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BIOL
413: PARASITOLOGY Dr.
Stan Eisen, |
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Topics for Parasite Rex |
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1a.
Parasites changing host behavior:
Polymorphus paradoxus altering
the behavior of infected gammarids. ·
Matt Jackson ·
Kristi Prevost ·
Rachel Haag
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7.
Living life in copula for 25
years – the schistosome species. ·
Suzi Ponnapula ·
Brooke Allen
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1b.
Parasites changing host behavior:
Toxoplasma gondii altering
the behavior of mice, and possibly changing the behavior of human males and
females. ·
Anmol Khan ·
Raelynn Pirtle ·
Meagan Lamica
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8. Trichinella spiralis as a “virus” of
striated muscle tissue. ·
Blake Jackson ·
Carrie Le
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2.
Why is there sex? (The Red
Queen Hypothesis) ·
Maegan Lytle ·
Caitlyn Ashley ·
Rachel Escue
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9.
Parasites are excellent immunologists:
antigen-shielding and antigen-shifting as immune evasion strategies. ·
Caitlin Clay ·
Melissa
McDonald
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3. Parasitic castration (Sacculina spp.) ·
Mary Austin
Mays ·
Coy Lock ·
Minh-Thao Pham
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10.
Endosymbiosis: Why is it that Plasmodium has chloroplast DNA? ·
Khoa Nguyen ·
Natalie Hurt
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4.
Viruses as “renegade DNA”. ·
Caitlyn Donegan ·
Kathleen Nelson ·
Hanna Orendorf
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11.
OMG! I can’t LIVE WITHOUT YOU!!
– Examples of mutualism: Green hydra,
lichens, termite flagellates, clownfish & sea anemones. ·
Natalie Hart ·
Huong Tran ·
Cameron Kasmai
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5. Brood parasitism (Females of one species
laying her eggs in the nest of another.) ·
Caleb Burrow ·
Ben Chism ·
Ting Wong
Natural World - Cuckoo Nature of the cuckoo duck -
David Attenborough - BBC wildlife |
12.
Is that a basketball in your pocket, or are you just happy to see
me? The pathology of Wuchereria bancrofti. ( http://www.youtube.com/watch?v=Q_BJJQ-nGkE
) ·
Adrienne Nalan ·
Terence Netzel
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6.
Sickle-cell anemia as a source of resistance to Plasmodium falciparum.
(You can tie this in to the pediatric SCA unit @ St. Jude Children’s
Research Hospital.) ·
Victoria Bujalski ·
Xiong Lin
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13.
Creation of an appropriate banner for the entrance to Sabbatini
Lounge, advertising, flyers on campus. ·
Mary Jane
Dickey ·
Kyle Hayes (A composite.) |
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T-shirt proposal:
http://www.cbu.edu/~seisen/TshirtProposalFa2009.htm
View of Veterinarian Rotation Sign-up sheet:
http://www.cbu.edu/~seisen/VeterinarianRotationSignUpWebVersion.htm
To the lecture notes for the course: http://www.cbu.edu/~seisen/ParasitesOnParade.htm
Details about the
lecture
Details about the lab
Supplemental reading list
Lecture schedule
Lab schedule
Dr. Stan Eisen
Office S203B
Tel. Ext.: 3447
e-mail: seisen@cbu.edu
Office Hours: T:
9 a.m. - 11 a.m., 2 p.m. - 5 p.m.
Th: 2 p.m. - 5 p.m.
F: 1 p.m. - 3
p.m.
·
For both lecture and
lab, you will need the following:
Schmidt, G., & Roberts, L.
.Foundations of Parasitology, edition 8.
Zimmer, C. 2003.
Parasite Rex. Arrow Books Ltd; New Ed edition (September 4, 2003), ISBN-10:
0099457997, ISBN-13: 978-0099457992
Eisen, S. (2009).
Parasites On Parade. Available from Print Shop.
·
In this course, we will be concentrating on
the biology of those parasitic organisms which are of medical or veterinary importance.
The emphasis of the course will be on natural history, so you will be
responsible for the following information pertaining to each parasitic organism
covered in class: 1)Binomial nomenclature; 2)Phylogenetic relationship; 3)Life
cycle; 4) Preferred definitive host(s); 5)Intermediate and/or vector host(s);
6)Geographical locations where the parasite is found and is endemic; 7)Organs
affected; 8)Symptoms associated with infection; and 9)Drugs used in treatment.
DETAILS ABOUT THE LECTURE
Your lecture grade will be based on the
following:
1. FOUR MIDSEMESTER EXAMS: Each of these exams will consist of essay and
identification/practical questions. Each of these exams will be comprehensive
and count 1/7 towards your final grade;
2. An essay describing techniques in parasite
surveillance and treatment, following a rotation through a veterinary clinic. You will be
asked to sign up for a 1-afternoon rotation through a veterinary clinic in the
Memphis area to observe techniques used in parasite surveillance and treatment. These rotations will be scheduled during a
3-week period, starting with Fall Break and the following 2 weeks. Upon completion of the rotation you are to
write a short report, not exceeding 3 double-spaced typewritten pages on the
types of animals examined and the parasites which were found. Participation in
the rotation and submission of the report will count 1/7 towards your final grade;
3. An exhibit open to the public, based on the
information contained in Parasite Rex,. This display will
substitute for a final exam during final exam week. The class will be divided into groups of 2-3, and each group will be responsible
for developing display materials pertaining to the species of parasites discussed in the outline shown
above. Grading will be on the basis
of clarity and accuracy of information, and attendance records. This exhibit, which will be scheduled for the
last Thursday of the semester in either TC-100 or Sabbatini Lounge, will count
2/7 towards your final grade.
You will be learning techniques which are
pertinent to the study of parasitology. These include and will not be limited
to the following:
1. Preparation of permanent slides of stained helminths;
2. Preparation of wet mount of fecal smears;
3. Removal of parasitic organisms from dissected host
animals;
4. Concentration of ova by fecal flotation
5. Preparation of a smear of parasitic protozoa from
earthworms.
These techniques will, in turn, be integrated
into 3 experiments.
1. Survey of parasites collected from freshwater fish
collected locally;
2. Conditions required for the excystation of Posthodiplostomum minimum metacercariae;
3. Life history and control of mosquitos
Generally, you will NOT be exposed to human
pathogens. Nonetheless, you will be required to follow rigorous safety
procedures.
Your final grade will be based on the
following:
1. Weekly quizzes of short answer and identification
questions, covering the previous lab’s specimens. Collectively, these quizzes will count 1/6
towards your final grade;
2. Two typewritten (word processed) laboratory reports
from the experiments described above. The papers are to be written in the same
format for articles appearing in the Transactions of the Tennessee Academy of
Science and will be due two weeks after the completion of data collection.
Grading of the laboratory reports will be on the basis of spelling, grammar,
clarity, and content. Each lab report will count 1/6 towards your final grade;
3. A midterm exam consisting of short answer and
identification questions. This exam will count 1/6 towards your final
grade.
4. A final exam consisting of short answer and
identification questions. This final exam will count 2/6 (1/3) towards your final grade.
Some of the experiments will require more
time than a single four-hour block of time allows. You will be expected to
collaborate in order to complete the work. Furthermore, participation in all
fishing and field trips is mandatory.
The grading scale for the lecture and the lab
will be the following:
A = 3.5 - 4.0
B = 2.75 - 3.49
C = 2.00 - 2.74
D = 1.00 - 1.99
F = <1
By the way, regarding laboratory reports:
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“Brevity is the soul of
wit.” |
CLASS ATTENDANCE POLICY
Every student is expected to attend classroom
and laboratory periods regularly. A student who has been absent, even for a
legitimate cause, is under obligation to make up the work. Any student who has
missed a total of eight hours of class may be dropped from the course, with a
mark of "F", at the discretion of the teacher.
Biol
413: Parasitology Lecture
Fall 2009
Lecture Schedule
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Date |
Day |
Lecture Topic |
Textbook Chapter(s) |
Parasites on Parade Pages |
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8/24 |
M |
Introduction: Types of interspecific interactions; Basic Principles in Ecology & Evolution Life cycle of Babesia microti |
1-2 |
1-18 |
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8/26 |
W |
The magnitude of the problem: Parasite Video |
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8/28 |
F |
Parasite Video, cont’d |
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8/31 |
M |
An introduction to the immune system: Basic Principles in Immunology & Pathology Oddly enough, infection with parasitic worms may reduce the risk of asthma: |
3 |
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9/2 |
W |
Phylum Arthropoda: Form, Function and Classification Insect life cycles Insect development |
33 |
198-240 |
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9/4 |
F |
34 |
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9/7 |
M |
Labor Day – No Class |
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9/9 |
W |
Mallophaga & Anoplura |
36 |
226-232 |
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9/11 |
F |
Hemiptera: True bugs |
37 |
214-216 |
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9/14 |
M |
Siphonoptera: The fleas |
38 |
219-231 |
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9/16 |
W |
LECTURE EXAM I – will include Chapters 1 and 2 from Parasite Rex. |
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9/18 |
F |
Diptera: Family Culicinae |
38 |
204-210 |
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9/21 |
M |
Diptera: Other flies |
39 |
211-213, 217-218 |
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9/23 |
W |
Strepsiptera, Hymenoptera, and others |
40 |
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9/25 |
F |
Parasitic Arachnids: Ticks |
41 |
233-237 |
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9/28 |
M |
YOM KIPPUR -- NO CLASS |
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9/30 |
W |
Parasitic Arachnids: Mites |
41 |
238-240 |
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10/2 |
F |
Basic Principles of Epidemiology |
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10/5 |
M |
Parasitic Protistans: Form, Function and Classification |
4 |
118-119 |
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10/7 |
W |
Order Kinetoplastida: The trypanosomes http://www.medicalecology.org/diseases/d_african_trypano.htm |
5 |
41-58 |
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10/9 |
F |
Tsetse: Guardian video |
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10/12 |
M |
Order Kinetoplastida: The leishmanias |
5 |
41-58 |
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10/14 |
W |
Other Flagellates (Chilomastix, Giardia, Trichomonas) Practice Lab Midterm Exam: Parasites Can Alter Host Behavior |
6 |
19,36-40 |
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10/16 |
F |
LECTURE EXAM II -- will include Chapters 3 and 4 from Parasite Rex. |
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10/19-10/25 |
M-Su |
FALL BREAK |
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10/26 |
M |
Subphylum Sarcodina: The Amebas (Entamoeba spp., Endolimax nana, Iodamoeba butschlii) |
7 |
19-28 |
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10/28 |
W |
http://www.cbu.edu/~seisen/BIOL413LcEx02IDAnswerKeyFa2005_files/frame.htm
Phylum Apicomplexa: Gregarines, Coccidia, and Related organisms. (Toxoplasma, Pneumocystis, Cryptosporidium) |
8 |
30-35, 99-105 |
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10/30 |
F |
Phylum Apicomplexa: Malaria Organisms and Piroplasms (Plasmodium spp.) http://www.cbu.edu/~seisen/Malaria.htm A description of sickle cell anemia:
etiology, pathology, treatment |
9 |
59-98 |
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11/2 |
M |
Phylum Ciliophora: Ciliated Protistan Parasites (Ichthyophthirus multifiliis, Balantidium coli) Phyla Myxozoa and Microspora: Protozoa with Polar Filaments |
10 11 |
106 |
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11/4 |
W |
Introduction to the Phylum Platyhelminthes Aspidobothrea |
13 14 |
107-108 |
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11/6 |
F |
Trematoda: Form, Function, and Classification of Digeneans Digeneans: Strigeiformes (The schistosomes) Granuloma Formation by Schistosoma |
15 16 |
117-124 |
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11/9 |
M |
Digeneans: Echinostomatiformes (Fasciola, Fasciolopsis) |
17 |
111-114 |
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11/11 |
W |
Digeneans: Plagiochiformes and Opisthochiformes (Paragonimus, Clonorchis) |
18 |
109-110, 115-116 |
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11/13 |
F |
LECTURE EXAM III -- will include Chapters 5 and 6 from Parasite Rex. Parasitism as a driving force for sex, or, The Red Queen Hypothesis: Endosymbiosis |
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11/16 |
M |
Monogenea Cestoidea: Form, Function, and Classification of the Tapeworms |
19 20 |
125 |
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11/18 |
W |
Tapeworms (Diphyllobothrium, Taeniarhynchus, Taenia) |
21 |
126-142 |
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11/20 |
F |
Tapeworms cont'd. (Echinococcus, Hymenolepis, Vampirolepis, Dipylidium) |
143-148 |
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11/23 |
M |
Phylum Nematoda Nematodes: Trichurida and Dioctophymatida (Trichuris, Trichinella) Optional initial draft of parasites
of Lepomis macrochirus lab report
due. |
22 23 |
149-150 151-153, |
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11/25 |
W |
Nematodes: Rhabditida, Pioneering Parasites (Strongyloides) Nematodes: Strongylida, Bursate Rhabditians (Ancylostoma spp., Necator) Final draft of parasites of Lepomis macrochirus lab report due. |
24 25 |
154-161 |
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11/27 |
F |
Thanksgiving |
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11/30 |
M |
Nematodes: Ascaridida, Intestinal Roundworms (Ascaris, Toxocara spp.) Nematodes: Oxyurida, the Pinworms (Enterobius) |
26 27 |
162-165 166 |
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12/2 |
W |
Nematodes: Spirurida, a Potpourri Nematodes: (Filaroidea), including Wuchereria, Onchocerca and Dirofilaria Maybe you shouldn't go here: Maybe you shouldn't go here either: Biology as art, or maybe it's art as biology: Nematodes: Camallanina, the Guinea Worms & others (Dracunculus) |
28 29
30 |
169-179 |
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12/4 |
F |
LECTURE EXAM IV -- will include Chapters 7 and 8 from Parasite Rex. |
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12/7 |
M |
Phylum Acanthocephala |
32 |
189-194 |
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12/9 |
W |
Phyla Mollusca, Annelida, Pentastomida Parasitic Chordates (brood parasitism among birds, lamprey) Natural World - Cuckoo Nature of the cuckoo duck -
David Attenborough - BBC wildlife |
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180-188, 195-196 241-247 |
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12/10 |
Th |
PARASITE REX exhibit |
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12/11 |
F |
Plants |
“1”-“6” |
Tentative Lab Schedule: Fall 2009
|
WEEK OF |
EXERCISE |
PAGE(S) in Lab book |
|
Aug 24 |
Studies in mutualism: |
“8”-“14” |
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Aug 31 |
Phylum Arthropoda Class Crustacea Argulus sp. (fish louse) adult, w.m. Class Insecta Order Anoplura Pediculus humanus (body louse)
Phthirus pubis (public louse)
Order Siphonoptera Pulex irritans (human flea)
Ctenocephalides canis (dog flea)
Order Hemiptera Cimex lectularis (bedbug)
|
226-232 219-231 214-216 |
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Sept 7 |
Monday: Labor Day Thursday: · Ova · Larvae · Pupae · Adult males · Adult females Glossina spp. · Adults |
204-210, 211-213, 217-218
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Sept 14 |
Monday: Insects: · Ova · Larvae · Pupae · Adult males · Adult females Glossina spp. · Adults Thursday: Arachnids: ·
Adult females ·
Adult males ·
Larvae Ixodes spp. · Adults Family Trombiculidae ·
Adult from Rana
pipiens Techniques in preparing permanent slides Setup up Mosquito lab |
204-210, 211-213, 217-218 237-240 “34-36” |
|
Sept 21 |
Monday: Arachnids: ·
Adult females ·
Adult males ·
Larvae Ixodes spp. · Adults Family Trombiculidae ·
Adult from Rana
pipiens Techniques in preparing permanent slides Set up Mosquito Lab Thursday: Finish Mosquito lab |
237-240 “34-36” |
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Sept 28 |
Monday – Lab will not met, but
please finish mosquito lab on Tuesday. Thursday: Optional first draft of Mosquito Bioassay experiment due. Flagellates Order Kinetoplastida: · T. cruzi trypomastigotes in blood smear · T. cruzi amastigotes in cardiac muscle c.s. ·
T.
brucei trypomastigotes in blood smear Leishmania spp. · L. tropica promastigotes in culture · L. donovani promastigotes in culture · L. donovani promastigotes in spleen smear |
41-58 |
|
Oct 5 |
Monday: Optional first draft of Mosquito Bioassay experiment due, Flagellates Order Kinetoplastida: · T. cruzi trypomastigotes in blood smear · T. cruzi amastigotes in cardiac muscle c.s. ·
T.
brucei trypomastigotes in blood smear Leishmania spp. · L. tropica promastigotes in culture · L. donovani promastigotes in culture · L. donovani promastigotes in spleen smear Thursday: NO LAB |
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Oct 12 |
COMPREHENSIVE MIDTERM EXAM Final draft of Mosquito Bioassay due. |
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Oct 19 through October 25 |
Fall Break |
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Oct 26 |
Other flagellates:
·
Trophozoites in fecal smear ·
Cysts in fecal smear Trichomonas vaginalis ·
Trophozoites in vaginal smear Chilomastix mesnili · Trophozoites in fecal smear · Cysts in fecal smear Phylum Sarcodina: · Trophozoites in fecal smear · Cysts in fecal smear ·
Trophozoites in liver tissue Entomoeba coli · Trophozoites in fecal smear · Cysts in fecal smear
· Trophozoites in fecal smear · Cysts in fecal smear
· Trophozoites in fecal smear · Cysts in fecal smear |
19, 36-40 19-28 |
|
Nov 2 |
Phylum Apicomplexa: · Tachyzoites in culture · Pseudocysts in brain tissue
Pneumocystis carinii** Plasmodium spp. · Oocysts outside mosquito stomach wall, c.s. · Blood stages Extraction of Monocystis spp. from earthworms, & preparation of permanent slides. |
30-35 99-105 59-98 |
|
Nov 9 |
Ciliophora: · Trophozoites in fecal smear · Cysts in fecal smear Aspidogastrea: Experiment II: Excystation of P. minimum metacercariae |
106 “27”-“30” |
|
Nov 16 |
Phylum Platyhelminthes: · Adult male, w.m. · Adult female, w.m. · Ova in fecal smear, w.m. · Ova in liver tissue, c.s. · Miracidia, w.m. · Cercariae, w.m. Schistosoma haematobium · Ova in fecal smear, w.m. Schistosoma japonicum · Ova in fecal smear, w.m. · Adult male, w.m. · Adult female, w.m.
· Adult, w.m. · Ova in fecal smear, w.m. · Metacercariae, w.m.
· Adult, w.m. · Adult in infected liver, c.s. · Ova in fecal smear, c.s. · Adults, c.s.
· Adult, w.m. · Ova in fecal smear, w.m. · Miracidia, w.m. · Redia, w.m. · Cercariae, w.m. Fasciolopsis buski · Adult, w.m. · Incubated ova, w.m. · Ova in fecal smear, w.m. Rough draft of Experiment II: Excystation experiment due Techniques for preparation of permanent slides |
67-84 |
|
Nov 23 |
Cestoda: Hymenolepis diminuta ·
Adult, w.m. ·
Ova in fecal smear ·
Cysticercoid larva, w.m. Diphyllobothrium latum ·
Scolex, w.m. ·
Mature proglottid, w.m. ·
Gravid proglottid, w.m. ·
Plerocercoid larva, w.m. ·
Ova in fecal smear Echinococcus granulosus ·
Adult, w.m. ·
Ova in fecal smear ·
Cysticercus larva, w.m. ·
Hydatid cyst, c.s. ·
Hydatid sand, w.m. Dipylidium caninum · Adult, composite, w.m. · Ova in fecal smear |
85-108 |
|
Nov 30 |
Nematodes: ·
Adults ·
3rd
stage larvae in muscle tissue Trichinella spiralis ·
Ova ·
Adult
females and males
·
Free-living
forms ·
Parasitic
larvae
·
Ova ·
Necator
americanus adult females ·
Necator
americans adult males ·
Ancylostoma
duodenale adult females ·
Ancylosotma
duodenale adult males ·
Ancylostoma
caninum adult females ·
Ancylostoma
caninum adult males Ascaris lumbricoides ·
Ova ·
Adult
females, whole ·
Adult
males, whole ·
Adult
females, c.s. ·
Adult
males, c.s. Toxocara spp. · Adult males and females · Ova Enterobius vermicularis ·
Adult
females ·
Ova Wuchereria bancrofti ·
Microfilariae
in blood, w.m.
·
Microfilariae
in nodules, c.s.
·
Microfilariae in dog blood ·
Adults |
109-136 |
|
Dec 7 |
Acanthocephala: Mollusca: Annelida: Chordata: |
141-154; 194-200 |
GUIDELINES
FOR LABORATORY REPORTS
1. Please DOUBLE-SPACE your laboratory report, and use
margin size to 1 inch.
2. Place a cover sheet at the front of your laboratory
report. The cover sheet should have your name, the report title, the course and
section numbers centered on the page. PLEASE
The laboratory report should include the
following sections:
1. An ABSTRACT section, in which you describe in briefest
form, the purpose, primary results and conclusions of the research report. By
convention, it is 200 words or 3% of the laboratory report, whichever is LESS;
2. An INTRODUCTION section, in which you provide
information pertaining to the problem as it is recognized and in which you
discuss background information which would be pertinent to the reader. The
purpose, in which you specify the questions to be addressed in THIS lab report,
should be in the LAST paragraph of the introduction section;
3. A MATERIALS AND METHODS section, in which you
discuss the organism(s) under study and the experimental protocol in
"text" form. PLEASE DO NOT INCLUDE A MATERIALS LIST. If there are
several parts to the experiment, each part should be described separately. In
text format, briefly describe the protocol you followed in conducting the
experiment. If there are several parts to the experiment, each part should be
described separately;
4. A RESULTS section, in which you discuss the
data from each part of the study in the same sequence as the parts were
described in the Materials and Methods section. Use a paragraph to tell the
reader what the main point is, and at the end of the sentence, refer to a
specific Table or FFigure, as in the following: "Seedlings exposed to either
.1% or .2% phosphate grew vigorously, but the controls did not (Figure
1)." It is essential to convert or present the data in an understandable
format. CHARTS OF RAW DATA ARE NEITHER NECESSARY NOR DESIRABLE!;
5. A DISCUSSION section, in which you relate the
results of your experiment to the general body of knowledge pertinent to this
area of research;
6. A REFERENCES section, in which you list the
references used for background information and/or protocol procedures,
including your laboratory textbook.
CRITERIA
FOR THE GRADING OF PAPERS AND EXPERIMENTAL REPORTS
The
maximum grade is a 4.0 and is a composite of three grades based on spelling
grammar, and content.
I. Spelling counts 10% of the total
grade. Each different spelling or typographical error will usually result in a
point deducted from the maximum. However, if one word is consistently
misspelled, it will be deducted only once. Low grades in spelling can be
avoided by keeping a dictionary on hand and proofreading your work before you
submit it for review.
II. Grammar counts 20% of the total
grade. Each grammar error (wrong tense, poor sentence of paragraph structure)
will usually result in a point deducted from the maximum. Low grades in grammar
can be avoided by proofreading your work before you submit it and by writing
practice essays.
III. Content counts 30% of the total
grade. The kinds of questions that are considered in evaluating content include
the following:
1. Is your information accurate?
2. Is your discussion logical?
3. Do you adequately support your argument?
4. Do you adequately correlate and contrast your data to
previous experience?
5. Do you support your conclusions with the appropriate
statistical test(s)?
IV. Format counts 40% of the total grade.
The kinds of questions that are considered in evaluating format include the
following:
1. Did you transform the raw data into a more useful and
appropriate format?
2. Do you follow the protocol for a laboratory report as
described in the Transactions of the Tennessee Academy of Sciences?
You should write your reports as if you were
submitting them to the Transactions of the Tennessee Academy of Sciences. I, in
turn, will review them as if I were an editor for the journal.
Name______________________
Grades
Spelling x 10% = .
Grammar x 20% = .
Content x 30% = .
Format x 40% = .
COMPOSITE GRADE .
Some
General Guidelines for Laboratory Reports
1. Use first-person past tense in the abstract, materials
& methods & results sections, since you are describing what you did.
In other words,
"we dissected the liver from Lepomis macrochirus" is clearer
than "Livers were dissected from Lepomis macrochirus".
2. Species names should be italicized or underlined.
For example,
"We studied the excystation behavior of Posthodiplostomum minimum"
or "We studied the excystation behavior of Posthodiplostomum minimum.
3. When a species has a long name, it is acceptable to
contract the genus name to one letter if you refer to it as such at the
beginning of your paper.
For example,
"We studied the excystation behavior of Posthodiplostumum minimum
(referred in this paper as P. minimum)."
4. The References Cited section should include those
articles or books from which you collected information and quote it in your
report. The citation in your paper should appear as (AuthorLastName,
YearOfPublication).
For example, "P.
minimum metacercariae become resistant to pepsin between days 26 and 44
(Eisen, 1999).
5. Each Figure should be numbered and referred to in the
text of your results section in parentheses.
For example,
"We observed maximal movement in the well where the larvae were first
exposed to acid saline with pepsin, followed by alkaline Tyrode's solution with
trypsin (Figure 1).