bluuebunny/crossref_metadata_2025 huggingface dataset:
Cefotaxime-resistant Nocardia asteroides strains are isolates of the controversial species Nocardia farcinica
Authors: Wallace R J , Tsukamura M , Brown B A , Brown J , Steingrube V A , Zhang Y S , Nash D R
Published: January 2020

A recent study of Nocardia asteroides revealed that 95% of clinical strains had one of five antibiotic resistance patterns. We found the pattern of resistance to cefotaxime and cefamandole in 19% of 200 clinical N. asteroides isolates. Isolates with this drug resistance pattern were from numerous geographic sources and were associated with significant clinical disease (56% of patients had disseminated infections). Phenotypic studies revealed that these isolates were relatively homogeneous and matched previous descriptions and reference strains of the controversial species N. farcinica. Growth at 45 degrees C, acid production from rhamnose, ability to utilize acetamide as a nitrogen and carbon source, and resistance to tobramycin and cefamandole were features of N. farcinica that could be tested in the clinical laboratory and allowed their distinction from N. asteroides. The serious nature of disease due to N. farcinica and its resistance to the newer cephalosporins suggest a clinical need for laboratory identification of this species. (Current tests used in clinical laboratories do not distinguish N. farcinica from N. asteroides.) This is the first recognition that N. farcinica has a specific drug resistance pattern and confirms the previously described concept that drug resistance patterns of N. asteroides may be associated with specific taxonomic groups.
Characterization of the sequence of colonization and nosocomial candidemia using DNA fingerprinting and a DNA probe
Authors: Reagan D R , Pfaller M A , Hollis R J , Wenzel R P
Published: January 2020

The objective of this hospital-based study was to determine the relationship between colonizing and infecting strains of Candida species and Torulopsis glabrata. Surveillance cultures from high-risk patients were paired with subsequent bloodstream isolates. Organisms were typed by using restriction endonuclease digestion of chromosomal DNA with BstNI and EcoRI, followed by Southern hybridization with a DNA probe (pBD4) derived from Saccharomyces cerevisiae. Sixteen patients for whom documented colonization preceded documented bloodstream infection were identified. The mean time between obtainment of surveillance isolates and obtainment of bloodstream isolates was 8 days, with a range of 1 to 423 days. For 15 (94%) of 16 patients, the DNA fingerprint pattern (using BstNI) of the surveillance isolate was identical to that of the bloodstream isolate. Isolates from 13 (81%) of 16 patients were unique to those patients. Typing by Southern hybridization with the pBD4 probe was less discriminating. We conclude that for a well-defined subset of hospitalized patients who were colonized by Candida species before developing nosocomial candidemia, the colonizing and infecting strains were identical, suggesting endogenous acquisition of infection. Restriction endonuclease digestion of chromosomal DNA was shown to be a discriminating and reproducible typing method for Candida species and T. glabrata.
Defining Staphylococcus epidermidis cell wall proteins
Authors: Patrick C C , Plaunt M R , Sweet S M , Patrick G S
Published: January 2020

Three Staphylococcus epidermidis isolates of differing bacteriophage types were studied to define proteins confined to the cell wall, which were surface exposed and thus available to interact with the host. Three major proteins of 37, 41, and 51 kDa were identified in all whole-cell lysates and cell wall extracts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Two additional proteins of 18 and 25 kDa became evident by using 125I labeling to delineate surface-exposed proteins. A classification scheme using P1 to P5 to delineate the 51-, 41-, 37-, 25- and 18-kDa proteins is proposed. Additionally, murine immune sera were used to identify two immunodominant proteins of 51 and 25 kDa (P1 and P4, respectively).
Mycobacterium gordonae pseudoinfection associated with a contaminated antimicrobial solution
Authors: Tokars J I , McNeil M M , Tablan O C , Chapin-Robertson K , Patterson J E , Edberg S C , Jarvis W R
Published: January 2020

At Yale-New Haven Hospital, 46 specimens submitted for mycobacterial culture during an 8-week period in 1989 were positive for Mycobacterium gordonae, a nontuberculous acid-fast bacterium (AFB) of low pathogenicity. The specimens were submitted from 34 patients who came from various inpatient and outpatient services. Four patients were begun on antimycobacterial therapy on the basis of an AFB isolate which was later identified as M. gordonae. Isolation of M. gordonae was associated with use of the BACTEC TB system (BACTEC TB; Becton Dickinson Diagnostic Instrument Systems, Towson, Md.) and an antimicrobial solution, BACTEC PANTA PLUS (PANTA; Becton Dickinson Diagnostic Instrument Systems). The manufacturer reported that two lots (B9K1 and C9K1) of PANTA kits containing a single production lot (N8C1) of PANTA, which had been shipped to 173 laboratories, had been contaminated with M. gordonae. A survey of mycobacteriology laboratories in the United States revealed that, during April to July 1989, the M. gordonae isolation rate was 5.8/1,000 AFB specimens processed at laboratories that did not use BACTEC TB, 11.4/1,000 AFB specimens at laboratories that used BACTEC TB but not the implicated lot of PANTA, and 23.5/1,000 AFB specimens at laboratories that used BACTEC TB and the lot of implicated PANTA. Intrinsic contamination of PANTA was attributed to ineffective sterilization of water used in the manufacturing process and was not detected prior to product shipment because cultures for AFB were not part of the quality control regimen. This episode emphasizes that clinical laboratories can detect pseudoepidemics promptly if they are alert to abrupt increases in isolation rates, especially of unusual or generally nonpathogenic organisms.
Detection of Mycobacterium leprae antigens in the sera of leprosy patients by sandwich immunoradiometric assay using monoclonal antibodies
Authors: Patil S A , Girdhar B K , Singh K P , Sengupta U
Published: January 2020

An immunological technique for demonstration of Mycobacterium leprae antigen in sera was developed by using specific as well as cross-reactive monoclonal antibodies. The sandwich immunoradiometric assay which we developed is a simple, robust assay that is sensitive to the nanogram level. Sera from 72 leprosy patients were screened for the presence of antigen by this assay. A total of 69% of untreated tuberculoid leprosy patients showed 35-kDa antigen positivity, and 45% of these patients showed anti-35-kDa antibody positivity. Consistently higher antigen positivity rates for the 35-, 12-, and 30- to 40-kDa components of M. leprae were observed in lepromatous leprosy patients than in tuberculoid leprosy patients. During the course of therapy the antigen positivity rate gradually declined, and the antigen could not be detected in any of the 15 patients with subsided cases of leprosy. As antigen is presumably in excess before the antibody response is evoked, our experimental approach for antigen detection is likely to be useful by itself or along with antibody detection for diagnosis of early leprosy.
Use of tri-gas incubator for routine culture of Campylobacter species from fecal specimens
Authors: Thompson J S , Hodge D S , Smith D E , Yong Y A
Published: January 2020

We evaluated a tri-gas incubator for Campylobacter isolation to be used instead of an anaerobic jar. Fecal specimens were cultured in duplicate onto charcoal selective medium and incubated at 43 degrees C for 48 h in two different environments: a tri-gas incubator (Forma Scientific) adjusted to provide an atmosphere of 10% CO2, 10% O2, and the balance N2; and evacuated anaerobic jars with a replacement gas mixture of 10% CO2, 5% O2, and 85% N2. A total of 106 Campylobacter jejuni and 8 Campylobacter coli isolates were obtained from 2,348 stool specimens. Of the positive specimens, 113 isolates came from the incubator and 111 isolates came from the anaerobic jars. An additional 32 previously positive specimens were replated onto charcoal selective medium and retested by both methods. We recovered 27 C. jejuni isolates, 26 isolates by each method. The isolates from the incubator typically produced discrete colonies, while the isolates from the anaerobic jar showed some degree of swarming in colony formation. The tri-gas incubator provided a cost-effective method for culturing Campylobacter spp.
Neutralization receptor-based immunoassay for detection of neutralizing antibodies to Escherichia coli verocytotoxin 1
Authors: Boulanger J , Petric M , Lingwood C , Law H , Roscoe M , Karmali M
Published: January 2020

The NeutRELISA, a modification of the receptor enzyme-linked immunosorbent assay developed for the detection of verocytotoxin 1 (VT1) which permits the rapid detection of neutralizing antibodies (NAb) against this toxin, has been performed. A standard concentration of VT1 was preincubated with VT-immune or nonimmune rabbit serum. The serum-toxin mixtures were then added to microtiter plates coated with deacylated globotriosyl ceramide (lyso-Gb3). The reduction of VT1 binding to lyso-Gb3 in the immune serum-toxin mixtures compared with the VT1-Gb3 binding in the nonimmune serum-toxin mixtures was detected by using mouse monoclonal antibody to VT1. After standardization of the NeutRELISA with rabbit sera, 57 human control serum samples were tested to establish a cutoff value below which NeutRELISA results would be considered positive. Thirty-three single serum samples known to demonstrate NAb to VT1 by biological assay reproducibly demonstrated VT1 NAb when tested by the NeutRELISA. There was a close correlation between the biological VT1 neutralization assay and the NeutRELISA. This assay offers a practical, rapid, and reliable approach for the detection of NAb to VT1 and other verocytotoxins.
Properties of group B streptococci with protein surface antigens X and R
Authors: Wibawan I W , Lämmler C
Published: January 2020

A total of 128 bovine and 134 human group B streptococci were serotyped by conventional methods. Among the bovine cultures, 60 (47%) had type antigen X, and among the human cultures, 53 (39%) had type antigen R. The occurrence of type antigens X and R was significantly related to the growth pattern of the bacteria in fluid media. Type X- and R-positive cultures and most of the nontypeable cultures predominantly formed long chains and grew as granular sediment with clear supernatant. In addition, group B streptococci with surface antigen X or R showed compact colony formation in soft agar and reacted positively in the salt aggregation test. These properties, possibly caused by the surface charges of the X- and R-positive cultures, might be related to bacterial pathogenicity.
Oligonucleotide probe for detection of the enteropathogenic Escherichia coli (EPEC) adherence factor of localized adherent EPEC
Authors: Jerse A E , Martin W C , Galen J E , Kaper J B
Published: January 2020

The enteropathogenic Escherichia coli adherence factor (EAF) probe detects isolates of enteropathogenic E. coli that exhibit localized adherence to HEp-2 cells. A 21-base oligonucleotide probe was constructed on the basis of a sequence from within the 1-kb EAF probe and was shown to have greater sensitivity and specificity than the EAF fragment probe in detecting localized adherent E. coli.
Fatal encephalitis caused by Dactylaria constricta var. gallopava in a snowy owl chick (Nyctea scandiaca)
Authors: Salkin I F , Dixon D M , Kemna M E , Danneman P J , Griffith J W
Published: January 2020

Dactylaria constricta var. gallopava (Cooke) Salkin et Dixon was found to cause fatal encephalitis in a 28-day-old, captivity-bred snowy owl chick (Nyctea scandiaca). The previously healthy bird suddenly developed ataxia, severe torticollis, and extensor rigidity of the legs. Since the animal did not improve with antibiotic or vitamin-mineral supplement therapy, the chick was euthanized 5 days after the onset of neurologic signs. At necropsy, all tissues except the brain were grossly normal. Cultures inoculated with blood from the brain and heart yielded a dematiaceous mould that subsequently proved to be D. constricta var. gallopava. This is the first report of natural central nervous system infection caused by D. constricta var. gallopava in a snowy owl.
Species, biotype, and bacteriophage type determinations compared with cell envelope protein profiles for typing Acinetobacter strains
Authors: Bouvet P J , Jeanjean S , Vieu J F , Dijkshoorn L
Published: January 2020

Species, biotypes, and phage types were determined for 120 Acinetobacter strains from clinical or environmental sources or from culture collections. These characteristics were compared with cell envelope protein profiles obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in previous studies. A considerable heterogeneity of species and types was observed by use of the various methods, in particular among strains from different sources. Acinetobacter baumannii was the most commonly found species in isolates from clinical sources, followed by Acinetobacter species 3. Nine biotypes were observed among A. baumannii strains. Further differentiation within most species and biotypes was achieved by protein profile typing and, to some extent, phage typing. Of 120 strains, 49 (41%) were not typeable by phages. Consistent results for the various classification methods were obtained for strains from common sources. Biotyping seemed an appropriate method for the screening of strains, whereas protein profile and phage typing could serve as additional methods to establish the identity or nonidentity of strains. Results of this study suggest that the combination of the typing methods is useful in epidemiological studies.
Serological response in Enterococcus faecalis endocarditis determined by enzyme-linked immunosorbent assay
Authors: Shorrock P J , Lambert P A , Aitchison E J , Smith E G , Farrell I D , Gutschik E
Published: January 2020

Enterococcus (Streptococcus) faecalis expresses three species-specific surface protein antigens of molecular weights 73,000, 40,000, and 37,000. On Western blotting (immunoblotting), they were detected strongly by immunoglobulin G (IgG) in sera from patients with E. faecalis endocarditis, but not in sera from patients with other E. faecalis infections or with endocarditis due to other streptococci. We developed an enzyme-linked immunosorbent assay system to measure IgG, IgM, and IgA levels to these antigens and evaluated its potential as a serodiagnostic test for E. faecalis endocarditis. The test correctly diagnosed E. faecalis endocarditis in 15 of 16 cases. Of 10 cases of endocarditis due to other streptococci and 10 E. faecalis infections other than endocarditis, 9 and 8, respectively, gave negative results. The test should prove particularly useful in culture-negative cases, for which choice of appropriate antibiotic therapy for E. faecalis endocarditis is vital.
Comparison of the staphylocoagulase activities of Staphylococcus aureus and Staphylococcus intermedius on Chromozym-TH
Authors: Raus J , Love D N
Published: January 2020

The staphylocoagulases of Staphylococcus intermedius (39 strains from clinical samples from dogs and 1 strain from a pigeon) and Staphylococcus aureus (40 strains from nine different animal species) were compared by using the chromogenic methods of Engels et al. (W. Engels, M. Kemps, and C. P. A. van Boven, J. Clin. Microbiol. 14:496-500, 1981). S. intermedius produced staphylocoagulase which resembled that of S. aureus in its rate and method of action on prothrombin, but S. intermedius produced it in lesser amounts. Therefore, chromogenic substrate tests such as Chromozym-TH (Boehringer GmbH, Mannheim, Federal Republic of Germany) (as described by Engels et al.) can be used for the detection of staphylocoagulase in both of these species. However, to detect accurately the presence of S. intermedius staphylocoagulase by this method, preconcentration of the extracellular proteins or an extension of the reaction time of the test would be required. The method described by Engels et al. was designed specifically for clinical laboratories that handle human samples. Under those circumstances the method could be regarded as specific, enabling an identification of S. aureus only. However, as both S. aureus and S. intermedius give positive results in this test, in veterinary diagnostic laboratories, heavy reliance must not be placed on this test for determination of organisms to the species level.
Klebsiella pneumoniae gastroenteritis masked by Clostridium perfringens
Authors: Rennie R P , Anderson C M , Wensley B G , Albritton W L , Mahony D E
Published: January 2020

An unusual food-borne outbreak of gastroenteritis associated with contaminated turkey occurred at a catered company meal. The average incubation period was 10 h, and the predominant symptoms were watery diarrhea and cramps. Vomiting did not occur. Initial epidemiological features and cultures from turkey and feces of infected patients suggested that the causative agent was Clostridium perfringens, but Klebsiella pneumoniae of capsular type K15 was also isolated in large numbers from both the turkey and feces of the same patients. Plasmid analysis and enterotoxin results supported the role of K. pneumoniae as the causative agent in this outbreak. Organisms other than commonly identified pathogens should not be ignored if present in high concentrations in both food and feces of infected persons.
Antigenic variation of Giardia lamblia in the feces of Mongolian gerbils
Authors: Moss D M , Mathews H M , Visvesvara G S , Dickerson J W , Walker E M
Published: January 2020

Enzyme-linked immunoelectrotransfer blot was used to study variations in Giardia lamblia antigens in extracts of feces from infected Mongolian gerbils. A 65-kilodalton antigen was found in feces that contained strain WB (ATCC 30957) cysts and in axenic culture of strains WB and CDC:0284:1 that contained trophozoites. The 65-kilodalton antigen from trophozoites of both strains was membrane associated. A 70-kilodalton antigen was found in feces that contained strain CDC:0284:1 cysts. It was persistent in 16 fecal collections and may be strain specific. Similar variations in antigens may occur in human feces. Coproimmunodiagnostic assays that use monoclonal antibodies will have to include all varieties of G. lamblia antigens present in the feces of giardiasis patients.
Impairment of Plasmodium falciparum-specific antibody response in severe malaria
Authors: Brasseur P , Ballet J J , Druilhe P
Published: January 2020

Serum antibody response to plasmodial antigens was investigated in 97 Thai patients with Plasmodium falciparum malaria. No difference in immunoglobulin G (IgG) antibody levels was detected between groups without or with cerebral manifestations of malaria (n = 40). In patients with the most severe form of the disease, i.e., those who died despite adequate therapy (n = 12), antibody detected in the immunofluorescent-antibody test was found at lower levels than in those who recovered (geometric means: IgG = 1/420 versus 1/3,800; IgM = 1/15 versus 1/70); similarly, precipitating malarial antibodies were present in only 1 of these 12 patients, while they were detectable in 65 of the remaining 85 patients (76.5%). In contrast, anticytomegalovirus antibody levels were similar in the different groups of patients. Results show that depression of antibody response may extend to antiplasmodial responses during severe malaria. The link between fatality and a low level of antibody production suggests that an appropriate immune response to malarial antigens may be required to achieve recovery with drug treatment and provides a new direction for malaria therapy research.
Growth of Vero E-6 cells on microcarriers in a cell bioreactor
Authors: White L A , Ades E W
Published: January 2020

Minimum essential medium supplemented with either 7% fetal bovine serum or 2% fetal bovine serum and 1.5% low protein serum replacement (Sigma Chemical Co., St. Louis, Mo.) factor was used to evaluate the growth of Vero 76 cells (E-6 clone) in a cell culture bioreactor. Three different microcarriers, collagen-coated polystyrene (Kontes Life Sciences Products, Vineland, N.J.), Gelibead (Hazelton Research Products, Inc., Lenexa, Kans.), and Cytodex 3 (Pharmacia Fine Chemicals, Piscataway, N.J.), were used to compare the propagation of these anchorage-dependent cells. The number of viable cells recovered during three harvests over an 11-day incubation period was evaluated. The Vero E-6 cells grew equally well with both media. When the collagen-coated dextran microcarriers (Cytodex 3) were used, better cell attachment and higher-density growth were obtained and 5.5 x 10(8) to 7.0 x 10(8) cells were recovered from the system.
Growth promotion of Bifidobacterium species by whey and casein fractions from human and bovine milk
Authors: Petschow B W , Talbott R D
Published: January 2020

An in vitro assay was used to study the growth-promotional activity of human milk (HM), cow's milk (CM), and whey and casein fractions of HM and CM for five strains of Bifidobacterium species isolated originally from stools of human infants. Whey- and casein-predominant CM-based infant formulas were studied as well. When compared on an equivalent protein basis, the growth promotion activity of HM was greater than that of CM for Bifidobacterium bifidum serovar pennsylvanicus and Bifidobacterium longum but comparable for B. bifidum, Bifidobacterium infantis, and Bifidobacterium breve. Pasteurization of HM and CM resulted in an increase of growth promotion activity for B. bifidum serovar pennsylvanicus and B. bifidum, a decrease for B. infantis, and no change for B. longum and B. breve. The growth promotion activity of HM whey was slightly higher than that of HM casein for four strains of bifidobacteria. When CM casein was a substrate, virtually no growth occurred for B. bifidum serovar pennsylvanicus, B. bifidum, B. infantis, and B. longum. The growth promotion activity of CM whey, however, was similar to that of HM whey. A similar trend was observed for CM-based infant formula. Whey-dominant formulas promoted better growth of B. bifidum serovar pennsylvanicus, B. bifidum, and B. infantis than casein-dominant formulas. The data suggest a direct relationship between amount of whey-specific factors and the ability to promote growth of clinically relevant strains of Bifidobacterium species by HM, CM, and CM-based infant formulas.
Evaluation of immunoglobulin M western blot analysis in the diagnosis of congenital syphilis
Authors: Lewis L L , Taber L H , Baughn R E
Published: January 2020

Western immunoblots of solubilized Treponema pallidum antigens were reacted with sera and cerebrospinal fluid (CSF) and developed with enzyme-conjugated antibodies to immunoglobulin M (IgM). A blot was considered positive if reactions included bands at the 47-, 17-, and 15.5-kilodalton positions along with a variable pattern at other low-molecular-weight positions. Sera from 23 of 25 symptomatic infants diagnosed with congenital syphilis yielded positive reactions. Of 80 asymptomatic infants considered at risk for developing symptomatic infection, 16 exhibited IgM patterns consistent with those seen in congenital syphilis, although 5 of these 16 gave reactions that were equivocal. To exclude false-positive reactions due to IgM rheumatoid factor, sera were fractionated and the IgM fractions were retested. Only the five initially equivocal sera gave nonreactive blots with the IgM fractions, whereas all others gave more prominent reactions that were qualitatively similar to those seen in serum samples. Sera from 18 normal infants failed to show any IgM reactivity to T. pallidum antigens on Western blots. The IgM Western blot was both more sensitive and more specific than the fluorescent treponemal antibody-absorbed (IgM) test using fractionated serum. Of the 17 CSF samples from infants with symptomatic congenital syphilis, 14 showed IgM reactivity in Western blots, whereas only 12 had a reactive CSF in the Venereal Disease Research Laboratory test. Our results indicate that this technique can be used to identify both symptomatic and asymptomatic infection in infants with T. pallidum, in some cases before standard serologic studies can confirm the diagnosis.
Species-specific oligodeoxynucleotide probes for the identification of periodontal bacteria
Authors: Dix K , Watanabe S M , McArdle S , Lee D I , Randolph C , Moncla B , Schwartz D E
Published: January 2020

Oligodeoxynucleotide probes were developed for identification of the periodontal bacteria Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, B. intermedius types I and II, B. forsythus, Eikenella corrodens, Fusobacterium nucleatum, Haemophilus aphrophilus, Streptococcus intermedius, and Wolinella recta. Probes were designed by sequencing the 16S rRNA for each bacterium, identifying hypervariable regions, and chemically synthesizing species-specific probes. These probes were specific when tested against a panel of nucleic acids from closely related bacteria.
Serum antibody response to Listeria monocytogenes, listerial excretion, and clinical characteristics in experimentally infected goats
Authors: Miettinen A , Husu J , Tuomi J
Published: January 2020

We induced an experimental listeriosis in five goats by oral inoculation of Listeria monocytogenes serovar 1/2a. The resulting generalized bacteremia was reflected by systemic illness in four of the five animals inoculated and by excretion of L. monocytogenes in milk by all five animals. By using an enzyme immunoassay, we recorded a markedly enhanced immunoglobulin G (IgG) antibody response in the two youngest and most seriously ill test animals. In these, the elimination of L. monocytogenes from the gastrointestinal tract occurred simultaneously with the development of the highest antibody levels at 14 to 15 days postinoculation. In the case of the oldest test animal, a preexisting, persistent IgG antibody response was recorded which was associated with the total absence of clinical symptoms and the shortest observed fecal carriage of L. monocytogenes, lasting for only 3 days. Two animals remained practically seronegative, and an IgM antibody response was not recorded for any of the animals. The findings suggest that an association exists between the humoral immune defense against Listeria infections, the clinical course of the infection, and the elimination of the Listeria organisms from the gastrointestinal tract.
Tuberculous anergic sera or purified protein derivative treatment induces modification in lymphocyte transformation of cells from patients with tuberculosis
Authors: Mejia-López H , Vaca-García O , Cristerna-Aguirre J M , Cruz-Chávez A , Selman-Lama M , Taylor M L
Published: January 2020

Peripheral blood mononuclear cells from 20 purified protein derivative (PPD)-reactive (PPD+) tuberculous patients were cultured in autologous, tuberculous anergic, or normal serum. After 12 h of incubation, the serum was eliminated and lymphocyte transformation with PPD was performed. Transformation was inhibited only in cells incubated with anergic serum. In contrast, cells from 11 anergic tuberculous (PPD-) individuals recovered the ability to respond to an optimal PPD dose after treatment with high PPD concentrations followed by several washings. The cells which recovered returned to their initial anergic state when incubated with sera from anergic patients. Under both conditions, incubation with sera did not abolish the response to the mitogen phytohemagglutinin. Cells from healthy PPD+ or PPD- individuals were used as controls. The most important finding derived from serum analysis was the increased levels of specific immunoglobulins G and A in anergic patients.
Isolation of salmonellae from dried rattlesnake preparations
Authors: Babu K , Sonnenberg M , Kathpalia S , Ortega P , Swiatlo A L , Kocka F E
Published: January 2020

Salmonella arizonae and other Salmonella serovars were isolated from four different rattlesnake preparations which were used for self-treatment of various diseases. A case of disseminated S. arizonae infection is reported in a patient who had acquired immunodeficiency syndrome and ingested dried rattlesnake.
Evaluation of a nonisotopic probe for detection of Chlamydia trachomatis in endocervical specimens
Authors: Woods G L , Young A , Scott J C , Blair T M , Johnson A M
Published: January 2020

A nonisotopic probe (Gen-Probe PACE; Gen-Probe, Inc., San Diego, Calif.) for detection of Chlamydia trachomatis in endocervical specimens was evaluated in 344 women attending a dysplasia clinic or an obstetrics clinic and 158 women who visited an emergency room. For each patient, the probe, a tissue cell culture, and a direct immunofluorescent-antibody test (DFA; MicroTrak; Syva Co., Palo Alto, Calif.) were used. C. trachomatis was detected in 54 specimens by at least one method. Forty-four, 44, and 37 specimens were positive by culture, probe, and DFA, respectively, and 31 were positive by all three methods. Considering culture-positive plus both probe- and DFA-positive results as the "gold standard," we determined the overall sensitivity, specificity, and positive and negative predictive values of the probe to be 80, 98, 82, and 98%, respectively. These values were 94, 98, 84, and 99%, respectively, in emergency room patients and 71, 98, 80, and 97%, respectively, in clinic patients. The sensitivities, specificities, and negative predictive values of the DFA and probe were comparable. The positive predictive values of the DFA in all patients and in emergency room and clinic patients were 97, 100, and 95%, respectively. Given the number of probe-positive results that were not confirmed by culture, we do not recommend using the Gen-Probe PACE to screen for C. trachomatis in women with a low to moderate risk for infection.
Purification and amino-terminal sequence analysis of the complement-fixing and precipitin antigens from Coccidioides immitis
Authors: Resnick S , Zimmer B , Pappagianis D , Eakin A , McKerrow J
Published: January 2020

Two proteins (21 and 48 kilodaltons) purified from endospore-spherule culture filtrates of Coccidioides immitis are identified as precipitin and complement-fixing antigens, respectively. To allow specific structural comparison to antigens identified by other investigators and as a first step to eventual serodiagnostic antigen production by recombinant DNA technology, amino-terminal amino acid sequences were determined for these antigens.
Pyrazinamidase activity as a phenotypic marker for several Aeromonas spp. isolated from clinical specimens
Authors: Carnahan A , Hammontree L , Bourgeois L , Joseph S W
Published: January 2020

Negative pyrazinamidase activity was significantly associated with Aeromonas sobria, and positive pyrazinamidase activity was associated with A. hydrophila and A. caviae (chi 2, P less than 0.0001). The absence of pyrazinamidase activity may be a potentially significant phenotypic marker for A. sobria.
Rapid detection of Pneumocystis carinii in bronchoalveolar lavage samples by using Cellufluor staining
Authors: Baselski V S , Robison M K , Pifer L W , Woods D R
Published: January 2020

Cellufluor (Calcofluor white) has been found to be a useful, rapid chemofluorescent stain for detection of Pneumocystis carinii cysts in bronchoalveolar lavage samples. When compared with toluidine blue O and Giemsa stains on 45 specimens (22 positive and 23 negative), the sensitivity and specificity of the Cellufluor stain were 95 and 100%, respectively.
Detection of human papillomavirus DNA in prostate gland tissue by using the polymerase chain reaction amplification assay
Authors: McNicol P J , Dodd J G
Published: January 2020

Human papillomavirus (HPV) is associated with specific benign and malignant lesions of the epithelial and mucosal surfaces. Of the sexually transmitted types, HPV type 16 (HPV 16) and HPV 18 are commonly associated with severe dysplasia and carcinoma of the uterine cervix. In men, genital HPV infections which have been studied are manifest as external lesions usually involving types other than 16 and 18. The nature of HPV 16 and 18 infections in men has not been explored. Since the most common neoplasias of the male genital tract involve the prostate gland, we assayed benign hyperplastic and cancerous prostate tumors for the presence of HPV DNA, using type-specific primers in polymerase chain reaction amplifications. Normal prostatic tissue obtained at autopsy was included in the survey. Amplified sequences specific for HPV 16 were found in 14 of 15 benign prostatic hyperplasias and in all of four carcinomas tested. In contrast, HPV 18 was identified in only three benign hyperplasias, which also contained HPV 16 DNA. Four of five normal prostates demonstrated no HPV infection; one contained HPV 16 sequences. The presence of these oncogenic HPV types in prostate tissues suggests a reservoir for sexual transmission; a potential role for the virus in the etiology of prostatic neoplasia remains to be demonstrated.
Sialidase (neuraminidase) activity among gram-negative anaerobic and capnophilic bacteria
Authors: Moncla B J , Braham P , Hillier S L
Published: January 2020

A filter paper spot test with 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid as a substrate was used to study the prevalence of sialidase activity among gram-negative anaerobic and capnophilic bacteria. A total of 567 isolates representing four genera of obligate anaerobes and four genera of capnophilic organisms was tested. Sialidase activity was detected in 94% of 66 isolates from the Bacteroides fragilis group, 98% of 66 B. bivius isolates, and all isolates of the following species (number of isolates follows species name): B. capillosus, 4; B. levii, 2; B. denticola, 22; B. loescheii, 23; B. melaninogenicus, 32; B. forsythus, 44; and B. buccalis, 2. However, sialidase activity was detected in only 29% of 7 B. buccae isolates, 79% of 14 B. disiens isolates, and 55% of 11 B. oralis isolates. Sialidase activity was not detected among any of 13 isolates of B. gracilis, 12 isolates of B. ureolyticus, 61 isolates of B. intermedius, or 26 isolates of B. corporis. Porphyromonas (Bacteroides) asaccharolytica (20 isolates) and P. endodontalis (8 isolates) did not demonstrate sialidase activity, while 25 isolates of P. gingivalis were sialidase positive. Sialidase activity was found in 10 (100%) of 10 isolates of Capnocytophaga ochracea of C. sputigena but not in any of 4 C. gingivalis isolates. Other gram-negative anaerobic or capnophilic bacteria, including the following, were negative for sialidase activity: Fusobacterium nucleatum, 39 isolates; Wolinella recta, 19 isolates; Eikenella corrodens, 17 isolates; Haemophilus aphrophilus, 10 isolates; and Actinobacillus actinomycetemcomitans, 10 isolates. These data demonstrate sialidase activity in several species of the genera Bacteroides and Porphyromonas and suggest that this characteristic may be useful for identification.
Rapid field inversion gel electrophoresis in combination with an rRNA gene probe in the epidemiological evaluation of staphylococci
Authors: Goering R V , Duensing T D
Published: January 2020

A rapid field inversion gel electrophoresis (FIGE) protocol was combined with an rRNA gene probe in the analysis of staphylococci that were difficult to study epidemiologically by conventional means. The following groups of clinical isolates were examined: (i) predominantly antibiotic-susceptible Staphylococcus aureus strains containing no detectable plasmids and unresponsive to bacteriophage typing and (ii) methicillin-resistant Staphylococcus epidermidis strains carrying a single plasmid ca. 30 kilobases in size. The results indicated that strain interrelationships could be established on the basis of SmaI-generated chromosomal restriction fragment length polymorphisms (RFLPs) analyzed by FIGE. RFLP analysis of strains known to be unrelated established the importance of minor differences in DNA banding patterns as indicators of strain dissimilarities. Hybridization studies with an rRNA gene probe confirmed this conclusion. These results suggest that FIGE analysis of chromosomal RFLPs (especially in combination with molecular probes) is an important addition to the armamentarium of molecular epidemiology.
Species identities of enterococci isolated from clinical specimens
Authors: Ruoff K L , de la Maza L , Murtagh M J , Spargo J D , Ferraro M J
Published: January 2020

Conventional tests and commercially available systems were used to determine the species identities of clinical isolates of enterococci. Strict adherence to the conventional test scheme of Facklam and Collins (R. R. Facklam and M. D. Collins, J. Clin. Microbiol. 27:731-734, 1989) resulted in the misidentification of lactose-negative Enterococcus faecalis isolates as Enterococcus solitarius, but this problem was overcome by the application of additional tests. The commercially available systems tested were unable to recognize some of the more recently described enterococcal species. E. faecalis accounted for 87.1% of 302 consecutive isolates. Enterococcus faecium (8.6%), Enterococcus avium (0.7%), Enterococcus durans (0.3%), Enterococcus gallinarum (1.0%), Enterococcus casseliflavus (1.0%), Enterococcus hirae (0.3%), and Enterococcus raffinosus (0.3%) isolates were also identified. None of the isolates produced beta-lactamase, but 15.4% of 235 isolates tested, including 1 strain of E. gallinarum, displayed high-level resistance to gentamicin.
Antibody responses to protein A in patients with Staphylococcus aureus bacteremia and endocarditis
Authors: Greenberg D P , Bayer A S , Turner D , Ward J I
Published: January 2020

To assess the significance of antibody to Staphylococcus aureus protein A (SpA) in human sera, we developed a modified enzyme-linked immunosorbent assay (ELISA). SpA antibody levels in 23 patients with S. aureus endocarditis (IE), 21 patients with non-IE S. aureus bacteremia, and 33 controls were measured. Geometric mean levels of antibody to SpA were significantly higher in S. aureus IE patients (134 ELISA units [EU]) than in uninfected controls (52 EU; P less than 0.01). Also, a significantly greater proportion of S. aureus IE patients (12 of 23) and S. aureus non-IE bacteremia patients (11 of 21) had antibody levels greater than an arbitrary threshold of 100 EU compared with uninfected controls (0 of 23; P less than or equal to 0.001). However, no significant differences in geometric mean SpA antibody levels between the bacteremic patients with and without IE were noted. The sensitivity and specificity of this ELISA to distinguish patients with S. aureus IE from those with non-IE bacteremia were low (52 and 48%, respectively). There was a significant association between SpA antibody levels and either immunoglobulin G or immunoglobulin M teichoic acid antibody levels (r = 0.406, P less than 0.05; r = 0.571, P = 0.002, respectively). For patients from whom multiple sera were available (13 IE and 5 non-IE patients), SpA antibody levels were measured over time and showed a wide temporal variation of immune responses. We conclude that antibody responses to SpA can be measured in many patients with invasive S. aureus disease but that the levels are of insufficient sensitivity or specificity to be of clinical use as a diagnostic or prognostic test.
Rotavirus serotypes 6 and 10 predominate in cattle
Authors: Snodgrass D R , Fitzgerald T , Campbell I , Scott F M , Browning G F , Miller D L , Herring A J , Greenberg H B
Published: January 2020

Calf fecal rotavirus strains were serotyped in enzyme-linked immunosorbent assays, using monoclonal antibodies to the VP7s of serotypes 1, 2, 3, 5, and 6 and to the VP4 of B223 (designated serotype 10). Sixty-six percent of 162 samples were typed as serotype 6, and 7% were serotyped as serotype 10. Most of the untyped strains did not react with a monoclonal antibody directed to a common VP7 epitope, indicating insufficient virus present in the samples. However, seven untyped samples that did react with this antibody were adapted to culture and typed, and six of these also proved to belong to serotype 6 or 10. Two of these viruses belonged to a monotype within serotype 6 that did not react with the serotype 6 monoclonal antibody. The seventh isolate reacted in cross-neutralization tests with serotype 8 viruses. Bovine rotaviruses from the United Kingdom, Federal Republic of Germany, and Japan that had been shown previously to be distinct from serotype 6 were compared in neutralization tests with B223 from the United States. These viruses proved to be a closely reacting group distinct from all other rotavirus serotypes, justifying the establishment of serotype 10 as the second major type of bovine rotavirus.
Novel gelatin particle agglutination test for serodiagnosis of leprosy in the field
Authors: Izumi S , Fujiwara T , Ikeda M , Nishimura Y , Sugiyama K , Kawatsu K
Published: January 2020

We developed a novel gelatin particle agglutination test (MLPA) for the serodiagnosis of leprosy; this test is especially useful for clinical practice and epidemiological surveys of leprosy in countries in which the disease is endemic. The antigen used in the test is the chemically synthesized trisaccharide moiety of Mycobacterium leprae-specific phenolic glycolipid I. MLPA is a simple and easy technique having sensitivity and specificity comparable to those of the conventional indirect enzyme-linked immunosorbent assay. The new technique was found to be useful for monitoring of chemotherapy and predictive diagnosis of high-risk individuals in contact with persons with leprosy and may be useful for the prediction of relapse. We are now preparing to supply a quality-controlled ready-to-use MLPA kit for leprosy control in countries in which leprosy is endemic.
Collagen binding, elastase production, and slime production associated with coagulase-negative staphylococci isolated from bovine intramammary infections
Authors: Watts J L , Naidu A S , Wadström T
Published: January 2020

Collagen binding, elastase production, slime production, and associated somatic cell counts were determined with 160 strains of coagulase-negative staphylococci isolated from bovine intramammary infections. Mean binding values for type I and II collagen with Staphylococcus epidermidis, S. chromogenes, and S. hyicus strains were 5.8, 6.6, and 7.4 and 4.3, 4.2, and 4.9%, respectively. Eleven of 28 (39.3%) S. epidermidis, 1 of 38 (2.6%) S. chromogenes, and 1 of 94 (1.1%) S. hyicus strains were elastase positive. Slime production was noted with 12 (42.9%) S. epidermidis, 1 (2.6%) S. chromogenes, and 11 (11.7%) S. hyicus strains. No differences in somatic cell counts were observed with type I or type II collagen binding, elastase production, or slime production with S. epidermidis or S. chromogenes. However, somatic cell counts associated with S. hyicus strains with collagen type I binding affinities of greater than 5 and type II binding affinities of greater than 3 were 320.7 x 10(3) compared with 163.9 x 10(3) for strains with lower binding affinities.
Is Candida stellatoidea disappearing from the vaginal mucosa?
Authors: Kwon-Chung K J , Wickes B L , Salkin I F , Kotz H L , Sobel J D
Published: January 2020

A total of 681 vaginal isolates of germ tube-positive or germ tube-untested white, yeastlike fungi obtained from patients in various cities of the United States were tested for the presence of Candida stellatoidea (type I). Only 1 of the 681 isolates was identified as C. stellatoidea.
Vibrio cholerae bacteremia associated with gastrectomy
Authors: Toeg A , Berger S A , Battat A , Hoffman M , Yust I
Published: January 2020

Bacteremia due to Vibrio cholerae is rare. Each of 15 cases previously reported in the English language literature occurred in the setting of immune deficiency. We describe an instance of non-serogroup O1 V. cholerae septicemia in an otherwise healthy patient. Susceptibility to such infection may have been enhanced by a prior gastrectomy for duodenal ulcer.
Complement fixation test for the diagnosis of Lyme disease
Authors: Artsob H , Huibner S
Published: January 2020

Sera from 43 patients were tested for complement-fixing antibodies to Borrelia burgdorferi; these patients included 8 with confirmed Lyme disease, 21 who were serologically positive but not likely to have Lyme disease, and 14 who were serologically negative. Seven individuals, all confirmed Lyme disease patients, had complement-fixing antibodies. Complement fixation may be a useful confirmatory test for Lyme disease.
Impact of the BACTEC NR system in detecting Candida fungemia
Authors: Muñoz P , Bernaldo de Quirós J C , Berenguer J , Rodríguez Créixems M , Picazo J J , Bouza E
Published: January 2020

As a result of the growing importance of candidemia, various techniques for the detection of Candida spp. in blood have been designed with a view to speeding up the laboratory procedure. We have performed a retrospective comparison of the efficiencies of the conventional VACUTAINER system (Becton Dickinson Vacutainer Systems) and the BACTEC NR system (Johnston Laboratories, Inc.). During a 4-year period, 88,300 blood cultures were processed, with growth of Candida species in 552. The mean times required for the detection of growth were 7.55 days with the VACUTAINER system and 4.1 days with the BACTEC NR system (P less than 0.05). During the first week of incubation, the VACUTAINER system detected 56.1% of the candidemias and the BACTEC NR detected 93.6% (P less than 0.05). Use of the BACTEC NR therefore permits reduction of the incubation period from the previously established 4 weeks to a more convenient 7 to 14 days.
Comparison of a serum replacement (Omni Serum) and fetal bovine serum in cell cultures used to isolate herpes simplex virus from clinical specimens
Authors: Johnston S , Siegel C
Published: January 2020

Traditionally, fetal bovine serum (FBS) has been the principal component in media used in the growth and maintenance of cell cultures. Recent shortages have affected the cost and availability of FBS to clinical laboratories. Furthermore, lot-to-lot variability can affect cell culture performance and growth. We evaluated a commercially available serum replacement (Omni Serum; Advanced Biotechnologies Inc., Columbia, Md.) for use in the growth of cell cultures and for use in maintenance media used for the isolation of herpes simplex virus from clinical specimens. Cells (rhabdomyosarcoma and mink lung) raised on 5% Omni Serum grew as well as those grown on 10% FBS. The sensitivity of the Omni-raised cells to herpes simplex virus that had been isolated from 111 clinical specimens was equal to that of the cells raised and maintained with FBS. Cells grown with 10% FBS and maintained with 2% Omni Serum displayed the same sensitivity and integrity in tubes (rhabdomyosarcoma and mink lung) and vials (MRC-5 cells) as cells grown with 10% FBS and maintained with 5% FBS. This study indicates that Omni Serum is an acceptable substitute for FBS in maintenance media for cell culture tubes and vials used for viral isolation from clinical specimens.