Counting low-copy number proteins in a single cell

We have designed a microfluidic device in which we can manipulate, lyse, label, separate, and quantify the protein contents of a single cell using single-molecule fluorescence counting. Generic labeling of proteins is achieved through fluorescent-antibody binding. The use of cylindrical optics enables high-efficiency (approximately 60%) counting of molecules in micrometer-sized channels. We used this microfluidic device to quantify beta2 adrenergic receptors expressed in insect cells (SF9). We also analyzed phycobiliprotein contents in individual cyanobacterial cells (Synechococcus sp. PCC 7942) and observed marked differences in the levels of specific complexes in cell populations that were grown under nitrogen-depleted conditions.     

A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins

The isolation of broadly neutralizing antibodies against influenza A viruses has been a long-sought goal for therapeutic approaches and vaccine design. Using a single-cell culture method for screening large numbers of human plasma cells, we isolated a neutralizing monoclonal antibody that recognized the hemagglutinin (HA) glycoprotein of all 16 subtypes and neutralized both group 1 and group 2 influenza A viruses. Passive transfer of this antibody conferred protection to mice and ferrets. Complexes with HAs from the group 1 H1 and the group 2 H3 subtypes analyzed by x-ray crystallography showed that the antibody bound to a conserved epitope in the F subdomain. This antibody may be used for passive protection and to inform vaccine design because of its broad specificity and neutralization potency.     

Aseptic suppurative meningitis in juvenile boxer dogs: retrospective study of 12 cases

Immune-mediated central nervous system inflammation is described in a series of 12 juvenile boxer dogs. A diagnosis of steroid-responsive meningitis-arteritis was made based on the clinical presentation and on diagnostic findings. The boxer breed was at a higher risk for this inflammatory condition than other breeds. Long-term follow-up (>2 years) confirmed a better prognosis in this breed than in the beagle and the Bernese mountain dog. Complete resolution of clinical signs without significant deficits or recurrences was obtained in all cases. Early clinical recognition and immunosuppressive treatment resulted in a better response and complete resolution of the disorder.     

UNIQUE TOPOGRAPHIC DISTRIBUTION OF GREYHOUND NONSUPPURATIVE MENINGOENCEPHALITIS

Greyhound nonsuppurative meningoencephalitis is an idiopathic breed‐associated fatal meningoencephalitis with lesions usually occurring within the rostral cerebrum. This disorder can only be confirmed by postmortem examination, with a diagnosis based upon the unique topography of inflammatory lesions. Our purpose was to describe the magnetic resonance (MR) imaging features of this disease. Four Greyhounds with confirmed Greyhound nonsuppurative meningoencephalitis were evaluated by MR imaging. Lesions predominantly affected the olfactory lobes and bulbs, frontal, and frontotemporal cortical gray matter, and caudate nuclei bilaterally. Fluid attenuation inversion recovery (FLAIR) and T2 weighted spin‐echo (T2W) sequences were most useful to assess the nature, severity, extension, and topographic pattern of lesions. Lesions were predominantly T2‐hyperintense and T1‐isointense with minimal or absent contrast enhancement.     

Echinococcus multilocularis in Belgium: prevalence in red foxes (Vulpes vulpes) and in different species of potential intermediate hosts

Echinococcus multilocularis causes a rare but potentially lethal zoonotic infection in humans. This tapeworm is known to be endemic in foxes in several countries of Western and Central Europe. In Western Europe, the common vole (Microtus arvalis) and the water vole (Arvicola terrestris) are considered to be the most important intermediate host species of this cestode whereas the red fox is by far the most important final host. The purpose of this study was to provide data on the prevalences in Wallonia (Southern part of Belgium) both in the red fox and in different potential intermediate hosts. A total of 990 red foxes were examined between January 2003 and December 2004 for the presence of E. multilocularis. The average prevalence was 24.55% (22.38-27.87). Out of 1249 rodents or insectivores belonging to the species Apodemus sylvaticus, Arvicola terrestris, Clethrionomys glareolus, Microtus arvalis, Microtus agrestris and Sorex araneus, only one M. arvalis (out of 914-0.11% (0.003-0.61) and one C. glareolus (out of 23-4.3% (0.1-21.9) were found to be infected. However, the muskrat (Ondatra zibethicus) seems to be a good intermediate host as 11.18% (9.72-12.76) of the animals (n=1718) were found to be infected. A positive correlation was found between the prevalences in foxes and in muskrats in each of the different geological regions. This study indicates that the muskrat is highly sensitive to this zoonotic tapeworm and could perhaps represent a good bioindicator when studying the epidemiology of this parasitic infection in Belgium and in other countries where the muskrat is present.     

Effects of blue shrimp Litopenaeus stylirostris and goldlined rabbitfish Siganus lineatus in mono‐ and polyculture on production and environmental conditions

This study was conducted to compare the effects of shrimp and rabbitfish in mono‐ and polyculture stocked at high biomass on production and environmental conditions in a mesocosm system. Shrimp (14 g) and/or rabbitfish (19 g) were stocked in four treatments with different density but with the same total biomass (236 g m^?2), including shrimp monoculture (SM) (17 shrimp m^?2), shrimp–fish polyculture (SF) (11 shrimp and 4 rabbitfish m^?2), fish–shrimp polyculture (FS) (6 shrimp and 8 rabbitfish m^?2) and fish monoculture (FM) (12 rabbitfish m^?2). After 10 weeks of experiment, shrimp survival and biomass were low in the treatments where shrimp were dominant (SM, SF), while rabbitfish survival and biomass were high in all the treatments. Shrimp mortality was assumed to be related to an excess of the system carrying capacity (CC). Results suggested that CC is linked to shrimp biomass/density rather than the system eutrophication level. The ecosystem became heterotrophic as daily feed supply was beyond 7 g m^?2 per day. This threshold corresponded to the environmental CC of the semi‐intensive shrimp culture system. Under these conditions, the combination of high fish biomass and low shrimp biomass appeared as the most valuable in terms of system performances.     

Effect of Crude Palm Oil Incorporation on Growth,Survival, Feed Efficiency,and Body Composition of Heterobranchus longifilis Fingerlings

An 8-week feeding trial was carried out to determine the effect of increasing dietary palm oil levels on African catfish juvenile growth and nutrient utilization. Six isonitrogenous diets were formulated to contain 0.5% to 15% crude palm oil with dietary lipid levels ranging from 5% to 20%. Growth performance and protein utilization efficiency of African catfish responded in a positive manner to palm oil addition up to 9% (diet 14% lipid). No further improvement in growth was observed with higher levels of palm oil in the diets, and lower survival was observed with inclusion of 15% palm oil (diet 20% lipid). Whole-body lipid content increased when fish were fed with increasing levels of dietary lipid. Under our experimental conditions, the best growth rate and nutrient utilization were observed with the inclusion of 9% crude palm oil in the diets for African catfish.