Activity 7: Isolation of Microorganisms a. Liquefy medium in a water bath.

- Microorganisms in nature are b. Obtain three 9-ml water blanks in

diverse and occur as a complex mixed screw cap tubes and label each tube from 1
population. to 3.
c. Secure a broth mix culture from your
A study of these microorganisms requires instructor.
that they be separated from each other and d. With a sterile pipet, transfer one ml
cultivated in an artificial environment. from the stock culture to tube #1. Cover and
Different techniques are employed to isolate shake vigorously.
the different species from the mixture and e. From tube #1, transfer 1 ml to tube #2.
cultivate them as pure cultures. Mix.
f. Again transfer 1 ml from tube #2 to
- consists of a population of cells tube #3. Shake and mix.
derived from a single cell. g. Using a pipet, transfer 1 ml from each
tube to a duplicate sterile plates. Pour
melted afar and mix as in the below.
is a very simple method of
h. Allow the agar to solidify and incubate
isolation if done properly.
as in A4.
i. Examine after 48 hours.
can also be used, and in
addition, can approximate the variable
organisms in the population. The use of a. Pour melted NA or CWA into each of six
enrichment or selective medium will sterile petri dishes and allow to
enhance growth of the microorganisms or solidify.
interest while inhibiting or killing the other b. Transfer to duplicate plates 0.1 ml
types of microorganisms. from each tube used in pour plate
technique.
c. Sterilize an L-shaped glass rod by
13 sterile petri dishes dipping in alcohol and flaming until alcohol
1 wire loop has burned off.
3 sterile 9-ml water blanks in screw cap d. Spread the inoculum evenly on the
tubes surface of the agar with an L-shaped rod.
1 water bath
in screw cap tubes
Nutrient agar
6 1-ml sterile pipets
culture broth Scanty
1 L-shaped sealed glass tubing moderate
abundant

Filiform
Echinulate
Beaded
a. Pour melted NA or CWA, following Spreading
aseptic technique in a petri dish and allow Rhizoid
the agar to solidify.
b. Sterilize the wire loop and get a loopful
of microorganisms of interest ffrom the petri
dish with mix culture done in the last
activity.
c. Streak very gently over the surface of
Punctiform circular
the agar, taking care not to destroy it, as in
Filamentous
figure below.
Rhizoid
d. Incubate the dish invertedly and
Irregular
examine the plate after 48 hours.
Effuse
e. Examine your previous petri dish
Flat
further. Choose another colony and get a
Raised
loopful.
Convex
f. This time streak this in an agar slant.
Entire
g. Incubate and examine tube after 48
Endulate
hours.
Erose

1. Compare the pour plate and streak
plate methods of isolating bacteria?
The pour plate and streak plate are methods
used in microbiology to mainly grow bacteria
and fungi in petri dishes with nutrient agar.
These two methods are similar in:
general purpose – plating methods to isolate
microorganisms.
principal materials – utilize the petri dish
and nutrient agar.
incubation method – incubate invertedly at
37 °C. Although these two methods share
quite the similarity, there are more
differences between these two than
similarities.
- is used to count the number of
colony-forming bacteria in a liquid specimen.
is used to isolate pure strain
from a single species of microorganisms.
Second, the order of microorganisms and
culture media – in pour plate, bacterial
broth is first poured in followed by the
melted nutrient agar meanwhile, in streak
plate, melted nutrient agar is first poured
in followed by the loopful of bacteria. Third,
amount of inoculum – in pour plate, the
inoculum is between 0.1-1.0 mL meanwhile,
in streak plate, the inoculum is only a
loopful of bacteria.
TYPE OF COLONIES
-produces surface and subsurface
colonies.
- produces surface colonies only.
METHOD DISADVANTAGES
Pour plate- microbes have to withstand the
temperature of the melted nutrient broth
during preparation.
Streak plate- there is a higher probability of
contamination.
2. In which plate do you find the surface
and the deep or buried colonies?
Three plating methods are widely used in
the isolation of bacteria: the streak plate
method, the pour plate method, and the
spread plate method.
Colonies can only be grown on the surface
using the streak plating and spread plate
methods. This is due to the fact that in the
streak plating method, the sample is
immediately diluted across the plate's
surface, while in the spread plate method,
the diluted sample is distributed
uniformly across the plate's surface. The
pour plate method, on the other hand, can
grow colonies on both the surface and a
deep or buried surface. Since melted agar is
applied to the bacterial dilute in the pour
plating process, bacteria may grow in and on
the solidified medium.
3. How do surface colonies differ from
deep or buried colonies?
Surface colonies and deep or buried colonies
differ in respiratory. The most fundamental
distinction between the two is that aerobic
bacteria expand on the surface, while
anaerobic bacteria may colonize deeper
areas.
- are bacteria that can
survive low levels of oxygen and live in an
anaerobic atmosphere.
- are exposed to air and thrive
above the surface of the medium.
4. Which is rapid in proving the purity of
a culture – on an agar or in a broth?
- In culturing bacteria growth, the most
commonly used nutrient media. It is
commonly used because it is rapid in
proving the purity of a culture.
- can be easily contaminated; under
the microscope, colonies are difficult to
observe, identify, and distinguish from one
another. To prove the purity of a culture, the
plate must have one kind of colony on the
media plate. All colonies must have the same
morphology (size, form, color, and so on).
5. Why are petri dishes incubated in an
inverted position?
Agar plates are incubated in an inverted
position for various reasons. First, the
accumulated water condensation can
disrupt the colonies and the microbial
count. Second, the evaporation of water
 from the media can cause dryness which Dissolve methylene blue in alcohol and then
can affect the microbial growth. Third, the adjust volume to 1000ml.
lid of the petri dish may contain
contaminants which could spread onto 2. Dorner’s nigrosin solution:
the media and grow along with the
sample microbes. Fourth and lastly, the Nigrosin, water soluble---10 grams
inverted positions allow for better handling.
Distilled water---100 ml
Conclusion
In this activity, we went over how to perform Immerse in boiling water bath for 30 minutes
different plating methods – streak plate in then add preservatives 0.5 ml formalin. Filter
both petri dish and test tube, pour plate, and
twice in double filter paper and store in test
spread plate and how to identify colony
morphology. There are five things that tubes.
fascinated us throughout the activity. First,
pour plate and streak plate differ in specific 3. Ziehl’s Carbon – fuchsin:
purpose – the pour plate is used to count
the number of colony-forming bacteria in Solution A:
a liquid specimen meanwhile the, streak
plate is used to isolate pure strain from a Basic fuchsin dye (90% dye content)-0.3 gram
single species of microorganisms. Second, Ethyl alcohol (95%)-10.0 ml
in streak and spread plate method,
colonies grow on the surface however, in Solution B:
pour plate method, colonies can grow both
on the surface and buried surface. Third, Phenol -5 grams
Surface colonies and buried colonies differ in Distilled water-95 ml
respiratory - aerobic bacteria expand on the Mix solution A and B
surface, while anaerobic bacteria may
colonize deeper areas. Fourth, Agar is the
preferred media as it is rapid in proving the
purity of a culture. Fifth, petri dishes are a. Gram 1. - Ammonium oxalate crystal
incubated invertedly because the violet
accumulated water condensation can disrupt
the colonies and the microbial count. Solution A:

Crystal violet (90% dye content)-2 grams Ethyl

Activity 8: Bacterial Staining alcohol (95%)- 20ml
Unstained bacterial cells are nearly
transparent when observed by light Solution B:
microscopy and hence are difficult to see.
However, most bacteria can be easily Ammonium oxalate-0.8 grams
stained. Distilled water- 80ml
Mix solutions A and B.
- involves spreading a drop of an
aqueous suspension of cells on a glass slide b. Gram 2 - Gram iodine
allowing it to dry in air, fixing it with gentle
heat, and then applying staining reagents. Iodine-1 gram
Potassium iodide-2 grams
Dyes as stains are used to make Distilled water -300 ml
microorganisms more easily seen, to show
certain cell structures, or to reveal their chemical
nature. c. Gram 3. - Ethyl alcohol (95%)

d. Safranin

2.5% solution in 95% ethyl alcohol-10 ml

1. Methylene blue in dilute alcohol
Distilled water-100 ml
Methylene blue (80%) dye content---0.3 gram)
5. Malachite green (Shaeffer and Fulton’s
Ethyl alcohol (95%) ---30 ml
spore stain 7.5% aqueous solution of malachite
Distilled water---1 Liter green oxalate.
Preparation of smears:

1. Wash the slide thoroughly with a Gram Ammoniu Gram’ Ethyl Safranin
detergent, rinse with water and dry with a clean Reactio m oxalate s Alcoho
n crystal iodin l
cloth or tissue paper. A drop of water placed on a
violet e
clean slide will spread out evenly. If the drop Gram Purple Purpl Purple Purplish/Bl
rounds up, clean the slide again. The slide Positive e ue
should be thoroughly cleaned and freed of grease. Gram Purple Purpl Colorle Reddish
Negativ e ss Pink
e
2. If the specimen comes from liquid culture,
remove a loopful as in exercise
1. Of what value is gram staining?
2. Spread the loopful evenly and thinly over
a 5 sq. mm area. If the bacterial specimen comes
from a solid medium, get a loopful by following - is a differential stain used to
aseptic technique, put drop of water in the slide differentiate gram positive and gram-negative
and mix until a turbid mixture is acquired before bacteria and is the simplest and most rapid test
spreading evenly. to characterize microorganisms, it will help
assess the adequacy of preliminary diagnosis and
3. Dry the film by air. Fix smear by heat or antimicrobial therapy selected after collecting the
by alcohol. In heat fixing, the slide is passed, film culture specimen and before the final
slide up quickly over the flame of the alcohol identification of the microorganisms.
lamp 2 or 3 times. Do not scorch. The slide
should feel hot to the fingers, but should not 2. Why are the steps in gram-staining so
burn them. The alcohol treatment consists of carefully standardized?
covering the smear with 95% alcohol for one
minute then draining off. Heat fixing is never Gram staining so carefully standardized because
used with preparations made of milk. it is the most important staining procedure in
microbiology. It is used to differentiate
The purpose of fixation is to kill the between gram positive and gram-negative
microorganisms, coagulate the protoplasm of the organisms.
cell and cause it to adhere to the slide.
Why is it necessary to employ a spore
The best bacterial smears are usually made by stain rather than rely on a diagnosis of
removing a small amount or surface growth from sporing from examination of gram stain?
agar culture and mixing with water. It is often
possible to use a drop of culture growing in a Bacteria can form spores in approximately 6 to 8
liquid medium, but such a smear is not always hours after being exposed to adverse conditions.
so satisfactory since certain constituents of the Spores are metabolically inactive and dehydrated.
medium may prevent the bacteria from adhering
to the slide or may interfere with staining. The - the primary stain in the
suspension used should always be diluted. endospore stain technique, is pushed into the
cells by heat. The primary stain is water-soluble
and does not bind well to cells, and rinses
quickly from the vegetative cells, causing the
Stain your smear with ammonium oxalate crystal counterstain to be readily absorbed.
violet for one minute. Wash thoroughly but gently
with tap water. Cover smear with iodine solution Of what importance are spore-forming
for one minute. Wash in tap water and shake off bacteria in the food industry?
excess moisture. Decolorize with 95% ethyl
alcohol for 15 to 20 seconds. Wash with water. In industrial food production, specific endo spore
Counterstain with safranin solution for 30 formers have been significant pollutants.
seconds. Wash with tap water and blot dry. The
B. cereus- is the most common pathogen or
gram (+) positive organisms appear blue or deep
spoilage organism in the dairy industry.
violet; the gram (–) negative, red or pink.
Alicyclobacillus acidoterrestris- has become a
significant quality-control organism in fruit
juice. Anaerobic spore formers such as non-
proteolytic Clostridium botulinum pose a risk
of protection and spoilage in chilled
processed foods.
Of what practical significance are capsule
forming bacteria in industry and
medicine?
Capsules increase bacteria's capacity to induce
disease. Capsules shield cells from eukaryotic
cells like macrophages engulfing them. Water is
also present in the capsules, which prevents the
bacteria from drying out. immunity to one kind
of capsule does not imply immunity to the others.
The capsule-specific antibody can prevent
phagocytosis.
Why is mordant necessary for staining of
bacterial flagella?
Ordinary stains are too small to visualize flagella
under a bright field microscope. A wet mount
technique is used to stain bacterial flagella.
The number and structure of flagella are
important for identifying species of motile
bacteria.
Differentiate differential stain from
simple stain.
Some staining methods need only one dye to be
applied to the sample, while others need several
dyes.
Single dye- is used in simple staining to
highlight specific structures in the specimen.
Differential staining identifies species based on
how they deal with various stains. Two species
can tend to be different colors in a differentially
stained sample.
Gram stain is a preliminary step in the initial
characterization and classification of
bacteria. This procedure differentiates the two
large groups the gram negative and gram
positive. Gram staining involves a four-part
process. The significance of endospore stain is
for detecting bacterial endospores by forming
spores, bacteria can survive in hostile
conditions. The number and structure of
flagella are important for identifying species of
motile bacteria. Gram negative and gram-
positive organisms are distinguished from each
other by differences in their cell walls. Gram
positive cells take up the crystal violet, cell walls
of gram-positive organisms include a thick layer
of protein-sugar complexes called
peptidoglycans. At the end of the gram staining
procedure, gram positive cells will be stained a
purplish-blue color. On the other hand, gram
negative cells also take up crystal violet.
Activity9: Antibiotics
The growth of microorganisms in nature might be
influenced by the presence of other
microorganisms.
- When the influenced is unfavorable or
harmful.
- substances produced by
microorganisms that inhibit the growth of other
microorganisms at very low concentrations. It
has been put to good use controlling the activities
of harmful bacteria.
1. Melt the sterile medium in the 2 test,
tunes and cool to 50 degree Celsius.
2. Pour into 2 separate petri dishes.
3. While the medium is still in its liquid
state, inoculate it with 1ml of the bacterial
culture by pour plate method. Follow aseptic
techniques.
4. When the agar has hardened using a
marketing pen, divide one of the plates into two
equal portions by marking the bottoms of the
petri dish.
5. Four small number of equal portions of
sterile water on 2 small beaker. To one dissolve a
small amount streptomycin and to the other a
small amount of penicillin.
6. Soak a piece of filter paper disk in
streptomycin solution. Aseptically transfer this
disk into one of the divisions on the plate marked
earlier and label as streptomycin. Do the same
with the other antitripic solution.
7. To the other plate mark four equal
divisions. Make four quadrants. Label each
quadrant as to the soap you are going to use.
8. Put small amount of sterile water in 4
small beakers. Make four different soap
solutions.
9. Soak a filter paper disks in each soap
solution. Aseptically transfer each of these disks
into each guardant and label according to the
soap name.
10. Incubate the plates at room temperature
for 48 hours.
11. After 48 hours measure the zone of
inhibition in mm and tabulate your observation.
Name of Antibiotic Zone of inhibition (mm)
Streptomycin 1 mm
Penicillin 8 mm
Name of Soap Zone of Inhibition
A. Meditex 14.1 mm
B. Duru 11.57 mm
C. Protex 14.97 mm
D. Roberts 19.27 mm
What substance is produced by a
microorganism that is capable of the growth
of other microorganisms?
The substance that a microorganism
produces that inhibits the growth of another
microorganism through killing is known as
antibiotics. Antibiotics are commonly used as
treatment of different bacterial infections.
Examples are penicillin, chloramphenicol, and
erythromycin.
In what way does the use of antibiotics
contributed the problem on the emergence of
drug resistant microorganisms?
Misuse and overuse of antimicrobials are the
main cause in the development of drug resistant
pathogens. This is because the increases in
antibiotic resistance are driven by a combination
of germs exposed to antibiotics and the spread of
those germs and their mechanisms of resistance.
As we use antibiotics, microorganisms develop
defense strategies against them. This makes the
drugs less effective. Also, when antibiotics are
not taken for the entire prescribed course,
pathogenic bacteria can adapt the process of.
Load those antibiotics. And eventually, for a
population that is completely resistant to the
antibiotic, regardless of the dosage.
What does the zone of inhibition imply? Does
the measurement of the zone of inhibition
implied that one antibiotic is better than the
other?
The size of the zone of inhibition is usually
related to the level of antimicrobial activity
present in the sample or product. A larger
zone of individual usually means that the
antimicrobial is more potent. The measurement
of the zone of individual implies that one of the
antibiotics is better than the other. Big cause
larger size of inhibitions indicates susceptibility
of the Organism. So, the antibiotic and smaller
size of in divisions indicates a resistance of the
microorganism to one antibiotic.
In conclusion, antibiotics are substances
produced by microorganisms that hinder the
growth of other microorganism at a very low
concentration. The inhibitor in the table one is e-
coli and it shows that streptomycin Has greater
innovation zone than penicillin. The inhibitor in
table two is also e-coli. Roberts brand has the
highest zone of inhibition compared to the others
with a mean of 19.27 zone of inhibition while
soap A which is Meditex has the smallest zone of
inhibition with the mean of 14.1. Larger
inhibitions indicate sensitivity of the organisms
to the antibiotic, and smaller innovations
indicate tolerance of the microorganisms to one
antibiotic. Measuring the zone of inhibition
shows that one antibiotic is stronger than the
other.
Activity 10: Nematodes
(ROUNDWORMS)
Many species of nematodes are free-living
forms however, several species are parasitic to
man. The target member of helminths parasites
of man belongs to the group of roundworms.
Among the species that are parasitic to man
include: Ascaris lumbriciodes, Trichinella
spiralis, Wuchereria bancrofti (or Bancroft’s
filarial worm), Enterobius vermicularis or pin
worms and Trichiuris trichiura.
• parasites that can live in your intestine.
• have long round bodies and range in
size.
• can live in or on humans and can cause
many problems.
• usually found in soil and stool and can
enter the body through the mouth or
direct contact with the skin.
1. Obtain a prepared slide of the following
roundworms:
a. Ascaris lumbriciodes
b. Trichinella spiralis
c. Wuchereria bancrofti
d. Enterobius vermicularis
e. Trichiuris trichiura.
2. Examine the prepared slides under the
 microscope using LPO and HPO.
3. Draw each organism/s observed. Place
your drawings on the space provided in the
worksheet.
4. Using your reference book, draw the
developmental stages (life cycle) of each
roundworm that you examined.
5. Describe and discuss the mode of entry
and method of reproduction of each
roundworm.
PHYSICAL DESCRIPTION:
- elongated, cylindrical body which tapers
at both ends; cream- colored,
sometimes with a pink tinge;
integument of the worm is a chitinous
layer of nonnucleated cuticula with
circular striations.
DIFFERENCE OF MALE AND FEMALE
Males: the tail curves ventrally; papillae
grouped pre- and pos- anally.
Females: have vulval opening located
ventrally at a point of constriction.
LENGTH OF ADULT WORM:
Females: 20-35 cm; 3-6 mm in diameter.
Males: 15-30 cm; 2-4 mm in diameter.
LIFE CYCLE
Females produce about 200,000 eggs per day
in the small intestine, where adult worms
mature and reproduce. In moist, warm soil,
where an embryo grows inside each egg, eggs
passed with feces will survive for years. The
larvae invade the intestinal wall after ingesting
eggs in water or on vegetables, allowing them
to enter the circulatory and lymphatic
systems. The larvae then travel to the lungs,
where they go through two more larval stages
in around two weeks. The worms are then
coughed up and swallowed in the pharynx. In
the intestine, the parasites mature into adults.
LABORATORY DIAGNOSIS
1. Looking for eggs in the stool under a
microscope
Procedure:
• Take a sample of your feces.
• Put the specimen in formalin or another
fixative to preserve it.
• Using the formalin–ethyl acetate
sedimentation technique, concentrate
the specimen.
• Examine at a wet mount of sediment.
2. Direct wet mount inspection-
adequate for detecting moderate to
heavy infections when concentration
procedures are not available.
3. Kato-katz or quantitative fecal
flotation- used to make quantitative
measurements of infection.
PHYSICAL DESCRIPTION:
- smallest nematode; body of the worm is
more slender at the anterior than at
the posterior end.
DIFFERENCE OF FEMALE AND MALE:
Males: larva has a long rectum, and the
anterior portion of the testis is posteriorly
angled.
Females: female larva has a shorter rectum
(about 25 m), a telogonic ovary, coiled uterine
and seminal receptacle primordia, and a
vaginal primordium.
LENGTH OF ADULT WORM
Females: 2.2 mm in length
Males: 1.2 mm in length
LIFE CYCLE
Humans are sometimes infected after
consuming fresh or insufficiently cooked meat
from infected animals. In most cases, the virus
is retained in a pig-pig or pig-rat-pig loop.
Since any mammal may be infected, the
disease may be acquired anywhere in the
world. This species ' life cycle starts with the
intake of the first stage juvenile from the
intermediate host. Within the first thirty
hours, the worm molts four times and then
mates.
LABORATORY DIAGNOSIS
1. Muscle biopsy- reveals the larvae
inside striated muscles.
PHYSICAL DESCRIPTION
- translucent white worm with a smooth
cuticle; head is rounded and separated
from the body by a neck-like
 constrictions.
DIFFERENCE OF MALE AND FEMALE
Males: have tail curves ventrally.
Females: have tail tapers gradually and is
rounded at the tip.
LENGTH OF ADULT WORM
Females: 80–100 mm; diameter ranging from
100-300 μm .
Males: 40 mm.
LIFE CYCLE
The microfilariae begin their life cycle as
larvae in the mosquito and then migrate to the
salivary glands. Parasites are injected into a
new individual when the mosquito feeds again.
Larvae migrate to deeper tissues through the
circulatory system, where they mature. Adults
can live and reproduce in lymphatic vessels
for up to 17 years. Microfilariae stay in
capillaries of internal organs, only swimming
freely in the bloodstream at night, which
coincides with most mosquitoes ' feeding
habits.
LABORATORY DIAGNOSIS
1. Peripheral blood smears (thick or thin)
stained with Giemsa or hematoxylin-
and-eosin - commonly used to
diagnose lymphatic filariasis .
2. Knott's technique - concentration
method that involve filtration through
a polycarbonate membrane or
centrifugation of a blood sample lysed
in 2% formalin.
PHYSICAL DESCRIPTION
- cylindrical body, and a cuticle with three
main outer layers made of collagen and
other compounds.
DIFFERENCE OF MALE AND FEMALE
Males: have a curled tail
Females: have a pointed tail
LENGTH OF ADULT WORM
Females: 8–13 mm; 0.3-0.5 mm wide
Males: 2–5 mm; 0.1-0.2 mm wide
LIFE CYLE
Female pinworms migrate to the anus at night
after mating in the colon and deposit eggs
perianally. Scratching dislodges eggs onto
clothing or bedding, where they dry out,
become aerosolized, and settle in water or on
food before being consumed. Scratching, on
the other hand, leaves eggs on the skin and
under the fingernails, allowing infected people
to re-infect themselves by ingesting the eggs
on their hands or in food. Adult worms mature
in a matter of weeks and live for about two
months in the intestine.
LABORATORY DIAGNOSIS
1. Microscopic identification of eggs
collected in the perianal area.
2. Scotch test- pressing clear cellulose
tape on the perianal skin and
inspecting the tape mounted on a
microscope slide.
3. Anal swabs or “Swube tubes”- a paddle
coated with adhesive material.
PHYSICAL DESCRIPTION
- elongated anterior end that contains the
mouth and esophagus that stretches
into a threadlike point; posterior end is
more blunt and contains the anus and
sex organs.
DIFFERENCE OF MALE AND FEMALE
Males: have tail curves ventrally
Females: straight posterior end
LENGTH OF ADULT WORM
Females: 35-45 mm long
Males: 30-40 mm long
LIFE CYCLE
T. Trichiura is spread via fecal-oral
transmission. If the infected person defecates
outside or if human feces as used as fertilizer,
eggs are deposited on soil. They can then
mature into a form that is infective.
Whipworm infection is caused by ingesting
eggs. This can happen when hands or fingers
that have contaminated dirt on them are put
in the mouth or by consuming vegetables or
fruits that have not been carefully cooked,
washed or peeled. Females may produce
pheromones to attract males. The mail coils
around a female with his curved area over the
female genital pore. The gubernaculum, made
of cuticle tissue, guides spicules which extend
through the cloaca and anus. Males use
spicules to hold the female during copulation.
LABORATOTY DIAGNOSIS
 1. microscopically identifying whipworm
eggs in a stool sample - the standard
method for diagnosing the presence of
whipworm
2. fecal concentration procedure -
recommended if eggs may be difficult to
find in light infections
3. Kato-Katz technique - can be used in
quantification of the latter because the
severity of symptoms depends on the
worm burden.
TREATMENT OF CHOICES
CONTROL ROUNDWORM INFESTATIONS
mebendazole, albendazole, pyrantel
pamoate.
- mebendazole or pyrantel pamoate.
diethylcarbamazine; ivermectin
(alternative).
No treatment.
- mebendazole
MODE OF ACTIONS AND CLINICAL
CONSIDERATIONS
Benzimidazole derivatives Representatives:
Albendazole and Mebendazole.
- inhibit microtubule formation and
glucose uptake.
Spectrum of action: Helminths, protozoa
Route of administration: Oral
Adverse effects: Possible diarrhea
Ivermectin- produce flaccid paralysis by
blocking neurotransmitters.
Spectrum of action: Helminths
Route of administration: Oral
Adverse effects: Allergic reactions may
result from antigens of dead helminths.
Pyrantel pamoate and
Diethylcarbamazine- bind to
neurotransmitter receptors, causing
complete muscular contraction.
Spectrum of action: Nematodes
Route of administration: Oral
Adverse effects: None
PREVENTIVE MEASURES
Aside from frequent handwashing, it is
recommended to clip fingernails; to wash
beddings and linens; to have good cooking
practices; to perform housecleaning and
dusting under beds, on windowsills and
over doors with damp mop.
It is advisable to use insect repellents or
mosquito netting; to wear loose, long,
light-colored clothes, or staying indoors;
and to eliminate standing water (a
requirement of mosquitoes ' life cycle)
around human dwellings.
At home, there must be a proper disposal
of human feces; and avoidance of using
human feces as fertilizer.
Activity 11: Nematodes
(HOOKWORMS)
Hookworms are several parasitic
roundworms that have hook-like appendages
surrounding their mouth. They usually bore
through the skin which attach itself to intestinal
walls. As intestinal parasites, they are
responsible for diseases in a number of animals.
And in humans they are marked by anemia,
abdominal pain, and diarrhea.
Hookworms belong to class Nematoda and
are classified as Ancylostoma, necator, and
Unicinaria. The species include Ancylostoma
duodenale, Ancylostoma ceylanicum,
Ancyclostoma braziliense, Ancylostom caninum,
and Necator americanus. Among the species
mentioned, the most common to humans are
Ancylostoma duodenale and Necator americanus.
• Grayish white or pinkish with the head slightly
bent • Possess well developed mouths with two
pairs of teeth
• Females are often longer and stouter compared
to men.
• Male A. braziliense have two broad lateral lobes
and a smaller dorsal lobe with rays on the
copulatory bursa.
• The lateral bursal rays are separated at the
tips.
• Females are more difficult to distinguish
between different species, and usually the teeth
are the only diagnostic tool that can be used.
• grey or reddish in color
• a buccal capsule that contains 3 pair of ventral
teeth
• eggs are oval
• male measures from 1-1.2 cm and female
measures from 1.4-1.6 cm
• smaller than A. duodenale • Position of the genital rudiment: Genital
• possesses a pair of cutting plates in the buccal rudiment is noticed behind the point mid-
capsule way between the end of the oesophagus
• males measures 5 to 9 mm long and females and the anus.
about 1 cm long
• Necator americanus adult worms size are
MODE OF ENTRY smaller in size and are more slender.
• Necator can only be transmitted through • The anterior end bends in opposite
penetration of the skin direction to the body curvature, buccal
• Ancylostoma can be transmitted capsule bears 4 chitinous plates -2 on
percutaneously, orally, and probably ventral surface and 2 on the dorsal
transplacentally. surface.
• Copulatory bursa the bursa is longer and
REPRODUCTION OF ANCYLOSTOMA narrower and it is distinguishable by the
DUODENALE split dorsal ray and approximation of two
of the lateral rays.
• Both males and females attach to the intestinal • The posterior end of female has no
walls during their life span but the male leaves at caudal spine.
one point to search for a female to mate with. • The Vulval opening is in front of the
• The average female life span is about one year, middle of the ventral side of the body.
during which it may lay from 10,000-30,000 eggs • Larvae of a Necator Americanus Round
a day during its adult life. in shape and is larger about 0.75 mm
• Females may produce a phermomone to attract long.
males. • The Buccal cavity is shorter lumen is
• Males use spicules to hold the female during noticed. • Oesophago-intestinal junction:
copulation. An apparent gap is noticed between the
oesophagus and the intestine.
REPRODUCTION OF NECATOR AMERICANUS • Posterior end of the intestine: No
refractile body is noticed.
• Egg production in females occurs five weeks or • Position of the genital rudiment: It is
more after the female matures. present in front of the point midway
• Mating occurs in the intestine of the host. between the end of the oesophagus and
• Males are required to find females and inject the anus. Cuticular striae Distinct visible.
their sperm into the females.
• Females are capable of producing 10,000 eggs
per day. DIAGNOSIS OF HOOKWORM
• The eggs are the passed out with the feces. INFESTATION CAN BE ESTABLISHED

COMPARE THE LARVAE AND ADULT • Identifying hookworm eggs in a sample of

STAGES OF ANCYLOSTOMA stool
DUODENALE AND NECATOR • Eosinophilia is often present in people
AMERICANUS
infected with hookworms
• The size of Ancylostoma duodenale • Blood tests for anemia and iron deficiency
adults worms are large in size and are are also done
coarser.
TREATMENT OR BEHAVIOR OR DRUG CAN
• The anterior end: Bends in the same
BE USED TO EVADE HOOKWORM
direction as the body curvature.
INFESTATIONS
• Buccal capsule: is armed with a pair of
chitinous ventro-lateral cutting plates
Anthelmintic medications, these medicines get
bearing teeth.
rid of parasitic worms in the body.
• Copulatory bursa: is broad and there is a
single dorsal ray and the nearly equal Common drugs:
spread of the three lateral rays • Albendazole
• The posterior end of male is provided • Mebendazole
with two spicules • Pyrantel pamoate
• Posterior end of female has a pointed • Thiabendazole
posterior end.
• Larvae in Ancylostoma Duodenale is 2 MEASURES THAT WILL PREVENT AND
Slightly conical in shape with a size of 0.5 CONTROL PARASITIC INFESTATION IN
mm long. MAN
• Oesophago-intestinal junction: There is
no distinguishable gap between the Wear shoes outside. Avoid skin contact with
oesophagus and the intestine. soil that may be contaminated. Avoid eating
food that could be contaminated. Avoid skin
contact with dog poop.
CONCLUSION
• Hookworms are several parasitic
roundworms that have hook-like appendages
surrounding their mouth. They usually bore
through the skin which attach itself to
intestinal walls.
• Ancylostoma duodenale worms are grayish
white or pinkish with the head slightly bent in
relation to the rest of the body. This bend
forms a definitive hook shape at the anterior
end for which hookworms are named.
• Ancyclostoma braziliense the lateral bursal
rays are separated at the tips, and the position
of attachment of the externodorsal ray is
unique in that it is closer to the beginning of
the dorsal trunk than in other species.
Females are more difficult to distinguish
between different species, and usually the
teeth are the only diagnostic tool that can be
used.
• Anyclostom caninum They are fairly rigid
Grey or reddish in color depending upon the
amount of blood in the intestine.
• Necator americanus is very similar in
morphology to A. duodenale. N. americanus is
generally smaller than A. duodenale with
males usually 5 to 9 mm long and females
about 1 cm long. Whereas A. duodenale
possess two pairs of teeth, N. americanus
possesses a pair of cutting plates in the buccal
capsule. Additionally, the hook shape is much
more defined in Necator than in Ancylostoma.
• The difference of Ancylostoma duodenale
and Necator americanus in terms of mode of
entry is that Necator can only be transmitted
through penetration of the skin whereas
Ancylostoma can be transmitted
percutaneously, orally, and probably
transplacentally.
• While for mode of reproduction, for
Ancylostoma duodenale both males and
females attach to the intestinal walls during
their life span, but the male leaves at one
point to search for a female to mate with. The
male coils around a female with his curved
area over the female genital pore. Males use
spicules to hold the female during copulation.
• While for Necator americanus, Mating occurs
in the intestine of the host. Males are required
to find females and inject their sperm into the
females. Females may produce a pheromone
to attract males. The male coils around a
female with his curved area over the female
genital pore.
• Hookworm infection is diagnosed by
identifying hookworm eggs in a sample of
stool. Stool should be examined within several
hours after defecation. Eosinophilia is often
present in people infected with hookworms.
Hookworm infection is an important
diagnostic consideration in people who have a
blood count done that shows eosinophilia,
especially if they are immigrants or travelers
returning from endemic regions where
sanitation is poor. Blood tests for anemia and
iron deficiency are also done.
• Possible treatment or behavior or drug can
be used to evade hookworm infestations are
Anthelmintic medications, these medicines get
rid of parasitic worms in the body. Common
drugs for intestinal hookworm include
albendazole, mebendazole, and pyrantel
pamoate.
• Lastly, here are some preventive measures to
control parasitic infection in man : Avoid skin
contact with soil that may be contaminated,
Avoid eating food that could be contaminated,
Avoid skin contact with dog poop (especially in
parks, where you can’t know if other people’s
pets have hookworm).
Activity 12: Trematodes
Many species of trematodes vary in size and they
are visible to the naked eye. The most distinct
characteristics in their external structures called
acetabula which means ”suckers”. This group is
also known as trematoda since these species
consist structurally “body with holes”. Human
trematode infections are confined largely in the
tropical regions and the orient. Their distribution
necessitates an appropriate molluscan
intermediate host in which the diseases may be
readily introduced and become endemic.
Human trematode infections are confined largely
in the tropical regions and the orient. Their
distribution necessitates an appropriate
molluscan intermediate host in which the
diseases may be readily introduced and become
endemic.
MORPHOLOGICAL CHARACTERISTICS OF
THE REPRESENTATIVE TREMATODES
● The male measures 12 mm by 0.5 mm
● Female measures 20 mm by 0.4 mm
● Both sexes have a strong sucker around the
mouth
● Tegument of the worms is coated with tiny
spines, ridges, and sensory
organs.
● Ovoidal, rounded or pear shape
● Thin shell pale yellow curve hook or spine or
lateral knob
● Eggs are shed in stool
● One of the largest flukes in the world
● Adult worm has a very characteristic leaf shape
with the anterior and
being broader than the posterior end and
anterior cone shaped projection
● Adult worm averaging 30 mm in length and 13
mm in width
● One of the largest flukes in the world
● Eggs are operculated and averaged 140 mm in
length and 75 mm in width
THE MODE OF ENTRY AND METHOD OF
REPRODUCTION OF EACH REPRESENTATIVE
TREMATODES
SCHISTOSOMA JAPONICUM
● Can penetrate the skin of the person who are
swimming, bathing, or washing in
contaminated water
● Within several weeks these parasites will evolve
into adult worms in the blood
vessel of the body where the female produces
eggs
● Some eggs may travel through the bladder or
intestine
● These species reproduces sexually, male and
female worm must mate in the veins of
the host for the female to lay eggs
FASCHIOLA HEPATICA
● No vector for transmission
● Occurs through ingestion of raw, freshwater
vegetation on which the flukes on their
metacercariae form are encysted
● Produce both sexually and asexually, adults
are hermaphrodite capable of both cross and self-
fertilization
DIAGNOSIS OF SHISTOSOMA JAPONICUM
AND FASCIOLA HEPATICA INFESTATION CAN
BE ESTABLISHED
SCHISTOSOMA JAPONICUM
● Can be diagnosed through the examination of
stool which will be examined
microscopically, but these eggs tend to be passed
intermittently and in small amount and may not
be detected so blood (serologic) test may be
necessary to perform
FASCHIOLA HEPATICA
●Most widely used form of diagnosis for fasciola
hepatica is either a stool sample, duodenal
aspirate or bilary aspirate. flukes do not begin to
produce eggs until 4 months after infection.
TREATMENT OF CHOICE
SCHISTOSOMA JAPONICUM
● Can be treated through Anthelmintics,
specifically Praziquantel it works by affecting the
worms motor work, spasmodic contraction of the
musculature and vesicle formation in tegument
FASCHIOLA HEPATICA
● Triclabendazole, a benzimidazole compound
active against immature and adult fasciola
hepatica. Like other benzimidazole the
mechanism of action of triclabendazole results
from inhibition of microtubule formation.
PREVENTIVE MEASURES
Outbreaks hymenolepiasis in children’s
institution can be controlled by chemotherapy
and prevented by improvement of hygiene.
● Intensive hookworm infections among
plantation workers may indicate the need for
better sanitary facilities on plantation and more
effective health education, as well as specific
treatment of those heavily infected. Some eggs
may travel through the bladder or intestine
● Foci of T. solium taeniasis and cysticercosis,
created by sale of infected pork products or the
importation of infected pigs, may need the
coordinated intervention of medical and
veterinarian services.
● In the case of waterborne outbreaks of
amoebiasis or giardiasis the source of infection
may need to be determined, followed by sanitary
measures to stop further spread of infection.
well as by having efficient sewage treatment
systems.
● In the case of waterborne outbreaks of
amoebiasis or giardiasis the source of infection
may need to be determined, followed by sanitary
measures to stop further spread of infection.
Intensive hookworm infections among plantation
workers may indicate the need for better sanitary
facilities.

● The control of outbreaks of ascariasis caused
by agricultural use of inadequately treated faecal
wastes (nightsoil) requires chemotherapy for
those affected and improvements in excreta
disposal methods.
● The control of outbreaks of ascariasis caused
by agricultural use of inadequately treated faecal
wastes (nightsoil) requires chemotherapy for
those affected.

CONCLUSION

Both sexes of Schistosoma japonicum have a

strong sucker around the mouth. The male
measures 12 mm by 0.5 mm and the female
measures 20. The female shape is ovoidal,
rounded or pear shape and eggs are shed in
stool.

● Faschiola hepatica is one of the largest flukes

in the world. Adult worm has a characteristic leaf
shape. Faschiola Hepatica is one of the largest
flukes in the world. Adult worm is 30 mm in
length and 13 mm in width

● Schistosoma japonicum can penetrate the skin

of the person who are swimming, bathing, or
washing in contaminated water. These parasites
will evolve into adult worms in the blood vessel of
the body where the female produces eggs.
Fasciola hepatica is a species of metacercariae.
Adults are hermaphrodite capable of both cross
and self-fertilization.

● Schistosoma japonicum can be diagnosed

through the examination of stool which will be
examined microscopically. These eggs tend to be
passed intermittently and in small amount and
may not be detected so blood (serologic) test may
be necessary.

● Schistosoma japonicum can be treated through

Anthelmintics, specifically Praziquantel.
Triclabendazole, a benzimidazole compound
active against immature and adult fasciola
hepatica.

● In regions where hookworm is prevalent, do not

go barefoot. Other skin-to-soil contacts should be
avoided, as should swallowing such dirt. People
can prevent infection by not defecating outside or
by not using human excrement as fertilizer, as