R 456

Technical Report :

OaLZT Yaw AUG 1 1966

DEEP-OCEAN BIODETERIORATION OF
MATERIALS — PART IV. ONE YEAR

AT 6,800 FEET

June 1966

NAVAL FACILITIES ENGINEERING COMMAND

U. S. NAVAL CIVIL ENGINEERING LABORATORY

Port Hueneme, California

Distribution of this document is unlimited.

DEEP-OCEAN BIODETERIORATION OF MATERIALS — PART IV. ONE YEAR
AT 6,800 FEET

Technical Report R-456
Y -RO11-01-01-042
by

James S. Muraoka

ABSTRACT

As Part IV of a series of reports on the biological deterioration of materials
in the deep ocean, this report covers the data obtained after exposing metallic and
nonmetallic specimens for 13 months on the floor of the Pacific Ocean at a depth
of 6,800 feet (Test Site 1). The specimens were attached to a Submersible Test Unit
that was retrieved in February 1965. Preliminary examination of the specimens was
made aboard ship, and the final examination, tests, and analyses were performed
at the Naval Civil Engineering Laboratory.

On recovery, most of the plastic materials and all of the rope specimens were
covered with bacterial slime. Cotton and manila rope specimens were severely
deteriorated by marine microorganisms. Wooden test panels, plastics, and Manila
ropes were attacked by molluscan borers. Glass, metals, natural and butyl! rubber,
and certain plastic materials were not adversely affected.

The results of breaking-strength tests on ropes, weight loss measurements of
wood panels, moisture absorption tests on plastics, and insulation resistance and
voltage breakdown tests on electrical insulating materials are presented.

Distribution of this document is unlimited.

The Laboratory invites comment on this report, particularly on the
results obtained by those who have applied the information.

CONTENTS

page

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0 0301 0040129

PREFACE

The U. S. Naval Civil Engineering Laboratory is conducting a research
program to determine the effects of the deep-ocean environment on materials. This
research will be of great value in establishing the best materials to be used in deep-
ocean construction in the Navy's conquest of "inner space."

A Submersible Test Unit (STU) was designed on which many test specimens
can be mounted. The STU can be lowered to the ocean bottom and left for long
periods of exposure. This report deals with STU I-4, in place for 13 months at a
depth of 6,800 feet. The test site is about 81 nautical miles southwest of Port
Hueneme, California.

INTRODUCTION

As part of a research program to determine the effects of the deep-ocean
environment on various materials, the U. S. Naval Civil Engineering Laboratory
(NCEL) in March 1962 placed the first of a series of Submersible Test Units,
designated STU I-1, on the ocean floor in 5,300 feet of water at Test Site | off
the coast of California (Figure 1). Since then additional STU's were placed on
the sea floor and these are listed in Table 1 with pertinent information about each
Sus

STU I-4 was recovered in July 1965 after 13 months on the sea floor in
6,800 feet of water. A sketch of the STU |-4 system as installed is shown in
Figure 2. The STU was loaded with 1,852 specimens of 567 materials (Figure 3).
The majority of these materials were metal and metal alloys for the study of corro-
sion. For biodeterioration study, nonmetallic materials, such as woods, ropes,
and plastics, were assembled in two aluminum racks (bio-racks) and then attached
to the side of the STU for exposure in the deep-ocean environment.

This report presents the materials and the methods used for attracting,
collecting, and evaluating deep-sea fouling and boring organisms and the results
of field and laboratory investigations of the materials recovered from STU I-4.

RESEARCH METHODS
Oceanographic Information

Concurrently with the STU program, numerous oceanographic and biological
data-collecting cruises to the STU sites have been conducted. These have produced
information about the environmental parameters, such as salinity, temperature,
oxygen content, and biological activity. Such information is essential in evaluating
changes in the materials exposed on the ocean floor, especially the corrosion of
metals. The environment for Test Site | (6,800-foot depth) is presented in Table 2
and in an unpublished report. *

Test Site | was selected because the area provides a nominal 6,000-foot depth
reasonably representative of the open-sea conditions in the eastern part of the Pacific
Ocean. It is located about 80 nautical miles southwest of Port Hueneme, California.

* U.S. Naval Civil Engineering Laboratory. Oceanographic data. No author.
Port Hueneme, Calif. In preparation.

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Santa Barbara Is.

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ort Hueneme
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Santa Catalina ls.

Figure 1.

(nominal depth of 2,500 feet).

Test Site | (nominal depth of 6,000 feet) and Test Site II

200 ft

aluminum buoy

hooks —>
5/16-in.-diam
stainless steel wire, —>
300 ft

pinger —*|

5/16-in.-diam
stainless steel wire, —™
400 ft

rope bale

acoustic release

aluminum buoy

l-in.-diam polypropylene
braided line, 300 ft

Pinger —e

l-in.-diam polypropylene
braided lift line, 6,000 ft

l-in.-diam polypropylene
braided line, 4,600 ft

grappling wire (polyethylene-
jacketed wire rope), 6,000 ft

1.3-in.-diam polypropylene
braided surge line, 600 ft

pinger/attitude sensor current meter and temp recorder ~

acoustic release __

concrete sinker 4

Figure 2. STU 1-4 complex as designed.

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Table 2. Environment at Test Site |

Surface
Water

Environmental Factors Test Site |

Depth, ft 6,800
oI
e2/1
34.52
7.84

3,000

Water temperature, OC

Dissolved oxygen content, ml/I
Salinity, °/oo (ppt)

pH

Hydrostatic pressure, psi

Current, knots

Green mud containing
glauconite, Foraminifera

Sediment
tests, quartz, etc.

Biological Activity

When NCEL first decided to place STU I-1 on the sea floor at Test Site | in
March 1962, neither the topography of the sea floor nor the kind of deep-sea
creatures that would be encountered were known. In order to obtain some informa-
tion, a deep-sea camera was lowered from the stern of a vessel while at the same
time STU I-1 was being lowered to the ocean floor from the bow of the vessel.
Numerous photographs were taken of the sea floor close to where STU I-1 was
placed.

The photographs revealed that the topography of the sea floor in the vicinity
of Test Site | was generally flat except for small mounds built by mud-dwelling
animals. The presence of these mounds indicates that there is considerable biolog-
ical activity in the area. While the photographs of the sea floor were being taken,
a 5- to 6-foot-long shark-like fish with a large dorsal fin came into view of the
camera and was photographed apparently drifting near the sea floor in 5,300 feet of
water (Figure 4).

Rock Samples
Rock specimens were desired from Test Site | so that the fouling organisms

attached to the rocks could be studied; these organisms could be expected to attach
themselves to other materials as well. A 10-inch-diameter, 36-inch-long steel pipe

with retaining rods welded across its lower end was employed for collecting rocks
from the ocean floor. The pipe dredge was lowered to the ocean floor from an
oceanographic vessel, USNS Davis, and the area was dredged for rock specimens.
Several passes were made across the area of Test Site | at a depth of 6,000 feet,
and rocks of various sizes were collected. Various organisms were attached to the
collected rock specimens (Figure 5), and these are listed in Table 3.

Several rocks had deep holes bored by pholads; neither live pholads nor their
empty shells were found, so it is not known whether these rock borers are active at
this depth. There was a deposit (as great as 1/4 inch thick) of manganese oxide on
the rock specimens. Chemical and instrumental analyses of these deposits were made
by U. S. Bureau of Mines, Tiburon, California. The results of these analyses are
presented in Table 4. Further information on the manganese deposits on the floor of
the Pacific is given in Reference 4. It is believed that bacteria play a major role
in the formation of nodules, for it is reported in Reference 5 that 36 pure bacterial
species were found and isolated from inside surface-sterilized manganese nodules.

Sediment Samples

Marine bacteria are among the major biological agents in the deterioration
and fouling of various materials submerged in the sea. To determine the type and
activity of bacteria in the deep sea, sediment samples were obtained and analyzed
in the laboratory by means of standard microbiological methods. Samples were also
obtained to determine the type of marine animals found in the sediment.

Table 3. Fouling Organisms on Rocks From 6,000-Foot Depth

Organisms Remarks

BiyOZOa res Li ceyegis . . . Six or more different species of encrusting
and erect forms; one encrusting form covered
about | square inch of rock surface

ForannlimitheCnesnemey en eer Many different species
Gllasstspongesi-weye a) oe Several different deep-sea species
Serpulid worm tubes. . . . Numerous large and small; the longest was

over 4 inches

VIG GINS 6 a 6 0 oe 6 One found in a pholad hole
Ghiittomes, socaters eek aston: One 1/2-inch long

Figure 4. A 5- to 6-foot-long shark-like fish with a large dorsal fin,
on the sea floor in 5,300 feet of water. (Courtesy of Carl
Shipek of the U. S. Navy Electronics Laboratory.)

Figure 5. Photomicrograph of colonial (encrusting) bryozoa, deep-sea
glass sponge, Foraminifera, and worm tubes on a rock
surface (magnified).

Table 4. Chemical and Instrumental Analyses of Manganese Incrustation

The samples were obtained about 30 miles south of San Miguel Island,
33° 46' N and 120° 49' W, at a depth between 5,000 and 6,000 feet.

Sample No. 1 (%)

Sample No. 2 (%)

0.64
1.10
0.010 0.013
12.4 Wed
16.42 19.08
0.26 0.24
0.05 0.07
0.13 0.22
0.07 0.05
0.02 0.03
0.05 0.05
0.05 0.04
Or35 0.42
0.56 0.44
1.03 1.20
10.1 8.9
0.74 0.78
3.67 2.61
0.38
0.09
0.8
46.0

1/ Calculated from percent water obtained by drying at 110°C.

The following samplers, previously described in Reference 2, were used to
collect sediment samples from the deep sea:

1. A gravity core sampler, which takes cores up to 4 feet long.

2. NCEL's scoop-type sampler, which collects about 225 cubic inches of
sediment from the soft bottom of the ocean.

3. A ZoBell bacteriological sampler with a sterile sampling unit (modified in
order to collect a mixture of seawater and sediment at the seawater -
sediment interface).

The results of the bacteriological analysis of the sediment samples are presented
in Tables 5, 6, and 7.

Table 5. Aerobic and Anaerobic Bacteria in Core Samples of Sediment

Aerobes | Anaerobes
(per gram | (per gram
of wet of wet
sediment) | sediment)

Presence (+) or
Absence (-) of
Sulfate Reducers

Depth of Sample
Below Ocean Floor

(in.)

Table 6. Aerobic Bacteria in Scoop Samples of Sediment

Sulfate-reducing bacteria present in all these samples.

Aerobes
(per gram of wet sediment)

Table 7. Aerobic and Anaerobic Bacteria in Sediment Obtained
With a ZoBell Bacteriological Sampler

Sulfate-reducing bacteria present in all these samples.

Aerobes Anaerobes
(per gram of (per gram of
wet sediment) wet sediment)

Z462a 300,000 not tested
Z462b 500,000 not tested

Z5a62 1,500,000 500,000
Z5b62 1,870,000 500,000
Z5d62 1,700,000 600,000
Z11a63 1,500,000 600,000
Z11b63 2,000,000 600,000

The number of aerobic, anaerobic, and sulfate-reducing marine bacteria was
determined in the laboratory on nutrient agar plates and in test tubes. Because of
the different nutritional requirements of marine bacteria and the limitation in the
enumeration procedure employed in this study, only a small percentage of the marine
bacterial population may have been determined from the bottom samples. The
following media were used for culturing marine microorganisms:

1. The nutrient medium used with pour plates for determining the general
aerobic bacterial population consisted of

Bacto =Pe PhOMe isn <Af- Nie nacitiie panemey 5.0 gm
FEMS [INCTANICHS 6 6 ba 6 6.6) 5) doc 0.1 gm
Measiiiextractaint sis es tor a deual aucenour 1.0 gm
Seawater (aged) ss ean nee 1,000 ml
Bacto saga isi) Gud noe Rupe ta ialae 20 gm
Diba ets. o: aacde ae ee eeree Ce ener een: TES

2. The general anaerobic bacterial population was determined by using the
Fisher-Spray anaerobic culture dishes with the above medium, which was
treated with sodium formaldehyde sulfoxylate (1.0 gm/I) to lower the
redox potential and with resazurin (0.001 gm/l) to serve as an indicator
of anaerobiosis. ”

3. The nutrient medium for determining the presence of sulfate-reducing
bacteria consisted of?

Potassium phosphate dibasic. ... . 0.2 gm
Magnesium sulfate hydrated... . . 0.2 gm
Selim Suite c\o’s oo so 6.5 eo ce 0.1 gm
Ferrous ammonium sulfate ...... 0.1 gm
Galeiumplactatepaeie ene ncn Cee 3.5 gm
Ascorbic: dcidiastwae ewes) Gene 0.1 gm
Bacto=peptome rissa rece 1.0 gm
Yeast-extracthm saps snes he e 1.0 gm
Seawaten(aged)) a eee 1,000 ml
oh gl es ir OR ll hadith bate Mest du ay one TED

For this study, agar was deleted from the above medium. The medium
was placed in test tubes with screw caps and sterilized. The screw caps
were tightened when the medium was still warm to exclude atmospheric
oxygen. Small samples of bottom sediment from various depths were
placed in these test tubes and incubated for several days at 15°C.

The sediment samples obtained with a scoop sampler were washed through a
plastic screen to collect mud-dwelling organisms. The animals were bottled and
preserved in a 5% glycerol -alcohol solution aboard ship for laboratory analysis.

A variety of animals was found in these sediment samples. Annelid worms were the
most abundant marine organisms collected in the vicinity of Test Site 1. Other mud
dwellers collected were nemertean (round worms), holothurian (sea cucumbers),
amphipods, molluscans, and Foraminifera tests. The Foraminifera tests found in the
sediment samples have been classified and are reported in Reference 2.

Test Materials

For evaluating deep-sea biological effects on nonmetallic specimens, two
bio-racks were attached to the STU. Each rack held several 3-foot-long plastic
rods and tubes and a 12x30x 1/8-inch laminated phenolic plastic sheet (Figure 6).
Numerous smaller test specimens were attached to the plastic sheet; one sheet was
secured to the upper section of a bio-rack, and an identical sheet was secured to
the bottom section. In order to expose the test materials to biodeterioration in mud
as well as in water, the two racks were attached to the STU so that the lower portions
would be either exposed near or buried in the bottom sediment and the upper portions
would be exposed to seawater 3 to 4 feet above the mud line.

The bio-rack test specimens are listed in the Appendix. The 2x6x1/2-inch
wooden panels were cut from sound lumber, and the surfaces were cleaned with
alcohol and then covered with plastic to avoid contamination. The plastic covers
were removed just before the test specimens were submerged. The wooden panels
were employed to collect specimens of any deep-sea fungi and marine borers which
may have been present on the ocean floor.

The sections of the 3-foot-long plastic rods, tubes, and pipe, and the rubber
tubes were treated in different ways. One section of each specimen was roughened,
a second section was wrapped in burlap, a third section was taped with plastic and
rubber electrical tape, and a fourth section was left smooth. The various wrappings
provided a favorable foothold for the attachment and growth of deep-sea fouling
and boring organisms. A large piece of untreated fir wood was fitted around both
ends of each specimen to act as bait to attract and lead borers into direct contact
with the specimen materials.

Four different kinds of rope, such as synthetic plastic fiber rope (nylon and
polypropylene) and natural fiber rope (cotton and Manila), were placed on the
bio-racks. Electrical cables covered with rubber or plastic insulation of various
thicknesses were also placed on the racks. A small piece of pine wood was fitted
around each cable specimen to act as bait for marine borers. Another group of
electrical conductors placed on the bio-racks consisted of 0.015-inch-thick insu-
lation over a No. 16 tin-coated copper wire. The materials used in the formulation
of the insulation are presented in Table 8. The wire specimens were 15 inches long;
some were stressed (coiled) and some were nonstressed (straight). Stress was applied
by coiling a 15-inch specimen lightly around a 1/4-inch-diameter glass rod and
then removing the rod. Both ends of each specimen were sealed with two coats of

rubber cement. The specimens were positioned so that one set of ropes and electrical
cables would be buried in the sediment (in which bacteria are ordinarily most active)
and an identical set would be exposed about 3 feet above the sediment.

Table 8. Materials Used in the Formulation of Insulating Materials Tested

Polyethylene (standard
polyethylene insulation)

Polyviny! chloride = s a

Government rubber Cumarone-indene | Hard clay and | Polymerized

styrene (styrene resin and micro- | water-ground | trimethyl
butadiene rubber) crystalline wax whiting dihydroquinoline

Silicone rubber — — —

Neoprene (Type W) Light process oil | Hard clay 4, 4 thiobis (6-
and petroleum tert-butyl m-
creosol)

Metal test specimens were also placed on the STU, but materials containing
antifouling paints or other toxic substances were excluded from exposure aboard the
STU. The current velocity at a depth of 2,340 feet (about 0.3 knot) was not great
enough to carry away any water-soluble toxic substance which might alter the
natural biological fauna found in the immediate vicinity of the STU.

RESULTS
Recovery of Test Materials

As soon as the recovered STU was placed on the deck of the ship, the test
panels were examined and any attached organisms were photographed. The animals
were then carefully lifted from the test specimens and preserved in a 5% glycerol -
alcohol solution for further examination and analysis in the laboratory.

As soon as the STU was secured to the deck, the bio-racks attached to the
side of the STU (Figure 7) were disassembled. The individual test specimens were
placed in plastic bags for later examinations, tests, and evaluation in the laboratory.

14

The wooden panels, riddled with marine borers, were soaked in an alcohol
solution to prevent decay and were placed in plastic bags. The wooden panels used
for fungi determination were placed directly into sterile plastic bags, sealed, and
stored in the ship's refrigerator. Rope specimens for the isolation of marine micro-
organisms were also placed in sterile plastic bags and stored ina refrigerator. In
order to determine moisture absorption, the 3-foot-long plastic rods and tubes and
other plastic materials were placed in plastic bags and sealed to retain any moisture
present in the plastics.

Marine Growth on Test Materials

Metal Panels. Generally there was very little fouling growth on the metal
panels. Species of deep-sea hydroids (Figure 8), including tubularia and sponges
(Figure 9), were the major attachment organisms found on test panels. Generally,
moderate amounts of hydroid growth were found attached to the surface of monel
and stainless steel panels, which were not covered with corrosion products. Very
little or no hydroid growth was evidenced on panels of mild steel and panels of
copper base alloys, which were covered with corrosion products.

Figure 6. Various kinds of materials assembled in bio-racks before
exposure.

15

*AyaAooas Ja4yo Ay apo!pewul
1S P JO pis ayy O} payoD44D syoDI-01q UO sjDIIEyDyWy */ 21NB14

16

*suawioeds jsa} jOJawW UO sploipAyy °*g a1nbi4

17

Figure 9. Species of 1/4-inch-long deep-sea sponges.

Typical fouling organisms found in shallow water, such as barnacles, tunicates,
and bryozoa, were not found attached to the panels exposed at a depth of 6,800 feet.

Wooden Panels. A total of twenty-six 1/2 x2x6-inch wooden test panels,
which included pine, fir, ash, maple, cedar, oak, and redwood, were exposed to
determine the effects of deep-sea animals on different woods. All the wooden
panels were attacked by marine borers and sustained light-to-heavy damage. The
severest damage was sustained by cedar and oak; moderate damage was sustained by
ash, pine, fir, and maple; and light damage was sustained by redwood. The external
sides of the panels exposed to seawater appeared to be relatively free of any borer
attack (Figure 10a); however, when the wooden panels were removed from the
phenolic laminated plastic sheet and the sides attached to the plastic sheet were
examined, it became evident that the interior of the wood was completely riddled
by marine borers (Figure 10b). The majority of the borer tunnels measured about
5/16 inch in diameter. The average borer tunnels in redwood measured about
1/8 inch in diameter. The loss in weight of each of these panels due to borer
damage is presented in the Appendix. The surface of the phenolic plastic sheet

to which the wooden panels were attached was also damaged by borer activity
(Figure 11).

ide.

a) External s

(

(b) Interior.

igure 10

Wooden Panels.

F

19

(a) Full view.

(b) Closeup view of borer damage.

Figure 11. Phenolic laminated plastic sheet where wood panels
were attached,

20

It is highly unlikely that the surfaces of the wooden panels were infested with
borers during descent aboard the STU because (1) the wooden panels were dry when
initially submerged and (2) the rate of descent to the sea floor was probably too
rapid (100 to 150 ft/min) for any borer attachment. In order to eliminate any doubt
about this matter, several panels were covered with a clean, white, bonded paper
and sealed with a plastic adhesive tape to prevent any borers present near the surface
from attaching themselves to the wood during descent (see Figure 6).

It is not known whether the paper coverings were detached from the wood
during descent to the bottom or at the sea floor. When the panels were recovered
and examined, they were also found to be riddled by borers identical to those found
on uncovered test panels.

The wood panels which were exposed about 0.5 feet above the sediment layer
were much more severely damaged by the borers than the set of panels exposed 3 to
4 feet above the sediment layer. This indicates that the deep-sea marine borers are
very active in large numbers immediately above the sediment layer and their number
tends to decrease in seawater as the distance from the sediment layer increases. A
similar phenomenon has been reported in Reference 10. It is possible that these
borers may not be present in seawater about 25 feet above the deep-sea floor.

The greatest borer population was found in the 2x4x 30-inch bait pieces
(untreated fir) fitted over the plastic rods and tubes (Figure 12). Some borers, about
3/16 inch in diameter, had penetrated about 1-1/2 inches into the center of the
wood from the cut end (Figure 13). Because of the deteriorated condition of the
wood, the 2x 4-inch bait pieces could easily be broken apart by hand (Figure 14).
Numerous white deep-sea sponges (Figure 9) were found attached to the surfaces of
these woods. Numerous amphipods (Figure 15) were found between the wooden
layers, and over 50 of the crustaceans were found trapped inside a plastic container.

The molluscan borers (Figure 16) in these wooden bait pieces have been
identified as Xylophaga washingtona Bartsch and Xylophaga duplicata Knudsen.
Complete descriptions of Xylophaga washingtona and Xylophaga duplicata are pre-
sented in References 11 and 12, respectively.

Marine bacteria and fungi were also responsible for the deterioration of the
wood. The surfaces of the test panels were softened by the action of bacteria and
fungi. Microscopic examination of sections through the wood revealed two types
of deterioration of wood cells:

1. Irregular tiny cracks and channels in the cell walls caused by bacteria.
This type of deterioration was the most commonly found on the surface of
the wood.

2. Wide channels in the secondary cell walls of the wood caused by higher
marine fungi. This type of deterioration is known as "soft rot" and was
less commonly found on the surface of the wood.

21

(b) Closeup view.

Figure 12. A 2x4x30-inch wooden bait piece riddled by borers.

22

3

Figure 14. Interior of wood completely riddled by borers. The
shells of the molluscan borers are visible.

23

Figure 15. Amphipods, a shrimp-like crustacean, from a

6,800-foot depth.

Figure 16. Closeup view of Xylophaga sp.

24

Brownish fungi hyphae (without the fruiting bodies) were detected in the wood.
An unsuccessful attempt was made in the laboratory to induce fructification by the
fungi for classification purposes.

Rope Specimens. A heavy growth of slime bacteria was present over the
surfaces of cotton and Manila ropes, and a light growth of slime was present over
the surfaces of nylon and polypropylene ropes (Figure 17).

The fibers of cotton rope were decayed considerably by bacterial action, and
as a result were easily pulled apart by tweezers (Figure 18). When the cotton rope
was handled after it had been dried, the rope fibers crumbled into small pieces.
Only a few borers were found on the surface of the cotton rope and none of these
had bored into the rope.

The 1/2-inch-diameter Manila rope specimens were severely damaged by
both borers and microorganisms. Borers ranging in size from 1/32-inch to 1/8-inch
diameter had completely infested the entire length of the 5-foot-long rope specimens
(Figure 19). The rope fibers were severed completely as the borers penetrated deep
into the rope. The slime bacteria were responsible for the decay and deterioration
of the fiber materials.

Examination of the nylon and polypropylene ropes under a stereoscopic
microscope showed that the fibers of these ropes were neither severed nor damaged
by marine borers. On the contrary, the fibers were in excellent condition
(Figure 20). Table 9 compares the breaking strength of rope specimens before and
after deep-sea exposure.

Plastic Specimens. The sections of 3-foot-long solid plastic rods and viny|
tubes recovered after 13 months on the sea floor in 6,800 feet of water are shown
in Figure 21.

There were numerous deep and shallow borer holes present underneath the
area where large wooden bait pieces were fitted over the lower and upper sections
of the plastics (Figure 22). It was found that the lower section exposed about
0.5 foot above the sediment layer had more borer holes than on the upper sections
exposed about 3 feet above the sediment. In other words, the borers were found
to be more active near the sediment than 3 feet above the sediment layer.

The number and size of the borer holes found on the surface of the plastic
specimens are presented in the Appendix.

A vinyl tube (NCEL No. 338) intended for low-temperature use had a heavy
bacterial slime growth over the entire tube and over the plastic, rubber, and burlap
wrappings. When recovered, the open tube was quite rigid and had decreased in
diameter by about 1/8 inch, perhaps because the plasticizer or some other chemicals
used in the tube's formulation may have been lost as a result of bacterial slime
activity or other effects of the deep-sea environment. The other three vinyl tubes
listed in the Appendix had a light slime growth over the surface; the diameters of
these tubes remained unchanged except where marine borers had penetrated the
surface of the plastic under the area of the wooden bait pieces.

25

Figure 17. Rope specimens secured to bio-rack, immediately after
recovery. Notice the dangling section of a cotton rope
on the left. Photographed at night.

Figure 18. Cotton rope deteriorated by bacterial action.

26

Figure 19. Manila rope deteriorated by marine microorganisms and borers.

Dif,

St T-4 Polypronylane Pope

Figure 20. Nylon and polypropylene rope specimens, in excellent
condition.

Figure 21. Plastic rods and tubes immediately after recovery. Borers
are visible in bait piece.

28

(b) Closeup view.

Figure 22. Numerous shallow and deep holes made by borers in plastic
rods and tubes underneath the area of wooden bait piece.

29

Table 9. Breaking Strength of Rope Specimens Before and
After Deep-Sea Exposure

Breaking Strength (Ib) L/

R Diameter
nee (in.) Before After
Exposure Exposure
Cotton V2 1,340
Manila 1/2 2,068
Nylon 1/4 1,900
Polypropylene 5/16 1,810

1,7 Average of two specimens for each test.
27 Estimated 100% of tensile strength of rope destroyed by microorganisms
and borers. :

Moisture absorption analysis was performed on the plastic rod and tube
specimens. In order to determine the amount of moisture the plastics had absorbed
during the deep-sea exposure, each of the 3-foot-long plastic specimens was
weighed before being submerged. The figures for weight could not be used in the
test because of the weight loss in the plastics as a result of marine borer attack.

For the moisture absorption study, a section about 3 inches long was cut from
each of the 3-foot-long plastic specimens which were stored in plastic bags aboard
ship; the section was then weighed in the laboratory. After being weighed, the
test specimens were dried at room ambiency for 1 month and were then reweighed.
The information about the amount of moisture gained by the plastic test specimens is
presented in the Appendix.

Wrappings Around Plastics. Except for the tape wrapped around vinyl tube
No. 338, the plastic electrical insulation tape (Figure 23), which was wrapped
around the plastic rods and tubes, was attacked by marine borers. The small
1/324inch-diameter holes were found along the edge of the overlap, but the borers
did not penetrate the plastic tape. The deepest holes showed that some borers had
penetrated approximately three quarters of the way through the 0.010-inch-thick
electrical tape. It is believed that this is the deepest the borers are able to pene-
trate directly into the tape.

30

Figure 23. Plastic electrical insulation tape wrapped around plastic rods.

The burlap wrappings (Figure 24) on all the plastic rods and tubes were
deteriorated by bacterial action, and the material could easily be torn apart by hand.
The rubber electrical tape wrappings were not affected by fouling organisms although
there were hydroids and sponges growing on the rubber material. Slight cracks
(Figure 25) were found on the rubber and were probably produced by the effects of
the deep-sea environment.

0.015-Inch Insulation on No. 16 Wire. The 15-inch-long nonstressed (straight)
wire specimens were lost because the plastic cable clamps which held them to the
phenolic laminated sheet were severed; however, the stressed (coiled) wire specimens
were recovered (Figure 26). In the laboratory, the insulations were examined under
a stereoscopic microscope for signs of any deterioration. Electrical tests, such as
insulation resistance and voltage breakdown tests, were also conducted on these
specimens. The results of the electrical tests are presented in Table 10.

The entire surface area of the silicone rubber insulation was roughened, and
the thickness of the insulation was reduced considerably by the nibbling and chewing
action of some marine organisms (Figure 27). The surfaces of other insulations, such
as polyvinyl chloride, polyethylene, Government rubber styrene (GR-S), and neoprene
rubber, were not affected. However, there were deep cracks in one area of a neo-
prene rubber insulation (Figure 28), exposing the conductor to seawater. The insulation
was exposed about 3 feet above the sediment.

31

Figure 25. Rubber electrical insulating tape over plastic rods.

32

pl oe IN, Ne

eer /

Figure 26. Coiled 0.015-inch insulation and single conductor and
multiconductor electrical cables. Glass microscope
slides are also attached to the phenolic sheet.

Figure 27. Silicone rubber insulation deteriorated by nibbling and
biting action of some marine animals.

33

Table 10. Deep-Ocean Effects on Insulation Resistance of
Electrical Insulating Materials

Insulation Resistance (meg)

After
Exposu re2/

Exposed Near Sediment

Insulating
Material for
Coiled Wires

Voltage
Brea kdown3/

Before
Exposure l/

Polyethylene 20,100,000 370,000 Did not fail
Polyvinyl chloride 4,400,000 480,000 Did not fail
Silicone rubber 6,200,000 60,400 Did not fail

Government rubber
styrene (styrene 5,500,000 200,000 Did not fail
butadiene rubber)

Neoprene 36,000 27,500 Did not fail

Exposed About 3 Feet Above Sediment
—

Did not fail
Did not fail
Did not fail

Polyethylene 20,100,000
Polyviny! chloride 4,400,000
Silicone rubber 6,200,000

1,370,000
800,000
32,000

Government rubber

styrene (styrene ~ 5,500,000 96,0004/ ae He
butadiene rubber)
mee 36,000 32,000 Did not fail

1) Average of eight tests.

2/ Average of two tests.

3/ Tested at 1,000 volts AC for 10 seconds.

4/7 Average of one test. Insulation of one wire deteriorated.

34

Figure 28. Deep cracks in neoprene rubber insulation. The wire
was exposed to seawater.

Electrical Cable Insulation for Single Conductors and Multiconductors. The
insulations of the 10-inch-long electrical cables (Figure 26) were examined under
a stereoscopic microscope for signs of any biodeterioration and for any physical
effects of the deep-sea environment. 5

Biodeterioration due to borer Gctivity was more pronounced (especially in the
area of a wooden bait piece) on a set of electrical cable insulations which were
exposed about 0.5 foot above the sediment layer than on an identical set of insula-
tions which were exposed about 3 to 4 feet above the sediment. The insulating
materials which were damaged by the borers under a wooden bait piece were Teflon
(Figure 29), polyvinyl chloride, silicone rubber, bakelite, polyethylene, nylon,
and fluorinated ethylene propylene. The insulating materials which were neither
affected by borer activity nor by the deep-sea environment were butyl! rubber,
neoprene rubber, and natural rubber.

In addition to damage by borer activity under the wooden bait piece, the
surfaces of the silicone rubber insulations were damaged by the nibbling and biting
action of some marine organisms. As a result of this type of damage over the entire
surface area (except underneath the area of cable clamps), the thickness of the
insulation was reduced considerably (Figure 30).

39

Figure 29. Borer holes on the surface of Teflon electrical insulation
in an area under the wooden bait piece.

Figure 30. Surface of silicone rubber damaged by nibbling and
biting action of some marine animals. The section
where the cable clamp was located was not damaged.

36

Laminated Plastics. The 1x6x1/8-inch laminated plastic materials (Figure 31)
which were exposed near the sediment and about 3 feet above the sediment were not
damaged. Percent moisture absorbed by the plastics was determined by weighing each
of the test specimens before and after exposure in the sea. The results are presented
in the Appendix.

FINDINGS

1. There is considerable biological activity occurring in the sediment at Test Site I.

2. Marine fouling organisms such as bryozoa, tube worms, and glass sponges were
found on rock samples collected in the vicinity of Test Site | in 6,000 feet of water.

3. Hydroids and sponges were found in small amounts on certain metal specimens
and on all of the nonmetallic specimens. Typical fouling organisms such as bryozoa
and barnacles were not found on these panels.

4. Marine microorganisms were responsible for the deterioration of cotton and
Manila rope specimens, burlap (jute fibers), and possibly a vinyl plastic tube

(NCEL No. 338).

5. Two species of marine borers were found in wood specimens and have been
identified as Xylophaga washingtona Bartsch and Xylophaga duplicata Knudsen.

The borers were responsible for the deterioration of the following materials: (1)
pine, cedar, oak, maple, ash, and redwood, (2) plastic rods (area under the wooden
bait pieces) such as delrin, nylon, polycarbonate, Teflon, cellulose acetate, poly-
ethylene, acrylics, and polystyrene, (3) vinyl tubes, and (4) Manila rope.

6. Electrical cable insulation composed of silicone rubber was deteriorated by the
nibbling and biting action of some marine animals.

7. The following materials in various forms and for various uses were not affected
by marine organisms: rubber vacuum tubing; nylon nuts and bolts; nylon and poly-
propylene ropes; ethy! cellulose cable clamps; neoprene, butyl, and natural rubber
electrical cable insulation; various plastic laminated sheets; and glass microscope
slides.

37

Figure 31. Laminated plastics after recovery from the sea.

38

CONCLUSIONS

1. The following materials are susceptible to total biological destruction and are
not suitable for use in the deep ocean: untreated pine, fir, redwood, cedar, ash,
oak, and maple; Manila and cotton ropes; and burlap (jute fibers).

2. The following materials are probably not susceptible to biological deterioration
in the deep ocean: rubber vacuum tubing; acrylic sheet; nylon nuts and bolts;
nylon and polypropylene rope; ethyl cellulose cable clamps; laminated plastic
sheets; glass slides; and electrical cable insulations composed of neoprene, buty|
rubber, natural rubber, and polyvinyl chloride.

3. The 3-foot-long plastic rods and tubes (except vinyl tube, NCEL No. 338), the
polyvinyl chloride pipe, and the electrical cable insulations (except silicone rubber)
may be suitable for use in the deep ocean if they are not placed in direct contact
with wood.

FUTURE PLANS

The research program to determine the effects of the deep-sea environment on
engineering materials is continuing.

A STU (I-2) which was exposed for a period of 2 years at a depth of 5,640 feet
was recovered in October 1965. The materials aboard this STU are currently being
examined, tested, and evaluated for biodeterioration.

Plans are being readied to retrieve STU II-2 in May 1966. It will have been
exposed for | year on the sea floor at a depth of 2,400 feet.

ACKNOWLEDGMENTS
Dr. Ruth D. Turner, Museum of Comparative Zoology, Harvard University,

identified the marine borer specimens. Dr. Jan Kohlmeyer, Institute of Fisheries
Research, University of North Carolina, examined the wood specimens for fungi.

39

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REFERENCES

1. U. S. Naval Civil Engineering Laboratory. Technical Report R-428: Deep-ocean
biodeterioration of materials — Part III. Three years at 5,300 feet, by J. S. Muraoka.
Port Hueneme, Calif., Feb. 1966.

2.———. Technical Report R-329: Deep-ocean biodeterioration of materials —
Part |. Four months at 5,640 feet, by J. S. Muraoka. Port Hueneme, Calif.,
Nov. 1964.

3.———.. Technical Report R-393: Deep-ocean biodeterioration of materials —
Part Il. Six months at 2,340 feet, by J. S. Muraoka. Port Hueneme, Calif.,
Aug. 1965.

4, R. S. Dietz. "Manganese deposits on the northeast Pacific sea floor," California
Journal of Mines and Geology, vol. 51, no. 3, July 1955, pp. 209-220.

5. H. L. Ehrlich. "Bacterial types associated with manganese nodules," unpublished
paper presented at the meeting of the Eastern New York Branch, American Society
for Microbiology, Oct. 26, 1961.

6. R. L. Starkey. "The general physiology of sulfate reducing bacteria in relation
to corrosion," Producers Monthly, vol. 22, no. 8, June 1958, pp. 12-30.

7. National Research Council. Prevention of Deterioration Center. Bibliography
on microbial corrosion of metals, compiled by R. W. H. Lee. Washington, D. C.,

July 1963. (PDC search no. 63-025, PDL 4807. )

8. U. S. Naval Applied Science Laboratory. Technical Memorandum 1 of
Laboratory Project 9400-72: Bibliography on microbiological corrosion (deep sea).
Brooklyn, N. Y., June 1965.

9. R. Y. Morita and C. E. ZoBell. "Occurrence of bacteria in pelagic sediments
collected during the Mid-Pacific Expedition," Deep-Sea Research, vol. 3, no. 1,
Oct. 1955, pp. 66-73.

10. J. S. Muraoka. "Deep-ocean boring mollusk," BioScience, vol. 15, no. 3,
Melo IRS, fs IMile

1]. R. D. Turner. "The family Pholadidae in the western Atlantic and the eastern
Pacific, Part I! — Martesiinae, Jouannetiinae and Xylophaginae," Johnsonia,
Museum of Comparative Zoology, Harvard University, vol. 3, no. 34, Mar. 29,
1955, pp. 65-160.

12. J. Knudsen. "The bathyal and abyssal Xylophaga," in Galathea report,
vol. 5, 1961, pp. 163-209.

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BINGHAM OCEANOGRAPHIC LABORATORY» YALE UNIVERSITY: Pe Oe BOX 20255 YALE STATIONs
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MRe WILLARD Je PIERSON JR» DEPARTMENT OF METEOROLOGY AND OCEANOGRAPHY s NEW
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DIRECTOR» INSTITUTE OF MARINE SCIENCEs THE UNIVERSITY OF TEXASs PORT ARANSAS»
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FISHERIES-OCEANOGRAPHY LIBRARY» 2U3 FISHERIES CENTER» UNIVERSITY OF WASHINGTON»
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CIVIL ENGINEERING DEPARTMENTs TEXAS A AND M UNIVERSITY» COLLEGE STATION» TEXe
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BOCA RATONs FLAe 33432

DRe EVELYN SINHAs EDITORs OCEANIC COGRDINATE INDEXs 7730 HERSCHEL AVENUE 9
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DRe KEN PREISS»s GEOLOGY AND CIVIL ENGINEERINGs DEPARTMENT OF GEOLOGY»
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CHIEFs BUREAU OF NAVAL WEAPONSs ATTN RESEARCH DIVISION,
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NEW MEXICO 87117

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COMMANDING OFFICERs BIOLOGICAL WARFARE LABORATORIES» FORT DETRICKs FREDERICKs

MARYLAND 21701

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WASHINGTONs De Ce 20396

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NORTH CAROLINA 28480

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MRe ED BEST» MARINE RESOURCES OPERATIONS» STATE OF CALIFORNIA s DEPARTMENT OF
FISH AND GAMEs MENLO PARKs CALIFe 94028

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Unclassified
Security Classification

DOCUMENT CONTROL DATA - R&D

(Security classification of title, body of abstract and indexing annotation must be entered when the overall report is classified)

1. ORIGINATIN G ACTIVITY (Corporate author) 2a. REPORT SECURITY CLASSIFICATION

U.S. Naval Civil Engineering Laboratory ST _Linellegsilige
Port Hueneme, California 93041 ee ee |
3. REPORT TITLE

Deep-Ocean Biodeterioration of Materials — Part IV. One Year at 6,800 Feet

4. DESCRIPTIVE NOTES (Type of report and inclusive dates)

March 1964 - March 1966

5. AUTHOR(S) (Last name, first name, initial)

Muraoka, James S.

6. REPORT DATE Ja. TOTAL NO. OF PAGES 7b. NO. OF REFS
June 1966 60 2

8a. CONTRACT OR GRANT NO. 9a. ORIGINATOR'S REPORT NUMBER(S)

TR-456

. PROJECT NO.

Y-RO11-01-01-042
9b. OTHER REPORT NO(S) (Any other numbers that may be assigned
this report)

-AVAILABILITY/LIMITATION NOTICES

Distribution of this document is unlimited.

- SUPPLEMENTARY NOTES 12. SPONSORING MILITARY ACTIVITY

Naval Facilities Engineering Command

- ABSTRACT

As Part IV of a series of reports on the biological deterioration of materials in the
deep ocean, this report covers the data obtained after exposing metallic and nonmetallic
specimens for 13 months on the floor of the Pacific Ocean at a depth of 6,800 feet (Test
Site 1). The specimens were attached to a Submersible Test Unit that was retrieved in
February 1965. Preliminary examination of the specimens was made aboard ship, and the
final examination, tests, and analyses were performed at the Naval Civil Engineering
Laboratory.

On recovery, most of the plastic materials and all of the rope specimens were covered
with bacterial slime. Cotton and manila rope specimens were severely deteriorated by marine
microorganisms. Wooden test panels, plastics, and Manila ropes were attacked by molluscan
borers. Glass, metals, natural and butyl rubber, and certain plastic materials were not
adversely affected.

The results of breaking-strength tests on ropes, weight loss measurements of wood panels,
moisture absorption tests on plastics, and insulation resistance and voltage breakdown tests
on electrical insulating materials are presented.

DD ieee 1473 0101-807-6800 Unclassified

Security Classification

Unclassified
Security Classification

KEY WORDS

Oceanography
Biology
Deterioration
Materials

Deep ocean
Environment
Fouling organisms
Marine borers
Seawater
Sediments

INSTRUCTIONS

1, ORIGINATING ACTIVITY: Enter the name and address
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all security classification of the report. Indicate whether
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ever, the suggested length is from 150 to 225 words.

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words but will be followed by an indication of technical con-
text. The assignment of links, rales, and weights is optional.

Unclassified
Security Classification

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