Reproduction disorders-malformed embryos of amphipods and eelpout

Brita Sundelin1 and Jakob Strand2

Stockholm University, ITM

AArhus University, dep of Bioscience

Background information for malformed embryos of amphipods

• Thirty years of field studies in contaminated and pristine areas, bioassays with contaminated sediments and chemicals spiked in sediments have resulted in increased occurrence of malformed embryos of the stenotherm key stone amphipod species Monoporeia affinis.

• The indicator is general and all types of contaminants cause malformations, while oxygen and food deficiency , and increased temperatures cause other types of embryo aberrations.

• A National Monitoring programme started in 1994 to use the indicator for assessment of contaminated sediments. During the last years also other amphipod species have been used. Assessment criteria have been calculated for the variables malformed embryos and proportion of females carrying malformed embryos.

• Method is recommended by ICES WGBEC

Contaminants result in malformed embryos of amphipods

Sundelin et al. 1998, 2008, MEPS , TIMES

Normal Malformed

Dead broods

Dead eggs

Arrested development

Other types of embryo aberrations

Background information for malformed embryos of Eelpout

• Reproductive success in eelpout is regarded as a general, i.e. non-specific, biological indicator of impaired fish reproduction, and a significant biological endpoint for assessing potential population-relevant effects.

• Several toxic substances including many endocrine disruptors can cause skeletal defects during fry development in fish.

• Other environmental factors, e.g. high temperature and oxygen depletion, seem rather to cause other types of abnormal fry development.

• Method is recommended by ICES WGBEC

Stage of development Indicator type

pre-core State/Impact

Primary importance Secondary importance

BSAP

Segment and Objective

Healthy wildlife

Viable populations of species

Concentrations of hazardous substance are

close to natural levels

MSFD

Descriptors and Criteria

8.2 Effects of contaminants (chronic and acute

pollution effects)

8.1. Concentrations of contaminants

1.2 Population size (abundance and

biomass)

1.3 Population condition (demography,

genetic structure)

Other relevant legislation: (e.g. WFD)

Legislative linkage:

Malformed Embryos of eelpout and amphipods

Concept/ design

Coordinated monitoring Assessment

Research needs for operationalization

(in relation to needs stated under the

coordinated monitoring and

assessment columns)

Data arrangements

Monitoring strategy (method, frequency, spatial resolution) in relation to relevant indicator parameters Technical guidelines Geographic scale Assessment method

GES / assessment criteria (currently all GES are provisional)

A ) in place B) under development C ) not available, what needs - action level?

A ) monitoring in place B ) monitoring needs revision C ) monitoring not available, what needs - action level?

A ) in place B ) needs revision, what needs doing C ) not available, what needs - action level?

HELCOM assessment units: A ) identified B) Identified not described C) not identified, what needs - action level?

A ) available and described B ) available not described C ) not available, what needs - action level?

A ) proposed and described B ) proposed but needs more supporting data C ) not available, what needs - action level?

A ) in place B ) needs revision, what needs doing C ) not available, what needs - action level?

B - rate of malformation related to type to be more explicitly described, several field studies in contaminated areas and laboratory assays with chemicals and all types of studies show the correlation between malformation rate and contaminants - TM

B - eelpout included in some CPs environmental monitoring programmes - TM

B - reproductive success of eelpout described in COMBINE and international guideline is under development and to be included -TM

B - identified on a general level, units to be defined, amphipods are stationary and reflect the contamination level at station, including several stations within an area allows for assessment of the area but not to transfer effects to other areas in the Baltic - TM

B - assessment information is available in TIMES 41 (the method description of the indicator) - TM

B - to be detailed, for amphipods 5-step scale to be turned into GES, Assessment criteria has been developed for MSFD and are available in report to SEPA and HAV (Reutgard and Sundelin 2013) - TM

Amphipod data currently only from Swedish area, wider sampling needed. Scientist in Denmark and Russia are working with amphipods reproduction

C - data pooling and collecting needs to be outlined - TM, HELCOM.

A/ amphipods concept is in place A/ eelpout concept is in place

A/ Monitoring in place A/ Monitoring in place

A/ technical guidance in place B/ National guidelines needs to go into international protocol

A/ identified for Monoporeia affinis,

A) Assessment method for amphipod is available and described A/Assessment method for eelpot is available and described

A/ Assesment criteria are proposed and described A/ Assesment criteria are proposed and described

Research is needed to be able to estimate population effects caused by differrent grade of embryo malformations.

B) National databases in SE and DK submit data to ICES

Malformed embryos of amphipods

• HELCOM Assessment Unit Level for M. affinis: level 2-3.

• The indicator is applicable in: all coastal areas around the Baltic except Gulf of Finland, Gulf of Gdansk and Belt Sea where the species has disappeared.

• Currently data is available : Bothnian Sea, Baltic proper.

Malformed embryos and eelpout

• HELCOM Assessment Unit Level for eelpout: 3-4

• The indicator is applicable in: all coastal areas around the Baltic except some parts of Bothnian Sea, and Gulf of Finland.

• Currently data is available : Kattegat, Belt Sea, western Baltic Sea, Swedish east coast and Gulf of GDansk.

GES values for malformed embryos

• The emphasis will be to develop GES-values based on the available monitoring data. A simple method to develop GES-values is to use percentiles of the resulting distributions .

• There are, however, limitations with GES-values based on percentiles. One limitation is that it assumes that the reference areas that are used to create the normal distribution are not affected. For the Baltic Sea, such reference sites may be difficult to find. Martin Reutgard, Matias Ledesma and Brita Sundelin were involved in the work with amphipods.

Calculation of assessment criteria for malformed embryos of Monoporeia affinis

• Data from the Monitoring program between 1994 and 2011 was used.

• 14 stations were included

• 8600 gravid females and 230 000 embryos were included.

• Repeated hazardous sampling, i.e. Bootstrapping, were we can control the sample size were used. We used a sample size of 50 females (about 1500 embryos) and repeated the hazardous sampling 100 000 times to get an even spread.

Proportion of malformed embryos of the amphipod Monoporeia affinis. Green line illustrates the background level and orange line the value that is significant increased from background.

Andel missbildade embryon

Fre

kve

ns

0.00 0.05 0.10 0.15

01

00

02

00

03

00

04

00

05

00

0

Proportion of malformed embryos

Proportion females with >1 malformed embryos. Green line illustrates mean value in background areas and orange line a significant deviation from background.

Andel honor med fler än ett missbildat embryo

Fre

kven

s

0.0 0.1 0.2 0.3 0.4 0.5

050

0010

000

1500

0

Proportion females with >1 malformed embryos

Assessment Criteria

Mean (GES?) Background level (BAC)

Increased level (EAC)

Proportion malformed embryos

2.6 % 0-3.8 % >3.8 %

Proportion females with >1 malformed embryos

15 % 0-22% >22 %

Reference: ICES WGBEC report 2012 based on input from BEAST and BALCOFISH projects

Revised Assessment criteria for abnormal fry development in eelpout

Jakob Strand, Aarhus University at HELCOM CORESET meeting in Roskilde, 11 -12. February 2013

<BAC/R

>BAC/R, <EAC

>EAC

Assessment Criteria BAC/R EAC

Prevalence of

malformed eelpout fry

1% 2%

Data from BALCOFISH, NOVANA and Toxicon

Jakob Strand, Aarhus University at HELCOM CORESET meeting in Roskilde, 11 -12. February 2013

2009 assessment for prevalence (%) of malformed fry in eelpout broods

<BAC/R

>BAC/R, <EAC

>EAC

Assessment Criteria BAC/R EAC

Prevalence of

early dead eelpout fry

2.5% 5%

Data from BALCOFISH, NOVANA and Toxicon

Jakob Strand, Aarhus University at HELCOM CORESET meeting in Roskilde, 11 -12. February 2013

2009 assessment for prevalence (%) of early dead fry in eelpout broods

Malformed embryos of amphipods and eelpout

List of issues that still need

to be solved for the

indicators

Describe what is hindering solving the issue

What does a specific rate

of malformation means

to the population?

It might be possible to use models to predict population

effects but then we need to collect amphipods and

determine the different age classes frequently during at

least one year to be able to estimate e.g. mortality rate.

Funding for data collection and analysis is still unsecure and

the amount of data needed for the analysis has not yet

been met.

Relationships to specific

group of substances and

other environmental

factors

Complex mixtures of contaminants (that often co-variate)

exist in all areas of the Baltic Sea, and it is from field studies

difficult to point at specific groups of contaminants that can

explain elevated levels of malformations in some areas.

Modelling impact on populations

A study of impact of malfored fry development in eelpout (Bergek et al. 2012) supports that induced malformation from environmental pollution can have a large effect on natural populations, and especially in sensitive eelpout populations with lower growth rates.

Consequently, even though a threshold value of 2 % malformations of fry may seems low, it could have serious consequences at the population level, especially then recognised that also other types of severe abnormal fry developments can occur in the same broods.

Monitoring-Sampling of gravid amphipod females

Winterbreeding Monoporeia affinis

Former program design to the left, present program to the right started in 2012.

The program design was changed in 2012 and the number of stations increased

Proportion of malformed embryos on monitoring stations in Baltic proper and Bothnian Sea

0

0,05

0,1

0,15

0,2

0,25

0,3

1994 1996 1998 2000 2002 2004 2006 2008 2010

Egentliga östersjön

Bottenhavet

Baltic proper

Bothnian Sea