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At least 487 records · Page 27Linked to original sources
USGS1967

Low-temperature incubation using a water supply

Cell and tissue culture has been concerned primarily with homiothermic vertebrate cells which require incubation at about 37 C, and there is a great variety of incubators designed to maintain temperatures which are usually above ambient. The culture of poikilothermic vertebrate cells--and invertebrate, plant, and some microbial cells--can often be carried out at ambient temperatures, but for some work cooler conditions must be provided. Variety among the so-called low-temperature incubators is somewhat restricted; there are no small units, and all require a power source to maintain temperatures below ambient. We have used a gravity-fed water supply for 5 years to provide trouble-free, constant, low-temperature incubation of stock cultures of fish and amphibian cells. Though it is but a small part of our low-temperature incubator capacity, it has no power requirements and it provides maximal protection against temperature rises which could be lethal to some of the cell lines. Though the system has limitations, there is a considerable likelihood that the domestic water supply in other laboratories can also be used to provide low-temperature incubation.

Applied Microbiology↗
USGS1968

Partial hydrolysis of dieldrin by Aerobacter aerogenes

Although dieldrin (1,2,3,4,10,10-hexachloro- 6,7-epoxy-1 ,4 ,4a ,5 ,6 ,7 ,8, 8a-octahydro-1 ,4-endo, exo-5, 8-dimethanonaphthalene) metabolism by mammals (F. Korte and H. Arent, Life Sci. 4:2017, 1965) and insects (D. F. Heath and M. Vanderkar, Brit. J. Ind. Med. 21:269, 1964) has been reported, little is known about the degradation of this important pesticide by microorganisms. Korte et al. (Ann. Chem. Liebigs 656:135, 1962) and Chacko et al. (Science 154: 893, 1966) reported that a number of ubiquitous microorganisms were incapable of degrading dieldrin; however, more recently Matsumura and Boush (Science 156:959, 1967) isolated several species of Pseudomonas and Bacillus which could degrade dieldrin, from a number of soil samples having similar activity. They did not specifically attempt to identify the dieldrin metabolites formed, but they did suggest, on the basis of an identical RF value with an authentic control that 6,7-trans-dihydroxydihydroaldrin (aldrin diol) might be a major product. Work carried out concurrently in this laboratory has shown that another ubiquitous bacterium, Aerobacter aerogenes , converts dieldrin in vitro to a compound chromatographically similar to 6,7-trans-dihydroxydihydroaldrin.

Applied Microbiology↗
USGS1969

Infectious pancreatic necrosis virus: Lyophilization and subsequent stability in storage at 4 C

Infectious pancreatic necrosis virus (ATCC strain VR # 299) was lyophilized with different additives and stored at 4 C. Ampoules were assayed at 2 days, at 1, 3, and 6 months, and at 1, 3, and 4 years. Processing losses were least and subsequent maintenance of infectivity was best in skim milk, lactalbumin hydrolysate, and lactose. Quantitative data are given in tabular form.

Applied Microbiology↗
USGS1973

Fish viruses: Buffers and methods for plaquing eight agents under normal atmosphere

A universal procedure was sought for plaque assay of eight fish viruses (bluegill myxovirus, channel catfish virus, eel virus, Egtved virus, infectious hematopoietic necrosis virus, infectious pancreatic necrosis virus, lymphocystis virus, and the agent of spring viremia of carp ( Rhabdovirus carpio ), in dish cultures of various fish cells. Eagle minimal essential medium with sodium bicarbonate-CO 2 buffer (Earle’s salt solution) was compared with minimal essential medium buffered principally with tris (hydroxymethyl)aminomethane or N -2-hydroxyethylpiperazine-N′-2′-ethanesulfonic acid at a pH or in the range of 7.6 to 8.0 depending upon temperature. Five fish cell lines collectively capable of replicating all fish viruses thus far isolated were tested and quantitatively found to grow comparably well in the three media. Two-phase (gel-liquid) media incorporating the various buffer systems allowed plaquing at 15 to 33 C either in partial pressures of CO 2 or in normal atmosphere, but greater efficiency and sensitivity were obtained with the organic buffers, and, overall, the best results were obtained with tris(hydroxymethyl)aminomethane. Epizootiological data, specific fish cell line response, and plaque morphology permit presumptive identification of most of the agents. At proper pH, use of organic buffers obviates the need for CO 2 incubators.

Applied Microbiology↗
USGS1974

Corynebacterial kidney disease of salmonids: Growth and serological studies on the causative bacterium

Comparative growth studies of 10 isolates of the Corynebacterium of salmonid kidney disease were quantified on four media. Greatest cell yield was obtained from Mueller-Hinton medium with 0.1% L-cysteine hydrochloride. Serological tests, in which rabbit antisera used against typical bacteria strains showed all to be antigenically homologous, provided a reliable method of identification.

Applied Microbiology↗
USGS1961

The identification and separation of Aeromonas liquefaciens from Pseudomonas fluorescens and related organisms occurring in diseased fish

Taxonomy of the motile species of the genus Aeromonas is briefly discussed. It is suggested that Aeromonas organisms, isolated from outbreaks of red mouth of trout, red sore of pike, infectious abdominal dropsy and hemorrhagic septicemia of warm water fish, and which show acid and gas in glucose broth, production of 2,3-butanediol hydrogen sulfide from motility sulfide medium, presence of cytochrome oxidase, and hydrolysis of starch, be designated as Aeromonas liquefaciens . These tests also serve to differentiate A. liquefaciens from fish-pathogenic Pseudomonas fluorescens and from enteric bacteria.

Applied Microbiology↗
USGS2003

Pathogenic human viruses in coastal waters

This review addresses both historical and recent investigations into viral contamination of marine waters. With the relatively recent emergence of molecular biology-based assays, a number of investigations have shown that pathogenic viruses are prevalent in marine waters being impacted by sewage. Research has shown that this group of fecal-oral viral pathogens (enteroviruses, hepatitis A viruses, Norwalk viruses, reoviruses, adenoviruses, rotaviruses, etc.) can cause a broad range of asymptomatic to severe gastrointestinal, respiratory, and eye, nose, ear, and skin infections in people exposed through recreational use of the water. The viruses and the nucleic acid signature survive for an extended period in the marine environment. One of the primary concerns of public health officials is the relationship between the presence of pathogens and the recreational risk to human health in polluted marine environments. While a number of studies have attempted to address this issue, the relationship is still poorly understood. A contributing factor to our lack of progress in the field has been the lack of sensitive methods to detect the broad range of both bacterial and viral pathogens. The application of new and advanced molecular methods will continue to contribute to our current state of knowledge in this emerging and

Clinical Microbiology Reviews↗
USGS2015

Temporal variation in viral hemorrhagic septicemia virus antibodies in freshwater drum ( Aplodinotus grunniens ) indicates cyclic transmission in Lake Winnebago, Wisconsin

Viral hemorrhagic septicemia virus (VHSV) is an emerging pathogen that causes mass mortality in multiple fish species. In 2007, the Great Lakes freshwater strain, type IVb, caused a large die-off of freshwater drum ( Aplodinotus grunniens ) in Lake Winnebago, Wisconsin, USA. To evaluate the persistence and transmission of VHSV, freshwater drum from Lake Winnebago were tested for antibodies to the virus using recently developed virus neutralization (VN) and enzyme-linked immunosorbent (ELISA) assays. Samples were also tested by real-time reverse transcription-PCR (rRT-PCR) to detect viral RNA. Of 548 serum samples tested, 44 (8.03%) were positive by VN (titers ranging from 1:16 to 1:1,024) and 45 (8.21%) were positive by ELISA, including 7 fish positive by both assays. Antibody prevalence increased with age and was higher in one northwestern area of Lake Winnebago than in other areas. Of 3,864 tissues sampled from 551 fish, 1 spleen and 1 kidney sample from a single adult female fish collected in the spring of 2012 tested positive for VHSV by rRT-PCR, and serum from the same fish tested positive by VN and ELISA. These results suggest that VHSV persists and viral transmission may be active in Lake Winnebago even in years following outbreaks and that wild fish may survive VHSV infection and maintain detectable antibody titers while harboring viral RNA. Influxes of immunologically naive juvenile fish through recruitment may reduce herd immunity, allow VHSV to persist, and drive superannual cycles of transmission that may sporadically manifest as fish kills.

Wisconsin↗
USGS2012

SYBR green-based real-time reverse transcription-PCR for typing and subtyping of all hemagglutinin and neuraminidase genes of avian influenza viruses and comparison to standard serological subtyping tests

Continuing outbreaks of H5N1 highly pathogenic (HP) avian influenza virus (AIV) infections of wild birds and poultry worldwide emphasize the need for global surveillance of wild birds. To support the future surveillance activities, we developed a SYBR green-based, real-time reverse transcriptase PCR (rRT-PCR) for detecting nucleoprotein (NP) genes and subtyping 16 hemagglutinin (HA) and 9 neuraminidase (NA) genes simultaneously. Primers were improved by focusing on Eurasian or North American lineage genes; the number of mixed-base positions per primer was set to five or fewer, and the concentration of each primer set was optimized empirically. Also, 30 cycles of amplification of 1:10 dilutions of cDNAs from cultured viruses effectively reduced minor cross- or nonspecific reactions. Under these conditions, 346 HA and 345 NA genes of 349 AIVs were detected, with average sensitivities of NP, HA, and NA genes of 10 1.5 , 10 2.3 , and 10 3.1 50% egg infective doses, respectively. Utility of rRT-PCR for subtyping AIVs was compared with that of current standard serological tests by using 104 recent migratory duck virus isolates. As a result, all HA genes and 99% of the NA genes were genetically subtyped, while only 45% of HA genes and 74% of NA genes were serologically subtyped. Additionally, direct subtyping of AIVs in fecal samples was possible by 40 cycles of amplification: approximately 70% of HA and NA genes of NP gene-positive samples were successfully subtyped. This validation study indicates that rRT-PCR with optimized primers and reaction conditions is a powerful tool for subtyping varied AIVs in clinical and cultured samples.

Journal of Clinical Microbiology↗
USGS2025

Fatal interactions: Pneumonia in bighorn lambs following experimental exposure to carriers of Mycoplasma ovipneumoniae

We hypothesized that bighorn sheep ewes with chronic nasal Mycoplasma ovipneumoniae carriage are the source of infection that results in fatal lamb pneumonia. We tested this hypothesis in captive bighorn ewes at two study facilities over a 5-year period, by identifying carrier ewes and then comparing lamb fates in groups that did (exposed pens) or did not (non-exposed pens) include one or more carrier ewes. Most (23 of 30) lambs born in exposed pens, but none of 11 lambs born in non-exposed pens, contracted fatal pneumonia. In addition, surviving lambs in exposed pens showed obvious signs of respiratory disease while lambs in non-exposed pens did not. In crossover experiments, individual non-carrier ewes had lambs that experienced fatal pneumonia in years when housed in exposed pens, but not in years when housed in non-exposed pens. The results of these studies clearly associate lamb pneumonia to exposure to M. ovipneumoniae carrier ewes, consistent with a necessary role for this agent in epizootic pneumonia of bighorn sheep. These data specifically highlight the role of chronic M. ovipneumoniae carriage by some bighorn ewes in the epidemiology of this population-limiting wildlife disease.

Idaho, Montana, Nevada, Oregon, South Dakota, Wash↗
USGS2016

Antemortem detection of chronic wasting disease prions in nasal brush collections and rectal biopsies from white-tailed deer by real time quaking-induced conversion

Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since spread to cervids in 23 states, two Canadian provinces, and the Republic of Korea. The expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of its spread; this is especially true in cases of relocation/reintroduction of farmed or free-ranging deer and elk or surveillance studies of private or protected herds, where depopulation is contraindicated. This study sought to evaluate the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay by using recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brush samples collected antemortem from farmed white-tailed deer ( n = 409). Antemortem findings were then compared to results from ante- and postmortem samples (RAMALT, brainstem, and medial retropharyngeal lymph nodes) evaluated by using the current gold standard in vitro assay, immunohistochemistry (IHC) analysis. We hypothesized that the sensitivity of RT-QuIC would be comparable to IHC analysis in antemortem tissues and would correlate with both the genotype and the stage of clinical disease. Our results showed that RAMALT testing by RT-QuIC assay had the highest sensitivity (69.8%) compared to that of postmortem testing, with a specificity of >93.9%. These data suggest that RT-QuIC, like IHC analysis, is an effective assay for detection of PrP CWD in rectal biopsy specimens and other antemortem samples and, with further research to identify more sensitive tissues, bodily fluids, or experimental conditions, has potential for large-scale and rapid automated testing for CWD diagnosis.

Journal of Clinical Microbiology↗
USGS2016

Seeded amplification of chronic wasting disease prions in nasal brushings and recto-anal mucosal associated lymphoid tissues from elk by real time quaking-induced conversion

Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since been detected across North America and the Republic of Korea. The expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of its spread; this is especially true in cases of relocation/reintroduction or prevalence studies of large or protected herds, where depopulation may be contraindicated. This study evaluated the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay of recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brushings collected antemortem. These findings were compared to results of immunohistochemistry (IHC) analysis of ante- and postmortem samples. RAMALT samples were collected from populations of farmed and free-ranging Rocky Mountain elk ( Cervus elaphus nelsoni ; n = 323), and nasal brush samples were collected from a subpopulation of these animals ( n = 205). We hypothesized that the sensitivity of RT-QuIC would be comparable to that of IHC analysis of RAMALT and would correspond to that of IHC analysis of postmortem tissues. We found RAMALT sensitivity (77.3%) to be highly correlative between RT-QuIC and IHC analysis. Sensitivity was lower when testing nasal brushings (34%), though both RAMALT and nasal brush test sensitivities were dependent on both the PRNP genotype and disease progression determined by the obex score. These data suggest that RT-QuIC, like IHC analysis, is a relatively sensitive assay for detection of CWD prions in RAMALT biopsy specimens and, with further investigation, has potential for large-scale and rapid automated testing of antemortem samples for CWD.

Journal of Clinical Microbiology↗
USGS1995

Detection and identification of aquatic birnaviruses by PCR assay

A reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for the detection and identification of aquatic birnaviruses. The four sets of primers (PrA, PrB, PrC, and PrD) that we used are specific for regions of cDNA coded by genome segment A of aquatic birnaviruses. PrA identifies a large fragment (1,180 bp) within the pVP2-coding region, and PrB identifies a 524-bp fragment within the sequence amplified by PrA. Primer set PrC frames a genome fragment (339 bp) within the NS-VP3-coding region, and PrD identifies a 174-bp sequence within the fragment identified by PrC. PrB and PrD amplified cDNAs from all nine recognized serotypes of aquatic birnavirus serogroup A as well as the N1 isolate that may represent a 10th serotype. These results indicate that these three primer sequences are highly conserved and can be used in PCR assays for group identification of these viruses. PrA routinely produced amplification products from eight serotypes but exhibited variable results with one serotype, and primer PrC identified 6 of the 11 virus isolates tested. The qualitative sensitivity of the RT-PCR assay was evaluated by comparison of the results with those of cell culture isolation assays. With the exception of one sample, the RT-PCR assay with primer PrD was as accurate as cell culture isolation for detecting virus in kidney and spleen tissues from naturally infected, asymptomatic carrier fish. These results indicate that the RT-PCR assay can be a rapid and reliable substitute for cell culture methods for the detection of aquatic birnaviruses.

Journal of Clinical Microbiology↗