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Hylebos Waterway Fish Injury Studies

Individual Data and Quality Assurance Results

CASE NARRATIVE

Reproductive Injury in Flatfish

Ovarian Histopathology

Histopathologist’s qualifications

Histologic sections of gonad from individual fish were examined by three

histopathologists (Mark Myers, Carla Stehr, Paul Olson) with at least three years of

experience in the histopathologic examination of English sole and rock sole. The latter two

examiners have completed a one-year training program under the direction of the chief

pathologist, Mark S. Myers. Each histopathologist examined approximately 33% of the total

number of cases.

Diagnostic criteria

Criteria for microscopic lesion diagnosis were adopted from comparative pathology

textbooks (Robbins, Cotran, and Kumar, 1984) and various publications in fish

histopathology (Myers et al., 1987; Moore and Myers, 1994; Johnson et al., 1991; Hinton et

al., 1992).

Histologic slide labeling

All histologic slides were labeled with a random number code so that the geographic

site of capture was not known to the examiner.

Internal quality control proceduresfor histologic slide ezamination

For histologic slides examined by Carla Stehr and Paul Olson affected with suspected

or diagnosed liver lesions, or where liver lesion diagnosis was not obvious, all tissues,

including ovaries, were re-exammed by the chief pathologist, Mark Myers. Additionally,

ovaries from 20 additional specimen were chosen at random for re-examination. Out of the

approximately 269 tissues sections of gonad examined (from 269 individual English sole), 38

tissue sections or 14% of samples were re-examined by the chief pathologist. The original

and final, confirmed diagnoses for these re-examined individual cases are set forth in Table

1.

Coding of diagnoses into database

Each final histopathologic diagnosis was entered into the database by the examining

histopathologist, using a lesion coding system developed at the Northwest Fisheries Science

Center. Lesion categories include the following:

-atresia of yolked oocytes

-atresia of non-yolked oocytes

-late stage atresia

-inflammation in the ovary
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Missing data

Occasionally the cassette used to hold tissues from individual fish will open up in the fixative

container, resulting in the loss of the tissue; also, in the necropsy procedure, occasionally

tissues are inadvertently not collected. Infrequent labeling errors may also occur during the

handling of a large number of fish. If the tissue is lost, or there is a question about the

labeling, lesion information is not used, and missing data for a particular tissue is indicated

by a blank record. Specimens collected for these studies, but where histopathology data is not

available are:

943374, 943531, 943562, 943742, 943806, 943843, 943281, 943293, 043296, 943572,

943651,943732,943761,943806,943843

Otoliths are sometimes lost or damaged during collection; therefore age data cannot be

obtained. This missing data is indicated by a blank record. Fish where we were unable to

obtain age data were specimens 943350 and 943649.

Sequential gaps in the specimen numbers do not represent missing fish. Those specimens

listed in Table 2 are female English sole reported in the Reproductive Injury study. Specimen

number not listed were either reported in the Toxicopathic Injury study, or assigned to other

projects.

Revisions 2/24/97

The data presented in this case narrative were previously included in with liver and kidney

data as part of the Toxicopathic Injury study case narrative submitted on 5/30/96. However,

since detailed data on the histopathology of ovarian lesions in English sole is only reported in

the Reproductive Injury Interpretive Report, this information now appears here in a separate

document. In addition to information on ovarian lesion occurrence, this case narrative also

includes ovarian lesion severity rankings. Data on ovarian Quality Assurrance has been

expanded; slides from 20 additional female English sole collected specifically for the

Reproductive Injury study were randomly selected and reread by the chief pathologist. No

additional changes to lesion diagnoses or other information contained in the case narrative

were made.
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Documentation and Abbreviations for Tables 1 and 2

Ovarian Histopathology Data

for Hylebos Flatfish Reproductive Injury Study

This case narrative includes ovarian histopathology for flatfish collected during the

Reproductive Injury Study of the Hylebos Fish Injury Studies, Phase I. Other histopathology

data for these fish are presented in for liver and kidney histopathology case narrative for

flatfish collected during the Toxicopathic and Reproductive Injury Studies.

Notes and Abbreviations

Date collected - data fish were caught, and tissue samples collected

Reader - indicates last name of the person who read the histopathology slides for that

specimen number. A blank record occurs when no tissue sections were available to be read

for that specimen.

Ovary lesions

YATR = early stage (alpha) atresia of yolked oocytes

YATRsev = severity rating for early stage atresia. of yolked oocytes

NYATR = early stage (alpha) atresia of non-yolked oocytes

NYATRsev = severity rating for early stage atresia. of non-yolked oocytes

LATR = late stage atresia, (can’t distinguish yolked from non-yolked at this stage)

LATRsev severity rating for late stage atresia

OVINFL = inflammation in the ovary

OVINFLsev = severity rating for inflammation in the ovary

Gonad stages

OVSTAGE = stage of maturation of the ovary

Age:

Actual years of age as indicated by growth rings on otoliths.

Blank records indicate no age data available; otoliths lost or damaged.

Sex:

1 = male

2 = female

Length: total length, reported in millimeters

Weight: total weight, reported in grams
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Explanation of codes

Presence or absence of lesions:

0= tissue examined, no lesion observed

1= tissue examined, lesion observed

blank = tissue not examined

Lesion severity:

0 = absent

1 = minimal, sparse, very few

2 = minimal to mild

3 = mild, few

4 = mild to moderate, several

5 = moderate, moderate number

6 = moderate to severe

7 = severe, abundant, numerous, dense

8 = excessive amount or number, excessively dense blank = tissue not examined

Stage of Maturation of the Ovary

1 = regressed; oogonia and primary oocytes

2 = late regressed; secondary oocytes

3 = previtellogenic; vacuolated secondary oocytes

4 = vitellogenic

5 = some hydrated oocytes; no post-ovulatory follicles (POFs)

6 = spawning; hydrated oocytes with POFs

7 = spawned out

8 = equivocal spawn out; characteristics of spent fish but no identifiable POFs
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