Purification and partial characterization of a cadmium-binding protein from the liver of rainbow trout (Onchorynchus mykiss).

This study describes the isolation and partial characterization of a low molecular weight (approximately 14 kDa), cadmium-binding protein from rainbow trout (Onchorynchus mykiss) liver. Rainbow trout were injected intraperitoneally with 3.5 mg/kg cadmium chloride (total body dose) twice weekly for 3 wk. Livers were removed and a cadmium-binding protein was isolated. Monoclonal antibodies produced against this protein were used in the affinity purification process. Amino acid analysis showed the protein contained 3.8 mol% cysteine, 3.5 mol% phenylalanine, 2.2 mol% tyrosine and 1.9 mol% histidine. The low cysteine content suggests that it was distinct from metallothionein. The monoclonal antibodies were also used to identify the protein in liver homogenates from both cadmium-exposed and control fish and in the testes of cadmium-exposed mice lacking the gene for both metallothionein-1 and metallothionein-II. The compound identified in this study represents a non-metallothionein cadmium-binding protein that appears to be highly conserved.     

Pathogenesis of liver lesions caused by experimental infection with Piscirickettsia salmonis in juvenile Atlantic salmon, Salmo salar L.

Piscirickettsia salmonis, the etiologic agent of salmonid rickettsial septicemia (SRS), or piscirickettsiosis, causes substantial economic losses to the salmon industry. The pathogenesis of the disease has not been fully characterized. The aim of this study is to describe the hepatic lesions associated with experimental P. salmonis infection in Atlantic salmon juveniles. Fish were maintained in fresh water and inoculated intraperitoneally (IP), orally, or on the gill surface with P. salmonis. A group of uninfected fish was kept as control. Liver samples from 5 fish in each inoculated group and 3 controls were collected weekly and processed for histological and immunohistochemical examination. Thickening of the liver capsule by inflammatory cells was a characteristic histologic feature of IP inoculated fish. Three weeks post-IP inoculation, 8 fish had died and 2 fish were sampled. Histological changes at this time consisted of vasculitis, presence of fibrin thrombi, vacuolated hepatocytes and focal areas of necrosis. Leukocytes containing intracytoplasmic basophilic microorganisms were seen within hepatic sinusoids. Vasculitis and intracytoplasmic vacuoles were prominent features in fish inoculated orally and on the gill surface. The presence of P. salmonis within hepatocellular vacuoles, endothelial cells, and leucocytes was confirmed by immunohistochemistry. The intracellular location of P. salmonis and the vascular damage seen in infected fish are characteristic of rickettsial infections. Histological lesions induced by experimental infection with P. salmonis using the oral and gill surface routes were similar to those observed in natural outbreaks of piscirickettsiosis. The tropism of P. salmonis for endothelial cells explains the vascular lesions observed in SRS, whereas hepatic lesions are due to ischemic necrosis and direct injury by intracytoplasmic organisms.     

Impacts of rising atmospheric carbon dioxide on model terrestrial ecosystems

In model terrestrial ecosystems maintained for three plant generations at elevated concentrations of atmospheric carbon dioxide, increases in photosynthetically fixed carbon were allocated below ground, raising concentrations of dissolved organic carbon in soil. These effects were then transmitted up the decomposer food chain. Soil microbial biomass was unaffected, but the composition of soil fungal species changed, with increases in rates of cellulose decomposition. There were also changes in the abundance and species composition of Collembola, fungal-feeding arthropods. These results have implications for long-term feedback processes in soil ecosystems that are subject to rising global atmospheric carbon dioxide concentrations.     

Bodyweights and growth rates of spring- and autumn-born Thoroughbred horses raised on pasture

AIM: To examine the growth of spring- and autumn-born Thoroughbred foals raised on pasture.

METHODS: Bodyweight and growth rates were measured in pasture-raised Thoroughbred horses, born in either spring (n=56) or autumn (n=7), from birth to approximately 13 and 17 months of age.

RESULTS: Birthweight tended to be lower in autumn- than spring-born foals (54.4, SD 7.92 kg vs 57.3, SD 5.90 kg; p=0.08). Between birth and 6 months of age, there was no difference in growth rate at equivalent ages between horses born in spring and autumn. Spring-born horses, which were weaned in the autumn, had lower post-weaning growth rates than autumn-born horses that were weaned in the spring. At time of the late yearling sales (March–April) in the Southern Hemisphere, unadjusted mean bodyweights of autumn-born horses (379.3, SD 24.8 kg) were lower (p=0.017) than those of the spring-born horses (437.2, SD 35.3 kg), although values in the autumn-born horses were all within two standard deviations (SD) of the mean of the spring-born animals. When adjusted for the covariates of birthweight and gender, the difference between spring- and autumn-born horses at that time was not significant (p=0.25).

CONCLUSIONS AND CLINICAL RELEVANCE: Some autumn-born foals could be marketed for late yearling sales in the Southern Hemisphere, on the basis of bodyweight. Furthermore, they might also be competitive in the Northern Hemisphere industry (sales or racing), as they would be competing against horses of the same official age.     

Isolation of a new virus, HBLV, in patients with lymphoproliferative disorders

A novel human B-lymphotropic virus (HBLV) was isolated from the peripheral blood leukocytes of six individuals: two HTLV-III seropositive patients from the United States (one with AIDS-related lymphoma and one with dermatopathic lymphadenopathy), three HTLV-III seronegative patients from the United States (one with angioimmunoblastic lymphadenopathy, one with cutaneous T-cell lymphoma, and one with immunoblastic lymphoma), and one HTLV-III seronegative patient with acute lymphocytic leukemia from Jamaica. All six isolates were closely related by antigenic analysis, and sera from all six virus-positive patients reacted immunologically with each virus isolate. In contrast, only four sera from 220 randomly selected healthy donors and none from 12 AIDS patients without associated lymphoma were seropositive. The virus selectively infected freshly isolated human B cells and converted them into large, refractile mono- or binucleated cells with nuclear and cytoplasmic inclusion bodies. HBLV is morphologically similar to viruses of the herpesvirus family but is readily distinguishable from the known human and nonhuman primate herpesviruses by host range, in vitro biological effects, and antigenic features.     

Monensin use against Neospora caninum challenge in dairy cattle

Using a randomized controlled trial design, a randomly allocated intervention group of 15 cows received a slow-release bolus that delivered 100 days of monensin. The negative control group of 15 cows received a placebo bolus that was identical to the monensin bolus, except without the monensin. Two weeks after bolus administration, all cows were challenged with a 2 ml subcutaneous injection of a live tachyzoite suspension. Whole blood and serum samples were collected from each cow every week for the first month post-challenge, and then every 2 weeks for the next 2 months. The extracted DNA from whole blood was tested for the Nc-5 gene fragment of Neospora caninum using a quantitative real time polymerase chain reaction. Serum was tested for antibodies to N. caninum using the IDEXX ELISA. Cows treated with monensin boluses had a significantly lower humoral immune response than cows treated with placebo boluses at one time point post-challenge (week 4 post-challenge). However, when adjusting for repeated measures within cows, the P value for this humoral difference was 0.098. No DNA for N. caninum was detected in either group, likely due to study design features.     

Isolation of human C-reactive protein complementary DNA and localization of the gene to chromosome 1

With a synthetic oligonucleotide mixture as probe, complementary DNA clones of C-reactive protein were isolated from an adult human liver complementary DNA library. The clones ranged in size from 700 to 1100 base pairs and were identified by partial DNA sequence analysis. One complementary DNA clone was used as a probe for hybridization with human-rodent DNA's isolated from somatic cell hybrids and bound to nitrocellulose filters (Southern blot analysis) to assign the human C-reactive protein gene to chromosome 1.     

Genomic diversity of human T-lymphotropic virus type III (HTLV-III)

The DNA genomes of human T-lymphotropic virus type III (HTLV-III) isolated from 18 individuals with AIDS or who were at risk for AIDS were evaluated for evidence of variation. Although all of the 18 viral DNA's hybridized throughout their entire genomes to a full-length cloned probe of the original HTLV-III isolate, each of the 18 isolates showed a different restriction enzyme pattern. The number of restriction site differences between isolates ranged from only 1 site in 23 to at least 16 sites in 31. No particular viral genotype was associated with a particular disease state and 2 of the 18 patients had evidence of concurrent infection by more than one viral genotype. Propagation of three different viral isolates in vitro for up to 9 months did not lead to detectable changes in their restriction patterns. These findings indicate that different isolates of HTLV-III comprise a spectrum of highly related but distinguishable viruses and have important implications regarding the pathogenicity of HTLV-III and attempts to develop effective diagnostic, therapeutic, and preventive measures for this virus.     

Floral flavonoids and pH in Dendrobium orchid species and hybrids

Anthocyanidins were identified in 28 Dendrobium species and hybrids selected for analysis based on colour and suitability in cut flower breeding. Flowers designated pink, red, maroon, orange, bronze, and brown in the trade were placed in RHS colour groups red-purple, purple-violet, violet on yellow, greyed-purple on yellow or yellow-orange, and brown. This colour range contained anthocyanins based on cyanidin, with peonidin occurring as a minor pigment. The colours of three blue genotypes, D. gouldii K280-6, D. biggibum ‘blue’, and D. Kultana ‘blue’, were light violet to purple by RHS standards and contained anthocyanins based on cyanidin. Peach-coloured flowers were classified as red or red-purple and included pelargonidin glycosides. Anthocyanin concentrations ranged from 0.13 to 0.18 μmoles/g FW in light lavender and peach, and up to 3.66 μmoles/g FW in brown. Combined cellular and vacuolar pH ranged narrowly from 4.67 to 5.09 among white, peach, lavender, and brown lines. Predominant copigments were flavonol glycosides based on kaempferol, quercetin, myricetin, and methylated derivatives. Flavonol aglycones and glycosylation sites differed little among two colour forms of D. gouldii and two D. Jaquelyn Thomas hybrids. Accumulation of quercetin, myricetin, and cyanidin indicated flavonoid 3' and 3',5' hydroxylation activities in several Dendrobium. Additional accumulation of isorhamnetin, syringetin, and peonidin indicated active flavonoid 3'- and 3',5'- O-methyltransferase enzymes. This revised version was published online in July 2006 with corrections to the Cover Date.