The Ohio Journal of - The Ohio Academy of Science

A-34 The Ohio Journal of Science
Vol. 112(1)
salamanders, turtles and fish, with a global distribution
and reported outbreaks in several countries and in at least
30 U.S. states. In frogs and salamanders, outbreaks are
characterized by a rapid die-off, particularly of larva and
recent metamorphs. Diagnosis requires amplification of
the major capsid protein gene (~1500 base pairs) using
polymerase chain reaction followed by sequencing and
comparison to reference sequences in GenBank. Sudden
die-offs of larval and metamorphosed American Bullfrogs
(Lithobates catesbeianus) in a 1-ha man-made wetland in
northwest Ohio, and of Wood Frogs (Lithobates sylvaticus)
in a reserve pond at North Chagrin Nature Center near
Cleveland, Ohio were reported. Samples were taken from
34 frogs and two turtles over the 26 September, the 11 and
12 October and 4, 6, and 9 November 2011. Diagnosis of
the FV3 strain from skin swabs, liver, and kidney biopsies
confirmed 17 Ranavirus cases in American Bullfrogs, and
one occurrence in an Eastern Painted Turtle (Chrysemys
picta picta) at the northwest Ohio site, and 12 cases (n =
17) in both adults and larva at the Cleveland site. This is
the first published account of amphibian die-offs caused by
FV3, and the first reported Ranavirus infection of a turtle,
in Ohio.
Poster Board No. 068 THE EFFECTS OF CHLORPYRIFOS ON
HIPPOCAMPAL GENE EXPRESSION IN MALE LONG-EVANS
RATS AFTER MULTIPLE SUBCUTANEOUS EXPOSURES.
Lynette Vana (lvana@ashland.edu), Mason Posner (mposner@
ashland.edu). Ashland University, Department of Biology/
Toxicology, 401 College Avenue, Ashland, OH 44805.
Chlorpyrifos is an organophosphate insecticide that is used
worldwide for crops and household purposes. It is sold under
the trade names of Dursban and Lorsban. Chlorpyrifos
functions as a cholinesterase inhibitor and is therefore
a neurotoxin proposed to detrimentally affect organisms
exposed to it. Organophosphates like chlorpyrifos have
also been used as weapons in warfare because of their
potent neurotoxicity to people; furthermore, detrimental
developmental effects have been recorded for children
who were exposed to this pesticide. Several studies
have suggested a link between chlorpyrifos exposure
and cognitive deficits, including effects on memory. A
previous study found changes in the expression of over
3,000 genes in the rat forebrain after a single dosing of
chlorpyrifos at 2mg/kg. Only a small number of these
changes in expression were subsequently confirmed by
real-time PCR, the genes of focus were not included in this
confirmation. The purpose of this present study was to
confirm the upregulation of two genes, Rpl19 and Synj1, in
a specific region of the rat brain, the hippocampus, to test
the hypothesis that chlorpyrifos can cause gene regulatory
changes in a region of brain related to memory. After
animal dosing, hippocampal brain tissue samples were
collected from the animals and preserved in RNAlater.
RNA was then extracted from these hippocampus tissues
for both control and dosed male Long Evans rats, and
oligonucleotide primers were developed to amplify each
gene. Reverse-transcriptase-polymerase chain reaction
(RT-PCR) analysis of purified RNA will test the hypothesis
that these two genes are greatly upregulated in the
hippocampus after multiple subcutaneous exposures to
chlorpyrifos.
Poster Board No. 069 THE PROTECTIVE EFFECTS OF THE
VIOLACEIN PIGMENT AGAINST UV-C IRRADIATION IN
CHROMOBACTERIUM VIOLACEUM. Andrew N. Abboud,
andrew.abboud9@gmail.com, 748 Oak Lea Dr., Tipp City,
OH 43571. (Tippecanoe High School and Central State
University).
Chromobacterium violaceum is a Gram-negative bacteria
found in tropical regions. C. violaceum has the distinct
phenotypic characteristic of a deep violet pigment called
violacein. Violacein has a high molar extinction in
methanol, suggesting that it is protective against visible
light. The purpose of this study was to establish the
protective effects of violacein against UV-induced cellular
damage. It was hypothesized that violacein protects DNA
and proteins (e.g. catalase) from UV-C induced damage.
Wild-type (WT) C. violaceum was mutagenized with
N-methyl-N’-nitro-N-nitrosoguanidine to produce mutants
with varying amounts of violacein. Mutants CV9, CV13,
and CV14 (non-pigmented) produced less pigmentation than
WT and retained colony morphology, while mutants H19,
H20, and H21 (hyper-producers) over-expressed violacein
but had an altered petite morphology. UV-induced DNA
damage was assayed through sub-culture post-irradiation
at 6,000μW*s -1 *cm -2 at λ=253.7nm. Sub-cultures of WT and
hyper-producers showed reduced viability after 48 hours;
nonpigmented mutants showed no growth, suggesting
violacein is protective against UV-induced DNA damage.
UV-induced catalase damage was assayed pre and post
irradiation. According to the paired t-test at the 5%
significance level (tvalue ±1.960), catalase activity in WT,
H19, H20 and H21 significantly decreased post-irradiation
and assumed the average negative t-values of 20.4058,
-15.9284, -12.7082 and 11.1229, respectively; catalase
activities of CV9, CV13 and CV14 significantly increased
post-irradiation and assumed the average positive t-values
of 16.2441, 27.0759 and 26.2194, respectively. Increased
catalase activity in non-pigmented mutants can potentially
be explained by the increased induction of catalase genes in
response to elevated reactive oxidative species, presumably
from lack of pigmentation. Taken together, these results
support the hypothesis that violacein is protective against
UV-induced cellular damage.
Poster Board No. 070 COMPARISON OF AN
IMMUNOCHROMATOGRAPHIC RAPID TEST, A MICROPLATE
ENzYME IMMUNOASSAY AND TRADITIONAL CULTURE
METHODS FOR DETECTION OF CAMPYLOBACTER SPP. IN
OUTPATIENT STOOL SAMPLES Karen Kruzer, kak123@case.
edu, Case Western Reserve University, Cleveland, OH.
Campylobacter spp. is a leading cause of bacterial
gastroenteritis, affecting over 2.4 million persons
annually. Campylobacteriosis infection is caused by
consuming unpasteurized milk, contaminated food or
water, or undercooked poultry. Food poisoning caused
by Campylobacter spp. can be debilitating, resulting in
diarrhea with varying severity from loose to bloody stools.
An analytical review of recent publications suggests a
problem with consistent detection of Campylobacter spp.,
therefore a comparison of antigen detection approaches
versus culture methods needs to be conducted. The
objectives were to compare antigen detection methods,
to compare sensitivity for recovery of Campylobacter
spp. using culture versus enzyme immunoassay, and
to tabulate incidence of bacterial, parasitic, and viral
pathogens. Three diagnostic methods were performed on
100 stool samples collected from outpatients. ProSpecT
EIA Test and ImmunoCard STAT! CAMPY® enzyme
immunoassays detected Campylobacter spp. antigens.
Traditional culture on Campylobacter spp. selective
medium and filtration on blood agar was also performed.
Campylobacter spp. is a seagull-shaped Gram negative
bacilli, catalase positive, oxidase positive, hippurate
positive, and motile. Disc diffusion susceptibility to
nalidixic acid, cephalothin, and erythromycin further
identified the species. Antigen detection tests recovered 7
positives, whereas culture methods recovered 3 positives.
The gold standard was two-fold. When culture served as
reference, sensitivity/specificity were high (both >65%);
ImmunoSTAT! positive predictive value was 28%. When
positive EIA or culture served as reference, ImmunoSTAT!
sensitivity decreased, but positive predictive value
increased. The highest incidence of enteric pathogens was
Campylobacter spp. and Clostridium difficile. Consistently
reliable identification of Campylobacter spp. is crucial for
diagnosis of the leading cause of enteritis globally.
Poster Board No. 071 ADHESION TO AND INVASION OF
EUKARYOTIC CELLS BY ISOLATED STAPHYLOCOCCUS AUREUS
ISOLATES. Darlene G. Walro 1 , dwalro@walsh.edu, Chris A.