Hepatic transcriptome analyses of juvenile white bass (Morone chrysops) when fed diets where fish meal is partially or totally replaced by alternative protein sources.

White bass (Morone chrysops) are a popular sportfish throughout the southern United States, and one parent of the commercially-successful hybrid striped bass (M. chrysops ♂ x M. saxatilis ♀). Currently, white bass are cultured using diets formulated for other carnivorous fish, such as largemouth bass (Micropterus salmoides) or hybrid striped bass and contain a significant percentage of marine fish meal. Since there are no studies regarding the utilization of alternative proteins in this species, we evaluated the global gene expression of white bass fed diets in which fish meal was partially or totally replaced by various combinations of soybean meal, poultry by-product meal, canola meal, soy protein concentrate, wheat gluten, or a commercial protein blend (Pro-Cision™). Six isonitrogenous (40% protein), isolipidic (11%), and isocaloric (17.1 kJ/g) diets were formulated to meet the known nutrient and energy requirements of largemouth bass and hybrid striped bass using nutrient availability data for most of the dietary ingredients. One of the test diets consisted exclusively of plant protein sources. Juvenile white bass (40.2 g initial weight) were stocked into a flow-through aquaculture system (three tanks/diet; 10 fish/tank) and fed the test diets twice daily to satiation for 60 days. RNA sequencing and bioinformatic analyses revealed significant differentially expressed genes between all test diets when compared to fish meal control. A total of 1,260 differentially expressed genes were identified, with major ontology relating to cell cycle and metabolic processes as well as immune gene functions. This data will be useful as a resource for future refinements to moronid diet formulation, as marine fish meal becomes limiting and plant ingredients are increasingly added as a reliable protein source.

Hepatic transcriptomic and metabolic responses of hybrid striped bass (Morone saxatilis×Morone chrysops) to acute and chronic hypoxic insult.

Striped bass (Morone saxatilis), white bass (Morone chrysops), and their hybrid are an important group of fish prized for recreational angling in the United States, and there and abroad as a high-value farmed fish. Regardless of habitat, it is not uncommon for fish of the genus Morone to encounter and cope with conditions of scarce oxygen availability. Previously, we determined that hybrid striped bass reared under conditions of chronic hypoxia exhibited reduced feed intake, lower lipid and nutrient retention, and poor growth. To better understand the molecular mechanisms governing these phenotypes, in the present study, we examined the transcriptomic profiles of hepatic tissue in hybrid striped bass exposed to chronic hypoxia (90days at 25% oxygen saturation) and acute hypoxia (6h at 25% oxygen saturation). Using high-throughput RNA-seq, we found that over 1400 genes were differentially expressed under disparate oxygen conditions, with the vast majority of transcriptional changes occurring in the acute hypoxia treatment. Gene pathway and bioenergetics analyses revealed hypoxia-mediated perturbation of genes and gene networks related to lipid metabolism, cell death, and changes in hepatic mitochondrial content and cellular respiration. This study offers a more comprehensive view of the temporal and tissue-specific transcriptional changes that occur during hypoxia, and reveals new and shared mechanisms of hypoxia tolerance in teleosts.

Two unusual presentations of urogenital histoplasmosis and a review of the literature.

Two unusual clinical presentations of urogenital histoplasmosis are described. A review of the literature on urogenital histoplasmosis is provided.

Clonal dissemination of vancomycin-resistant Enterococci and comparison of susceptibility testing methods.

One hundred fifty clinical isolates of Enterococcus faecalis (88 isolates) and Enterococcus faecium (62 isolates) were tested in vitro for their susceptibility to vancomycin and high-level aminoglycosides (HLA). Remel's Synergy Quad Plates (RSQ) were used as the reference method and compared to Kirby-Bauer disc diffusion test, Vitek GPS-TA card, MicroScan Panel (GP-6), and Etest. Streptomycin susceptibility results for MicroScan GP-6 and RSQ were recorded at 24 and 48 h and all other methods and antibiotics were read at 24 h or less. When compared with the agar screen method, all of the methods demonstrated > 99% agreement. One isolate was falsely sensitive to gentamicin at 24 h, but resistant at 48 h, when tested on both MicroScan and RSQ agar screen. Thirty-nine isolates showed resistance to vancomycin with all methods. These isolates were from three different local hospitals and were identified as E. faecium. Pulse-field gel electrophoresis demonstrated that all of the vancomycin-resistant isolates were derived from the same clone. Of interest is the observation that high-level resistance to aminoglycosides varied between the clonally related isolates.

Infection due to Paecilomyces lilacinus: a challenging clinical identification.

We describe a case of noninvasive sinusitis caused by Paecilomyces lilacinus in a patient with diabetes mellitus. Cure was achieved by endoscopic drainage and aspiration of the fungal mass. We discuss the difficulty in and clinical importance of distinguishing Paecilomyces from Aspergillus.

A pseudoepidemic of Rhodotorula rubra: a marker for microbial contamination of the bronchoscope.

Rhodotorula rubra was isolated from bronchoscopy specimens from 11 patients. An investigation of the bronchoscopy equipment and the bronchoscopy suite revealed contamination of the suction channel with R rubra, as well as potentially pathogenic bacteria. Disinfection control methods included gas sterilization of the bronchoscope and the institution of an alcohol and air flush through the suction channel to allow complete drying of the scope between each patient use. We have had no further isolates of R rubra from bronchoscopy specimens since these measures were instituted, and repeat cultures from the suction channel have been negative.

Are serum levels of vancomycin useful in the first week of therapy?

Controversy exists regarding the need to monitor serum concentrations of vancomycin with some investigators recommending measurement of peak and trough concentrations in the first week of therapy and regularly thereafter, whereas others contend that empiric dosing produces safe and effective drug concentrations so that testing is unnecessary. Since vancomycin concentrations are measured, routinely in our hospital in the first week of therapy, we conducted a 12 month study to assess their clinical value in patients who were treated when gram positive cocci were detected in blood culture smears. One-hundred-five patients had gram positive cocci on blood culture smears. These bacteria were pathogens in 15 patients with Staphylococcus aureus and in 18 with coagulase negative staphylococci based on microbiologic criteria and a chart review confirming their clinical significance. Ten patients with S. aureus and 8 patients with coagulase negative staphylococci that were pathogens and 10 patients with coagulase negative staphylococci that were contaminants were treated with vancomycin. Serum peak and trough concentrations of vancomycin obtained within the first 5 days of therapy in these 28 patients were 14 to 40 micrograms/ml and 4.8 to 20 micrograms/ml. These concentrations were much above the MIC's of the microorganisms (< 4 micrograms/ml). Five patients had increases of serum creatinine of more than 0.6 mg% and in each patient the increases were attributable to other causes-shock, heart failure, and preexisting renal failure. Fifty five peak and trough concentrations 19 of which were drawn in patients with contaminated cultures were measured at a cost of $2,475.(ABSTRACT TRUNCATED AT 250 WORDS)

Mycobacterium chelonae (M. chelonae subspecies chelonae): report of a patient with a sporotrichoid presentation who was successfully treated with clarithromycin and ciprofloxacin.

We describe a sporotrichoid presentation of infection with Mycobacterium chelonae (M. chelonae subspecies chelonae). The disease occurred in a patient receiving corticosteroid therapy and was cured by the use of clarithromycin and ciprofloxacin.

Comparison of two culture approaches, blind passage and dual observation, for detecting Chlamydia trachomatis in various prevalence populations.

Chlamydia trachomatis diagnosis in our laboratory consisted of dual inoculation of shell vials and detection of inclusions by using fluorescein-conjugated monoclonal antiserum; the second culture vial was conventionally used for blind passage when the first vial was negative. We compared the increase in positivity using blind passage with that of a strategy utilizing observation of two stained monolayers (dual observation) without blind passage, in an effort to reduce the reporting time and labor associated with the conventional approach. A total of 4,329 specimens were obtained from an obstetrics and gynecology (OB-GYN) clinic (2,563 specimens) and the sexually transmitted disease clinic (1,766 specimens). These specimens were used to compare the two strategies. Blind passage of 1,269 initially culture-negative specimens from the OB-GYN clinic resulted in an additional 6 positive chlamydial diagnoses. In comparison, a similar number of specimens (1,294) from the OB-GYN clinic collected subsequently to the first group were tested by dual observation. There were five additional positive findings. A similar evaluation of specimens from the sexually transmitted disease clinic was performed. Blind passage of 313 initially culture-negative specimens yielded 3 additional positive diagnoses, whereas dual observation of 1,435 similar specimens resulted in 9 positive diagnoses. On the basis of analysis of 4,332 specimens, sensitivity of dual observation is comparable to that of blind passage; labor, cost, and reporting time of dual observation are reduced in comparison to those of blind passage.

Standardization of ELISA for the detection of anti-cardiolipin antibodies--effect of non-specific IgG binding.

Various frequencies of anti-cardiolipin antibodies reported in patients with systemic lupus erythematosus and other autoimmune and infectious diseases necessitates the need for standardization of the immunoassay. We report here the usefulness of "no-antigen" control for each serum in determining the actual value of the anti-phospholipid antibodies. This non-specific "no-antigen" binding was quite variable from serum to serum and in general was higher in the patient population than in healthy individuals which served as controls. This non-specific binding may be associated with the increased IgG content of the sera as shown by the linear association of the absorbance of the back-ground readings to increases in IgG concentration (correlation coefficient 0.98).

Evaluation of the modified Visuwell Strep-A enzyme immunoassay for detection of group-A Streptococcus from throat swabs.

The modified Visuwell Strep-A enzyme immunoassay (EIA) was compared with culture for detection of group-A Streptococcus from throat swabs. Throat swabs in modified Stuarts medium obtained after culture at two institutions were tested in Visuwell. Cumulative results were n = 417, sensitivity 87.8%, specificity 89.9% predictive value positive (PVP) 67.9%, predictive value negative (PVN) 96.8%, and accuracy 89.5%. At another site, swabs were delivered to the laboratory without transport medium, cultured, and subsequently tested by Visuwell (n = 202, sensitivity 79.6%, specificity 84.5%, PVP 65.2%, PVN 91.9%, accuracy 83.2%). When 1+ culture-positive specimens were considered negative, the sensitivity and PVN increased from 79.6% to 90.2% and 91.9% to 97.1% respectively. Overall performance of the modified Visuwell was comparable with that of the initial assay for throat swabs transported with or without modified Stuarts medium. Cross reaction with organisms other than group-A Streptococcus normally found in the oropharynx was negligible in Visuwell and the limit of detection of group-A Streptococcus was 5 x 10(4) colony-forming units.

Evaluation of a visual, rapid, membrane enzyme immunoassay for the detection of herpes simplex virus antigen.

We evaluated a 12-min, direct, monoclonal antibody-based enzyme immunoassay (EIA) (SureCell; Kodak, Rochester, N.Y.) which aids in the detection of herpes simplex virus infection; the assay system is also approved for culture confirmation. The test was evaluated from direct clinical samples and compared with conventional culture methodology by using a single swab. A total of 265 specimens from 180 female cervical-urogenital sites, 62 male urogenital sites, 4 rectal sites, 3 skin sites, 6 oral sites, and 10 colposcopy sites were collected on Dacron or cotton swabs and placed in viral transport medium (VTM). Within 6 h of receipt, 0.2 ml of the vortexed VTM was inoculated into each of two replicate cell cultures. Cell monolayers were observed daily for ten days, and cytopathic effect was confirmed by using an indirect immunoperoxidase reagent. The procedure for the SureCell assay conformed to the manufacturer's recommendations. When conventional culture was compared with EIA results, the overall sensitivity, specificity, positive predictive value, negative predictive value, and agreement were 64.4, 98.9, 96.7, 84.4, and 87.2%, respectively. Variables affecting the EIA sensitivity are the stage of the lesion and conventional culture methodologies. A review of culture results for 32 EIA false-negative tests indicated that 15 were detected after 48 h of incubation. Cytopathic effect observed at 48-, 72-, and 96-h cutoffs altered the sensitivity for the EIA. To ensure detection of SureCell herpes simplex virus-negative specimens, it is recommended that an unused aliquot of VTM be tested in cell culture.

A comparison between erythrocyte sedimentation rate (ESR) and selected acute-phase proteins in the elderly.

The erythrocyte sedimentation rate (ESR) and selected acute-phase proteins (APPs) were studied in 101 elderly people (mean age, 72 years) to determine their utility as diagnostic aids in subjects with underlying infections or inflammation. ESR and values for serum immunoglobulin A (IgA), the fourth component of complement (C4), haptoglobin, and alpha-1-antitrypsin (AAT) all correlated with infection or inflammation. C4 was the only test predictive of mortality at six months. Neither ESR nor any of the APPs demonstrated concomitantly high sensitivity, specificity, and positive predictive values. Receiver-operating characteristic curve analysis revealed low true positive to false positive ratios for all of the tests studied. In the elderly, measurement of APPs as a guide to underlying infection or inflammation has limited utility and offers no advantage over the traditional low-cost ESR.