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Mycology and Virology > MODULE 1 UNIT 1 > Flashcards

MODULE 1 UNIT 1 Flashcards

1
Q

study of fungi, termed specifically as

A

“Mycology”

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2
Q

Once in the scientific history, fungi were classified as “plants”. This was derived from the axiom attributed to Carl Linnaeus (father of modern taxonomy) “Plants grow and live, while animal grow, live and [?]”. This attribution begets misleading to the future discovery
and identification of plants.

A

feel

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3
Q

The term Mycology was derived from the Greek word [?] which means fungus (plural form fungi).

A

“mykes”

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4
Q

is the study of fungi encompassing environmental impact, genetic and biochemical properties

A

Mycology

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5
Q

is devoted to the study of fungi, its impact and relationship to human disease.

A

Medical Mycology

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6
Q

Approximately, [?] fungal pathogens out of more than 100,000 known fungal species are medically important, and about [?] of these species were often associated with mycosis.

A
  • 200

- 50 to 60

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7
Q

Fungi play a very important role in various fields including but not limited to environmental, bioeconomy, etc.

A
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8
Q

is devoted primarily to the relationship of fungi to human.

A

Medical mycology

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8
Q
  1. Due to increasing number of [?] environmental molds.
A

ubiquitous

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9
Q
  1. Implicated as [?] pathogens, capable of producing serious or lethal disease in hosts that are immunocompromised (debilitated).
A

opportunistic

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10
Q
  1. One of the leading causes of [?] infection.
A

nosocomial

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10
Q
  1. Often mistaken as [?] infection that produces complicated and fatal consequences.
A

bacterial

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11
Q
  1. Increase [?] (travel to a geographical area where a fungus exists as part of the communal flora of the local population, or is endemic to the area).
A

morbidity

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12
Q
  1. [?] population.
A

Aging

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13
Q

an asexual form of a fungus

A

Anamorph

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14
Q

a cell that produces and extrudes conidia; the tip tapers lengthens and acquires a ring of cell wall material as each conidium is released; oil immersion magnification may be required to see the rings.

A

Annellide

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15
Q

ability of fungus to use a specific carbon or nitrogen source for growth; assimilation is read by the presence or absence of growth

A

Assimilation

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16
Q

enlarged, dome-shaped tip of a sporangiophore that extends into the sporangium.

A

Columella

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17
Q

the cell that produces conidia

A

Conidiogenous cell

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18
Q

a specialized hyphal structure that serves as a stalk on which conidia are formed. The shape and arrangement of the conidiophores and the conidia are generally characteristic of a genus. The suffix phore means “carrying” and is added to the word that denotes what it is carrying; e.g., conidiophores bear conidia and sporangiophores bear sporangia.

A

Conidiophore

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19
Q

An asexual propagule that forms on the side or the end of the hypha or conidiophore.

A

Conidium (plural conidia)

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20
Q

may consist of one or more cells, and the size, shape, and arrangement in groups are generally characteristic of the organism.

A

Conidium

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21
Q

Conidium is always borne [?], i.e., not enclosed within a saclike structure such as a sporangium.

A

externally

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22
Q

If a fungus produces two types of conidia, those that are small and usually single celled are referred to as

A

microconidia

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23
Q

whereas the larger [?] are usually segmented into two or more cells.

A

macroconidia

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24
Q

prefix meaning dark (brownish or blackish)

A

Pheo-

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25
Q

a subcutaneous or systemic disease caused by a variety of black fungi that develop in tissue or dark hyphae and(or) yeast-like cells.

A

Paeohyphomycosis

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26
Q

cut off sharply; ending abruptly with a flattened edge.

A

Truncate

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27
Q

fungi were used as an antiseptic and anesthesia due to the “magical and spiritual properties.

A

35,000 B.C.

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28
Q

people were convinced that association with fungi with entail the formation of disease(s).

A

Middle Ages

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29
Q

fungi were plants with no fruit nor seed

A

Renaissance period

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30
Q

Birth of the first mycological studies. Pier Anton Micheli (founder of modern mycological studies). It is in this period where mycology was separated from botany

A

18th Century

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31
Q

Fungi were recognized as a potential causative agents of diseases that are usually fatal in nature.

A

Mid-20th Century

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32
Q

TAXONOMY, (Gk.: [?]= arrangement; [?] = method).

A
  • taxis

- anomia

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33
Q

Branch of science that deals with systematic biological classification of all living organisms.

A

TAXONOMY

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34
Q

Taxonomy is divided into three (3) disciplines viz.

A

Classification, Nomenclature, and Identification.

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35
Q

Synonym of “systematic” or biosynthetic

A

TAXONOMY

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36
Q

In 2011, a breakthrough in the field of fungal taxonomy occurred: the concept of “One Fungus = One Name” concept.

A
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36
Q

In 2011, a breakthrough in the field of fungal taxonomy occurred: the concept of [?] concept.

A

“One Fungus = One Name”

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37
Q

Throughout the past century, pleomorphic fungi had been defined with two different names based upon the phenotypic identification of the sexual stage (?) or the asexual form (?).

A

teleomorph

anamorph

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38
Q

For example, the anamorph form Blastomyces dermatitidis was known as [?] in the teleomorph form.

A

Ajellomyces dermatitidis

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39
Q

With the advent of numerous advances in molecular taxonomy, the teleomorphic and anamorphic stages of several medically important fungi were confirmed to be the

A

same organism

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40
Q

Recognizing the unique dilemma posed by the identification of pleomorphic fungi having two names, the [?] in Melbourne, Australia, in July 2011 recommended the discontinuation of the dual nomenclature system for fungi with anamorphic and teleomorphic forms

A

Nomenclature Section meeting of the International Botanical Congress

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41
Q

Known as the [?] this recommendation resulted in a historic revision of the International Code of Botanical Nomenclature.

A

Amsterdam Declaration on Fungal Nomenclature

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42
Q

Becoming effective on January 1, 2013, the principle of “One fungus = One name” assigned priority to the [?] independently of whether the organism was originally described as the anamorph or the teleomorph.

A

oldest genus (species) name

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43
Q

is the orderly arrangement of organisms into taxonomic groups on the basis of similarity.

A

Classification

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43
Q

is the orderly arrangement of organisms into taxonomic groups on the basis of similarity.

A

Classification

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44
Q

There are three criteria for taxonomical classification:

A

(i) Cell type
(ii) Level of cellular organization,
(iii) Nutritional status

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45
Q

this pertains whether a define organism is classified as Eukaryotic or Prokaryotic

A

(i) Cell type

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46
Q

discusses whether an organism is a unicellular or multicellular

A

(ii) Level of cellular organization

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47
Q

Nutritional status is divided further according to:

A

a. Food acquisition

b. Energy and carbon source

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48
Q

is the labelling of the units defined.

A

Nomenclature

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49
Q

is the process of determining whether an unknown belongs to one of the units defined in and labeled in.

A

Identification

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50
Q

Taxonomical classification of fungi: Domain

A

Eukarya

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51
Q

Taxonomical classification of fungi: Kingdom

A

Fungi

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52
Q

Taxonomical classification of fungi: Phylum

A

Zygomycota

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53
Q

Taxonomical classification of fungi: Class

A

NA

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54
Q

Taxonomical classification of fungi: Order

A

Mucorales

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55
Q

Taxonomical classification of fungi: Family

A

Mucoraceae

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56
Q

Taxonomical classification of fungi: Genus

A

Rhizopus

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57
Q

Taxonomical classification of fungi: Species

A

arrhizus

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58
Q

Taxonomical classification of fungi: Sub-species

A

NA

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59
Q

is one among the numerous causative agents of Mucormycosis, a fungal infection caused by ubiquitous environmental molds. This mold is one of the causative agents isolated from COVID-19-associated Mucormycosis epidemic in India started last May 2021.

A

Rhizopus arrhizus

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60
Q

The Cell wall structure is rigid contains primarily [?] of polysaccharide plus [?] protein and glycoproteins

A

90%

10%

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61
Q

is the major carbohydrate content consist of repeating monomers of N-acetyl-glucosamine (NAG) that provides shape and protection from external stress factors such as osmotic lysis (unaffected by antibiotics).

A

Chitin

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62
Q

Minor carbohydrates are also present viz.

A

Glucan, Mannan, Chitosan, and Galactosan.

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63
Q

There are various functional properties of the cell envelope. Simply provision of its shape and interference between the fungus and its external environment, acts as [?] for some enzymes, and processes [?] which allow interaction with other organisms.

A
  • binding site

- antigenic properties

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64
Q

Phospholipid structure arranged in a two-layered fashion scattered randomly.

A

Cell Membrane

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65
Q

is the major type of sterol present that regulates solute intake and secretion (transport system) via selective permeability and target of some antibiotics such as Nystatin.

A

Ergosterol

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66
Q

Functions of CM include

A

facilitate biosynthesis of cell wall and capsular material

protects the cytoplasm

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67
Q

This is an external coating covering the cell wall, and few of the fungi exhibits this structure such as Cryptococcus neoformans

A

Capsule

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68
Q

an opportunistic fungi and the causative agent of Cryptococcosis among individuals who are immunosuppressed (eg, People living with HIV, PLHIV).

A

Cryptococcus neoformans

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69
Q

Considered as a virulence factor and compose of amorphous
polysaccharide and may influence the fungal growth by preventing
dissociation of buds and dispersion yeast.

A

Capsule

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70
Q

Bounded by a nuclear membrane and may vary in size, shape, and number. Contains usually 1 nucleolus of mostly RNA.

A

Nucleus

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71
Q

The [?] are linear in configuration, compared to bacterial chromosomes that are often circular.

A

chromosomes

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72
Q

are present at all times for protein synthesis.

A

Ribosomes

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73
Q

functions in energy generation.

A

Mitochondrion

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74
Q

associated cellular organelles of cytoplasm

A

(i) Nucleus
(ii) Ribosomes
(iii) Mitochondrion
(iv) Endoplasmic Reticulum
(v) Vacuoles

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75
Q

all fungi are non-motile, except those groups that belong to

A

Phylum Chytridiomycota (Chytrids)

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76
Q

The ecologically-derived modes of nutrition, fungi are considered as [?] (Gk., chemo = chemical; + hetero = an(other); troph = nourishment) and [?], utilize various organic compounds as sources of nutrients via absorption and capable of growing on dead and living organic matter respectively.

A

chemoheterotrophic

saprophytic

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77
Q

In terms of their atmospheric requirements, fungi are mostly [?] and to some extent few are [?].

A

aerobic

facultative anaerobic

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78
Q

Fungi produce [?], that is utilized for the synthesis of the amino acid lysine de novo through α-aminoadipate pathway and as a precursor for penicillin. This pathway is used as the target for the development of new antibiotics.

A

α-aminoadipate

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79
Q

plays a very important role in the identification of fungi for both macroscopic and microscopic methods.

A

Fungal morphology

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80
Q

Today with the luxury of molecular methods; with the appropriate selection of [?] plus the [?] of isolated fungi remains the key points in differentiating and in identification of fungi.

A

biochemical tests

traditional microscopic and macroscopic observation

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81
Q

Unicellular; Microscopic, characterized by spherical to ellipsoidal, varies in size, with a diameter of 3-5µ.

A

Yeasts

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82
Q

Mode of reproduction of yeast is through an asexual process known as the process of

A

budding

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83
Q

Budding comes in three sequential stages viz,

A

bud emergence, protuberance, and separation.

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84
Q

Yeasts are macroscopic, often observed in culture media (eg, Saboraud’s dextrose, Potato dextrose agar) and exhibits [?] colonies that measure 0.5 to 3.0mm in diameter. Optimum temperature for growth is 35-37ºC.

A

opaque, creamy to pasty

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85
Q

Microscopic, composed of a cylindrical structure called hypha (plural hyphae) which is the basic unit in a fungus.

A
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85
Q

Multicellular, 2-10µ in diameter; Microscopic, composed of a cylindrical structure called hypha (plural hyphae) which is the basic unit in a fungus.

A

Molds

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86
Q

The entire body of the hyphae referred to as [?] (colony surface) or the vegetative body of a fungus

A

thallus

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87
Q

the cross walls that cross sectionally divide a hypha is called

A

septum

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88
Q

is a mat of an intertwined hyphae that constitutes the colony surface of a mold.

A

Mycelium

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89
Q

Macroscopically, molds can be observe via

A

obverse (surface) or reverse (back).

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90
Q

creamy, bright or light gray to brown

A

obverse (surface)

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91
Q

non pigmented to yellow, orange, or red

A

reverse (back)

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92
Q

CLASSIFICATION OF HYPHAE

A
  1. Acc. to existence of septa
  2. Acc. shape and morphology
  3. Acc. to pigment production
  4. Acc. to hyphal growth
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93
Q

(fungus with cross walls), in most cases dichotomous occurs when a hyphae branches into two equal diameter to the hyphae from which they originated

A

Septate

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94
Q

(fungus without cross walls), Coenocytic hyphae results from nuclear division within a cell without division in the cytoplasm

A

Aseptate

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95
Q

the thin walls and lack of regular septation decreases the internal support of these broad hyphae and allows them to become characteristically twisted, collapsed, and folded in a ribbonlike fashion (Mucormycosis).

A

pauciseptate hyphae

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96
Q

characterized by multiple projections in a hypha resembling an old comb hyphae appearance (eg, Microsporum audouinii)

A

Pectinate

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97
Q

marked by hyphae with club-shaped cells, in which the layer end of one cell being attached to the smaller end of an adjacent (eg, Coccidioides sop.).

A

Racquet

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98
Q

the hypha forming a coiled or corkscrew like turns (eg, Trichophyton mentagrophytes)

A

Spiral

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99
Q

terminal hyphae branches that are irregular, broad and antler-like in appearance. (eg, Trichophyton schöenleinii).

A

Favic Chandelier

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100
Q

this is specified by a round knot-like structure formed by intertwined hyphae and seen among dermatophytes.

A

Nodular

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101
Q

produces root-like structure along the vegetative hyphae especially observed among Zygomycetes (eg, Rhizopous and Absidium)

A

Rhizoids

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102
Q

basically transparent, clear and colorless hyphae, as shown from Figure 1.6 among agents of mucormycosis (ie, Saksenaea vasiformis)

A

Hyaline

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103
Q

in comparison to hyaline, the structures of dematiaceous hyphae are brown to black in color, due to melanotic pigment in the cell walls.

A

Dematiaceous

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104
Q

forms filamentous (fuzzy) colonies

A

Apical elongation (macroscopic)

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105
Q

this is when mycelium buried down the culture medium for water exchange and nutrient absorption.

A

Vegetative

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5
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106
Q

— projects on the surface of the medium, with reproductive structures known as spores which can be sexual or asexual.

A

Aerial

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107
Q

move towards the tip of the hyphae (towards the plasma

membrane) where they release various enzymes and other compounds.

A

Cell vesicles

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108
Q

Enzymes release by these particles involved in the

A

lysis and synthesis of the cell wall

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109
Q

Once the new cell has formed, enzymes at the tip start synthesizing a new cell wall around the new cell to protect it. Followed by strengthening of the [?]. Then disappearance of [?]

A

actin cap

Spitzenkörper

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110
Q

play an important role in organizing (regulating) hyphal growth. Found behind hyphal tip (apex)

A

Spitzenkörper

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111
Q

is triggered by stems for the sub apical accumulation of wall precursors (presumable vesicles) reaching a critical concentrations

A

Lateral elongation

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112
Q

is the ability of some fungi to grow into two (2) forms, specifically yeast and mold forms in which dependent on temperature.

A

FUNGAL DIMORPHISM

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113
Q

Yeast

Temperature:
Growth:
Form:

A

35-37ºC
In vivo
Pathogenic

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114
Q

Mold

Temperature:
Growth:
Form:

A

25-30ºC
In vitro
Infective

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115
Q

Some of the notable examples of fungi that exhibits dimorphism include:

A
  1. Sporothrix schenkii
  2. Blastomyces dermatitidis
  3. Histoplasma capsulatum
  4. Coccidioides immitis
  5. Paracoccidioides brasiliensis
  6. Penicilium notatum (P. chrysogenum)
  7. Talaromyces (Penicillium) marneffei
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116
Q

Requires the formation of special clusters for fertilization and nuclear fission

A

Sexual reproduction

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117
Q

— sex organ, gametes (sex cells) and either monoecious and dioecious. It involves mitosis and meiosis

A

Gametangium

118
Q

are developed as a result of a primary nuclear fusion with reduction in chromosome number during their formation. Germinates and forms hyphae and mycelium

A

Sexual spores

119
Q

•2-8 ascospores within an ascus (sac). the shape of each aids in
identification of fungus.

A

Ascospores (eg,, Saccharomyces spp., Ascomycetes)

120
Q

•2-8 ascospores within an ascus (sac). the shape of each aids in
identification of fungus.

A

Ascospores (eg,, Saccharomyces spp., Ascomycetes)

121
Q

2-4 basidiospores on the surface of a club-shaped called

basidium

A

Basidlospores (eg., Basidiomycetes)

122
Q

When two sporangiophores sexually fuse to form a large thick-walled bodies

A

Zygospores (eg.. Mucor & Rhizopus)

123
Q

contains zygospores along a non-septate hyphae

A

Zygosporangium

124
Q

Produced from a fusion of 2 non-identical hyphae

A

Oospores (eg., Phythophthora. phythium, etc.)

125
Q

are preferred terms used when there is a merging of nuclear material or genes combined.

A

Sporulation and spores

126
Q

are formed by budding, no fusion of nuclei takes place in the formation of spore.

A

The asexual spores

127
Q 
Either unicellular (do not form mycelium often produced pasty type of colonies) or multicellular (form mycelium, 
produce
A

filamentous colony

128
Q

Spores contained in sporangia or sacs are produced [?] (at the end)

A

terminally

129
Q

specialized stalk which bears the sporangia

A

Sporangiophore

130
Q

Sporangiophore are observed among

A

Zygomycetes, Rhizopus

131
Q

Abundant club-shaped microconidia and oval-shaped macroconidia containing 5-8 cells.

A

Microsporum gypseum

132
Q

Results from the transformation of a vegetative yeast or specialized hypha called [?].

A

conidiophores

133
Q

Often referred to as macro- (?) and microcondidia (?)

A

multicellular

unicellular

134
Q

Freed by abstraction at the point of attachment

A

Mitospores (Conidiospores)

135
Q

Observed among Ascomycetes and Deuteromycetes

A

Mitospores (Conidiospores)

135
Q
  • Asexual spore formed from a budding process or blowing out along the mycelium or from another blastospore — blastogenesis
  • Blastoconidia
A

Blastospores (Candida spp.)

136
Q

•Formed by fragmentation (segmentation) or disarticulation of hyphae (mycelium) which results to cuffing of rectangular thick-walled spore

A

Arthrospores (eg., Dermatophytes, Coccidioides)

137
Q
  • Formed by budding from a pseudohypha
A

Chlamydospores (eg.. Candida spp.)

138
Q

Chlamydospores are thick-walled resting spores either within (?) the segments: hyphal sides (?); or hyphal tip (?)

A
  • intercallary
  • sessile
  • terminal
139
Q
  • Vesicle. enlarged swollen cell often at the end of a conidiophore or sporongiophore and ¡t bears conidia
  • The conidiospores are formed usually at the sides (tips) of a hyphae and are usually ¡n chains
A

Phialospores (eg., Aspergillus spp.. Penicillium spp.)

140
Q

Perfect fungi**

A

Ascomycetes
Basidiomycetes
Zygomycetes

141
Q

Imperfect fungi

A

Deuteromycetes

142
Q

Ascomycetes

Sexual spore:
Asexual spore:
Hyphae:

A

Ascospore
Any conidiospore
Septate

143
Q

Basidiomycetes

Sexual spore:
Asexual spore:
Hyphae:

A

Basidiospore
Conidiospore
Sepate

144
Q

Sexual spore:
Asexual spore:
Hyphae:

A

Zygospore
Conidiospore
Coenocytic

145
Q

Deuteromycetes

Sexual spore:
Asexual spore:
Hyphae:

A

None
Conidia
Septate

146
Q

are those capable of reproduction both sexual and asexual

A

Perfect fungi

147
Q

reproduced via asexual type only (sporogenesis)

A

Imperfect fungi (fungi imperfecti)

148
Q

CLASSIFICATION OF FUNGAL INFECTIONS

A

Superficial
Cutaneous
Subcutaneous
Systemic

149
Q

Superficial

Tissue Involved (Site of Infection):
Examples:
A

Outer skin layer, hair

Malassezia furfur

150
Q

Cutaneous

Tissue Involved (Site of Infection):
Examples:
A

Skin, hair, nails (keratinized area)

Trichophyton spp.

151
Q

Subcutaneous

Tissue Involved (Site of Infection):
Examples:
A

Connective tissue, muscles, bones

Sporothrix schenkii

152
Q

Systemic

Tissue Involved (Site of Infection):
Examples:
A

All body systems

Histoplasma capsulatum

153
Q

Do not penetrate tissues

A

Superficial

154
Q

Do not penetrate tissues but can cause inflammation

A

Cutaneous

155
Q

Penetrate tissues; transmitted via traumatic inoculation

A

Subcutaneous

156
Q

Thermal dimorphic

A

Systemic

157
Q

Opportunistic

Tissue Involved (Site of Infection):
Examples:
A 
Any part (localized or systemic)
Aspergillus, spp.,; Candida spp.
158
Q

Infects immunosupressed hosts

A

Opportunistic

159
Q
  1. [?] (ie, aseptic technique)
A

Proper collection

160
Q
  1. Never use [?] for fungi
A

anaerobic transport media or anaerobic containers

161
Q
  1. [?] is recommended for transport
A

Room temperature

162
Q
  1. [?] delivery of the samples to the laboratory
A

Rapid

163
Q
  1. [?] into proper and appropriate medium (eg, SDA, Cornmeal)
A

Inoculation

164
Q
  1. [?] at a suitable temperature
A

Incubation

165
Q

Optimum Temperature

Blood and CSF samples
Dermatological (ie, skin, hair, nails)
Others

A

30-37ºC
15-30ºC
4ºC

166
Q

Pretreatment of Clinical Specimens prior to inoculation — importance

  1. Releases fungi [?] within cells
  2. [?] fungal material in the specimen
  3. Help reduce or eliminate bacteria present in contaminated specimens because of the action of mucolytic agents such as [?]
A
  1. enclosed
  2. Concentrate
  3. N-acetyl-L-cysteine, 5%\ oxalic acid, or dithiothreitol (sputolysin)
167
Q

Biosafety Considerations:

  1. All work must be carried out in a certified [?] or higher, whenever is possible
  2. [?] are recommended for personnel working with clinical specimens that may contain dimorphic fungi as well as other potential pathogenic fungi.
  3. [?] must be worn for processing specimens and culture.
A
  1. type 2 laminar-airflow BSC
  2. BSL2 procedures
  3. Gloves
168
Q

I. Skin and interspaces

(i) Wipe lesions and interspaces between toes with [?] to remove surface bacterial contaminants.
(ii) Using the edge of the glass slide or a blunt scalpel blade, firmly scrape the lesion, particularly at the [?]. If multiple lesions are present, choose the most recent for scrapings as old loose scale is often not satisfactory.
(iii) Place scrapings between [?] or place in a clean envelope labeled with the patient’s data.

A

70% alcohol
growing margin
two glass slides

169
Q

II. Nail

(i) Clean nail with [?]
(ii) [?] Scrape outer surface and discard; scrape the deeper portion.
(iii) Remove a portion of debris from under the nail with a [?].
(iv) Collect [?] or nail clippings.
(v) Place all material in a [?] labeled with the patient’s data.

A
  • 70% alcohol.
  • Dorsal plate-
  • scalpel
  • whole nail
  • clean envelope
170
Q

III. Hair

(i) No cleaning of [?] is needed.
(ii) Select infected areas (with scaling or alopecia) and with forceps epilate at least [?]
(iii) For hairs broken off at the scalp level, use a [?]
(iv) Place hairs between [?] or in a clean envelope labeled with the patient’s data.

A
  • scalp
  • 10 hairs.
  • scalpel or a blade knife.
  • two clean glass slides
171
Q

IV. Sputum [tracheal lavage, bronchial lavage, (endo)tracheal aspirate]

(i) Sputum should be [?] and collected in the early morning.
(ii) Have patient remove [?] and rinse vigorously with water.
(iii) Sputum should be the result of a [?] or induced by aqueous aerosol.
(iv) Collect [?] in a sterile container

A
  • fresh
  • dentures
  • deep cough (not saliva)
  • 5-10 ml
172
Q

V. Urine

(i) The urine specimen most suitable for making a diagnosis of mycoses of the urinary tract is a [?] specimen. [?] should be collected when aspiration or cystoscopy cannot be done.
(ii) Early morning specimens are aseptically collected in . [?] have no value. If a delay in processing beyond [?] is anticipated the urine should be placed in a 4°C refrigerator for up to [?] to inhibit the overgrowth of rapidly growing bacteria.

A
  • catheterized; clean-catch, midstream

- sterile containers; Twenty- four-hour collections; 2h; 12-14 hours

173
Q

VI. Blood

(i) Blood is collected aseptically to avoid [?]. The collection site is cleaned with a disinfectant at the time of collection.
(ii) Uses sodium polyanethol sulfonate (SPS, Liquid) as an anticoagulant. Collect 8 ml blood in a yellow vacutainer tube (contains [?] SPS; final concentration with blood is 0.05%).
(iii) The [?] system is ideal for fungal blood culture. Ten ml of blood is directly collected in an isolator tube.
(iv) NOTE: [?] is the method of choice for patients with disseminated Histoplasmosis.

A
  • microbial contamination
  • 1.7 ml of 0.35%
  • Lysis Centrifugation Isolator
  • Blood culture
173
Q

VII.Exudates

(i) The skin over [?] should be disinfected.
(ii) Using a sterile needle and syringe, aspirate material from [?]
(iii) Place the material in a sterile container. The syringe also serves as a transport container if the needle is [?]

A
  • pustular lesions
  • undrained abscesses
  • capped
174
Q

VIII.Vaginal material

(i) Using a [?], collect material from the vagina.
(ii) Insert swabs into a

A
  • sterile swab

- sterile tube

175
Q

IX. Tissue

(i) Tissue is aseptically collected from the [?] of the lesion.
(ii) Place between moist gauze squares, add a small amount of sterile water [?] to keep tissue from drying out and send immediately to the laboratory. Keep refrigerated not exceeding 8-10 hours at 4°C until processed.

A
  • center and the edge

- 0.85% NaCl

176
Q

X. Cerebrospinal Fluid

(i) As much spinal fluid as possible is collected and the placed in a [?]
(ii) Generally, a [?] is used.
(iii) If processing is to be delayed, samples should be stored at [?] and not be refrigerated is an adequate fluid culture in which fungal elements can survive until subcultured.

A
  • sterile container
  • number 3 tube
  • 30-37 °C
177
Q

XI. Bone Marrow

(i) Aspirate approximately [?] of bone marrow and place it in a sterile container.
(ii) [?] can be added as an anticoagulant.
(iii) The [?] may be used.

A
  • 3-5mL
  • SPS or heparin (sodium)
  • pediatric isolated blood collection system
178
Q

XII. Transudates (pleural fluid, synovial, peritoneal)

(i) Specimens are collected aseptically through [?]
(ii) Place in [?]

A
  • thoracentesis, arthrocentesis, and paracentesis.

- sterile containers.

179
Q

DIAGNOSIS OF HUMAN MYCOSIS: TWO APPROACHES

A

I. Clinical Diagnosis

II. Laboratory Diagnosis

180
Q

based on demographic information, patient history (eg, travel, occupation, etc.), predisposing factors, and merely focused on manifestations (ie, signs and symptoms)

A

I. Clinical Diagnosis

181
Q

usually microbiologic (ie, fungal CS), serologic, and molecular approach.

A

II. Laboratory Diagnosis

182
Q

LABORATORY EVALUATION: TWO APPROACHES

A
183
Q

Most organic substances might be confused with fungi are digested in the presence of moderately strong alkaline solutions.

A

Potassium hydroxide (KOH) preparation

184
Q

Digestive capabilities can be enhanced with gentle heating or the addition of 40% dimethyl sulfoxide.

A

Potassium hydroxide (KOH) preparation

185
Q

Digestive capabilities can be enhanced with gentle heating or the addition of 40% dimethyl sulfoxide.

A

Potassium hydroxide (KOH) preparation

186
Q

Used to distinguish fungi in thick mucoid specimens or in specimens that contain keratinous material viz, skin, nails, and hairs.

A

Potassium hydroxide (KOH) preparation

187
Q

10% concentration for skin and soft tissues, body fluids and 20% concentration for nails and hard tissues.

A

Potassium hydroxide (KOH) preparation

188
Q

The polysaccharide capsule of organism is refractory to the particles of ink, and capsules appear as halo around the organism.

A

Colloidal carbon wet mounts (ie, India ink, Nigrosin)

189
Q

Used for visualization of Cryptococcus spp.

A

Colloidal carbon wet mounts (ie, India ink, Nigrosin)

190
Q

Nonspecific, non-immunological fluorochromes that bind to the β-(1⇥3) & β-(1⇥4) polysaccharides, specifically cellulose and chitin of fungal cell walls and fluoresces when exposed to long-wave UV light

A

Calcofluor white (CFW), syn. Blankofluor and Uvitex

191
Q

• The fluorochrome can be mixed with KOH to clear the specimen for easier observation of fungal elements (eg., P. jirovecii) that appear bluish white agains dark background.

A

Calcofluor white (CFW), syn. Blankofluor and Uvitex

192
Q

• Fluorescence microscope is required.

A

Calcofluor white (CFW), syn. Blankofluor and Uvitex

193
Q

Basic mounting medium for fungi that consist of phenol, lactic acid, glycerol, and aniline (cotton) blue dye.

A
194
Q

Basic mounting medium for fungi that consist of phenol, lactic acid, glycerol, and aniline (cotton) blue dye.

A

Lactophenol Cotton Blue (LCB)

195
Q

• Used as both a mounting fluid and stain.

A

Lactophenol Cotton Blue (LCB)

196
Q

Lactic acid acts as a clearing agent and aids in preserving the fungal structures, phenol acts as a killing agent, glycerol prevents drying, and cotton blue (Poirrier’s blue and aniline blue are analogous to cotton blue) gives color to the structures.

A

Lactophenol Cotton Blue (LCB)

197
Q

Fungi (spores and hyphae) are often gram positive but some are stain poorly.

A

Gram stain (Hucker Modification)

198
Q

Differentiates Nocardia form other aerobic actinomycetes.

A

Modified acid-fast stain

199
Q

Fungus are usually seen as small, oval yeast cells is often contained with macrophages, stains blue and has a hyaline halo that represents poorly staining cell wall.

A

Wright’s (Giemsa) stain

200
Q

• For the detection of H. capsulatum in bone marrow and Buffy coat specimens.

A

Wright’s (Giemsa) stain

201
Q

• Also used to visualize trophozoite of P. jirovecii

A

Wright’s (Giemsa) stain

202
Q

• Hydrolyzes the cell with aldehydes, which are often then able to combine with the modified Schiff reagent (Basic Fuchsin) coloring the cell wall carbohydrates a bright pink-magenta.

A

Periodic Acid-Schiff (PAS)

203
Q

• Demonstrates the double-contoured refractile walls of Blastomyces dermatitidis that may not be

A

Periodic Acid-Schiff (PAS)

204
Q

• Fungi, including P. jirovecii, are gray to black; background is green. It also stains Nocardia and other actinomycetes.

A
205
Q

• Silver nitrate precipitates on the fungal cell wall

A

Gomori Methenamine Silver (GMS)

206
Q

• Its drawback is that it often stains fungi too densely to observe structural details.

A

Gomori Methenamine Silver (GMS)

207
Q

• Generally the preferred method for tissue sections

A

Gomori Methenamine Silver (GMS)

208
Q

• Chromic acid oxidizes adjacent hydroxyl groups of cell-wall polysaccharides to aldehydes.

A

Gridley stain

209
Q

• Cell walls purple to magenta, yeasts rose to purple, and capsules deep purple. Background yellow

A

Gridley stain

210
Q

• Enhances, visualization of fungi and their morphology. Natural color of cell walls masked. Tissue response cannot be studied.

A

Gridley stain

211
Q

Specific method of detecting fungi in body fluids.

A

Fluorescent antibody stain (immunofluorescence, IF)

212
Q

• Direct technique: fluorescein-labeled Ab reacts with fungal antigen in cell wall.

A

Fluorescent antibody stain (immunofluorescence, IF)

213
Q

• Indirect technique: unlabeled Ab complexes with fungal antigens. Fluorescein-labeled conjugate reacts with globulins attached to fungal antigens. Cell walls yellow-green.

A

Fluorescent antibody stain (immunofluorescence, IF)

214
Q

• Some use the immunoperoxidase technique. Specific and highly sensitive. Can be used to detect and measure antibodies.

A

Fluorescent antibody stain (immunofluorescence, IF)

214
Q

The mucopolysaccharide in the capsular material of the fungus stains deep rose to red, whereas the other tissue elements stains yellow.

A

Mayer Mucicarmine stain (others: Southgate’s, Alcian blue)

215
Q

• Useful for differentiating C. neoformans (gattii) from other fungi of similar size and shape when found in samples of tissue.

A

Mayer Mucicarmine stain (others: Southgate’s, Alcian blue)

216
Q

• B. dermatitidis and Rhinosporidium seeberi may also react positive.

A

Mayer Mucicarmine stain (others: Southgate’s, Alcian blue)

217
Q

• Good for differentiation of dimorphic fungi and works well on sputum smears

A

Papanicolaou (PAP) stain

218
Q

Isolation of saprophytic & pathogenic fungi from sterile sites. Bacteria may grow in the medium.

A

Brain Heart Infusion Agar (BHIA)

218
Q

(inhibits saprophytic molds),

A

Cycloheximide

219
Q

(inhibits bacteria).

A

Chloramphenicol

220
Q

Best for isolation of pathogenic fungi exclusive of dermatophytes.

A

BHIA + antimicrobials

221
Q

Recovery of fungi from blood or bone marrow

A

BHI Biphasic Blood culture bottles

222
Q

[?] comprises both liquid and solid medium.

A

Biphasic— culture system

223
Q

Isolation of dermatophytes from keratinized material.

A

Dermatophyte test

medium (DTM)

224
Q

produce alkaline metabolites that changes the pH indicator from yellow to red.

A

Dermatophytes

225
Q

inhibit saprophytic fungi and bacteria.

A

Antibiotics

226
Q

SDA + chloramphenicol, cycloheximide, and 1% glucose for the Isolation of dermatophytes.

A

Mycosel (Mycobiotic)

227
Q

Primary recovery of saprobic and pathogenic fungi; also dermatophytes

A

Sabouraud Dextrose Agar (SDA)

228
Q

SDA is consist of pancreatic

A

digest of casein, peptic digest of animal tissue, & dextrose at 4% conc’n & buffered at a pH of 5.6

229
Q

Contain chloramphenicol; Similar to caffeic acid agar.

Breakdown of the medium by the phenol oxidase resulting to the production of melanin

A

Birdseed (Niger) Agar

230
Q

For the isolation of C. neoformans that appear as dark brown to black colonies

A

Birdseed (Niger) Agar

231
Q

Tween 80, a surfactant, isspecifically incorporated in lieu of dextrose for the demonstration of pseudohyphae, chlamydospores, and arthrospore formation

A

Cornmeal agar with Tween 80

232
Q

Use for the cultivation and differentiation of Candida spp. on the basis of morphological characteristics. Used in micro (slide) culture

A

Cornmeal agar with Tween 80

233
Q

Conversion of mold phase of B. dermatitidis to yeast phase

A

Cottonseed agar

234
Q

Sucrose is the sole carbon source, with NaNO3 serving as the sole nitrogen source

A

Czapek-Dox Agar

235
Q

Use for the differentiation of Aspergillus spp.

A

Czapek-Dox Agar

236
Q

Stimulate production of spores and sporulating bodies

A

Potato Dextrose Agar (PDA)

237
Q

Demonstrates pigment production of Trichophyton rubrum. Used in slide (micro) culture

A

Potato Dextrose Agar (PDA)

238
Q

Identification of Microsporum audouinii

A

Rice medium

239
Q

Nutritional requirement tests for the differentiation of Trichophyton spp. Contains either Casein agar base of NH4NO3 agar + inositol, thiamine, or nicotinic acid

A

Trichophyton Agars

240
Q

Confirmation of nitrate reduction by C. neoformans

A

Nitrate reduction medium

241
Q

Detection of urease production by C. neoformans and for differentiation of T. mentagrophytes from T. rubrum

A

Urea Agar

242
Q

Detection of carbohydrate (eg., dextrose, maltose, lactose, etc.) assimilation through utilization of carbon or nitrogen of yeast in the presence of oxygen

A

Yeast Assimilation Media (CTA assimilaiton Sugars)

243
Q

Identification of yeast by fermentation reactions with various carbohydrates

A

Yeast Fermentation Broth

244
Q

Large (7,500mm2)

A

Petri DIsh

245
Q

Small (1,500mm2)

A

Test Tube

246
Q

Good Oxygen supply

A

Petri DIsh

247
Q

Poor Oxygen supply

A

Test Tube

247
Q

Relatively fast Rate of drying

A

Petri DIsh

248
Q

Relatively slow Rate of drying

A

Test Tube

249
Q

Poor (lid is easily displaced) Security of closure

A

Petri DIsh

250
Q

Good Security of closure

A

Test Tube

251
Q

Relatively large Probability of dissemination

A

Petri DIsh

252
Q

Relatively small Probability of dissemination

A

Test Tube

253
Q

Relatively easy Detection of mixed culture

A

Petri DIsh

254
Q

Relatively difficult Detection of mixed culture

A

Test Tube

255
Q

OTHER MATERIALS and CONSIDERATIONS for FUNGAL CULTURE

A
  1. Teasing needle (inoculating needle)
  2. Incinerator
  3. Sand overplayed with disinfectant
  4. Incubation of culture
256
Q

Incubation of culture:
• [?], mold forms, most common temperature for incubation.
• [?], opportunistic and dimorphic organisms
• [?], conversion to yeast phase for dimorphic fungi.

A

25-30ºC
30ºC
25-37ºC

257
Q

< 5 days

Saprobes, opportunistic fungi, yeast

A

Rapid growers

258
Q

6 - 10 days

Subcutaneous & opportunistic fungi, dermatophytes

A

Intermediate growers

259
Q

> 11 days

Systemic & subcutaneous fungi

A

Slow growers

260
Q

observed at the entire verse or reverse side of the colony.

A

Pigment production

261
Q

have different pigment on each side, both colors should be recorded.

A

Dermatophytes

262
Q

Most are [?], having olive- green to dark brow —black color.

A

dematiaceous

263
Q

Some are hyaline (?) pastel or colorless colonies.

A

moniliaceous

264
Q

Appears leather-like, or waxy little mycelium, seems to merge with the agar

A

Glabrous

265
Q

Resembles plush or suede, short aerial hyphae of equal length

A

Velvety

266
Q

Resembles colonies of other Staphylococcus spp. (formerly CoNS), “bacteria like” but more dry and dull (waxy-pasty). No aerial mycelium, with a delicate fringe around the colonies in BAP

A

Yeast like

267
Q

Wooly or “Floccose”, large quantities of long aerial hyphae that becomes entangled and may fill the entire petri dish

A

Cottony

268
Q

Powdery due to heavy conidiation or sporulation, has even hyphae and abundant conidia

A

Granular

269
Q

Presence of radial groves from the center of the culture toward the rim

A

Rugose

270
Q

Random folds (long, short, parallel at right angles or combination)

A

Folded

271
Q

Have central depression (concavity) surrounded by raised edges. Resembles S. pneumoniae colonies

A

Crateriform

272
Q

Have many warts or rough knobs on the surface

A

Verrucose

273
Q

Brain-like convolutions

A

Cerebriform

274
Q
  1. Tease mount
    (i) Place [?] of the mounting fluid (eg., LCB) on the slide.
    (ii) Gently remove a [?] from the agar and place it on the slide
    (iii) Using two [?], tease the strands apart.
    (iv) Place a [?] and examine under a microscope
A
  • 1-2 drops
  • portion of the colony
  • teasing needles
  • coverslip
275
Q
  1. Scotch tape method
    (i) Wrap a piece of [?] around a tongue blade, with the sticky side out.
    (ii) Touch the colony with the [?]
    (iii) Lay the tape, sticky tape down on a [?] on a slide.
A
  • clear cellophane tape
  • sticky tape.
  • drop of mounting fluid
276
Q

Slide culture

(i) Cut a [?] form an agar (eg., SDA, Cornmeal, PDA), then transfer on a sterile glass slide.
(ii) Inoculate all sides of the agar block, then cover with a cover slip. (?)
(iii) Place the set-up in a [?] (wet house) and incubate until growth appears.
(iv) Prepare a sterile glass slide with [?] of mounting fluid (LCB).
(v) Carefully remove the coverslip from the block by [?]
(vi) [?] the coverslip carefully onto the mounting fluid.
(vii) [?] and examine under a microscope.

A
  • square block (1cm2)
  • 22x22mm
  • moist chamber
  • 1-2 drops
  • lightly twisting.
  • Lower
  • Set aside
277
Q

Conidia and spores form a fresh culture are washed off in sterile water and placed in labelled vials

A

Storage in water

278
Q

Labelling glass tubes with a screw-capped and placed in a freezer (-70ºC)

A

Freezing

279
Q

Layering an entire slant with mineral oil, capping the tube tightly and storing at room temperature

A

Mineral oil overlays

280
Q

Freeze drying with the use of special equipment (lyophilizer)

A

Lyophilization

281
Q

Antibody detection may be useful particularly when they occur in immunocompetent individuals.

A

Immunologic Methods

282
Q

Several immunological techniques have been used to detect antibodies during fungal infections viz.:

A

(i) Immunodiffusion (ID)
(ii) Countercurrent immunoelectrophoresis (CIE)
(iii) Enzyme-linked immunosorbent assay (ELISA)
(iv) Complement-fixation test (CFTs)
(v) Fluorescent-enzyme immunoassay (FEIA)

283
Q

Ab detection is helpful in diagnosing allergic bronchopulmonary aspergillosis (ABPA), aspergilloma, and Chronic cavitary aspergillosis

A

Aspergillus spp.

284
Q

Limitation: lack of sensitivity due to cross-reactivity with dimorphic fungi

A

Blastomyces spp.

285
Q

Sensitivity 62%

Specificity 53%

A

Candida spp.

286
Q

Ab detection can be a marker for reactivation of disease in solid organ transplant recipient

A

Cryptococcus spp

287
Q

Aspergillus spp.

Assay:
Antibod(ies)/Antigen(s) detected:

A
  • CIE, FEIA, EIA

- Aspergillus IgG Aspergillus IgG

288
Q

Blastomyces spp.

Assay:
Antibod(ies)/Antigen(s) detected:

A
  • Commercial kits

- Antigens (eg., A Ag, WI1 Ag)

289
Q

Candida spp.

Assay:
Antibod(ies)/Antigen(s) detected:

A
  • EIA

- Mannan Ag Candida IgG, IgM, & IgA

290
Q

Cryptococcus spp.

Assay:
Antibod(ies)/Antigen(s) detected:

A
  • EIA

- Cryptococcus IgG, IgM, IgA; Ag: Glucuronoxylmannan

291
Q

• A polysaccharide present in the cell wall of many fungi viz., Aspergillus, Candida, Fusarium with little or none on the following viz., Cryptococcus, Blastomyces, Mucoraceous molds.

A

1,3-β-D-glucan detection

292
Q

• Found in the blood of patients with invasive fungal infections, and its ability to activate factor G of the horse-shoe crab coagulation pathways that allows it to be measured and quantified

A

1,3-β-D-glucan detection

293
Q

Molecular Methods
Advantages:
(i) Has the ability to detect organisms that are present in [?] of that cannot be cultured.
(ii) Removal of the risks assoc. with culturing category 3 (BSL 3 fungi)— especially for
(iii) Decrease time for identification of the agent, & the potential to detect [?] in primary clinical specimens.
(iv) Offers the potential to identify pathogens in biopsy samples that have been [?] and even [?]

A
  • small numbers
  • H. capsulatum
  • anti fungal resistance
  • formalin-fixed; wax-embedded.
293
Q

Molecular Methods
Advantages:
(i) Has the ability to detect organisms that are present in [?] of that cannot be cultured.
(ii) Removal of the risks assoc. with culturing category 3 (BSL 3 fungi)— especially for
(iii) Decrease time for identification of the agent, & the potential to detect [?] in primary clinical specimens.
(iv) Offers the potential to identify pathogens in biopsy samples that have been [?] and even [?]

A
  • small numbers
  • H. capsulatum
  • anti fungal resistance
  • formalin-fixed; wax-embedded.

Mycology and Virology (20 decks)

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