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Chapter 1. Red Blood Cells and Platelet Preservation: Historical Perspectives and Current Trends
Multiple Choice
Identify the choice that best completes the statement or answers the question.
____ 1. Which metabolic pathway is responsible for generating 90% of the ATP for the RBC?
a. Pentose phosphate shunt c. Glycolysis
b. Luebering-Rapoport shunt d. Methemoglobin reductase
____ 2. A unit of blood was returned to the blood bank before it was spiked. Apparently the patient’s IV failed. The
unit of blood was outside the blood bank for 35 minutes. Which of the statements below is most accurate?
a. The unit of blood should be discarded immediately.
b. The unit of blood can be returned to inventory.
c. The unit of blood must be transfused within 4 hours or be discarded at the end of that time.
d. The unit of blood must be transfused with 24 hours.
____ 3. What effect does storage have on platelets?
a. Shrinking c. Repulsion
b. Lysis d. All of these.
____ 4. In the normal hemoglobin-oxygen dissociation curve, what percentage of oxygen is released to the tissues
when PO2 averages 40 mm Hg?
a. 75% c. 100%
b. 25% d. 50%
____ 5. What factors are known to influence platelet metabolism and function?
a. Storage temperature c. Platelet count
b. Initial pH d. All of the above
____ 6. Which of the following red blood cell morphologies may be present on the peripheral blood smear as a result
of loss of RBC membrane?
a. Spherocytes c. Burr cells
b. Target cells d. Schistocytes
____ 7. What does the term autologous transfusion refer to?
a. A parent donating blood for his or her child
b. An individual donating blood for a friend
c. An individual donating blood for a relative
d. An individual donating blood for his or her own transfusion
____ 8. What is the primary function of hemoglobin?
a. Iron metabolism c. Oxygen transport
b. Porphyrin synthesis d. Signal transduction
____ 9. All of the following areas of red blood cell biology are crucial for normal erythrocyte survival except:
a. cellular metabolism. c. site of the ABO antigen attachment.
b. RBC membrane. d. hemoglobin structure.
____ 10. What is the correct biochemical composition of the RBC membrane?
a. 52% protein, 40% lipid, 8% carbohydrate
b. 40% protein, 8% lipid, 52% carbohydrate
c. 8% protein, 52% lipid, 40% carbohydrate
d. 8% lipid, 40% carbohydrate, 52% protein
____ 11. All of the following biochemical changes are associated with loss of red blood cell viability upon storage
except:
a. decreased pH. c. increased ATP level.
b. loss of red blood cell function. d. decreased glucose consumption.
____ 12. Which red blood cell preservative has a storage time of 35 days?
a. ACD c. AS-1
b. CPDA-1 d. CPD
____ 13. The RBC membrane is relatively permeable to all of the following except:
a. chloride. c. bicarbonate.
b. sodium. d. water.
____ 14. Red blood cells frozen using the high-concentration glycerol technique are usually stored at
a. 0oC c. -65oC
o
b. -20 C d. -80oC
____ 15. What is the major biochemical consideration in platelet storage?
a. Glucose metabolism c. Production of carbon dioxide
b. Oxygen supply d. Regulation of pH
____ 16. What would the hemoglobin-oxygen dissociation curve depict in a patient exhibiting clinical signs of
alkalosis?
a. Normal c. Shift to the right
b. Shift to the left d. None of the above
____ 17. Name the main lipid components of a red blood cell membrane.
a. Phospholipid c. Glycolipid
b. Sphingomyelin d. Glycophorin A
____ 18. The ABO blood groups were discovered in 1901 by whom?
a. Charles Drew c. Loutit and Mollison
b. Karl Landsteiner d. Edward Lindeman
____ 19. A standing order of platelets was shipped to your facility by your supplier. It was inadvertently left in the
corner of the department until discovered 36 hours later. What would the appropriate action be for the blood
banker?
a. If the temperature in the box was 22 ± 2°C and the platelet swirl seemed OK, it would be
OK to accept the unit into inventory.
b. The platelets have fallen outside the supplier’s quality assurance. The unit should be
discarded because the pH has probably dropped too low and platelet activation has been
compromised.
c. If the temperature was 1°C to 6°C and the platelet swirl seemed OK, it would be OK to
accept the unit into inventory.
d. If the platelets appeared OK and passed the platelet swirl test after being placed on the
agitator, they could be accepted into the inventory.
____ 20. Which metabolic pathway permits the accumulation of 2,3 diphosphoglycerate (2,3-DPG)?
a. Glycolysis c. Pentose phosphate shunt
b. Luebering-Rapoport shunt d. Methemoglobin reductase
____ 21. All of the following are consistent with a "shift to the right" of the hemoglobin-oxygen dissociation curve
except:
a. increased 2,3-DPG.
b. 50% O2 saturation to tissues.
c. decreased 2,3-DPG.
d. decreased hemoglobin affinity for O2.
____ 22. Why are platelet transfusions performed?
a. Therapeutically to stop bleeding c. Both reasons.
b. Prophylactically to prevent bleeding d. Neither reason.
____ 23. What cryoprotective agent is added to red blood cells upon freezing?
a. Dextrose c. Glycerol
b. Adsol d. All of the above
____ 24. If platelets are to be stored for 5 days on a rotator, what is the optimal storage temperature?
a. 1°C to 6°C c. 35°C to 37°C
b. 20°C to 24°C d. 1°C to 10°C
____ 25. Platelets are transfused to play which role in hemostasis?
a. Maintenance of vascular integrity
b. Initial arrest of bleeding by platelet plug formation
c. Stabilization of the hemostatic plug
d. All of the above
____ 26. Which of the following best describes "integral" membrane proteins?
a. Reside at the cytoplasmic surface of membrane
b. Span the entire membrane surface
c. Form the red blood cell cytoskeleton
d. None of the above
____ 27. How is stroma-free hemoglobin solution prepared?
a. Outdated red blood cells are concentrated, and stroma is removed.
b. Outdated red blood cells are diluted with saline, and stroma is removed.
c. Outdated red blood cells are lysed, and stroma is removed.
d. None of the above
____ 28. What is the normal life span of an RBC?
a. 100 days c. 120 hours
b. 120 days d. 2 days
____ 29. Regarding loss of RBC membrane deformability, all of the following are true except:
a. increase in ATP level.
b. decrease in ATP level.
c. increase in calcium level.
d. decrease in spectrin phosphorylation level.
____ 30. One of the most important controls of hemoglobin's affinity for oxygen is:
a. glucose. c. K+.
b. 2,3-diphosphoglycerate (2,3-DPG). d. Ca++.
____ 31. The normal position of the oxygen dissociation curve depends on three ligands normally found within the
RBC. Which one of the following is not one of these ligands?
a. H+ ions c. 2,3-diphosphoglycerate (2,3-DPG)
b. CO2 d. Na+
____ 32. Which of the following events does not occur while RBCs are stored?
a. 2,3-DPG levels increase.
b. Potassium levels increase.
c. Hgb has a decreased affinity for oxygen carrying capacity.
d. 2,3-DPG and potassium levels increase.
____ 33. In order to maintain ATP levels in stored blood, ______________ can be added to CPD to extend the shelf-
life of stored RBCs from 21 days to 35 days. This new preservative is designated as CPDA-1.
a. mannitol c. adenine and glucose
b. adenine saline d. Rejuvenix
____ 34. Which type of blood storage container is no longer available for use in the United States because it may limit
the viability of RBCs?
a. Glass bottles c. DEHP-free polyolefin containers
b. PVC plastic bags with DEHP d. Latex-free plastic containers
____ 35. A rare unit of blood became outdated 48 hours ago but is needed for a patient. Which of the following
concepts applies to this situation?
a. The blood could be rejuvenated by adding Rejuvesol, being washed appropriately, and
being transfused within 48 hours.
b. The blood could be rejuvenated with Rejuvesol, washed, and given immediately to the
patient.
c. Once a unit is outdated, it is no longer available for use.
d. The unit can be rejuvenated immediately, washed, and stored in the appropriate
refrigerator until needed later in the week.
____ 36. FDA-approved rejuvenation solution contains all of the following EXCEPT:
a. adenine c. inosine
b. glycerin d. phosphate
____ 37. Rejuvenated RBCs may be prepared up to three days after expiration when stored in all of these
EXCEPT:
a. ACD
b. AS-1
c. CPD
d. CPDA-1
____ 38. When is the corrected count increment (CCI) of platelets is usually determined?
a. Immediately prior to transfusion. c. Ten to 60 minutes after transfusion.
b. During the transfusion procedure. d. One to two days after transfusion.
____ 39. Generally, the quality control measurements required by various accreditation organizations for platelet
concentrates include:
a. platelet concentrate volume and platelet count.
b. leukocyte count if claims of leukoreduction are made.
c. pH of the unit.
d. All of the above
____ 40. Which of the following is not a major factor that influences platelet shape and activation while the platelet is
in storage?
a. pH c. Agitation
b. Volume d. Temperature
____ 41. Proper agitation of platelets while they are being stored is:
a. important because when not agitated properly the platelets will stick together and not
perform properly when transfused.
b. important because the pH of the stored platelets will increase and the platelets will lose
functionality.
c. important because the pH of the stored platelets will decrease and the platelets will lose
functionality.
d. not important because it has been deemed unnecessary by the FDA.
____ 42. Which of the following is not a commercial system approved by the FDA for screening for bacterial
contamination in platelet collections?
a. BacT/ALERT c. BACTEC
b. eBDS d. Scansytems
____ 43. Which of the following is a possible future method in pathogen reduction to treat platelet components?
a. UV light and amotosalen c. Vitamin B12 and UV light
b. Amphotericin B d. Penicillin
____ 44. Which of the following is licensed additive solutions approved for the storage of red blood cells for 42 days?
a. Adsol (AS-1) c. Optisol (AS-5)
b. Nutricel (AS-3) d. All of the above
Chapter 1. Red Blood Cells and Platelet Preservation: Historical Perspectives and Current
Trends
Answer Section
MULTIPLE CHOICE
1. ANS: C PTS: 1 KEY: Taxonomy Level: 1 LO: 1-3

3. ANS: B PTS: 1 KEY: Taxonomy Level: 1 LO: 1-14
5. ANS: D PTS: 1 KEY: Taxonomy Level: 2 LO: 1-14
6. ANS: A PTS: 1 KEY: Taxonomy Level: 2 LO: 1-7
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gleaming, murderous eye and pierced to the brain.
The brute gave a few convulsive struggles and then straightened out
beside its mate.
The situation had changed with the quickness of a kaleidoscope.
The would-be killers had themselves been killed just at the moment
that their victory had seemed assured.
But by whom had their death been brought about? Were these
newcomers friendly or hostile to Bomba? The thought came to the
boy that they might be of the party of headhunters. If so, he might
have escaped death in one form to meet it in another even more
terrible.
But as he hesitated, he heard their voices more clearly and
recognized them. Then he waited no longer.
He slid hurriedly to the ground and came face to face with Ashati and
Neram, the former slaves of Jojasta, the medicine man of the Moving
Mountain!
CHAPTER XIII
THE MAD MONKEY
Ashati and Neram dropped to their knees before Bomba, clasping

his hands and bowing their black heads before him. Neram, bending
lower, took one of Bomba’s sandaled feet and placed it on his neck
as a sign that he was slave and Bomba master.
The heart of the lonely boy swelled at this sign of gratitude and
affection. He stooped and raised the kneeling men, and made them
stand on their feet before him.
“You have saved Bomba’s life,” he said with deep feeling. “If you had
not come when you did the jaguars would have killed him. Bomba
will not forget.”
“Ashati and Neram would have been nothing now but bones buried
in the heart of the Moving Mountain if you had not come to their
help,” replied Ashati, who seemed to be the spokesman for the two.
“You saved them from death, and freed them from the cruel yoke of
Jojasta, the medicine man. Ashati and Neram have no master but
Bomba, and will go anywhere in the jungle at the side of Bomba as
his slave. Their lives belong to Bomba.”
“You shall go with me wherever I go,” replied Bomba. “But you shall
go as my friends and not as my slaves. Bomba has nothing to offer
you but friendship. If you will take that and go with him, he will be
glad.”
So it was settled, and with many more expressions of gratitude and
devotion on the part of the ex-slaves of Jojasta, Bomba and the two
men set to work to skin and quarter one of the dead jaguars that had
unwittingly furnished them a feast.
While engaged in this and the building of a fire for the roasting of the
best portions of the meat, Ashati and Neram imparted to Bomba
news that he was anxious to hear.
In their wanderings they had noted the bands of Nascanora and his
half-brother, Tocarora, heading in the direction of the Giant Cataract.
This had been only two days before and not far from the place where
they were now standing. From their hiding place in the brush, the
slaves had seen that each party had with it a number of captives.
“Was there a white man among them?” asked Bomba eagerly.
“Yes,” replied Ashati; “an old man, very thin and with white hair.”
Bomba’s heart leaped. Casson then was still alive! He had not
succumbed to the hardships of the journey.
“Can you take Bomba to the place where you saw them and point
out to him the way they were going?” he asked.
“Yes,” replied Ashati. “But they must be a long way from there now,
for they were going fast. And they kept looking behind them as
though they thought men were coming after them.”
The news set Bomba on fire with impatience to be off. Under his
urging and example, the roasting of the meat was swiftly completed.
He divided the food equally between the three of them. Whatever
might happen to them in the future, it was certain that they would not
starve.
After they had got fairly started, with Ashati as guide, toward the spot
where the path of the slaves had crossed that of the savages,
Bomba narrated to his companions some of his experiences since
he had left the hut of Pipina.
When he came to the part that concerned the loss of his bow and
arrows, Ashati insisted that the boy should take his and that he,
Ashati, could do very well with his hunting knife. Besides, he would
make another set of weapons at their first resting place.
Bomba would have refused, but seeing that he could not do so
without hurting the feelings of the devoted fellow, at last accepted the
gift.
“It was an arrow from that bow that found the heart of the jaguar that
would have sunk its claws in your flesh and ripped it from the bones,”
said Ashati, as he handed it over.
“Bomba has not asked you yet how it was you came just in time to
save his life,” said the lad.
“We hunted Bomba day and night,” Ashati replied. “Our lives were
yours, for you had saved them. We came on your trail in the jungle,
and followed after. Ashati saw you in the tree as the jaguar lifted its
paw to strike. Then Ashati prayed to the Spirit of the Jungle and shot
his arrow. The Spirit made it go straight, and the jaguar died.”
It was now late afternoon, and Bomba and his followers had not
traveled far before the swift tropic night descended on them and
forced them to rest for the night.
Bomba chafed at the necessity, though he himself was almost
exhausted in mind and body by the stirring events of the day that
had taxed both to the utmost.
He was up with the dawn, however, and, rousing Ashati and Neram,
summoned them to share his hasty breakfast and start on the day’s
journey. They obeyed with willingness, though no more than half
awake. Their bodies wasted and their strength sapped by years of
deprivation and torments at the hands of Jojasta could not throw off
the fatigue as readily as the healthy jungle lad, whose veins were
pulsing with vitality.
But Bomba’s quest of Nascanora could not wait. Delay, however
slight, might result in the death of his friends, if indeed they still lived.
If Ashati and Neram could travel at Bomba’s pace, he would be glad
to have them with him, for they were companions in his loneliness
and allies in case of danger.
But if they could not keep up with him, he would have to go on
ahead, leaving them to join him when they could.
But once roused, they seemed as eager as himself to continue the
journey. Even the prospect of an encounter with Nascanora and his
braves did not deter them, as long as they were under Bomba’s
leadership. What they had already seen of him had led them to
attribute to him almost magical power. Their anxiety to please and
serve the lad in every way they could deeply touched Bomba, in
whose life loyalty and service of any kind had been so sadly lacking.
They stamped out the embers of the fire they had built to keep off
the jungle beasts during the night, ate of their supply of jaguar meat,
and struck onward through the forest in the direction of the Giant
Cataract.
The sun rose higher, and with full daylight came a fresh burst of
speed on the part of Bomba. If Ashati and Neram found it hard to
keep up with him, they did not murmur. They would have suffered
any hardship rather than be left behind by him whom they had
chosen as their master.
They traveled all day without meeting with any unusual adventure,
pausing only briefly at noon to roast some jaboty eggs they found in
the forest that gave a welcome variation to their meat diet.
The shadows were beginning to gather when they came at last to the
spot where the paths of the slaves and the two parties of
headhunters had crossed.
The trail of the Indians was cold, but it was not difficult for one so
versed in woodcraft as Bomba to pick it up. There was still a little
daylight remaining, and he persisted in utilizing every moment of it to
gain another mile or two before he called a halt for the night.
On and on they went, although by this time they were nearly
stumbling with fatigue. They were penetrating a part of the jungle
that was new to Bomba. Pools, swelled by the recent rain, were
frequent, some of them so deep that it was necessary to cross them
by notched trunks of trees, the crude bridges of the jungle.
Crossing one of these, Ashati, wearied almost to fainting, stumbled
and would have fallen had not Bomba seized him and dragged him
to the safety of the further bank.
They had gone but a few yards farther, Bomba’s eyes straining to
detect the faintly marked trail, when there was a thud, and on the
ground before them, directly in their path, appeared a figure so
grotesque in form and ugly in face that Bomba took a startled step
backward and the two slaves fell to the ground in a fit of shuddering
terror.
“The mad monkey!” chattered Neram, and then, as the creature
advanced on them, uttered an ear-piercing shriek.
Gibbering and mouthing ferociously, froth slavering from its jaws, the
huge ape sprang toward Bomba and the cowering slaves.
Bomba was paralyzed at first by the hideous appearance of the
beast and infected to some degree with the superstitious terror that
animated Ashati and Neram. He seemed bereft of the power of
movement.
Then gathering together his forces, he sprang backward swiftly and
fitted an arrow to his bow.
CHAPTER XIV
BESET BY ENEMIES
Even as Bomba drew his arrow to its head he found that he felt a
strange unwillingness to inflict injury on this antagonist.
The monkeys were his friends. Often they had helped him when
other foes, much more like Bomba in form and appearance, had
sought to take his life. He could not forget how the swarm of
monkeys had turned the tide of battle in his fight with Nascanora and
his braves when they had attacked the cabin.
So Bomba called to this slavering, hideous object in the language of
the monkeys, trying to show that he was not an enemy.
But if anything, the aspect of the ape became still more fierce and
threatening. It uttered a shrill cry and sprang at the lad with hairy
arms outstretched to grasp him.
Ashati and Neram gave a shout of warning, and seeing that their
young leader was in grave danger, conquered their fear and sprang
to his help.
The bow of the jungle boy sang with a twanging sound as the arrow
sped from it. But the attack of the monkey was so swift that the
arrow, instead of entering the heart as Bomba had intended, pierced
the flank of the brute.
With a howl of pain and rage, the great ape plucked out the arrow,
and swung around upon Bomba with the bloody point of the weapon
upraised.
But Bomba was quicker than the ape. He sprang aside and, drawing
his machete from his belt, struck the animal’s arm a blow that cut
deep and caused the blood to spurt into the distorted face of his
assailant.
The arrow dropped clattering to the ground, and with a weird and
terrible howl the ape swung itself with its uninjured arm into the
branches of a tree and vanished into the depths of the jungle.
Trembling, Ashati and Neram faced Bomba in that uncanny twilight.
For a while they said nothing, but stood staring solemnly at each
other.
It was Bomba who first broke the silence, and in the deep stillness of
the shadow-filled jungle his voice sounded hollow in his own ears.
“The mad monkey! If he is alone, all will be well. But if there are
others——”
“There are others,” interrupted Ashati. “One mad monkey needs but
bite another, and that one too will become mad. It is in that way the
evil spirits get possession of a flock of monkeys and set loose a
thousand demons upon the jungle.”
“And the bite of one sets loose an evil spirit within ourselves, and we
become even as the mad monkeys,” said Neram, his teeth
chattering.
Bomba knew that he spoke truly. A kind of hydrophobia would
sometimes be communicated by a snake-bitten monkey to its mates,
and by them to any human being that came within their reach while
the epidemic was raging. He had known of whole flocks of monkeys
having been decimated before the terrible disease had run its
course. And at such times there was no inhabitant of the jungle to be
so much feared as a mad monkey.
“We cannot stay here,” said Bomba, looking about him. “We must go
on. Perhaps we shall find shelter, a cave or an abandoned hut of a
caboclo, where we can spend the night and leave this terrible place
with the daylight.”
“Yes, we must go on,” agreed Ashati, and Neram nodded his head in
agreement. “To stay here would be to bring that hurt monkey back
for his revenge.”
“With good fortune we may escape the notice of others of the flock,
since it is night and they may sleep,” suggested Neram.
“Then we must make no more noise than the foot of the jaguar,”
warned Bomba, and, turning, he sped silently and swiftly from the
place, followed closely by his companions.
They had proceeded only a short distance when a horrible chattering
overhead caused them to look up, and in the branches of a tree they
saw two big apes gibbering and grinning at them, with the same
awful look in their eyes that had marked the first one they had
encountered.
As the little party moved swiftly on, a large castanha nut struck
Ashati on the shoulder with such force that he was felled to the
ground.
Bomba lifted him to his feet and hurried him onward just as a
bombardment of the heavy missiles came pelting down. A hideous
wailing and a sharp, crackling sound like a crazy laugh followed the
three fugitives as they raced onward.
By this time, superstitious terror had taken complete possession of
Ashati and Neram, and Bomba himself could feel the hair rising on
his head. The swift-falling darkness, the knowledge that danger was
all about them, that insane enemies were skulking perhaps behind a
tree, leering at them from branches overhead, crouching in ambush
behind a concealing bush or shrub, at any moment to reach out a
hairy arm—all these things combined to fill them with terror.
Their flight was unreasoning. They plunged through thorn bushes
that tore at their flesh, and felt no pain. They stumbled and fell into
black ooze that might hold writhing snakes, and scarcely thought of
it. To put distance between themselves and this nightmare became
their only aim.
Once a terrible figure dropped upon the neck of Ashati from the
branches above. The native gave a dreadful shriek and threw
himself upon his face.
The mad beast catapulted from the shoulders of Ashati and fell at
the feet of Bomba. It was the work of a moment for the lad to sink his
machete to the handle into the hairy body. The thing crouched as
though to spring, then gave a ghastly screech and sprawled upon
the ground.
“He is dead?” asked Neram, coming forward, unbelieving.
At the words, the prostrate Ashati raised himself on hands and knees
and crawled over to where the lifeless brute was lying.
“Dead!” he gasped, and dragging himself to his feet stared hard at
Bomba.
For it was a superstition among the natives that a mad creature was
possessed of an evil spirit that made it immune to death. The fact
then that Bomba had killed one of the mad monkeys so easily filled
them with surprise and hope. If he had killed one, he might kill many.
Their weapons were not as powerless as they had feared.
But their relief was swiftly changed to fear by a strange, weird
rustling among the trees that swelled in volume as they listened, as
though a great storm had entered the jungle and was sweeping all
before it. But there was no storm nor sign of one.
“The mad monkeys!” whispered Bomba. “They have told each other
that we are here. They know that we are few and they are many.
Come!”
If their flight had betrayed panic earlier, it was nothing to the fear that
now gave wings to their feet. They might hold their own against a
few. They could not face an army, such as, from the sounds, now
seemed to be in pursuit of them.
But even then Bomba had no hate against his pursuers, such as he
felt toward jaguars and snakes. He pitied them as the suffering
victims of a terrible disorder. None the less, he knew that they were
bent on taking his life and that if it came to a combat it was a case of
kill or be killed.
There was but one hope, and that a slender one; the hope that they
might find some cave or other shelter in which they could barricade
themselves and hold the maddened animals at bay. He knew of no
such place in that vicinity, and even if he had, it would have been
difficult to find it in the dark.
Ashati and Neram, with what breath they had, were muttering
prayers to the Spirit of the Jungle. If ever help was needed, it was
needed then.
Onward they plunged through the black night of the jungle, that
terrible rustling as of a mighty wind coming closer and closer with
every moment.
As the pursuing monkeys drew closer to their prey, they began to
howl and jabber horribly. It seemed to Bomba that the whole jungle
was one hideous jangle of sound.
Neram screamed. A hairy arm had reached from a low-hanging
branch, encircling his neck. He struck at it with his knife and
stumbled after Bomba and Ashati.
They were panting, spent. In another moment that awful swarm of
maddened beasts would descend upon them.
Stumbling blindly on, Bomba felt his foot slip into a hollow at the
same time that his body struck violently against a hard substance.
He stretched out his hands and felt rather than saw in the darkness
that he had come to the entrance of a cave.
“Quick!” he gasped, stepping back and nearly falling over the
cowering form of Neram. “Into the cave! Quick!”
CHAPTER XV
LYING IN AMBUSH
The two slaves seemed not to hear Bomba.

Perhaps they were half dead from fright, or perhaps they thought
that the malady that had attacked the monkeys had disordered
Bomba’s mind as well.
Despite his command, they continued to crouch stupidly close to the
ground, striving to keep out of reach of the clutching hands that
swung from the branches overhead.
One great ape dropped to the ground and came swiftly toward them
on all fours, uttering howl after howl, so wild and eerie that it froze
the blood in the veins of the adventurers.
Finding that the slaves were too terrified to heed his orders, Bomba
seized Neram, who was nearer to him, and flung him bodily through
the opening of the cave. He took Ashati by the arm and dragged him
forward, shouting:
“The cave! The cave! Do you hear what I say? Get inside quick, and
I will follow.”
From inside the cave Neram reached out a hand and drew Ashati
within the sheltering blackness.
As Bomba was about to follow, the monkeys, following the
leadership of the great ape that had begun the actual attack,
dropped to the ground and swarmed toward Bomba.
The boy leaped for the gaping mouth of the cave just as the first of
the monkeys grazed his arm with its paw.
Neram and Ashati dragged him further into the depths of the cavern,
while, with a shriek of rage, the baffled monkey, seeking to follow,
dashed instead into the rock surface at the side.
Bomba stubbed his foot, bruising it as he fell forward into the cave.
He stooped down to examine the object and found that it was a great
rock.
“Come, help me!” he cried to the two slaves, who were fully awake
now to the fact that there was hope of escape.
They stooped to aid him, and as the monkeys recovered from their
surprise at the sudden disappearance of their enemies and began to
search out the entrance to the cave, those within rolled the great
stone to the entrance, fairly blocking it.
While they held that stone in place they were safe.
Bomba and his companions sank down on the ground, gasping for
breath, and pressed their weight against the rock.
The weird howling of the enraged monkeys outside their shelter
made them tremble even now, lest their cunning foes should find
some other entrance to the cave and come pouring in to indulge in
massacre.
But as time passed and they remained unmolested in their retreat,
while the awful howling of the monkeys died down to a mere jumble
of sound, they began to take courage and hope that all would still be
well with them.
Bomba had feared at first that the cave might be the lair of some wild
beast of the jungle.
But the fact that the stone was so near the mouth of the cave and
was evidently intended to block up the entrance seemed to argue
human occupation. Some native, perhaps, had become an outlaw
from his tribe and had chosen the cave as his home.
When it became certain that the monkeys had tired of their quest
and were straggling off again into the jungle, Bomba ventured to
make a cautious tour of the cave to make sure that his guess had
been correct.
Even then he was afraid that at any moment his fingers might touch
something warm and alive but not human.
However, the cave was a small one, and he soon found, to his great
relief, that, save for Ashati, Neram and himself, it was unoccupied.
Tired beyond words, but jubilant at their escape, Bomba returned to
the two, who still squatted on the ground close to the stone that
guarded the entrance to the cave.
Ashati was inclined to indulge in prophecy.
“The Spirit of the Jungle is good,” he stated, as Bomba threw himself
down beside them. “Twice it has saved the life of Bomba. The
captives of the wicked Nascanora and his half-brother Tocarora will
be spared. Bomba will live to seek out Sobrinini, and find from her
the secret that the poor old white man cannot tell. It is for that that
the Spirit of the Jungle has twice saved the life of Bomba.”
“Ahma,” said Neram, which meant “so be it,” as he bowed his head
reverently.
It was not long before all three were asleep, stretched on the ground
against the great rock, so that the slightest push on it would be
certain to rouse them.
But they were undisturbed all through the long hours of the night,
and at dawn awoke, refreshed and ravenous for the great chunks of
roasted jaguar meat they had prepared the day before.
They ate quickly, listening meanwhile for any sounds from the jungle
which might warn them of a further attack on the part of the mad
monkeys.
But, aside from the ordinary jungle noises, everything was as quiet
as they could wish. Unless they were still slyly watched by their
enemies, their escape from the cave should be a matter of no great
difficulty.
Nevertheless, both Neram and Ashati were reluctant to leave the
friendly shelter of the cave, and urged that Bomba rest there another
day and night until all danger should be gone.
But Bomba would not listen to this.
“You stay,” he said. “But Bomba must go. If, as you say, it was the
Spirit of the Jungle that twice saved his life yesterday, the Spirit
might be angry if Bomba were afraid and perhaps the next time
would not save him from the hairy arm of the great ape or the sharp
claws of the jaguar. No, Bomba must go.”
So, seeing that he was in earnest and thinking that if the lad were
under the protection of the Spirit of the Jungle they would be also,
Neram and Ashati said nothing more and helped Bomba roll the
great stone from the entrance to the cave.
“Where you go we will go,” said Ashati gently, as the welcome
sunlight streamed in. “Though you will not have us for your slaves,
you are our master. Let Bomba start and we will follow.”
Making no sound, they went swiftly through the jungle and did not
pause or stop to rest until they had put a great distance between
them and the scene of the fantastic nightmare adventure of the
evening before.
When the sun was high above their heads and all the jungle seemed
to fry and crackle beneath the heat of it, Bomba and his companions
sat down to eat the last of the jaguar meat and some berries and
nuts they had gathered by the way. Farther back they had found a
stream of clear, cool water, where they had slaked their thirst.
They resumed their route and had not gone far before they heard the
sound of rushing waters. The sound lent wings to Bomba’s feet, and
the faithful slaves kept pace with him, no matter how fast he went.
They came out soon on the banks of a river. The noise of the
foaming waters had been growing louder and louder until now it
smote upon their ears like thunder. A torrent of black water dashed
along the river bed and leaped angrily against the rocks that studded
its course, flinging a shower of spray upon Bomba and his
companions where they stood at the edge of the fringe of trees that
bordered the river.
“The River of Death!” muttered Ashati in hushed tones. “It is so that
our people call it.”
“And it is well named,” said Neram, making a cabalistic sign as
though to ward off evil.
“Beyond the River of Death,” said Bomba in a voice of impatience,
“though so far away that the eye cannot see it, is the Giant Cataract.
Come, let us be quick. Nascanora and his braves cannot be far
away.”
They started again, following the course of the stream. Suddenly
Neram paused with his head to one side as though he were
listening.
“Hark!” he said, when Bomba would have questioned him. “Someone
comes.”
As soon as the words had fallen from his lips, Bomba threw himself
upon the ground and put his ear to it.
Instantly he was on his feet again, drawing Ashati and Neram within
the shelter of the trees, which at that point were growing in great
profusion.
“We will watch as silently as the jaguar watches for his prey,” he
hissed. “Let no one move or make a sound.”
Hidden by the rank marsh grass that formed a perfect covert, the
three lay motionless, not a breath betraying their hiding place.
For what seemed a long time to the intent watchers, no one
appeared in sight. For all that could be seen, the jungle was free
from all human presence and given over to its animal inhabitants.
Unseen himself from the bank of the river, only a few yards away,
Bomba could yet have a perfect view of any living thing that might
pass by. Absolutely still, as motionless as though he were an image
of stone, his eyes alone moved to show that he lived and breathed.
The faint sound of footsteps that they had heard gradually became
more distinct, and at last a solitary traveler came into view. The man
was an Indian, but of a tribe with which Bomba was not familiar. He
was journeying swiftly when those concealed in the long grass
caught their first glimpse of him, glancing again and again over his
shoulder as though he feared pursuit.
Motioning to Neram and Ashati to remain silently where they were,
Bomba wriggled through the underbrush as sinuously as a snake.
Not a motion of the grass betrayed his presence.
Then, without a sound, as though he had risen from the heart of the
earth, Bomba leaped to his feet directly in the path of the
unsuspecting native.
The fellow started back and opened his mouth to yell, but before a
sound could issue from it Bomba had clapped one lean, brown hand
over his mouth and with the other had thrown him prone on the
ground.
Kneeling on the chest of his captive, who was too paralyzed by the
suddenness of the attack to make more than a feeble resistance,
Bomba called softly to Ashati and Neram. As though, like Aladdin, he
had conjured them by the rubbing of a lamp, they were instantly at
his side.
At their unexpected appearance, the feeble resistance of the native
ceased altogether. He seemed to be stupefied with terror, and stared
from one to the other of his captors with red and watery eyes that
begged for mercy.
“We will not hurt you,” said Bomba, taking his hand from his
prisoner’s mouth while at the same time he motioned Neram and
Ashati to grasp the fellow, should he seek to break away. “Only tell
us what you know and you shall go free. But lie to us,” he added,
with a significant motion toward his machete, “and you die!”
CHAPTER XVI
THE ISLAND OF SNAKES
The native writhed and twisted, the eyes bloodshot with terror at the
threat.
Ashati and Neram came a step nearer with menace in their eyes,
and the captive subsided again, gasping and groaning beneath the
pressure of Bomba’s knee upon his chest.
“Do not kill me! Only do not kill me!” he jabbered. “I will answer you!
You have only to ask.”
“Then listen to me, and listen well.” The fierce, impatient note was in
Bomba’s voice again. “Nascanora, the great chief, the chief with a
black heart, is the man I want to find. Give me news of him and you
shall go free.”
A gleam of hope came into the prisoner’s eyes.
“Yes, yes, I will tell. I will tell you anything I know. But you must not
let Nascanora know that I have told, or he will cut my heart and fry it
over a fire.”
“Nascanora shall not know,” broke in Bomba, as he lightened
somewhat his pressure on the man’s chest and bored him through
and through with his eyes as though to wrench the truth from him.
“But if you speak with a forked tongue your place will be with the
dead.”
“I will make straight talk,” asseverated the native. “I do not want to
die.”
“Where is Nascanora? Quick!” demanded Bomba.
“In two days’ time,” replied the trembling native, “he will pass this
spot on the other side of the river.”
“I thought he had already passed,” said Bomba.
“He has,” returned the native. “But he turned back to burn another
village and take some more captives. He will move slowly, for he has
other enemies that he wants to capture near Snake Island, where
lives the old witch, Sobrinini——”
Bomba cried out in astonishment and quick hope.
“You know Sobrinini?” he asked eagerly.
The native marked his change of tone, and seemed encouraged by
it. His terror abated, and he spoke so swiftly that the words stumbled
over each other and Bomba could scarcely make out what he said.
“I know of Sobrinini. Who does not know of her who lives near the
Giant Cataract?” he cried. “But I do not go near her island, for it is full
of snakes and Sobrinini is a woman of evil whose frown means
death.”
“Where is this island?” Bomba asked in a fever of eagerness.
“It is called Snake Island. But to go there is to die.”
The native shivered with a superstitious terror as he spoke the
words.
Bomba glanced at Ashati and Neram, who had been listening with
absorbed interest to the story of the native and on whose faces was
reflected the same look of fright.
Bomba released his hold upon the prisoner and arose to his feet.
“There are two things I must know,” he said.
“Command, and you shall be obeyed,” returned the native humbly.
“Was there a white man among Nascanora’s captives?”
“I cannot say. He had many captives and he will return as I have
said. That is all that the jungle has told me.”
“How do you go to this island where the witch woman, Sobrinini,
lives with her snakes?” asked Bomba, feeling that the native had
spoken the truth.
The native looked at him with curiosity dawning in his reddened
eyes.
“You will not go there?” he cried in horror. “I tell you it would be better
for you to go to the giant anaconda and let him wind his coils about
your body than to seek out Sobrinini on her island that lies under the
curse of the gods. I tell you again that to go there is to die.”
“That is for me to say,” replied Bomba. “I do not fear Sobrinini. I do
not fear her snakes. I have come a far way to see her and I will see
her. Tell me what I want to know. Bomba does not like to ask a thing
twice.”
In response to this imperative demand, the native jabbered out
directions, telling Bomba the course he must take to reach Snake
Island if he did not wish to be swept to destruction by the rapids of
the River of Death.
When Bomba had extracted all the information he could, he
motioned the native to rise to his feet.
“Listen!” he said. “If I were Nascanora I would kill you, so as to be
sure that your tongue would be still. But Bomba’s heart is not black
like that of the chief of the headhunters. I am going to let you go free.
But if you tell anyone that you have seen me you will find that
Bomba’s knife is sharp and his vengeance is swift. Go now and
remember what I say.”
After the man had disappeared, Bomba wasted little time in
discussing the matter with his companions. For if he were to
accomplish all he hoped to before Nascanora should pass that way,
he would have to work hard and fast.
He was in a ferment of eagerness to visit Sobrinini and wrest from
the strange woman the secret of his birth. He was sure she knew.
Jojasta had said she could tell him. Casson’s queer actions when the
name of Sobrinini had been mentioned and his statement that “Nini
would know” had further confirmed his conviction.
As for the terrors that, according to the native, barred access to her,
he cared not at all. He was used to meeting and overcoming danger.

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Ashati and Neram dropped to their knees before Bomba, clasping

The two slaves seemed not to hear Bomba.

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