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Card 05 - Parasitology: Entamoeba histolytica (AMCB-08)

Reference: Medical Microbiology 9e (Murray), Jawetz Medical Microbiology 28e

Q1. Morphological forms and identifying characteristics of E. histolytica

Answer (Short): E. histolytica has 3 morphological forms:

Form	Size	Key Features
Trophozoite	12-50 µm	Motile, ameboid; cytoplasm has outer hyaline + inner granular zone; contains ingested RBCs (pathognomonic); nucleus has central karyosome + fine peripheral chromatin
Cyst (mature)	10-20 µm	4 nuclei; chromatoid bars with rounded ends; glycogen vacuole
Precyst	Intermediate	1-2 nuclei; transitional form

বাংলায়: E. histolytica-র ৩টি রূপ আছে। Trophozoite হলো সক্রিয় রূপ - এটি মলের মধ্যে দেখা যায়, এর ভেতরে লাল রক্তকণিকা (RBC) থাকা এর সবচেয়ে বড় পরিচয়। Cyst হলো গোলাকার, ৪টি নিউক্লিয়াস আছে, chromatoid bar-এর মাথা গোল।

Q2. Which form is infective, pathogenic, and diagnostic? Why?

Role	Form	Reason
Infective	Mature Cyst (4-nucleated)	Acid-resistant wall survives stomach, releases trophozoite in duodenum
Pathogenic	Trophozoite	Invades intestinal wall, causes ulcers and tissue destruction
Diagnostic	Cyst (in formed stool) / Trophozoite (in diarrheal stool)	Trophozoite with RBC in liquid stool = pathognomonic of active disease

বাংলায়: Cyst হলো infective কারণ এটি পেটের এসিড থেকে বাঁচতে পারে। Trophozoite হলো pathogenic কারণ এটাই টিস্যু ক্ষতি করে। Diagnosis-এ cyst (formed stool) বা trophozoite (দাস্তের মল) দেখা যায়।

Q3. Definitive host, Intermediate host, Mode of infection

Definitive host: মানুষ (Human) - E. histolytica-র কোনো intermediate host নেই
Intermediate host: None (নেই)
Mode of infection: Feco-oral route - contaminated water/food-এ mature cyst গিলে ফেলা; মাছি ও তেলাপোকাও cyst বহন করে

বাংলায়: মানুষই একমাত্র host। দূষিত পানি বা খাবারের মাধ্যমে cyst খেয়ে ফেললে infection হয়।

Q4. Life cycle stages & how E. histolytica produces intestinal ulcer

Life Cycle:

Ingestion of cyst (contaminated food/water)
Cyst passes stomach → reaches duodenum
Gastric acid triggers → excystation → quadrinucleated cyst → 4 trophozoites
Trophozoites colonize large intestine (cecum, ascending colon)
Trophozoites → form precysts → cysts → passed in feces (infective cycle continues)

How ulcer forms:

Trophozoite binds intestinal epithelium via galactose-inhibitable lectin
Releases cytotoxins and proteases
Lysis of epithelial cells → flask-shaped ulcer (small entry point, wide necrotic base in submucosa)
Flask shape = narrow neck through mucosa, expanded necrotic area in submucosa

বাংলায়: Cyst খাওয়ার পর পেটে acid-এ উদ্দীপিত হয়ে trophozoite বের হয়, বড় অন্ত্রে যায়, intestinal wall-এ আটকে cytotoxin ছাড়ে, কোষ নষ্ট করে। Flask আকৃতির ঘা তৈরি হয় - মুখ ছোট কিন্তু ভেতরে বড় necrotic এলাকা।

Q5. Virulence factors of E. histolytica

Gal/GalNAc Lectin - host cell-এ attach করে
Amebapores - pore-forming proteins, host cell membrane ফুটো করে
Cysteine proteases - tissue degrade করে, complement inactivate করে
Phospholipase A - membrane lipid ভাঙে
Host cell lysis - neutrophil lysis করে তাদের toxic contents ছাড়িয়ে দেয়

বাংলায়: E. histolytica রোগ করে কারণ এর lectin দিয়ে কোষে ধরে, তারপর amebapore দিয়ে কোষ ফুটো করে, protease দিয়ে টিস্যু কাটে।

Q6. Lesions produced by E. histolytica + Pathogenesis of Amoebiasis

Lesions:

Flask-shaped ulcers - cecum ও ascending colon-এ বেশি
Secondary bacterial invasion → inflammatory cells, neutrophils
Coalescence of ulcers → large mucosal destruction
Ameboma - granulomatous tumor-like mass on intestinal wall
Liver abscess (most common extraintestinal) - right lobe
Lung, brain, spleen abscess (rare)

Pathogenesis:

Cyst ingested → trophozoite released
Trophozoite attaches via lectin
Cytotoxin release → lytic necrosis
Flask ulcer forms → spreads laterally under mucosa
May penetrate muscularis → serosa → peritoneal perforation
Portal circulation → liver abscess (anchovy paste pus)

বাংলায়: প্রথমে cecum-এ ঘা হয়, এরপর ছড়িয়ে পড়ে। গুরুতর হলে liver-এ abscess হয় যেখানে "anchovy sauce" ধরনের তরল থাকে।

Q7. What is E. dispar? Differentiation from E. coli and E. dispar

E. dispar:

E. histolytica-র morphologically identical non-pathogenic species
Commensal - intestinal tissue invade করে না
RBC contains করে না cytoplasm-এ
PCR বা isoenzyme (zymodeme) analysis দিয়ে আলাদা করতে হয়

E. histolytica vs E. coli vs E. dispar:

Feature	E. histolytica	E. coli	E. dispar
Trophozoite size	12-50 µm	20-30 µm	Same as E.h
Karyosome	Central, sharp	Eccentric, coarse	Central
Peripheral chromatin	Fine, even	Coarse, irregular	Fine, even
RBC in cytoplasm	Yes	No	No
Cyst nuclei	1-4	1-8	1-4
Chromatoid bars	Rounded ends	Splintered/frayed	Rounded
Pathogenic	Yes	No	No

বাংলায়: E. dispar দেখতে একদম E. histolytica-র মতো, কিন্তু রোগ করে না, RBC খায় না। E. coli-র cyst-এ ৮টা পর্যন্ত নিউক্লিয়াস হয়, chromatoid bar-এর মাথা ভাঙা থাকে।

Q8. Amoeboma + Extraintestinal sites + Diagnosis of Hepatic Amoebiasis

Amoeboma:

Intestinal wall-এ granulomatous inflammatory tumor-like mass
Large bowel lumen block করতে পারে
X-ray তে carcinoma-র মতো দেখায়

Extraintestinal sites:

Liver (most common) - right lobe abscess
Lungs - pleuropulmonary amoebiasis
Brain - cerebral abscess
Spleen
Skin/perineum - cutaneous amoebiasis

Diagnosis of Hepatic Amoebiasis:

Imaging: Ultrasound / CT scan / MRI - right lobe abscess (elevation of right diaphragm)
Serology: EIA (Enzyme Immunoassay) - strongly positive
Aspiration: "Anchovy paste/sauce" colored pus - but amebae rarely found in pus (only at abscess wall)
Stool exam: Cyst/trophozoite in stool

বাংলায়: Amoeboma মানে অন্ত্রের দেয়ালে tumor-এর মতো গোটা। Liver abscess diagnose করতে ultrasound + blood test (serology) করা হয়। Abscess-এর পুঁজ anchovy sauce-এর মতো দেখতে।

Q9. "Anchovy Sauce Pus" - কী, কোন parasite, কি isolate করা যায়?

Anchovy sauce pus = হেপাটিক amoebiasis-এ liver abscess থেকে drain করা পুঁজ যা চকলেট/অ্যাঞ্চোভি সস-এর মতো গাঢ় বাদামী রঙের
Parasite: Entamoeba histolytica
Isolate করা যাবে কি? - সাধারণত না। কারণ:
- Trophozoites abscess-এর wall-এ থাকে, center-এ না
- Abscess-এর ভেতরের necrotic material ব্যাকটেরিওলজিক্যালি sterile
- সরাসরি pus-এ ameba পাওয়া যায় না / খুব কম পাওয়া যায়
- Wall থেকে নেওয়া biopsy-তে পাওয়া সম্ভব

বাংলায়: Anchovy sauce পুঁজ হলো liver abscess-এর বৈশিষ্ট্যপূর্ণ তরল। কিন্তু এই পুঁজ থেকে E. histolytica isolate করা যায় না, কারণ ameba পুঁজের মাঝখানে না থেকে abscess-এর দেয়ালে থাকে।

Q10. Amoebic vs Bacillary Dysentery (Macroscopy + Microscopy)

Feature	Amoebic Dysentery	Bacillary Dysentery (Shigella)
Macroscopy
Stool	Scanty, blood-stained mucus; no pus	Copious, mucus + pus + blood
Odour	Offensive (foul)	Less offensive
Ulcer shape	Flask-shaped, discrete	Superficial, diffuse

Microscopy
Exudate	Mainly RBCs, few pus cells	Pus cells (neutrophils) predominant
Charcot-Leyden crystals	Present	Absent
Organism	E. histolytica trophozoites with RBC	Shigella bacilli
WBC in stool	Few	Many (neutrophils)

Most reliable finding: Microscopic - finding of E. histolytica trophozoites containing RBCs in stool is pathognomonic for amoebic dysentery. এটা সবচেয়ে reliable কারণ এটি specific - শুধু E. histolytica-র তে দেখা যায়।

বাংলায়: Amoebic dysentery-তে মলে বেশি RBC থাকে, পুঁজ কম। Bacillary dysentery-তে মলে পুঁজ (neutrophil) বেশি। কিন্তু সবচেয়ে নির্ভরযোগ্য পরীক্ষা হলো microscope-এ মলে E. histolytica trophozoite দেখা যেখানে RBC আছে - এটা দেখলেই নিশ্চিত।

References:

Murray's Medical Microbiology 9e - Chapter 72 (Intestinal Protozoa)
Jawetz, Melnick & Adelberg's Medical Microbiology 28e - Chapter 46

Create a quick reference PDF on E. histolytica pathology

Reading File

Running Command

mkdir -p /tmp/workspace/entamoeba-pdf && ls /tmp/workspace/

Writing File

~/entamoeba-pdf/generate_pdf.py

from reportlab.lib.pagesizes import A4
from reportlab.lib import colors
from reportlab.lib.styles import getSampleStyleSheet, ParagraphStyle
from reportlab.lib.units import cm, mm
from reportlab.platypus import (
    SimpleDocTemplate, Paragraph, Spacer, Table, TableStyle,
    HRFlowable, KeepTogether
)
from reportlab.lib.enums import TA_CENTER, TA_LEFT, TA_JUSTIFY
from reportlab.platypus import PageBreak

OUTPUT = "/tmp/workspace/entamoeba-pdf/Entamoeba_histolytica_Quick_Reference.pdf"

# ── Colors ──────────────────────────────────────────────────────────────────
DARK_BLUE   = colors.HexColor("#1A3A5C")
MED_BLUE    = colors.HexColor("#2E6DA4")
LIGHT_BLUE  = colors.HexColor("#D6E8F7")
ACCENT_RED  = colors.HexColor("#C0392B")
ACCENT_GRN  = colors.HexColor("#1E8449")
ACCENT_ORG  = colors.HexColor("#D35400")
PALE_YELLOW = colors.HexColor("#FFFDE7")
PALE_GREEN  = colors.HexColor("#E8F5E9")
PALE_RED    = colors.HexColor("#FDEDEC")
PALE_GREY   = colors.HexColor("#F4F6F7")
WHITE       = colors.white
BLACK       = colors.black
DARK_GREY   = colors.HexColor("#2C3E50")

# ── Styles ───────────────────────────────────────────────────────────────────
styles = getSampleStyleSheet()

def make_style(name, parent="Normal", **kwargs):
    return ParagraphStyle(name, parent=styles[parent], **kwargs)

TITLE_ST = make_style("TITLE_ST", fontSize=22, textColor=WHITE,
                       fontName="Helvetica-Bold", alignment=TA_CENTER,
                       leading=28, spaceAfter=2)
SUBTITLE_ST = make_style("SUBTITLE_ST", fontSize=11, textColor=LIGHT_BLUE,
                          fontName="Helvetica", alignment=TA_CENTER,
                          leading=16)
SECTION_ST = make_style("SECTION_ST", fontSize=12, textColor=WHITE,
                          fontName="Helvetica-Bold", alignment=TA_LEFT,
                          leading=16, leftIndent=6)
BODY_ST = make_style("BODY_ST", fontSize=9, textColor=DARK_GREY,
                      fontName="Helvetica", leading=13, spaceAfter=3)
BODY_B_ST = make_style("BODY_B_ST", fontSize=9, textColor=DARK_GREY,
                        fontName="Helvetica-Bold", leading=13)
BULLET_ST = make_style("BULLET_ST", fontSize=9, textColor=DARK_GREY,
                        fontName="Helvetica", leading=13,
                        leftIndent=12, firstLineIndent=-8, spaceAfter=2)
SMALL_ST = make_style("SMALL_ST", fontSize=8, textColor=colors.HexColor("#555555"),
                       fontName="Helvetica", leading=11)
TH_ST = make_style("TH_ST", fontSize=8.5, textColor=WHITE,
                    fontName="Helvetica-Bold", alignment=TA_CENTER, leading=11)
TD_ST = make_style("TD_ST", fontSize=8, textColor=DARK_GREY,
                    fontName="Helvetica", alignment=TA_CENTER, leading=11)
TD_L_ST = make_style("TD_L_ST", fontSize=8, textColor=DARK_GREY,
                      fontName="Helvetica", alignment=TA_LEFT, leading=11)
WARN_ST = make_style("WARN_ST", fontSize=8.5, textColor=ACCENT_RED,
                      fontName="Helvetica-Bold", leading=12)
KEY_ST = make_style("KEY_ST", fontSize=9, textColor=ACCENT_RED,
                     fontName="Helvetica-Bold", leading=13)

# ── Helpers ───────────────────────────────────────────────────────────────────
def section_header(title, color=DARK_BLUE):
    tbl = Table([[Paragraph(title, SECTION_ST)]], colWidths=[17.6*cm])
    tbl.setStyle(TableStyle([
        ("BACKGROUND", (0,0), (-1,-1), color),
        ("TOPPADDING",    (0,0), (-1,-1), 5),
        ("BOTTOMPADDING", (0,0), (-1,-1), 5),
        ("LEFTPADDING",   (0,0), (-1,-1), 8),
        ("ROUNDEDCORNERS", [4,4,4,4]),
    ]))
    return tbl

def bullet(text, bold_prefix=None):
    if bold_prefix:
        return Paragraph(f"<bullet>&#x2022;</bullet> <b>{bold_prefix}</b> {text}", BULLET_ST)
    return Paragraph(f"<bullet>&#x2022;</bullet> {text}", BULLET_ST)

def kv_row(k, v, bg=WHITE):
    return [Paragraph(f"<b>{k}</b>", TD_L_ST), Paragraph(v, TD_L_ST)]

def hr():
    return HRFlowable(width="100%", thickness=0.5, color=colors.HexColor("#BDC3C7"), spaceAfter=4, spaceBefore=4)

# ── Page template ─────────────────────────────────────────────────────────────
def add_page_decorations(canvas, doc):
    canvas.saveState()
    w, h = A4
    # Header bar
    canvas.setFillColor(DARK_BLUE)
    canvas.rect(0, h - 18*mm, w, 18*mm, fill=1, stroke=0)
    canvas.setFont("Helvetica-Bold", 8)
    canvas.setFillColor(WHITE)
    canvas.drawString(15*mm, h - 12*mm, "Entamoeba histolytica  |  Quick Reference")
    canvas.drawRightString(w - 15*mm, h - 12*mm, "Parasitology  |  AMCB-08")
    # Footer bar
    canvas.setFillColor(DARK_BLUE)
    canvas.rect(0, 0, w, 10*mm, fill=1, stroke=0)
    canvas.setFont("Helvetica", 7)
    canvas.setFillColor(LIGHT_BLUE)
    canvas.drawString(15*mm, 3.5*mm, "Reference: Murray's Medical Microbiology 9e  |  Jawetz Medical Microbiology 28e")
    canvas.drawRightString(w - 15*mm, 3.5*mm, f"Page {doc.page}")
    canvas.restoreState()

# ── Build content ─────────────────────────────────────────────────────────────
story = []

# ===== COVER / TITLE BLOCK =====
cover_data = [[
    Paragraph("Entamoeba histolytica", TITLE_ST),
], [
    Paragraph("Quick Reference Guide  |  Parasitology (AMCB-08)  |  Rhizopoda", SUBTITLE_ST),
]]
cover_tbl = Table(cover_data, colWidths=[17.6*cm])
cover_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,-1), DARK_BLUE),
    ("TOPPADDING",    (0,0), (-1,-1), 14),
    ("BOTTOMPADDING", (0,0), (-1,-1), 14),
    ("LEFTPADDING",   (0,0), (-1,-1), 10),
]))
story.append(cover_tbl)
story.append(Spacer(1, 8))

# ===== Q1 — MORPHOLOGICAL FORMS =====
story.append(section_header("1.  Morphological Forms of E. histolytica"))
story.append(Spacer(1, 4))

morph_data = [
    [Paragraph("Form", TH_ST), Paragraph("Size", TH_ST), Paragraph("Key Identifying Features", TH_ST)],
    [Paragraph("Trophozoite\n(Vegetative)", TD_ST),
     Paragraph("12–50 µm", TD_ST),
     Paragraph("Motile ameboid form; outer hyaline ectoplasm + inner granular endoplasm; "
               "nucleus with CENTRAL karyosome + fine even peripheral chromatin; "
               "RBCs in cytoplasm (PATHOGNOMONIC)", TD_L_ST)],
    [Paragraph("Pre-cyst", TD_ST),
     Paragraph("10–20 µm", TD_ST),
     Paragraph("Transitional form; glycogen vacuoles present; chromatoid bars appear; "
               "1–2 nuclei; loses motility", TD_L_ST)],
    [Paragraph("Mature Cyst\n(Quadrinucleate)", TD_ST),
     Paragraph("10–20 µm", TD_ST),
     Paragraph("Spherical, thick wall; 4 nuclei (each with central karyosome); "
               "chromatoid bars with ROUNDED ends; glycogen vacuole disappears when mature", TD_L_ST)],
]
morph_tbl = Table(morph_data, colWidths=[2.8*cm, 1.8*cm, 13*cm])
morph_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), MED_BLUE),
    ("BACKGROUND",    (0,1), (-1,1), PALE_YELLOW),
    ("BACKGROUND",    (0,2), (-1,2), PALE_GREEN),
    ("BACKGROUND",    (0,3), (-1,3), PALE_BLUE := colors.HexColor("#EBF5FB")),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 6),
    ("VALIGN",        (0,0), (-1,-1), "TOP"),
    ("ROWBACKGROUNDS",(0,0), (-1,-1), [None]),
]))
story.append(morph_tbl)
story.append(Spacer(1, 6))

# ===== Q2 — INFECTIVE / PATHOGENIC / DIAGNOSTIC =====
story.append(section_header("2.  Infective, Pathogenic & Diagnostic Forms", MED_BLUE))
story.append(Spacer(1, 4))

ipd_data = [
    [Paragraph("Role", TH_ST), Paragraph("Form", TH_ST), Paragraph("Reason", TH_ST)],
    [Paragraph("INFECTIVE", TD_ST), Paragraph("Mature Cyst (4-nucleated)", TD_ST),
     Paragraph("Acid-resistant cyst wall survives gastric acid; excystation occurs in duodenum releasing trophozoites", TD_L_ST)],
    [Paragraph("PATHOGENIC", TD_ST), Paragraph("Trophozoite", TD_ST),
     Paragraph("Only form that invades intestinal wall; produces cytotoxins, proteases; causes flask-shaped ulcers and tissue destruction", TD_L_ST)],
    [Paragraph("DIAGNOSTIC", TD_ST), Paragraph("Cyst (formed stool)\nTrophozoite (diarrheal stool)", TD_ST),
     Paragraph("Trophozoite with RBC in liquid stool = pathognomonic for active invasive disease; cyst in formed stool = carrier state", TD_L_ST)],
]
ipd_tbl = Table(ipd_data, colWidths=[2.6*cm, 4.5*cm, 10.5*cm])
ipd_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), MED_BLUE),
    ("BACKGROUND",    (0,1), (0,1), colors.HexColor("#FDEDEC")),
    ("BACKGROUND",    (0,2), (0,2), colors.HexColor("#FEF9E7")),
    ("BACKGROUND",    (0,3), (0,3), colors.HexColor("#E8F8F5")),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 6),
    ("VALIGN",        (0,0), (-1,-1), "MIDDLE"),
    ("FONTNAME",      (0,1), (0,-1), "Helvetica-Bold"),
    ("TEXTCOLOR",     (0,1), (0,1), ACCENT_RED),
    ("TEXTCOLOR",     (0,2), (0,2), ACCENT_ORG),
    ("TEXTCOLOR",     (0,3), (0,3), ACCENT_GRN),
]))
story.append(ipd_tbl)
story.append(Spacer(1, 6))

# ===== Q3 — HOST & MODE =====
story.append(section_header("3.  Host & Mode of Infection", ACCENT_GRN))
story.append(Spacer(1, 4))

host_data = [
    [Paragraph("<b>Definitive Host</b>", TD_L_ST), Paragraph("Humans only — no animal reservoir needed for life cycle", TD_L_ST)],
    [Paragraph("<b>Intermediate Host</b>", TD_L_ST), Paragraph("NONE", TD_L_ST)],
    [Paragraph("<b>Mode of Infection</b>", TD_L_ST), Paragraph("Feco-oral route — ingestion of mature cysts via contaminated water, food, vegetables; mechanical vectors: flies, cockroaches; oral-anal sexual contact", TD_L_ST)],
    [Paragraph("<b>Infective Dose</b>", TD_L_ST), Paragraph("As few as 1 cyst can cause infection", TD_L_ST)],
]
host_tbl = Table(host_data, colWidths=[4*cm, 13.6*cm])
host_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,-1), PALE_GREEN),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 6),
    ("VALIGN",        (0,0), (-1,-1), "TOP"),
    ("ROWBACKGROUNDS",(0,0), (-1,-1), [PALE_GREEN, WHITE, PALE_GREEN, WHITE]),
]))
story.append(host_tbl)
story.append(Spacer(1, 6))

# ===== Q4 — LIFE CYCLE =====
story.append(section_header("4.  Life Cycle & Flask-Shaped Ulcer Formation", DARK_BLUE))
story.append(Spacer(1, 4))

lc_steps = [
    ("1", "Cyst ingested", "Contaminated food/water; cyst survives gastric acid"),
    ("2", "Excystation", "In duodenum; gastric acid stimulates release of trophozoites; quadrinucleate cyst → 4 trophozoites"),
    ("3", "Colonization", "Trophozoites reach large intestine (cecum, ascending colon); multiply by binary fission"),
    ("4", "Invasion", "Trophozoite attaches via Gal/GalNAc lectin → releases cytotoxins → lytic necrosis of epithelium"),
    ("5", "Flask Ulcer", "Small entry point → narrow neck through mucosa → expanded necrotic area in SUBMUCOSA (flask shape)"),
    ("6", "Spread", "Lateral spread under mucosa; may penetrate muscularis → serosa → peritoneal cavity; portal vein → LIVER"),
    ("7", "Encystation", "Trophozoites → precysts → mature cysts → excreted in feces → complete cycle"),
]
lc_data = [[Paragraph(s, TH_ST), Paragraph(n, TH_ST), Paragraph(d, TH_ST)] for s,n,d in [("Step","Stage","Details")]]
for step, name, detail in lc_steps:
    bg = PALE_YELLOW if int(step) % 2 == 0 else WHITE
    lc_data.append([
        Paragraph(f"<b>{step}</b>", TD_ST),
        Paragraph(f"<b>{name}</b>", TD_L_ST),
        Paragraph(detail, TD_L_ST),
    ])
lc_tbl = Table(lc_data, colWidths=[1*cm, 3*cm, 13.6*cm])
lc_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), MED_BLUE),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 4),
    ("BOTTOMPADDING", (0,0), (-1,-1), 4),
    ("LEFTPADDING",   (0,0), (-1,-1), 5),
    ("VALIGN",        (0,0), (-1,-1), "TOP"),
    ("BACKGROUND",    (0,4), (-1,4), colors.HexColor("#FEF9E7")),
    ("TEXTCOLOR",     (0,4), (1,4), ACCENT_RED),
]))
story.append(lc_tbl)
story.append(Spacer(1, 8))

# ===== Q5 — VIRULENCE FACTORS =====
story.append(section_header("5.  Virulence Factors", ACCENT_RED))
story.append(Spacer(1, 4))

vf_data = [
    [Paragraph("Factor", TH_ST), Paragraph("Mechanism", TH_ST)],
    [Paragraph("Gal/GalNAc Lectin", TD_L_ST), Paragraph("Mediates attachment to host colonic epithelium; essential for cytolysis to occur", TD_L_ST)],
    [Paragraph("Amebapores", TD_L_ST), Paragraph("Pore-forming proteins that insert into host cell membranes → ion leakage → cell death", TD_L_ST)],
    [Paragraph("Cysteine Proteases", TD_L_ST), Paragraph("Degrade extracellular matrix (collagen, fibronectin); inactivate complement; cleave IgA", TD_L_ST)],
    [Paragraph("Phospholipase A", TD_L_ST), Paragraph("Degrades membrane phospholipids contributing to tissue damage", TD_L_ST)],
    [Paragraph("Neutrophil Lysis", TD_L_ST), Paragraph("Trophozoites kill neutrophils → release toxic granule contents → amplifies tissue destruction", TD_L_ST)],
    [Paragraph("Ca²⁺ Dysregulation", TD_L_ST), Paragraph("Irreversible increase in intracellular calcium → host cell membrane permeability changes → cell death", TD_L_ST)],
]
vf_tbl = Table(vf_data, colWidths=[4.5*cm, 13.1*cm])
vf_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), ACCENT_RED),
    ("ROWBACKGROUNDS",(0,1), (-1,-1), [PALE_RED, WHITE]),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 6),
    ("VALIGN",        (0,0), (-1,-1), "TOP"),
    ("FONTNAME",      (0,1), (0,-1), "Helvetica-Bold"),
]))
story.append(vf_tbl)
story.append(Spacer(1, 8))

# ===== PAGE BREAK =====
story.append(PageBreak())

# ===== Q6 — LESIONS & PATHOGENESIS =====
story.append(section_header("6.  Lesions & Pathogenesis of Amoebiasis", DARK_BLUE))
story.append(Spacer(1, 4))

les_data = [
    [Paragraph("Site", TH_ST), Paragraph("Lesion", TH_ST), Paragraph("Notes", TH_ST)],
    [Paragraph("Cecum / Ascending colon", TD_L_ST), Paragraph("Flask-shaped ulcer (primary)", TD_L_ST),
     Paragraph("Most common primary site; small mucosal opening, wide submucosal necrosis", TD_L_ST)],
    [Paragraph("Sigmoid / Rectum", TD_L_ST), Paragraph("Secondary ulcers", TD_L_ST),
     Paragraph("Later lesions due to caudal spread", TD_L_ST)],
    [Paragraph("Intestinal wall", TD_L_ST), Paragraph("Ameboma", TD_L_ST),
     Paragraph("Granulomatous tumor-like mass; may obstruct bowel lumen; mimics carcinoma on imaging", TD_L_ST)],
    [Paragraph("Liver (right lobe)", TD_L_ST), Paragraph("Amebic liver abscess", TD_L_ST),
     Paragraph("Most common extraintestinal site; ANCHOVY SAUCE (chocolate) pus; via portal vein; serologically positive", TD_L_ST)],
    [Paragraph("Lung", TD_L_ST), Paragraph("Pleuropulmonary amoebiasis", TD_L_ST),
     Paragraph("Direct extension from liver abscess through diaphragm; right-sided", TD_L_ST)],
    [Paragraph("Brain / Spleen", TD_L_ST), Paragraph("Cerebral / Splenic abscess", TD_L_ST),
     Paragraph("Rare; haematogenous spread; high mortality", TD_L_ST)],
]
les_tbl = Table(les_data, colWidths=[4*cm, 4*cm, 9.6*cm])
les_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), MED_BLUE),
    ("ROWBACKGROUNDS",(0,1), (-1,-1), [WHITE, PALE_GREY, WHITE, PALE_YELLOW, WHITE, PALE_GREY]),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 6),
    ("VALIGN",        (0,0), (-1,-1), "TOP"),
    ("BACKGROUND",    (0,4), (-1,4), colors.HexColor("#FEF9E7")),
    ("FONTNAME",      (0,4), (1,4), "Helvetica-Bold"),
]))
story.append(les_tbl)
story.append(Spacer(1, 6))

# ===== Q7 — DIFFERENTIATION =====
story.append(section_header("7.  E. histolytica vs E. coli vs E. dispar", DARK_BLUE))
story.append(Spacer(1, 4))

diff_data = [
    [Paragraph("Feature", TH_ST), Paragraph("E. histolytica", TH_ST), Paragraph("E. coli", TH_ST), Paragraph("E. dispar", TH_ST)],
    [Paragraph("Trophozoite size", TD_L_ST), Paragraph("12–50 µm", TD_ST), Paragraph("20–30 µm", TD_ST), Paragraph("12–50 µm", TD_ST)],
    [Paragraph("Karyosome", TD_L_ST), Paragraph("Central, SHARP", TD_ST), Paragraph("Eccentric, coarse", TD_ST), Paragraph("Central, sharp", TD_ST)],
    [Paragraph("Peripheral chromatin", TD_L_ST), Paragraph("Fine, even ring", TD_ST), Paragraph("Coarse, irregular/clumped", TD_ST), Paragraph("Fine, even ring", TD_ST)],
    [Paragraph("RBC in cytoplasm", TD_L_ST), Paragraph("YES ✓ (pathognomonic)", TD_ST), Paragraph("NO", TD_ST), Paragraph("NO", TD_ST)],
    [Paragraph("Cyst nuclei (max)", TD_L_ST), Paragraph("4", TD_ST), Paragraph("8", TD_ST), Paragraph("4", TD_ST)],
    [Paragraph("Chromatoid bars", TD_L_ST), Paragraph("ROUNDED ends", TD_ST), Paragraph("Splintered/frayed", TD_ST), Paragraph("Rounded ends", TD_ST)],
    [Paragraph("Pathogenic", TD_L_ST), Paragraph("YES", TD_ST), Paragraph("NO", TD_ST), Paragraph("NO", TD_ST)],
    [Paragraph("Differentiation from E. dispar", TD_L_ST), Paragraph("PCR, isoenzyme (zymodeme), EIA antigen detection", TD_L_ST), Paragraph("—", TD_ST), Paragraph("Morphologically identical to E.h.", TD_ST)],
]
diff_tbl = Table(diff_data, colWidths=[4.2*cm, 4.5*cm, 4.5*cm, 4.4*cm])
diff_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), MED_BLUE),
    ("ROWBACKGROUNDS",(0,1), (-1,-1), [WHITE, PALE_GREY]),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 4),
    ("BOTTOMPADDING", (0,0), (-1,-1), 4),
    ("LEFTPADDING",   (0,0), (-1,-1), 5),
    ("VALIGN",        (0,0), (-1,-1), "MIDDLE"),
    ("TEXTCOLOR",     (1,4), (1,4), ACCENT_GRN),
    ("FONTNAME",      (1,4), (1,4), "Helvetica-Bold"),
    ("TEXTCOLOR",     (1,7), (1,7), ACCENT_RED),
    ("FONTNAME",      (1,7), (1,7), "Helvetica-Bold"),
]))
story.append(diff_tbl)
story.append(Spacer(1, 6))

# ===== Q8 — AMOEBOMA & HEPATIC DIAGNOSIS =====
story.append(section_header("8.  Amoeboma + Extraintestinal Sites + Hepatic Amoebiasis Diagnosis", ACCENT_ORG))
story.append(Spacer(1, 4))

amoeboma_box = [
    [Paragraph("AMOEBOMA", TH_ST)],
    [Paragraph("• Granulomatous, tumor-like mass on intestinal wall\n"
               "• Due to chronic infection → fibrosis + inflammatory reaction\n"
               "• May block bowel lumen → intestinal obstruction\n"
               "• Mimics carcinoma on X-ray / colonoscopy → biopsy needed to differentiate", TD_L_ST)],
]
amoeboma_tbl = Table(amoeboma_box, colWidths=[17.6*cm])
amoeboma_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0), (-1,0), ACCENT_ORG),
    ("BACKGROUND",    (0,1), (-1,1), colors.HexColor("#FEF5E7")),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 8),
]))
story.append(amoeboma_tbl)
story.append(Spacer(1, 4))

ext_col1 = [
    Paragraph("<b>Extraintestinal Sites</b>", BODY_B_ST),
    Paragraph("1. Liver (right lobe) — MOST COMMON", BULLET_ST),
    Paragraph("2. Lung — pleuropulmonary", BULLET_ST),
    Paragraph("3. Brain — cerebral abscess", BULLET_ST),
    Paragraph("4. Spleen — splenic abscess", BULLET_ST),
    Paragraph("5. Skin — cutaneous amoebiasis", BULLET_ST),
]
ext_col2 = [
    Paragraph("<b>Hepatic Amoebiasis Diagnosis</b>", BODY_B_ST),
    Paragraph("• Ultrasound / CT / MRI: right lobe abscess (elevation of right diaphragm)", BULLET_ST),
    Paragraph("• Serology (EIA): strongly positive in >90% cases", BULLET_ST),
    Paragraph("• Aspiration: ANCHOVY PASTE pus (bacteriologically sterile)", BULLET_ST),
    Paragraph("• Stool microscopy: cysts/trophozoites (often negative)", BULLET_ST),
    Paragraph("• Note: amebae are in abscess WALL, NOT in the pus", BULLET_ST),
]
ext_tbl = Table([[ext_col1, ext_col2]], colWidths=[8.5*cm, 9.1*cm])
ext_tbl.setStyle(TableStyle([
    ("VALIGN", (0,0), (-1,-1), "TOP"),
    ("LEFTPADDING", (0,0), (-1,-1), 0),
    ("RIGHTPADDING", (0,0), (-1,-1), 4),
]))
story.append(ext_tbl)
story.append(Spacer(1, 6))

# ===== Q9 — ANCHOVY SAUCE PUS =====
story.append(section_header("9.  Anchovy Sauce Pus — Key Concepts", MED_BLUE))
story.append(Spacer(1, 4))

anch_data = [
    [Paragraph("What is it?", TD_L_ST),
     Paragraph("Chocolate-brown / dark reddish-brown pus from amebic liver abscess, resembling anchovy sauce in color and consistency", TD_L_ST)],
    [Paragraph("Caused by?", TD_L_ST),
     Paragraph("Entamoeba histolytica trophozoites — lytic necrosis of hepatic tissue produces liquefied necrotic material", TD_L_ST)],
    [Paragraph("Can you isolate parasite from pus?", TD_L_ST),
     Paragraph("NO — trophozoites are located at the WALL of the abscess, not in the central necrotic pus. Aspirated pus is bacteriologically sterile and ameba-free.", TD_L_ST)],
    [Paragraph("How to isolate?", TD_L_ST),
     Paragraph("Biopsy of abscess wall may demonstrate trophozoites; serology (EIA) is the practical diagnostic test", TD_L_ST)],
]
anch_tbl = Table(anch_data, colWidths=[4.5*cm, 13.1*cm])
anch_tbl.setStyle(TableStyle([
    ("ROWBACKGROUNDS",(0,0), (-1,-1), [LIGHT_BLUE, WHITE]),
    ("GRID",          (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0), (-1,-1), 5),
    ("BOTTOMPADDING", (0,0), (-1,-1), 5),
    ("LEFTPADDING",   (0,0), (-1,-1), 6),
    ("VALIGN",        (0,0), (-1,-1), "TOP"),
    ("FONTNAME",      (0,0), (0,-1), "Helvetica-Bold"),
    ("TEXTCOLOR",     (0,2), (0,2), ACCENT_RED),
    ("FONTNAME",      (0,2), (0,2), "Helvetica-Bold"),
]))
story.append(anch_tbl)
story.append(Spacer(1, 6))

# ===== Q10 — AMOEBIC vs BACILLARY DYSENTERY =====
story.append(section_header("10.  Amoebic vs Bacillary Dysentery", DARK_BLUE))
story.append(Spacer(1, 4))

dys_data = [
    [Paragraph("Feature", TH_ST), Paragraph("Amoebic Dysentery\n(E. histolytica)", TH_ST), Paragraph("Bacillary Dysentery\n(Shigella)", TH_ST)],
    [Paragraph("MACROSCOPY", TH_ST), Paragraph("", TH_ST), Paragraph("", TH_ST)],
    [Paragraph("Stool volume", TD_L_ST), Paragraph("Scanty", TD_ST), Paragraph("Copious", TD_ST)],
    [Paragraph("Stool contents", TD_L_ST), Paragraph("Blood + mucus; little pus", TD_ST), Paragraph("Blood + mucus + PUS", TD_ST)],
    [Paragraph("Odour", TD_L_ST), Paragraph("Offensive (foul)", TD_ST), Paragraph("Less offensive", TD_ST)],
    [Paragraph("Ulcer shape", TD_L_ST), Paragraph("FLASK-shaped, discrete", TD_ST), Paragraph("Superficial, diffuse", TD_ST)],
    [Paragraph("MICROSCOPY", TH_ST), Paragraph("", TH_ST), Paragraph("", TH_ST)],
    [Paragraph("Predominant cells", TD_L_ST), Paragraph("RBCs > pus cells", TD_ST), Paragraph("Pus cells (neutrophils) predominant", TD_ST)],
    [Paragraph("Charcot-Leyden crystals", TD_L_ST), Paragraph("PRESENT", TD_ST), Paragraph("Absent", TD_ST)],
    [Paragraph("Organism seen", TD_L_ST), Paragraph("E. histolytica trophozoite with RBC", TD_ST), Paragraph("Shigella bacilli (Gram -ve)", TD_ST)],
    [Paragraph("WBC in stool", TD_L_ST), Paragraph("Few", TD_ST), Paragraph("Many (neutrophils)", TD_ST)],
    [Paragraph("MOST RELIABLE FINDING ★", TH_ST), Paragraph("Trophozoite with RBC in cytoplasm (PATHOGNOMONIC)", TH_ST), Paragraph("Culture of Shigella", TH_ST)],
]
dys_tbl = Table(dys_data, colWidths=[4.5*cm, 6.5*cm, 6.6*cm])
dys_tbl.setStyle(TableStyle([
    ("BACKGROUND",    (0,0),  (-1,0),  MED_BLUE),
    ("BACKGROUND",    (0,1),  (-1,1),  DARK_BLUE),
    ("BACKGROUND",    (0,6),  (-1,6),  DARK_BLUE),
    ("BACKGROUND",    (0,11), (-1,11), ACCENT_GRN),
    ("ROWBACKGROUNDS",(0,2),  (-1,5),  [WHITE, PALE_GREY]),
    ("ROWBACKGROUNDS",(0,7),  (-1,10), [WHITE, PALE_GREY]),
    ("GRID",          (0,0),  (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",    (0,0),  (-1,-1), 4),
    ("BOTTOMPADDING", (0,0),  (-1,-1), 4),
    ("LEFTPADDING",   (0,0),  (-1,-1), 5),
    ("VALIGN",        (0,0),  (-1,-1), "MIDDLE"),
    ("TEXTCOLOR",     (0,1),  (-1,1),  LIGHT_BLUE),
    ("TEXTCOLOR",     (0,6),  (-1,6),  LIGHT_BLUE),
    ("FONTNAME",      (0,1),  (-1,6),  "Helvetica-Bold"),
]))
story.append(dys_tbl)
story.append(Spacer(1, 8))

# ===== QUICK MEMORY AIDS =====
story.append(section_header("★  Quick Memory Aids", ACCENT_GRN))
story.append(Spacer(1, 4))

mem_data = [
    [Paragraph("Flask ulcer", BODY_B_ST), Paragraph("= small mucosal entry → narrow neck → wide submucosal necrosis", BODY_ST)],
    [Paragraph("RBC in trophozoite", BODY_B_ST), Paragraph("= pathognomonic for E. histolytica (E. coli & E. dispar do NOT eat RBCs)", BODY_ST)],
    [Paragraph("Cyst rule: 4 vs 8", BODY_B_ST), Paragraph("= E. histolytica max 4 nuclei; E. coli max 8 nuclei", BODY_ST)],
    [Paragraph("Chromatoid bars", BODY_B_ST), Paragraph("= E. histolytica ROUNDED ends; E. coli SPLINTERED/frayed ends", BODY_ST)],
    [Paragraph("Anchovy pus + no ameba", BODY_B_ST), Paragraph("= ameba at abscess WALL, not in the pus — cannot isolate from aspiration", BODY_ST)],
    [Paragraph("E. dispar vs E. histolytica", BODY_B_ST), Paragraph("= morphologically identical; differentiate by PCR / zymodeme / EIA antigen stool test", BODY_ST)],
    [Paragraph("Most reliable microscopy", BODY_B_ST), Paragraph("= trophozoite with ingested RBCs in diarrheal stool = pathognomonic for amoebic dysentery", BODY_ST)],
]
mem_tbl = Table(mem_data, colWidths=[4.2*cm, 13.4*cm])
mem_tbl.setStyle(TableStyle([
    ("ROWBACKGROUNDS", (0,0), (-1,-1), [PALE_GREEN, WHITE]),
    ("GRID",           (0,0), (-1,-1), 0.4, colors.HexColor("#BDC3C7")),
    ("TOPPADDING",     (0,0), (-1,-1), 5),
    ("BOTTOMPADDING",  (0,0), (-1,-1), 5),
    ("LEFTPADDING",    (0,0), (-1,-1), 6),
    ("VALIGN",         (0,0), (-1,-1), "MIDDLE"),
]))
story.append(mem_tbl)
story.append(Spacer(1, 6))

# ===== BUILD =====
doc = SimpleDocTemplate(
    OUTPUT, pagesize=A4,
    topMargin=22*mm, bottomMargin=15*mm,
    leftMargin=15*mm, rightMargin=15*mm,
)
doc.build(story, onFirstPage=add_page_decorations, onLaterPages=add_page_decorations)
print(f"PDF created: {OUTPUT}")

Running Command

cd /tmp/workspace/entamoeba-pdf && python generate_pdf.py 2>&1

Running Command

ls -lh /tmp/workspace/entamoeba-pdf/Entamoeba_histolytica_Quick_Reference.pdf

Your PDF is ready! Download it here:

Entamoeba histolytica Quick Reference

PDF Document · PDF

PDF-এ কী কী আছে (2 pages, colour-coded):

Section	Content
Q1	Morphological forms table (Trophozoite, Pre-cyst, Mature Cyst) with sizes & features
Q2	Infective / Pathogenic / Diagnostic forms - colour coded
Q3	Host & mode of infection
Q4	Life cycle 7-step table + flask ulcer formation
Q5	All 6 virulence factors (Lectin, Amebapores, Proteases etc.)
Q6	All lesions table (flask ulcer → ameboma → liver abscess → lung/brain)
Q7	E. histolytica vs E. coli vs E. dispar - full comparison table
Q8	Amoeboma definition + extraintestinal sites + hepatic amoebiasis diagnosis
Q9	Anchovy sauce pus - what it is, why ameba can't be isolated
Q10	Amoebic vs Bacillary dysentery (Macro + Micro comparison)
★ Memory Aids	7 key mnemonics for exam (flask ulcer, RBC rule, 4 vs 8 nuclei, etc.)

Blue/red/green colour-coding, header/footer with page numbers, and references from Murray's & Jawetz included.

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