Journal of Biological Research, Vol. 3 (2002)
The Induction of Hypothyroidism and Interstitial Cystitis in Mice Fed a Soy-based Diet.
Rebecca L. Ambrose
Faculty Advisor: Dr. Allen Crooker
It has been hypothesized that a soy-based diet can induce hypothyroidism, which may be linked to a chronic inflammation of the bladder called interstitial cystitis. Evidence that an increased soy intake can cause hypothyroidism was demonstrated in mice fed a 19% soy diet. All mice fed the soy diet showed an increase in weight gain and body mass index (BMI) within the first two weeks of starting the diet. After eight weeks all six treated mice had gained an average of ten grams more weight and had a higher BMI than the six untreated controls, indicating that hypothyroidism was induced. Bladder tissues from the mice fed the soy diet were examined by both light microscopy and scanning electron microscopy. The tissue samples did not reveal the presence of interstitial cystitis. Based on these results, it can be presumed that soy-induced hypothyroidism does not cause interstitial cystitis in mice.
Effects of Fish Predation on Species Replacement of a Native Crayfish: Orconectes propinquus versus an Invader, Orconectes rusticus.
Faculty Advisor: Dr. Mark Kuhlmann
The rusty crayfish, Orconectes rusticus, was introduced into upstate New York by fishermen as bait. Since its arrival, the rusty crayfish has been displacing other native species of crayfish, such as Orconectes propinquus. Differential fish predation is thought to be a key cause of the displacement. To test this hypothesis, I placed equal numbers of size matched O. rusticus and O. propinquus into small pools with a single small-mouthed bass, a natural predator, or without a predator as a control. After 3-4 days, I removed the fish and counted the surviving number of crayfish. Survival with the predator did not differ significantly between the two species of crayfish, indicating that predation is not differential for equal-sized individuals. However, since O. rusticus grows larger than O. propinquus, and body size affects predation risk, O. rusticus adults may be less vulnerable to predation.
The Utility of Microsatellite DNA Loci from the Blue Tit (Parus caeruleus) for Studies of North American Resident Passerines.
Craig F. Barrett
Faculty Advisors: Dr. Peter Fauth and Dr. Andrew Ray
DNA microsatellite repeats are being used widely in studies on ecology, evolution, and behavior in animals. A series of PCR primers have been designed for a European hole-nesting bird, the Blue Tit (Parus caeruleus). These primers amplified microsatellite loci in 20 bird families globally, suggesting that they would do so in North American members of the same genus (Parus)—namely the Black Capped Chickadee (Parus atricapillus) and the Eastern Tufted Titmouse (Parus bicolor). Three different Blue Tit primers were tested on the two aforementioned North American species, plus on the closely related White-Breasted nuthatch (Sitta carolinensis)—to investigate the utility of these primers for future studies on resident birds. Genomic DNA was collected from the birds’ red blood cells and incorporated into a Polymerase Chain Reaction using each of three selected primers. The products were then electrophoresed to visualize evidence of polymorphism among each microsatellite locus. The number of primers yielding PCR product and an estimate of the number of alleles per locus will be presented for each species.
Mutagenicity of Pure and Altered Tobacco in Salmonella typhimurium Strain TA100.
Faculty Advisor: Dr. F. A. Ray.
The mutagenicity of homegrown tobacco and consumer brand cigarette tobacco extracts were tested and compared using the Ames Test. Both of the tobacco extracts were tested in Salmonella typhimurium TA100, and the mutagenicity was determined by the number of revertant colonies that were able to grow in the presence of the potential mutagen. Because the store brand tobacco contains added chemicals, it was predicted that this tobacco would cause more revertants than the homegrown tobacco. Because the homegrown tobacco does not have added chemicals, the mutagenicity of the store brand tobacco should also be higher. After a 48-hour incubation period, it was found that the Winston tobacco caused significantly more revertants than the homegrown tobacco when both were tested in the absence of the S9 rat liver enzymes. However, in the presence of S9 liver enzymes, there was not a significant difference in the mutagenicity of either type of tobacco. Also, within each tobacco type, the S9 did not appear to effect the number of revertant colonies found.
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The Prevalence of Borrelia burgdorferi in White-Footed Mice, Peromyscus leucopus, and Deer Ticks, Ixodes scapularis, at Pine Lake in West Davenport, New York.
Faculty Advisor: Dr. Mary Allen
Lyme disease is a zoonosis, an inadvertent infection of humans with an animal pathogen, and is the most common arthropod vector-borne disease in the United States. The pathogen, Borrelia burgdorferi, is transmitted from ticks to humans during blood meals. Little is known about the presence or existence of the Lyme disease spirochete and its arthropod vector, Ixodes scapularis, at Pine Lake in West Davenport, New York. The objective of this research was to determine the infection rate of deer ticks and their most competent host, white-footed mice, with the Lyme disease causative bacteria. Ticks and blood samples were collected from white-footed mice, Peromyscus leucopus trapped at Pine Lake. Host seeking ticks were quested by dragging vegetation. To analyze the presence of the B. burgdorferi spirochete in ticks and animal sera, direct immunofluorescence microscopy and ELISA were used respectively. Lyme disease is a growing concern to the general public, especially to those people who are active outside during the summer and fall months. Many students, faculty, and staff use Pine Lake, and should be informed of possible health risks, such as Lyme disease.
In Vivo and In Vitro Heterochronic Transplantations and Cellular Growth in Mouse CNS.
Faculty Advisor: Stanley K. Sessions
The use of neurotransplants in human subjects as a treatment of neurodegenerative disorders has increased greatly as techniques have advanced. Although the transplantation of fetal neurons into an adult can alleviate symptoms of such diseases as Parkinsons disease, little is known about the specifics of its success. As an attempt to uncover the cause of this new growth in the affected individuals brain, in vitro and in vivo studies were conducted. Fetal cells were transplanted into the brain of a sub-adult mouse and, in tissue culture, cephalic cells of a mouse embryo in very early development were transplanted into established sub-adult age mouse cells. It was expected that the fetal donor cells would simply grow into the adult brain, which would serve as the etiology for such new growth. Immunocytochemistry was employed to detect grafted cells (in vivo) and to compare cellular growth (in vitro). In vivo, the donor cells survived and were detectable, while the in vitro experiment yielded amazing cellular growth that has yet to be conclusively interpreted. The results are interpreted in terms of the known effects of fetal brain transplants.
Transfection of Plasmid DNA into Human Cells Using Electroporation.
Faculty Advisor: Dr. F. A. Ray
The inducible promoter is a recent discovery that offers molecular biologists and geneticists a powerful tool in determining the impact of specific genes on a mammalian host cell. This tool allows us to see what results when a single gene's expression is turned on in the presence of a chemical trigger. Unfortunately, this highly effective tool is utterly useless if researchers cannot transfect it into mammalian cells. In this experiment I attempted to transfect the viral SV40 T antigen gene into human fibroblasts to perpetuate a stable cell line of SV40 T antigen positive cells that can be induced into expression. When the initial attempt failed it was determined that the electroporation was unsuccessful because the voltage and microfarad levels were not adjusted for optimum transfection efficiency. The pSV3neo plasmid, a general plasmid that contains the T antigen gene, was used in two courses of transfections to determine the optimal transfection voltage and microfarad levels at which to perform the transfection. Once the data were collected a final attempt was made to complete the transfection of pINDT, the plasmid containing the T antigen gene, and pVgRXR, containing genes required for the expression of the T antigen gene. Valuable information was ascertained regarding both the voltage and microfarad levels for a successful transfection. Also, the results demonstrated which conditions of the cells as well as proper amounts and cleansing techniques of the plasmids were required to achieve optimal transfection efficiency. It was found that the electroporator at Hartwick College works optimally at 240 V and 800mF for a single selected plasmid transfection. The results of this experiment may benefit future research being conducted in this area, utilizing T antigen harboring plasmids.
Half The Frog I Should Have Been:Making Haploid Frogs For Teaching High School Biology.
Faculty Advisor: Dr. Stan Sessions
I am interested in creating unusual labs to stimulate scientific inquiry in high school biology students. The phenotypic consequences of haploidy (only one set of chromosomes instead of two) were studied in frogs (Rana pipiens) by irradiating sperm with ultraviolet irradiation. Ploidy was initially examined by comparing nuclear sizes in diploid and haploid tadpoles. Chromosome squashing technique using silver-stained Nucleolar Organizer Regions (NOR) was used to confirm ploidy in these animals. I found that UV light has extreme effects on the viability and mobility of sperm cells. Nuclear size of the haploids was significantly smaller than that of the diploids, reflecting the differences in chromosome number and DNA amount. This difference could be used in the classroom as a convenient way to distinguish haploids from diploids for studies of things like behavior, development rate, viability, and the effects of ploidy on phenotype. These results indicate the potential for induced haploidy in frogs as a useful hands-on lab project for 7-12 biology classes.
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Educating the Visually Impaired in Biology.
Kristy C. Kulas
Faculty Advisor: Dr. Stanley K. Sessions
This study examines the effects of teaching style and class composition on the education of visually impaired students in Biology. Two methods of instruction, traditional and constructivist, were compared, as well as two class compositions, segregated and inclusive, or mainstreamed. The study involved students in four sixth grade science classes at Sidney Middle School. The effectiveness on learning of four different combinations of variables was investigated, (1) traditional segregated, (2) traditional mainstreamed, (3) constructivist segregated, and (4) constructivist mainstreamed. In each of the four treatment groups, students were given a pre-test, were artificially blinded using blindfolds during a lesson on a topic in Biology, and were then given a post-test. The Analysis of Variance (ANOVA) and the Least Significant Difference (LSD) statistical tests were used to compare the mean gain scores of the four treatment groups. The hypothesis tested was that constructivist teaching in a mainstreamed setting would work best for visually impaired students. If this is true, then students in the constructivist mainstreamed group would show the most improvement. The results of this study show that constructivist teaching is more effective than traditional teaching, and that student performance in mainstreamed and segregated classrooms is indistinguishable.
The Effects of Caffeine and Ephedrine on Laboratory Rats.
NicolIe M. Madigan
Faculty Advisor: Dr. Allen Crooker
The effects of herbal ecstasy on rats were evaluated by feeding them standard rat chow containing .003125% ephedrine and .001% caffeine. There were two treatment groups with 4 rats each, a low and a high, a well as a control group of 3 rats. The high dosage of ephedrine was 0.75ml per day (equivalent to 400mg in a human), the low dosage was 0.2ml per day (equivalent to 100mg in a human); the caffeine dosage was 0.6ml per day for both the high and low dosage groups. Weight, water intake, urine, and pulse were measured. The control group gained 13.3g (per rat) during week 1, 1.6g during week 2, and 4.3g during the recovery period. The low dosage group gained 3.6g (per rat), then lost 3.6g, and during recovery gained 7.3g. The high dosage group originally lost 2.3g (per rat), then an additional 6g during week two, and then gained 6.3 during the recovery period. Appetite was lowered for both the high and low dosage groups, but returned to normal after the two-week administration period. There was little difference in the water intake and urine excreted between the three groups. Pulse rate increased immediately after consumption of the treated food by 30 beats per minute in the high dosage group and 15 beats per minute for the low dosage group. Pulse rate then dropped 15 beats per minute for the high dosage group after consumption and returned to normal for the low dosage group. The control pulse rate remained the same regardless of feeding time. Overall, there was a decrease in appetite and weight gain due to the consumption of caffeine and ephedrine; pulse rate was increased soon after administration, and lowered after 12 hours.
Effects of Andrographis paniculata and Monopril on Blood Pressure in Rats.
Faculty Advisor: Dr. Linda Swift
Andrographis paniculata and Monopril were both tested on rat aortas to see if they lower blood pressure. Andrographis paniculata belongs to the family Acanthaceae, is a medicinal plant grown in Thailand, and they are unsure of how it works. Monopril is a proven vasodilator and works by preventing the conversion of a hormone called angiotensin I to angiotensin II which is a potent blood vessel constrictor. The constriction and relaxation of the rat aortic rings was measured with a force transducer connected to a power lab in a 10 mL organ bath. 10-6 M phenylephrine was used to make the aortic rings contract. Andrographis paniculata relaxed the aorta 8% +/- 1.006 while Monopril relaxed the aorta 1.9% +/- .3834. This was a significant change with the mean significant difference of p<0.05. At the same concentrations, Andrographis paniculata had more of a vasodilator effect than Monopril.
Effect of Coccinia indica on Blood Glucose Levels in Alloxan-induced Diabetic Mice.
Faculty Advisor: Dr. Linda Swift.
Diabetes is a condition in the body where the pancreas does not produce enough insulin to process glucose or the body does not use the glucose properly. In this study, the anti-diabetic effects of the medicinal plant Coccinia indica were tested on alloxan-induced diabetic mice and were compared to the reference drug, Tolbutamide. The mechanism and the ability to lower blood glucose levels are known for Tolbutamide. This study will try to determine how effective Coccinia indica is in lowering blood glucose levels over a short amount of time in moderately and severely diabetic mice. The mice were categorized as either moderately (100-300 mg/dl) or severely diabetic (over 300 mg/dl) according to their blood glucose levels. The dosage for the moderately diabetic mice was 200 mg/kg per body weight and 400 mg/kg for the severely diabetic group. Blood glucose levels were tested before the treatment and two and four hours after it. Coccinia indica and Tolbutamide decreased the blood glucose levels for the moderately diabetic mice by 77.17 and 61.05 percent and for the severely diabetic mice, 21.7 and 47.97%.
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Relationships between Pitcher Morphology and Inquiline Community in the Pitcher-Plant Sarracenia purpurea.
Faculty Advisor: Dr. Mary Allen
I studied the relationships between pitcher morphology and inquiline community in the purple pitcher plant Sarracenia purpurea, in Upstate New York. The cup-shaped leaves called pitchers, collect rainwater and trap insect prey. The inquiline community resides in the fluid of the pitcher plant leaves. It includes larvae of the pitcher plant mosquito Wyeomyia smithii and the midge larvae Metriocenemus knabi, and a diverse bacterial community. In 64 pitchers, each from one of two populations located in different geographical locations, I measured pH, temperature, and volume of pitcher fluid. I also measured three morphological characteristics of pitcher leaves: height, width, and opening area. The following relationships were observed in both populations used for this study. I found a positive correlation between the abundance of midge larvae and pitcher height, and a positive correlation between the abundance of mosquito larvae and pitcher width. Pitcher width was also positively correlated with pitcher fluid volume. It appears as though female mosquitoes may have selected pitchers that were wider and consequently contained more fluid to deposit eggs. The relationship between midge larvae and height may be a result of female midges depositing eggs in pitchers that were taller; taller pitchers may be more accessible than shorter ones. I also found a positive correlation between midge and mosquito larval abundances. This is evidence of a commensal relationship and may be a result of midge larvae breaking down decaying matter into small particles, providing greater substrate availability for bacteria, which are consumed by mosquito larvae.
Comparative Cytogenetic Analysis of the Phylogenetic Relationships of Desmognathine Salamanders, Family Plethodontidae.
Amy K. Samuels
Faculty Advisor: Dr. Stanley K. Sessions
Phylogenetic relationships among the species of the Plethodontid salamander subfamily Desmognathinae were examined using cytogenetics. The analysis indicates that the monotypic species Leurognathus marmoratus should be placed in the genus Desmognathus. Previous cytogenetic and molecular data identify a "core" group consisting of D. ochrophaeus, D. monticola, D. fuscus, D. aeneus, D. auriculatus, including D. marmoratus, while D. quadramaculatus and D. wrighti each have unique cytogenetic features. This study confirmed these results using D. quadramaculatus, D. marmoratus, D. ochrophaeus, and D. wrighti. The cytogenetic characters were used to construct a hypothesis of phylogenetic analyses within the subfamily.
Morphological Rules and Patterns of Tetrapod Limb Development Determined by Examination of the Salamander Hynobius.
Faculty Advisor: Dr. Stanley Sessions
There is much controversy surrounding the subject of evolution of from fins to limbs. In this study, regeneration in primitive Hynobius salamanders was examined. Hynobius limb development is unique because of the formation of a long fin-like projection between the first and second digits. Some experts believe that this is a larval adaptation for swimming while others argue that this is leftover "baggage" from a finned ancestor. Adult Hynobius limbs were severed at the wrist and allowed to regenerate. Since adults would not need a fin to help them swim like larva would, it would be unnecessary for its formation during regeneration. However, if it is indeed evolutionary baggage, the prolonged tip should form regardless. Of the six regenerated limbs, not one showed any sign of a fin-like projection. Histological examination revealed dead and dying cells present between the first and second digit which is of interest since, while cell death is a factor in amniotes, it has not been thought to be a player in amphibian limb development. These findings suggest that the interdigital membrane in Hynobius is a larval adaptation only.
Alternative reproductive strategies of the male bluegill sunfish, Lepomis macrochirus, at Pine Lake.
Jill B. Sullivan
Faculty Advisor: Dr. Mark Kuhlmann
The bluegill sunfish, Lepomis macrochirus, has been observed to exhibit two male reproductive strategies, parental and cuckolder. Parental males construct nests within colonies in lakes in order to attract females during the breeding season from late May to the end of June. Cuckolder males distribute themselves around the colony and attempt to fertilize the eggs of females spawning in a parental males' nest. There are two varieties of cuckolder males: sneaker (smaller males) and satellite (larger, resemble females). I conducted observations of mating behavior and collected males (n=14) at Pine Lake in West Davenport, NY, to determine what male reproductive types are present in that population. Each captured fish was weighed, measured, milked of an ejaculate, then sacrificed and dissected in order to determine gonad weight. A cluster analysis of body weight and gonadosomatic index (GSI = [gonad weight/total body weight]*100) identified two groups of males (cluster coefficient = 6.191), small males with high GSI (probably satellites) and large males with low GSI (probably parentals). GSI, absolute gonad weight, and sperm concentration were significantly negatively correlated to body size. This study suggests that there are both parental and cuckolder males in Pine Lake and cuckolder males exhibit some favorable reproductive characteristics over parental L. macrochirus males.
The Effect of Oil of Oregano on Escherichia coli Numbers in Vitro and in the Laboratory Mouse (Mus musculus).
Faculty Advisor: Dr. A. Crooker
Escherichia coli is one of the main etiologic agents in persistent diarrhea in children of developing countries and in 20-40% of all travelers diarrhea. Due to its prevalence in diarrhea deaths throughout underdeveloped countries it is important to find a safe substance that can inhibit this bacteria. In this study, the antibacterial effects of oil of oregano were evaluated in vivo and in vitro. The in vitro study consisted of using filter paper discs saturated with 9 ul and 4ul of oregano to evaluate the inhibition of Escherichia coli growth on agar plates. The in vivo study consisted of infecting nine laboratory mice with E. coli and administering mice .05 ml or .03 ml of oil of oregano daily for twenty days. The numbers of E. coli were measured both before the treatment and every two days until the end of the experiment. In vitro results indicated that the two concentrations of oil of oregano inhibited E. coli but not in a concentration dependent manner; oil of oregano had a zone of inhibition similar to that of penicillin. In vivo the numbers of E. coli were substantially reduced but not in a concentration dependent manner. In vitro and in vivo use of oil of oregano inhibited E. coli growth and therefore may help control diarrhea. Further study of the effect of oil of oregano on bacteria in vivo is needed to confirm the oral usage of oil of oregano to inhibit bacteria.