A new real-time PCR assay for improved detection of the parasite Babesia microti.

Babesiosis is an emerging zoonosis with important public health implications, as the incidence of the disease has risen dramatically over the past decade. Because the current gold standard for detection of Babesia is microscopic examination of blood smears, accurate identification requires trained personnel. Species in the genus cannot be distinguished microscopically, and Babesia can also be confused with the early trophozoite stage (ring forms) of Plasmodium parasites.

Thermogenesis, blood metabolites and hormones, and growth of lambs born to ewes supplemented with algae-derived docosahexaenoic acid.

Neonatal lamb mortality is a major factor affecting profitability in the sheep industry, and lamb thermogenesis is a key element in neonatal lamb survival. Increased lamb vigor has been reported when ewes were supplemented during late gestation with algae-derived docosahexaenoic acid (DHA); however, the effects of DHA on lamb thermogenesis and immunocompetence have not been investigated. Eighty twin-bearing Targhee ewes (ages 2 to 5 yr; 68.

Ecological risk assessment of zinc from stormwater runoff to an aquatic ecosystem.

Zinc (Zn) risks from stormwater runoff to an aquatic ecosystem were studied. Monitoring data on waterborne, porewater, and sediment Zn concentrations collected at 20 stations throughout a stormwater collection/detention facility consisting of forested wetlands, a retention pond and first order stream were used to conduct the assessment. Bioavailability in the water column was estimated using biotic ligand models for invertebrates and fish while bioavailability in the sediment was assessed using acid volatile sulfide-simultaneously extracted metal (AVS-SEM).

Characterization of S-adenosylhomocysteine hydrolase from Cryptosporidium parvum.

The S-adenosylhomocysteine hydrolase from the apicomplexan Cryptosporidium parvum (CpSAHH) has been characterized. CpSAHH is a single-copy, intronless gene of 1479 bp encoding a protein of 493 amino acids with a molecular mass of 55.6 kDa.

Divergent polyamine metabolism in the Apicomplexa.

The lead enzymes of polyamine biosynthesis, i.e. ornithine decarboxylase (ODC) and arginine decarboxylase (ADC), were not detected in Toxoplasma gondii [the limit of detection for ODC and ADC was 5 pmol min(-1) (mg protein)(-1)], indicating that T.

Localization of pyruvate:NADP+ oxidoreductase in sporozoites of Cryptosporidium parvum.

Cryptosporidium parvum contains a unique fusion protein pyruvate:NADP+ oxidoreductase (CpPNO) that is composed of two distinct, conserved domains, an N-terminal pyruvate:ferredoxin oxidoreductase (PFO) and a C-terminal cytochrome P450 reductase (CPR). Unlike a similar fusion protein that localizes to the mitochondrion of the photosynthetic protist Euglena gracilis, CpPNO lacks an N-terminal mitochondrial targeting sequence. Using two distinct polyclonal antibodies raised against CpPFO and one polyclonal antibody against CpCPR, Western blot analysis has shown that sporozoites of C.

Electron tomographic and ultrastructural analysis of the Cryptosporidium parvum relict mitochondrion, its associated membranes, and organelles.

Sporozoites of the apicomplexan Cryptosporidium parvum possess a small, membranous organelle sandwiched between the nucleus and crystalloid body. Based upon immunolabelling data, this organelle was identified as a relict mitochondrion. Transmission electron microscopy and tomographic reconstruction reveal the complex arrangement of membranes in the vicinity of this organelle, as well as its internal organization.

Acute and chronic toxicity of nickel to rainbow trout (Oncorhynchus mykiss).

Of the fish species tested in chronic Ni exposures, rainbow trout (Oncorhynchus mykiss) is the most sensitive. To develop additional Ni toxicity data and to investigate the toxic mode of action for Ni, we conducted acute (96-h) and chronic (85-d early life-stage) flow-through studies using rainbow trout. In addition to standard toxicological endpoints, we investigated the effects of Ni on ionoregulatory physiology (Na, Ca, and Mg).

Acute and chronic toxicity of nickel to a cladoceran (Ceriodaphnia dubia) and an amphipod (Hyalella azteca).

This study evaluated acute and chronic nickel (Ni) toxicity to Ceriodaphnia dubia and Hyalella azteca with the objective of generating information for the development of a biotic ligand model for Ni. Testing with C. dubia was used to evaluate the effect of ambient hardness on Ni toxicity, whereas the larger H.

Cryptosporidium parvum mitochondrial-type HSP70 targets homologous and heterologous mitochondria.

A mitochondrial HSP70 gene (Cp-mtHSP70) is described for the apicomplexan Cryptosporidium parvum, an agent of diarrhea in humans and animals. Mitochondrial HSP70 is known to have been acquired from the proto-mitochondrial endosymbiont. The amino acid sequence of Cp-mtHSP70 shares common domains with mitochondrial and proteobacterial homologues, including 34 amino acids of an NH2-terminal mitochondrion-like targeting presequence.

Expression and functional characterization of a giant Type I fatty acid synthase (CpFAS1) gene from Cryptosporidium parvum.

A 25-kb CpFAS1 gene from Cryptosporidium parvum has been engineered and expressed as five individual maltose-binding protein (MBP)-fusion proteins: an N-terminal loading unit, three fatty acyl elongation modules, and a C-terminal reductase. Enzymatic activities of all domains (except the reductase) were individually assayed as recombinant proteins. The preferred substrate for the fatty acyl ligase (AL) domain in the loading unit was palmitic acid (C16:0).

Mitochondrial-type iron-sulfur cluster biosynthesis genes (IscS and IscU) in the apicomplexan Cryptosporidium parvum.

Several reports have indicated that the iron-sulfur cluster [Fe-S] assembly machinery in most eukaryotes is confined to the mitochondria and chloroplasts. The best-characterized and most highly conserved [Fe-S] assembly proteins are a pyridoxal-5'-phosphate-dependent cysteine desulfurase (IscS), and IscU, a protein functioning as a scaffold for the assembly of [Fe-S] prior to their incorporation into apoproteins. In this work, genes encoding IscS and IscU homologues have been isolated and characterized from the apicomplexan parasite Cryptosporidium parvum, an opportunistic pathogen in AIDS patients, for which no effective treatment is available.

A Narf-like gene from Cryptosporidium parvum resembles homologues observed in aerobic protists and higher eukaryotes.

Here we report a Narf-like gene from the apicomplexan Cryptosporidium parvum (CpNARF). CpNARF is an intronless, single-copy gene of 1680 bp which encodes a putative protein of 560 amino acids with a calculated molecular mass of 63.1 kDa.

Characterization of S-adenosylmethionine synthetase in Cryptosporidium parvum (Apicomplexa).

The S-adenosylmethionine synthetase gene of the apicomplexan Cryptosporidium parvum (CpSAMS), an agent of diarrhea in immunocompromised and healthy humans and animals is described. CpSAMS is a single-copy, intronless gene of 1221 bp encoding a polypeptide of 406 amino acids with a molecular mass of 44.8 kDa.

Cryptosporidium parvum Cpn60 targets a relict organelle.

Chaperonin 60 (Cpn60) is a well-established marker protein for eukaryotic mitochondria and plastids. In order to determine whether the small double-membrane-bounded organelle posterior to the nucleus in the apicomplexan Cryptosporidium parvum is a mitochondrion, the Cpn60 gene of C. parvum sporozoites ( CpCpn60) was analyzed and antibodies were generated for localization of the peptide.

Transfusion-associated babesiosis after heart transplant.

We describe a 54-year-old spleen-intact man with transfusion-associated Babesia microti infection after a heart transplant. Adult respiratory distress syndrome developed in the patient, and he required mechanical ventilation. Our experiences with this patient suggest that babesiosis should be considered in the differential diagnosis of transplant patients who have fever and hemolytic anemia.

Evaluation of in vitro and in vivo activity of benzindazole-4,9-quinones against Cryptosporidium parvum.

A series of benzindazole-4,9-quinones was tested for growth-inhibitory effects on Cryptosporidium parvum in vitro and in vivo. Most compounds showed considerable activity at concentrations from 25 to 100 micro M. For instance, at 25 micro M the derivatives 5-hydroxy-8-chloro-N1-methylbenz[f]-indazole-4,9-quinone and 5-chloro-N2-methylbenz[f]indazole-4,9-quinone inhibited growth of C.

Cryptosporidium parvum: the first protist known to encode a putative polyketide synthase.

We are reporting a putative multifunctional Type I polyketide synthase (PKS) gene from the apicomplexan Cryptosporidium parvum (CpPKS1). The 40 kb intronless open reading frame (ORF) predicts a single polypeptide of 13,414 amino acids with a molecular mass of 1516.5 kDa.

Alpha-proteobacterial relationship of apicomplexan lactate and malate dehydrogenases.

We have cloned and sequenced a lactate dehydrogenase (LDH) gene from Cryptosporidium parvum (CpLDH1). With this addition, and that of four recently deposited alpha-proteobacterial malate dehydrogenase (MDH) genes, the phylogenetic relationships among apicomplexan LDH and bacterial MDH were re-examined. Consistent with previous studies, our maximum likelihood (ML) analysis using the quartet-puzzling method divided 105 LDH/MDH enzymes into five clades, and confirmed that mitochondrial MDH is a sister clade to those of y-proteobacteria, rather than to alpha-proteobacteria.

Characterisation of a novel transporter from Cryptosporidium parvum.

P-ATPases are transmembrane proteins that hydrolyse ATP to drive cations or other substances across biomembranes. In this study we present the characterisation of a novel P-ATPase from the apicomplexan parasite Cryptosporidium parvum (CpATPase3), an opportunistic pathogen in autoimmune deficiency syndrome patients, for which no treatment is available. The single copy gene encodes 1488 amino acids, predicting a protein of 169.

Evaluation of in vitro activity of aurones and related compounds against Cryptosporidium parvum.

The efficacy of a series of aurones, auronols and 4-methoxy-alpha-pyrones has been screened for the ability to inhibit the intracellular growth of the parasitic protist Cryptosporidium parvum using an in vitro enzyme linked immunosorbent assay (ELISA). All aurones of this series were active at 25 to 100 microM. 10 of 19 aurones inhibited the intracellular growth of C.

Pyruvate : NADP+ oxidoreductase from the mitochondrion of Euglena gracilis and from the apicomplexan Cryptosporidium parvum: a biochemical relic linking pyruvate metabolism in mitochondriate and amitochondriate protists.

Most eukaryotes perform the oxidative decarboxylation of pyruvate in mitochondria using pyruvate dehydrogenase (PDH). Eukaryotes that lack mitochondria also lack PDH, using instead the O(2)-sensitive enzyme pyruvate : ferredoxin oxidoreductase (PFO), which is localized either in the cytosol or in hydrogenosomes. The facultatively anaerobic mitochondria of the photosynthetic protist Euglena gracilis constitute a hitherto unique exception in that these mitochondria oxidize pyruvate with the O(2)-sensitive enzyme pyruvate : NADP oxidoreductase (PNO).

Characterization of a heavy metal ATPase from the apicomplexan Cryptosporidium parvum.

P1-ATPases are transporters which pump heavy metals across membranes, either to provide enzymes with essential cofactors or to remove excess, toxic metal cations from the cytosol. The first protist P1-ATPase (CpATPase2) has been isolated from the apicomplexan Cryptosporidium parvum, an opportunistic pathogen of AIDS patients. This single copy gene encodes 1260 amino acids (aa), predicting a protein of 144.