma marginale has recently been shown to infect endothelial cells in vitro, but it remains unknown as to whether endothelial infection also occurs in vivo. In this report, we demonstrate through dual fluorescence microscopy that A marginale, detected by the monoclonal antibody ANAF16C1, co-localizes with the endothelial cell marker, von Willebrand factor, in tissue sections from an experimentally inoculated calf. The results indicate that A marginale infection includes endothelial cells and has implications for both pathogenesis and immune mechanisms.     

Relationship between Mycobacterium avium subspecies paratuberculosis, IL-1alpha, and TRAF1 in primary bovine monocyte-derived macrophages

Mycobacterium avium subspecies paratuberculosis (MAP) is a facultative intracellular pathogen that resides in host macrophage cells. Presently, little is known about how MAP is able to subvert the normal bacteriocidal functions of infected macrophages. Previously, we reported that ileal tissues from MAP infected cattle contained high levels of interleukin-1 alpha (IL-1alpha) and tumor necrosis factor receptor-associated factor 1 (TRAF1), relative to ileal tissues from uninfected cattle. High-level expression of these two proteins could have profound effects on macrophage function, intracellular signaling, and apoptosis. We now demonstrate that high levels of TRAF1 protein are located primarily within macrophages infiltrating areas of MAP infection. We have also utilized cultured bovine monocyte-derived macrophage cells (MDM) either infected with live MAP or stimulated with recombinant IL-1alpha (rIL-1alpha) to determine if there is a relationship between IL-1alpha and TRAF1 expression. These studies have identified a dose dependent increase in TRAF1 protein levels in bovine MDM in response to infection with live MAP or following treatment with rIL-1alpha. Sustained TRAF1 protein expression was dependent upon interaction of rIL-1alpha with it's receptor and rIL-1beta was also able to enhance TRAF1 gene expression. Our results suggest that MAP may use the IL-1-TRAF1 system to enhance TRAF1 protein expression in infected bovine MDM. These novel results provide evidence for a new avenue of research on the effect of MAP and other intracellular pathogens on macrophage signaling and apoptosis.     

Predicting canopy macrolichen diversity and abundance within old-growth inland temperate rainforests

Windward slopes of the inland mountain ranges in British Columbia support a unique inland temperate rainforest (ITR) ecosystem. Increasing fragmentation and the loss of old ITR stands have highlighted the need for determining conservation biology priorities among remaining old forest stands. We have addressed this issue by surveying foliose macrolichens within 53 old ITR stands in British Columbia's 135,000 ha very wet-cool interior-cedar hemlock (ICHvk2) biogeoclimatic subzone in the upper Fraser River watershed. Study plots were stratified by leading tree species and by “wet” versus “dry” relative soil moisture conditions. Other plot variables included: temperature, precipitation, incident solar loading, and canopy openness. Ordination plots showed a distinct assemblage of foliose cyanolichens, including Lobaria pulmonaria, Lobaria retigera, Sticta fuliginosa, Nephroma isidiosum, Nephroma occultum, and Pseudocyphellaria anomala, whose abundance was correlated with increasing relative soil moisture, temperature, canopy openness, precipitation, and basal area of spruce. Logistic regression models similarly identified relative soil moisture and temperature in all parsimonious models. Leading tree species, in combination with “wet” relative soil moisture and/or temperature, were important factors explaining the presence or absence of five (Cavernularia hultenii, L. retigera, N. occultum, Platismatia norvegica, and Sticta oroborealis) of the eight modeled old-growth associate lichen species. This combination of conditions favouring the development of canopy lichen communities in old forests was best expressed in low elevation water receiving sites. We hypothesize that groundwater availability in these sites promotes species richness and abundance of canopy lichens by creating more favourable conditions for growth, and by reducing fire return intervals which allows for the accumulation of rare species over time. Historically, forests in these wet “toe-slope” positions were disproportionately targeted for logging. Their conservation should now be a high priority, given their disproportionate significance to maintaining canopy lichen diversity in the present-day landscapes.     

From government to governance in forest planning? Lessons from the case of the British Columbia Great Bear Rainforest initiative

Much has been written about a supposed shift ‘from government to governance’ in many policy areas, including forest policy. However, the idea remains very much at the level of hypothesis as few empirical studies have confirmed the transition. Part of the problem is the multi-dimensional character of governance itself, which includes traditional ‘government’ as one of many possible governance modes. By providing a three dimensional picture of these potential governance modes, including overlapping institutional, political and regulatory dimensions, this article analyses the complex and incomplete character of moves towards any new governance mode in a high-profile land use planning exercise in British Columbia, that of the “Great Bear Rainforest” protected area strategy on the province's mid-coast region in 2006. Little evidence of such a shift is uncovered, despite much rhetoric to the contrary.     

Laboratory evaluation of fipronil,a phenylpyrazole insecticide,against adult Anopheles (Diptera: Culicidae) and investigation of its possible cross‐resistance with dieldrin in Anopheles stephensi

Adult mosquitoes from two strains of Anopheles gambiae and from three strains of Anopheles stephensi were exposed to 0.25% fipronil‐treated papers in WHO test kits or to 500 mg fipronil m⁻² impregnated mosquito netting in bioassay spheres. For comparison, tests were also carried out with the pyrethroid permethrin, using the same methods and doses, and on papers treated with 0.4 and 4% of the cyclodiene insecticide dieldrin. Compared with the same doses of permethrin, fipronil showed less and delayed activity. Two of the An stephensi strains were resistant to fipronil and dieldrin. To investigate whether this was due to a resistance mechanism in the An stephensi strains acting against both insecticides, the most fipronil‐ and dieldrin‐tolerant strain was further selected in two separate lines with one of the insecticides, followed by tests with the insecticide that the line had not been selected with. This indicated a concomitant rise of resistance to dieldrin in the fipronil‐selected line and vice versa. Repeated back‐crossing of the two lines with a susceptible strain and re‐selection with either dieldrin or fipronil gave evidence for the involvement of a single resistance mechanism to both insecticides. Permethrin resistance in both lines declined with selection for dieldrin or fipronil and confirms the absence of cross‐resistance between fipronil and pyrethroids. © 2001 Society of Chemical Industry     

Biochemical composition of New Zealand geoduck clam broodstock (Panopea zelandica) conditioned under different temperature and feeding regimes

Geoduck clams are amongst the most valuable cultured shellfish available on the world market, driving interest in the establishment of the native New Zealand species, Panopea zelandica (Quoy & Gaimard, 1835), as a candidate for aquaculture. A critical element of domestication is the effective management of breeding stock to optimize gamete and offspring quality. To develop a reliable broodstock conditioning protocol for P. zelandica, we investigated the biochemical composition of geoducks exposed to nine factorial treatment combinations of temperature (7.5, 11.5 and 16.5°C) and feeding ration [10 000, 50 000 and 100 000 cells mL^?1 of a (1:1 cell count) mixture of Tisochrysis lutea and Chaetoceros muelleri]. In addition, a reference group of geoducks was conditioned in an aquaculture pond‐nursery system, providing a dilute, mixed phytoplankton culture at ambient temperature. Glycogen, protein, lipid and fatty acid contents of flesh and viscera were measured at days 0, 36 and 73 of conditioning. Glycogen, protein and lipid analyses indicated that geoducks within all treatments achieved a positive energy balance, except for those in the treatment combining the highest temperature and lowest feeding ration. Comparisons of fatty acid profiles of animals among treatments and with the reference group revealed that eicosapentaenoic (EPA, C20:5n‐3), docosahexaenoic (DHA, C22:6n‐3) and arachidonic (ARA, C20:4n‐6) fatty acids were important contributors to gametogenic development for geoduck conditioning.     

Do protected areas mitigate the effects of fisheries‐induced evolution on parental care behaviour of a teleost fish?

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Using low‐cost drones to monitor heterogeneous submerged seaweed habitats: A case study in the Azores

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Market‐led options to scale up legume seeds in developing countries: Experiences from the Tropical Legumes Project

There are several hurdles to ensure sustainable seed production and consistent flow of improved legume varieties in sub‐Saharan Africa (SSA) and South Asia (SA). The unreliable demand, autogamous nature of most of the grain legumes, and slow variety replacement rate by smallholder farmers do not provide strong incentive for private seed companies to invest in legume seed business. Unless a well thought‐out and comprehensive approach to legume seed delivery is developed, current seed shortages will continue, eroding emerging market opportunities. The experiences reported here are collated through a 10‐year partnership project, the Tropical Legumes in SSA and SA. It fostered innovative public–private partnerships in joint testing of innovative market‐led seed systems, skills and knowledge enhancement, de‐risking private sector initiatives that introduced in new approaches and previously overlooked entities in technology delivery. As new public and private seed companies, individual seed entrepreneurs and farmer organizations emerged, the existing ones enhanced their capacities. This resulted in significant rise in production, availability and accessibility of various seed grades of newly improved and farmer demanded legume varieties in the target countries.