Widespread episodic thiamine deficiency in Northern Hemisphere wildlife.

Many wildlife populations are declining at rates higher than can be explained by known threats to biodiversity. Recently, thiamine (vitamin B1) deficiency has emerged as a possible contributing cause. Here, thiamine status was systematically investigated in three animal classes: bivalves, ray-finned fishes, and birds. Thiamine diphosphate is required as a cofactor in at least five life-sustaining enzymes that are required for basic cellular metabolism. Analysis of different phosphorylated forms of thiamine, as well as of activities and amount of holoenzyme and apoenzyme forms of thiamine-dependent enzymes, revealed episodically occurring thiamine deficiency in all three animal classes. These biochemical effects were also linked to secondary effects on growth, condition, liver size, blood chemistry and composition, histopathology, swimming behaviour and endurance, parasite infestation, and reproduction. It is unlikely that the thiamine deficiency is caused by impaired phosphorylation within the cells. Rather, the results point towards insufficient amounts of thiamine in the food. By investigating a large geographic area, by extending the focus from lethal to sublethal thiamine deficiency, and by linking biochemical alterations to secondary effects, we demonstrate that the problem of thiamine deficiency is considerably more widespread and severe than previously reported.

Evaluation of the 5α-reductase inhibitor finasteride on reproduction and gonadal development in medaka, Oryzias latipes.

5-α reductase (5αR) inhibitors have an anti-androgenic effect in mammals because they inhibit the conversion of testosterone to the potent androgen, dihydrotestosterone. Finasteride is a type-2 5αR inhibitor that is used as a human pharmaceutical for the treatment of prostate cancer, benign prostate hyperplasia and male pattern baldness. This study evaluated the impacts of finasteride (50, 500 and 5000µg/L) on the development and reproduction of medaka (Oryzias latipes) exposed continuously over multiple generations (F0, F1 and F2). The exposure was initiated with reproductively mature fish (F0 generation) and continued until the hatching of the F2 generation. There were no significant effects on survival, fecundity or fertility in the F0 (50, 500, 5000µg/L) and F1 (50, 500µg/L) generations. The F1 generation exposed to 5000µg/L exhibited significant mortality. Histopathology of the gonads demonstrated that medaka and pre-clinical species respond similarly to finasteride exposure. Intersex condition and maldeveloped gonads were observed in F0 generation males exposed to 5000µg/L and F1 generation males exposed to 500µg/L. F1 generation males exposed to 500µg/L displayed reduced gonadosomatic index with an increased incidence of testicular degeneration. Males in both generations exhibited an increased incidence of Leydig cell hyperplasia at concentrations ⩾500µg/L. F0 generation females exposed to 5000µg/L exhibited increased gonadosomatic index. An increased prevalence of accelerated post-ovulatory follicle involution was observed in females at concentrations ⩾500µg/L in both generations. The gonadal changes induced by finasteride support the idea that 5-α reductase inhibition impacts androgen signaling in fish. Results from this study are discussed in the context of differential expression of the androgen receptor between species of fish.

Spatial trends of dioxin-like compounds in Atlantic anguillid eels.

Several temperate freshwater eel stocks have experienced unsustainable declines, yet to be explained. The decline of lake trout (Salvelinus namaycush) in Lake Ontario has been linked to aryl-hydrocarbon receptor agonists such as polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), dioxin-like polychlorinated biphenyls (dl-PCBs), and polychlorinated naphthalenes (PCNs), and the question remains whether eels are affected similarly by these compounds. Concentrations of PCDD/Fs, dl-PCBs, and PCNs were determined in eels collected at seven locations in eastern Canada including L. Ontario, one location in New York, USA, and one location in Flanders, Belgium. Concentrations varied greatly among origins, indicating dissimilar historic loadings to local areas. The risk to eel reproduction was evaluated with 2,3,7,8-TCDD toxic equivalents, and increased by 10-fold from the least to most contaminated site. The risk to eel recruitment from dioxin-like compounds in American eel using available guidelines is low. The development of a more comprehensive model for eel recruitment risk assessment due to dioxin-like compounds, using eel-specific guidelines, is recommended. Toxic equivalents were 5-fold higher when based on mammalian toxic equivalency factors compared to fish values. About half of the eels captured in L. Ontario exceeded the Canadian guideline for fish consumption (20pg TEQ g(-1) ww), but there were no other exceedances in Canada. The current risk to eel consumers in Canada is low overall, except for highly urbanized and industrialized areas.

Damaged intestinal epithelial integrity linked to microbial translocation in pathogenic simian immunodeficiency virus infections.

The chronic phase of HIV infection is marked by pathological activation of the immune system, the extent of which better predicts disease progression than either plasma viral load or CD4(+) T cell count. Recently, translocation of microbial products from the gastrointestinal tract has been proposed as an underlying cause of this immune activation, based on indirect evidence including the detection of microbial products and specific immune responses in the plasma of chronically HIV-infected humans or SIV-infected Asian macaques. We analyzed tissues from SIV-infected rhesus macaques (RMs) to provide direct in situ evidence for translocation of microbial constituents from the lumen of the intestine into the lamina propria and to draining and peripheral lymph nodes and liver, accompanied by local immune responses in affected tissues. In chronically SIV-infected RMs this translocation is associated with breakdown of the integrity of the epithelial barrier of the gastrointestinal (GI) tract and apparent inability of lamina propria macrophages to effectively phagocytose translocated microbial constituents. By contrast, in the chronic phase of SIV infection in sooty mangabeys, we found no evidence of epithelial barrier breakdown, no increased microbial translocation and no pathological immune activation. Because immune activation is characteristic of the chronic phase of progressive HIV/SIV infections, these findings suggest that increased microbial translocation from the GI tract, in excess of capacity to clear the translocated microbial constituents, helps drive pathological immune activation. Novel therapeutic approaches to inhibit microbial translocation and/or attenuate chronic immune activation in HIV-infected individuals may complement treatments aimed at direct suppression of viral replication.

Distribution, prevalence, and intensity of the swim bladder parasite Anguillicola crassus in New England and eastern Canada.

In the summer of 2005, yellow phase American eels Anguilla rostrata were examined for the swim bladder parasite Anguillicola crassus from 26 locations in New England, USA, ranging from the Pawcatuck River, Rhode Island, to the East Machias River, Maine. An additional 12 sites were sampled within Canada during the summers of 2006 and 2007: 7 sites in southern Nova Scotia and 5 sites within the St. Lawrence River system. In 2007, eels were also obtained from New Brunswick, northern Nova Scotia, Prince Edward Island, and Newfoundland through the commercial eel fishery. All locations in Rhode Island (n = 3) and Massachusetts (n = 10) and 7 in Maine (n = 13) had infected eels, with parasite prevalence ranging from 7 to 76%. No eels sampled from southern Nova Scotia or the St. Lawrence River system were infected with the parasite. New Brunswick and northern Nova Scotia had infected eels ranging from 3 to 30% parasite prevalence, with Cape Breton, Nova Scotia, being the furthest north the parasite has been reported in American eels. There was no significant relationship between parasite prevalence and latitude. The present study supports the hypothesis that the parasite is capable of expanding its range further into the Maritimes and could potentially reach the St. Lawrence River system.

Direct identification of Staphylococcus aureus from positive blood culture bottles.

Fluorescence in situ hybridization (FISH) using peptide nucleic acid (PNA) probes targeting Staphylococcus aureus 16S rRNA is a novel method for direct identification of S. aureus from positive blood culture bottles. The test (S. aureus PNA FISH) is performed on smears made directly from positive blood culture bottles with gram-positive cocci in clusters (GPCC) and provides results within 2.5 h. A blinded comparison of S. aureus PNA FISH with standard identification methods was performed in collaboration with eight clinical microbiology laboratories. A total of 564 routine blood culture bottles positive for GPCC recovered from both aerobic and anaerobic media from three different manufacturers (ESP, BACTEC, and BacT/Alert) were included in the study. The sensitivity and specificity of S. aureus PNA FISH were 100% (57 of 57) and 99.2% (116 of 117), respectively, with 174 GPCC-positive ESP blood culture bottles, 98.5% (67 of 68) and 98.5% (129 of 131), respectively, with 200 GPCC-positive BACTEC blood culture bottles, and 100% (74 of 74) and 99.1% (115 of 116), respectively, with 190 GPCC-positive BacT/Alert blood culture bottles. It is concluded that S. aureus PNA FISH performs well with commonly used continuously monitoring blood culture systems.

Fluorescence in situ hybridization with peptide nucleic acid probes for rapid identification of Candida albicans directly from blood culture bottles.

A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described. The test (the C. albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C. albicans 26S rRNA. The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55 degrees C. Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy. The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C. albicans (n = 72), C. dubliniensis (n = 58), C. glabrata (n = 5), C. krusei (n = 2), C. parapsilosis (n = 4), and C. tropicalis (n = 3). The performance of the C. albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity. It is concluded that this 2.5-h method for the definitive identification of C. albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management.

Rapid identification of Staphylococcus aureus directly from blood cultures by fluorescence in situ hybridization with peptide nucleic acid probes.

A new fluorescence in situ hybridization (FISH) method with peptide nucleic acid (PNA) probes for identification of Staphylococcus aureus directly from positive blood culture bottles that contain gram-positive cocci in clusters (GPCC) is described. The test (the S. aureus PNA FISH assay) is based on a fluorescein-labeled PNA probe that targets a species-specific sequence of the 16S rRNA of S. aureus. Evaluations with 17 reference strains and 48 clinical isolates, including methicillin-resistant and methicillin-susceptible S. aureus species, coagulase-negative Staphylococcus species, and other clinically relevant and phylogenetically related bacteria and yeast species, showed that the assay had 100% sensitivity and 96% specificity. Clinical trials with 87 blood cultures positive for GPCC correctly identified 36 of 37 (97%) of the S. aureus-positive cultures identified by standard microbiological methods. The positive and negative predictive values were 100 and 98%, respectively. It is concluded that this rapid method (2.5 h) for identification of S. aureus directly from blood culture bottles that contain GPCC offers important information for optimal antibiotic therapy.