BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

NEISSERIA SPP. VIRULENCE FACTOR OF NEISSERIA

• Aerobic, Nonmotile, Gram negative • Receptors for human transferrin
diplococci o Allow it to compete for iron
• All are diplococci except N. elongata, N.
weaver and N. bacilliformis that are rod • Capsule (N. meningitidis)
shape. o Prevent phagocytosis
• All species are catalase (+), cytochrome
oxidase (+) except: N. elongata and N. • Pili or Fimbrae
baciliformis o Exhibits hairline projection
• Capnophilic o Five distinct colony types
• Natural habitat: Mucous membrane of RT & ▪ T1 and T2 posses pili and
UGT. virulent while;
• N. gonorrhea and N. meningitides are human ▪ T3 to T5 do not have pili
pathogen o Aid in attachment to host tissues
• N. weaver are commensal in URT of dogs. o Help prevent phagocytosis
o Aid in exchange of genetic material
GENERAL CHARACTERISTICS from cell to cell (sex pili)
• Neisseriacease
o Neisseria, Kingella, Eikenella, • Antigenic variation of pili
Simonsiella, Alysiella o Have T1 though T2 upon isolation,
• Neisseria but subculture they lose pili,
o Aerobic, capnophilic, gram-negative becoming T3 through T5
diplococci o Functions to evade the immune
o Oxidase- and catalase-positive system
• N. elongata and N. bacilliformis are
exceptions • Cell-outer membrane proteins
o Catalase-negative and rod-shaped o Antigenic variation
• N. weaver is also an exception o Causes production of useless
o Catalase-positive and rod-shaped antibodies
• Exist as usual flora in the upper respiratory ▪ Bind to pathogen and prevent
and urogenital tracts binding of effecttive
o Meaning most isolates are not antibodies
pathogenic and are routine normal
flora • Lipooligosaccharide (LOS) or Endotoxin
o N. mengitidis can be commensal o Lipid A moiety
inhabitant in carriers o Core LOS
• Primary pathogens o Factor that mediates damage to body
o N. gonorrhoeae and N. meningitidis tissue and elicits inflammatory
response
• Characterisic of the Family Neisseriaceae
o Contains blebs
o Tissue damage

• Immunoglobulin A (IgA) protease

o Cleaves IgA on mucosal surfaces
o Disseminated infections
o Anal canal

MAJOR OUTER MEMBRANE PORIN PROTEINS

• Porin Protein A/B
o Effective against inflammatory
• response and complement killing

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 BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

o Channel for nutrient to pass into o Epididymis: inflammation of small

waste product exit the cell. coiled tube of testicle
o Por A & B for N. meningitidis, Por B: o Urethral structure: narrowing of the
N. gonorhhea urethra
o Prostatis: inflammation of the
• Protein II (opacity) prostate gland
o Facilitate adherence of phagocytosis
and epithelial cells CLINICAL MANIFESTATIONS IN WOMEN
• DISEASE IN THE FEMALE
• Protein III (reduction modified protein, o As many as 50% are asymptomatic
Rmp) o Endocervix is most common site of
o Blocks the host serum bactericidal infection
action (IgG) • SYMPTOMS (if symptomatic)
o Dysuria
NEISSERIA GONORRHEAE o Cervical discharge
o Lower abdominal pain
• Gonorrhea is acute pyogenic infection of
• COMPLICATIONS
noncilliated columnar and transitional
o Pelvic inflammatory disease (PID)
epithelium.
o Sterility
• Acquired thru sexual contact
o Ectopic pregnancy
(transmission)
o Perihepatits (Fitz-Hugh-Curtis
• Occur in urethra, endocervix, anal canal, syndrome)
pharynx, conjunctiva of the eye
• Less likely infections – disseminated VIRULENCE FACTOR OF NEISSERIA
infections, anal canal GONORRHEAE
• Secondary cause of STD. 1. Pili (T1-T2 virulent, T3- T5 avirulent)
• Men are symptomatic while women are • Exchange of genetic material
asymptomatic • Attachment to human mucosal cell
• SPS inhibits N. gonorrheae (Gelatin is invasion
added) • Inhibits phagocytosis
• Newborn: Ophthalmia neonatorum: 2. Capsule, Lipooligosaccharide Protein II
gonococcal eye infection during vaginal (Opa)
delivery • Adhere to phagocytic and epithelial
• Humans are the only natural host cell
o Gonorrhea: mean “flow of seed” 3. Protein II (RMP)
(purulence mistaken for semen) • Block bactericidal effect of IgG
o Also called clap, from french word 4. Outer membrane Porin(PorB)
brothel (clapoir) • Protection from the host immune
• Reservoir: asymptomatic carriers response
CLINICAL MANIFESTATIONS IN MEN
• DISEASE IN THE MALE
o Incubation period: 2 to 7 days
o Transmitted only by intimate
sexual contact
o 95% of infected men show
symptoms of acute infection
• SYMPTOMS
o Dysuria, urethral discharge
• COMPLICATIONS

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 BACTERIOLOGY LECTURE MIDTERMS
NEISSERIA AND MORAXELLA
LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022
DISEASE AND INFECTION OF
GONORRHEAE
• Acute purulent urethritis, prostatitis, and
epididymitis: Males
• Acute cervicitis: Females
• pharyngitis, anorectal infections,
• conjunctivitis (e.g., ophthalmia
neonatorum of newborns acquired
during birth from an infected
mother).
• Pelvic inflammatory disease
(PID) may cause sterility
ectopic pregnancy or perihepatitis
also referred to as
Fitz-Hugh–Curtis syndrome.
INFECTIONS IN OTHER SITES
DISSEMINATED GONOCOCCAL DISEASE
(LESS THAN 1%)
• Blood-borne dissemination
o Fever, intermittend bacteremia,
and rash on the extremities may be
present
NEISSERIA & MORAXELLA
o Mostly from aginine, hypoxanthine,
and uracil (AHU) strains affecting
women
o Blood-borne media containing
sodium polyanethol sulfonate
(SPS) inhibits N. gonorrhoeae
• Gonococcal arthritis
o Purulent arthritis
o Result of disseminated gonococcal
bacteremia
• Extragenital infections
o Pharyngitis
o Anorectal infections
▪ Most common in men who
have sex in men
▪ 30% to 60% of womn with
genital infection can get
N. anorectal infection
• Disease in newborns
o Ophthalmia neonatorum
▪ Occurs during vaginal
delivry
o Infection is preventable with the
application of eye drops at birth
(erythromycin)
LABORATORY DIAGNOSIS
MICROSCOPY
• Presence of intracellular Gram negative
diplococci, coffee or kidney shaped with
presence of PMN or pus cells.
• Pharyngeal specimen unnecessary to do
gram stain
• >5 PMN: Non gonococcal urethritis
CLINICAL SPECIMENS
• Genital sites
• Normal handling
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 BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

• Urethra in males VERIFY BACTERIA ARE GRAM-NEGATIVE

o Collect purulent discharge DIPLOCOCCI
o No discharge, insert swab (Dacron • Penicillin disk test
or rayon) 2cm in erthra and slowly
rotate CULTURE MEDIA
• Endocervix in females • Chocolate Agar is media of choice
• Thayer Martin
CLINICAL SPECIMENS FROM AREAS OTHER • Modified Thayer Martin
THAN GENITAL SOURCES • Martin Lewis
• Rectal culture • New York City
o Insert swab 4 to 5 cm into the anal • GC LECT
canal
• Oral/pharyngeal OXIDASE TEST
• Eye • Test on filter paper or directly in plate
• Blood/joint fluids • Oxidase reagent used: 1% dimethyl-p-
phenylenediamine dihydrochloride &
CULTURE OF N. GONORRHOEAE tetramethyl-p-phenylenediamine
• Chocolate agar supplemented with dihydrochloride
inhibitors for gram-positive, gram- • Pink initially turning black
negative, and yeast • (+): Purple color/ blue color
o Prevent overgrowth of normal flora
• Inoculated culture media must be
incubated at 35 degrees in 3% and 5%
carbon dioxide (CO2)
o Candle extinction jar
• Colony morphology on modified Thayer-
Martin (MTM) agar
o Small, gray, translucent, raised

TRANSPORT SYSTEM
• Transgrow James E Martin Biological
Environmental Chamber (JEMBEC),
Gonopak. Bio-Bag, Amies medium
• If direct plating is impossible: Amies
medium w/ charcoal (6 hours)
• Special media with capnophilic
atmosphere
o Specimen rolled in a Z patterm on CARBOHYDRATE UTILIZATION TEST
media Cystine Trypticase Agar (CTA)
o Contain 1% of a single carbohydrate
DIRECT MICROSCOPIC EXAMINATION ▪ Glucose, maltose, lactose,
MORPHOLOGY sucrose
• Gram-negative, kidney bean-shaped • Indicator: Phenol red
diplococci o Read in 24 to 72 hours
o In symptomatic males – 89% likely
to be gonococcal infection
o In women – 50% to 70%, less
diagnostic because of similar
vaginal flora

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 BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

OTHER TEST
• Multitest Method
o Enzyme method (substrate +
biochemistry test)
o Also identify Haemophilus

Immunologic Assay
• Coagglutination
o Use monoclonal antibody against
N. gonorrhea. Attached to killed S.
aureus
• Fluorescent Antibody Test
o Monoclonal antibodies to Por
protein – fluorescent tags
o No cross rxn: N. cinerea but w/
cross rn: N. lactamica
• Matrix Assisted Laser Desorption/
Ionization time of flight (MALDI-TOF)
Mass Spectrophotometry
o Identification by unique protein
signatures of the organism
o Protein signatures and based on
size & charge on electric field

Nucleic Acid Assays Method

• Advantages
o Rapid
o Sensitive
• Disadvantages
Rapid Test o Nonamplified probe tests are only
• 2 to 4 hours from pure culture marginally more sensitive than
cervical culture in women
Problems o Not approved for pharyngeal or
rectal specimens
• Weak acid production from glucose
o Do not allow for recovery of an
o Some strains of N. gonorrhoeae
organism to be used for
• Misidentification of N, subflava and N.
suspectibility testing
meningitidis
o Sucrose-negative strains of N.
subflava
• Strains of N. cinerea give positive glucose
reactions

CHROMOGENIC SUBSTRATES
• Detects enzymes that hydrolyze colorless
substrates to produce colored end
products
• Only test isolates from selective media
• Selective media inhibit nonpathogenic
species that can yield false positives for
pathogenic species

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NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

ADDITIONAL METHODS o CHOC: Colorless to gray, convex

• Superoxol test colonies
• Auxotypes – require special additives • Glucose & Maltose (+)
• Rapid O-nitrophenyl-B D
ANTIMICROBIAL RESISTANCE galactopyranoside
• Plasmid-mediated penicillinase-producing o Detects Lactose utilization
N. gonorrhoeae (PPNG) – beta lactamase • Gamma Glutamyl Aminopeptidase (+)
production
• Chromosome-mediated penicillin
resistance (PenR) – alterations in penicillin
binding proteins (PBPs)
• Plasmid-mediated tetracycline resistance
(TRNG)
• Chromosomal-mediated resistance to both
penicillin and tetracucline (CMRNG)
• Chromosomal-mediated spectinomycin
resistance
• Chromosomal-mediated fluoroquinolone
resistance (QRNG) CLINICAL MANIFESTATIONS
• Incubation period is 1 to 10 days by
colonizing mucosa
NEISSERIA MENINGITIDIS o Small number have bacteria enter
• Commensal and invasive in the bloodstream and potentially the
nasopharynx in up to 30% of populaton central nervous system (CNS)
o Transmitted by close contact with o Leads to septicemia and/or
respiratory secretions meningitis
o Primarily affects the infants, • Characterization of meningitis
adolescents, and o Frontal headache
immunocompromised o Stiff neck
o Increased risk with crowded living o Nausea and vomiting
conditions o Sometimes fever
• Site: Rectal and urogenital o Purulent meningitis
• Highest incidence found on the infants • Meningococcemia
• Adhere at the nasopharyngeal mucosa o Appearance of skin petechiae
• Meningococcemia/ sepsis o Purpura
o Occur without meningitis o Tachycardia
o Hypotension
• Disseminated Intravascular Coagulation
• Fulminant disease
• Septic Shock
o Hemorrhage in the adrenal glands
• Waterhouse Friderichsen Syndrome
▪ Waterhouse-Friderichsen
(Hemmorhage of the Adrenal Gland)
syndrome
o Increase in Defieciency in C5 & C8
• Other complications
• Highly fatal (25% even if treated)
o Arthritis, pericarditis, pneumonia,
conjuctivitis, and urethritis
LABORATORY DIAGNOSIS
• Treatment
• Gram stain of CSF: Cytocentrifuge
o Penicillin for confirmed meningitis
• SBA and CHOC agar (72 hours) o Third-generation cephalosporins
o SBA: Medium size, gray and for meningococcemia
convex, encapsulated strains are ▪ Prophylaxis with rifampin
mucoid and green tinge or ciprofloxacin

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NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

• Vaccine available for some subtypes but • Sinusitis and otitis media
not very effective (only for A, C, Y and W- ▪ Third most common cause in
135) children
• Rarely: endocarditis, meningitis, and
tracheitis
MORAXELLACEAE
• Moraxella catarrhalis PREDISPOSING FACTORS
• Acinetobacter • Advanced age
• Psychorobacter • Immunodeficiency
▪ Can have severe infections
MORAXELLA CATARRHALIS • Neutropenia
• Family Moraxellaceae
SPECIMEN COLLECTION AND
• Normal commensal of the respiratory tract IDENTIFICATION
o Opportunistic pathogen of the URT
• Middle ear effusion
• Can cause LRTI in adults with chronic
• Nasopharynx
obstructive pulmonary dse.
• Sinus aspirates, sputum, or bronchial
• Predisposing factor: Age,
aspirates
Immunodeficiency, Neutropenia and
chronic debilitating disease
IDENTIFICATION
• Produce Beta Lactamase
• Oxidase and catalase positive
• Cause of Otitis media and sinusitis
• Asaccharolytic
• Family Moraxellaceae
• Positive DNase and butyrate esterase
• Normal commensal of the respiratory tract reactions
o Important opportunistic pathogen
CULTURE AND DIRECT EXAMINATION
LABORATORY DIAGNOSIS
• SBA and CHOC: smooth opaque, gray to
white colonies
• “Hockey puck”
• Can tolerate 28 Celsius
• Inhibited on gonococcal agar with colistin
• Dnase (+)
• Butyrate esterase (+)

CLINICAL INFECTIONS
• Pneumonia COMMENSAL NEISSERIA SPECIES

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NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

• Normal inhabitants
• Rarely cause disease as opportunistic
infections
• Groups
o Group 1: pathogens
o Group 2: commensals that grow on
selective media
o Group 3: commensals that do not
grow on selective medi

DISEASES OF NONPATHOGENIC NEISSERIA SPECIES

COLONIAL MORPHOLOGY AND PRIMARY SITES OF NEISSERIA AND RELATED ORGANISMS

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 BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

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 BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

CHARACTERISTICS OF SIGNIFICANT SPECIES OF NEISSERIA, MORAXELLA, AND KINGELLA

PATHOGENECITY AND HOST RANGE FOR NEISSERIA AND MORAXELLA

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 BACTERIOLOGY LECTURE MIDTERMS

NEISSERIA AND MORAXELLA

LOVELAINE Z. ROBERTO | NOVEMBER 4, 2022

REVIEW QUESTIONS 9. What will be the solution to prevent the

1. What will be the result of Neisseria inhibition of N. gonorrheae in a SPS
meningitidis in Superoxol test? anticoagulant?
✓ NO BUBBLING ✓ ADD GELATIN
2. Neisseria gonorrheae can grow on the 10. Which of the following is rod shape, gram
following media except negative cocci and produce catalase positive
✓ BLOOD AGAR PLATE result?
3. During the carbohydrates utilization test, ✓ WEAVERI
Glucose, Maltose and Sucrose turn into 11. Women is asymptomatic in Gonorrhea
Yellow. What is the probable organism? infection
✓ NEISSERIA SICCA ✓ TRUE
4. Treatment of choice in patient with Opthalmia 12. Penicillin is the drug of choice for Neisseria
neonatorium? meningitidis
✓ 1% SILVER NITRATE ✓ TRUE
5. What is the reason of using Trimetophrim 13. Moraxella catarrhalis produce Beta
lactate in the Modified Thayer Martin? lactamase.
✓ INHIBIT SWARMING ✓ TRUE
6. All the following are characteristic of 14. NYC is used also for Mycoplasma hominis
Moraxella catarrhalis except and Ureaplasma urealyticum.
✓ CAN GROW ON MTM ✓ TRUE
7. Which among the following Neisseria is 15. Neisseria gonorrheae is nonfastidious
encapsulated? organism
✓ NEISSERIA MENGITIDIS ✓ FALSE
8. All Neisseria are Catalase and Oxidase
Positive except
✓ NEISSERIA ELONGATA

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