INFORME TÉCNICO · 2019-03-25 · 1 INFORME TÉCNICO RESPUESTA A SIC 14 – ECOSISTEMAS ACUÁTICOS II CUECAR S.A. Y BLANVIRA S.A. Exp. 2018/14000/011210 El presente informe tiene

1

INFORME TÉCNICO

RESPUESTA A SIC 14 – ECOSISTEMAS ACUÁTICOS II

CUECAR S.A. Y BLANVIRA S.A.

Exp. 2018/14000/011210

El presente informe tiene como objetivo dar respuesta a la Solicitud de Información Complementaria 14 (en adelante SIC) realizada por la DINAMA con fecha 4 de feberero del 2019 por informe de la División Emprendimientos de Alta Complejidad respecto a los documentos presentados con la Solicitud de Autorización Ambiental Previa (en adelante SAAP), presentadas por las firmas Cuecar S.A. y Blanvira S.A. para la instalación de una Zona Franca y una Planta de Producción de Celulosa a instalarse en la zona Norte del departamento de Durazno a orillas del río Negro. Dicha SIC es una segunda solicitud referente a la información de Ecosistemas Acuáticos presentada en el Estudio de Impacto Ambiental. A continuación se presenta la respuesta a esta SIC.

Muestreos de peces e invertebrados bentónicos

Solicitud:

“(...)””se evidencia la necesidad de acordar entre el titular y DINAMA un diseño de muestreo, el conjunto de variables a medir y los indicadores ambientales para establecer la línea de base de biota acuática previo a seguir realizando actividades de muestreo, so pena de que el esfuerzo que la actividad conlleva no produzca información de utilidad para el objetivo de esta línea de base.”

Respuesta:

En el “Anexo I – Fish community Field Program” se adjunta un documento como adelanto de la respuesta a este punto, documento que ya fue presentado por Ecometrix oralmente ante técnicos de DINAMA en reuniones mantenidas en marzo de 2019. Más adelante se ampliará información para completar esta respuesta.

2

Identificación de zonas de relevancia para la biota

Solicitud:

“La identificación de estas áreas, solicitada mediante la SIC 05 de fecha 20 de noviembre de 2018, es de importancia para la valoración de los impactos de la planta industrial sobre los ecosistemas acuáticos. Asimismo es un insumo fundamental para el diseño de medidas de mitigación, en caso de ser necesario, y para el diseño metodológico del monitoreo de la línea de base de biota acuática y de los monitoreos en la fase de operación.

Para identificar las áreas de relevancia para la biota es necesario contar con un mapa de ambientes acuáticos del medio receptor que permita, al menos, definir potenciales áreas de reproducción, cría y alimentación de especies amenazadas de peces (anexo III en Loureiro et al., 2013) y/o de interés comercial (pesquería).

Para la elaboración del mapa de ambientes acuáticos se entiende necesario que en él se integren, al menos, los siguientes componentes:

1. Profundidad

2. Morfología del margen (ej. playa arenosa, playa pedregosa, barranca)

3. Sustrato/tipo de fondo

4. Velocidad del agua.

5. Vegetación en el margen del curso de agua (con las categorías por ejemplo: plantas emergentes, sumergidas, flotantes, monte ripario, vegetación herbácea, pajonales, juncales, totorales).

En cuanto a la extensión geográfica del mapa de ambientes es necesario que este se extienda, al menos, 5 km aguas abajo y 4 km aguas arriba del punto previsto para la descarga de efluentes. De este modo se pretende relevar las singularidades de todos los sitios a ser utilizados para el monitoreo de línea de base y posterior seguimiento, así como de todos aquellos lugares potencialmente más afectados por tal descarga.

Se deberá presentar a DINAMA una memoria descriptiva de los sitios de relevancia identificados que ameriten alguna actividad específica en cuanto a monitoreo de línea de base y posterior seguimiento, incluyendo su justificación. Asimismo, se deberá incluir una memoria descriptiva del mapa de ambientes acuáticos de todo el tramo de 9 km de extensión antedicho y que comprenda los componentes antes mencionados (1 al 5).

La información cartográfica será presentada también en formato kml, kmz y/o shape incluyendo archivos con las distintas capas de información (profundidad, morfología del margen, sustrato/tipo de fondo, velocidad del agua, vegetación en el margen).

Adicionalmente, y como fuera solicitado en la SIC 05, en base al mapa de zonas de relevancia para la biota se deberá valorar los potenciales efectos de la planta industrial sobre las áreas identificadas, considerando las distintas condiciones hidrológicas posibles del río Negro.”

Respuesta:

En el “Anexo II - Fish Habitat Assessment” se presenta la respuesta a esta solicitud. La traducción de este documento al idioma español será presentada más adelante.

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Disruptores endócrinos

Solicitud:

“El abordaje realizado por el titular para la evaluación de la potencial afectación del ecosistema acuático por efecto de los efluentes de la planta de celulosa actuando como disruptor endócrino (ED), se centró sobre los efectos en la reproducción de peces.

Por una parte ese análisis concluye que es de esperar un muy bajo riesgo de potenciales efectos reproductivos en peces, en base a una serie de argumentos presentados como “hallazgos clave” que dan contexto y que sustentan en gran parte el análisis realizado, pero sin embargo no se incluyen ni las citas ni las referencias bibliográficas para la mayoría de tales argumentos, lo que dificulta la valoración de la evidencia expuesta. Por lo expuesto se solicita que se incluyan las citas y las referencias bibliográficas que sustentan el análisis de riesgo por contaminación por EDs que ha sido hasta ahora realizado.

Sin perjuicio de la solicitud precedente, corresponde anotar que los EDs actúan también alterando otras funciones en peces, como el desarrollo, funciones tiroideas y alteraciones morfológicas (Pait & Nelson, 2002, Parrott et al., 2004, Mills & Chichester, 2005), y también sobre otros grupos taxonómicos (Matthiessen et al., 2018), cuestiones que no han quedado cubiertas con el análisis hasta ahora presentado. Por ello se deberá ampliar el análisis sobre potenciales interferencias de los EDs a funciones distintas a la reproducción en peces, y se deberá considerar también la potencial afectación sobre otro tipo de organismos.

Por otra parte, para el análisis de riesgo se entiende necesario incluir una descripción de la situación actual del medio receptor en cuanto a la contaminación por EDs, identificando potenciales fuentes de contaminación y vacíos de información existentes. Asimismo se deberá identificar los compuestos presentes en el efluente de la planta con potencial de actuación como EDs, así como las concentraciones esperadas de ellos en la descarga.”

Respuesta:

En el “Anexo III - Potential for EDC Effects in Modern Mill Effluents” y en el “Anexo IV - Review of EDC References Provided by DINAMA” se presenta la respuesta a esta solicitud. La traducción de estos documentos al idioma español será presentada más adelante.

Ing. Civil H/S Carlos Amorín Por Estudio Ingeniería Ambiental

»ANEXO I

SIC 14 - ECOSISTEMAS ACUÁTICOS II

Fish Community Field Program

1

Environmental IntelligenceUPM Pulp Mill 18th March 2019

Fish Community Field Program

(2019)

2

Environmental IntelligenceAgenda

• Area Overview

• Field Methods

• Preliminary Results

• Questions and Discussion

3

Environmental IntelligenceSampling Area of the Baygorria Reservoir

Zone 1

(NF)

Zone 2

(FF)

Zone 3

(Ref)

Study area is located in

the upstream area of

the reservoir, as such, it

behaves as a river.

4

Environmental IntelligenceFish Community Sampling

• Three primary habitat types were fished in each

zone; main channel, island channel, and lagoons.

• In zone one the main channel was sampled in the

area of the proposed location for the intake and

discharge pipe locations.

• All areas were sampled in triplicate during the day.

Additionally, night sampling was conducted within

the lagoon areas of each zone.

• Each net set included a NORDIC Net and a larger

mesh gillnet.

• Due to a change in weather, the fishing program

was stopped. It will be completed in March.

5

Environmental IntelligenceFish Sampling Areas

Zone AreaID Day/Night Habitat Other Net Type# of Net

Sets

# of Netsets

Completed

NORDIC 3 2

Large Mesh 3 2

NORDIC 3 3

Large Mesh 3 2

NORDIC 3 3

Large Mesh 3 2

NORDIC 3 2

Large Mesh 3 3

NORDIC 3 3

Large Mesh 3 3

NORDIC 3 3

Large Mesh 3 3

NORDIC 3 3

Large Mesh 3 3

NORDIC 3 3

Large Mesh 3 3

NORDIC 3 3

Large Mesh 3 3

NORDIC 3 0

Large Mesh 3 0

NORDIC 3 0

Large Mesh 3 0

NORDIC 3 0

Large Mesh 3 0

NORDIC 3 0

Large Mesh 3 0

NORDIC 3 0

Large Mesh 3 0

Main Channel

Lagoon

Island Channel

-

-

Intake

Dischage

-

-

Island Channel

Lagoon

Main Channel

Lagoon

Main Channel

Island Channel

K

N

D

D

N

D

D

N

D

E

F

G

H

I

J

3

1

2

D

A

B

C

D

Zone 1

Zone 2

Zone 3

AB

C

D

E

FG

Fish sampling to be conducted in March identified by grey highlight.

6

Environmental IntelligenceFish Community (Preliminary Results)

Zone 1

Zone 2

Zone 3

AB

C

D

E

FG

• Main Channel – Areas C (Ref Zone 3), F (NF Zone

1, intake), and G (NF Zone 1, discharge).

• Lagoon – Areas B (Ref Zone 3), D (NF Zone 1, US

intake), and E (NF Zone 1, DS intake).

• Island Channel – Area A (Ref Zone 3)

For each of the three comparisons the

following endpoints were calculated and

graphed; CPUE, BPUE, and Richness.

Species lists are also presented.

7

Environmental IntelligenceFish Community Main Channel (Preliminary Results)

NORDIC LargeMesh

NORDIC LargeMesh

NORDIC LargeMesh

Ref Intake Discharge

0

5

10

15

20

25

30

35

40

45

50

CP

UE

(#/h

ou

r)

* *

Zone 1

Zone 2

Zone 3

C

FG

(C) (F) (G)

* Identifies when symbols overlap.

8


NORDIC LargeMesh

NORDIC LargeMesh

NORDIC LargeMesh


0

200

400

600

800

1000

1200

1400

1600

BP

UE

(g

/ho

ur)

*

*

*

Zone 1

Zone 2

Zone 3

C

FG

(C) (F) (G)


9


NORDIC LargeMesh

NORDIC LargeMesh

NORDIC LargeMesh


0

2

4

6

8

10

12

14

16

18

Ric

hn

ess

* * *

Zone 1

Zone 2

Zone 3

C

FG

(C) (F) (G)


10


Area

# of Net Sets 2 3 3 3 3 3

Net Type NORDIC Large Mesh NORDIC Large Mesh NORDIC Large Mesh

Apareiodon affinis x x x

Astyanax rutilus x x x

Astyanax sp. x

Bryconamericus iheringii x x x

Characidium aff. Zebra x x

Characidium sp. x

Characidium sp. / Apareiodon affinis x

Crenicichla minuano x x

Crenicichla sp. x x

Cyanocharax alburnus x x x

Cyphocharax voga x x

Diapoma terofali x x x

F. Cichlidae x

Gymnogeophagus sp. x x

Gymnogeophagus tiraparae x

Hoplias lacerdae x

Hypostomus aspilogaster x x

Hypostomus commersoni x x x

Mojarra x

Mojarra tipo 1 x

Mojarra tipo 2 x

Mojarra tipo 3 x

Mojarra tipo 4 x

Odontesthes humensis x x x

Oligosarcus jenynsii x

Oligosarcus oligolepis x x x

Platanichthys platana x x x

Vieja del agua (no ID) x

- Identifes priority speies for conservation


bold - identifies a species that is threatened in Uraguay

11

Environmental IntelligenceFish Community Lagoons (Preliminary Results)

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

Ref Day Ref Night US Day DS Day DS Night

0

10

20

30

40

50

60

70

CP

UE

(#/h

ou

r)

*

* *

Zone 1

Zone 2

Zone 3

B

D

E

(B) (B) (D) (E) (E)


12


NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh


0

1000

2000

3000

4000

5000

6000

7000

BP

UE

(g

/ho

ur)

*

*

*

Zone 1

Zone 2

Zone 3

B

D

E

(B) (B) (D) (E) (E)


13


NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh

NO

RD

IC

La

rge M

esh


0

2

4

6

8

10

12

14

Ric

hn

ess

**

**

**

Zone 1

Zone 2

Zone 3

B

D

E

(B) (B) (D) (E) (E)


14


Area

Day/Night

# of Net Sets 3 2 3 2 3 3 3 3 3 3

Net Type NORDIC Large Mesh NORDIC Large Mesh NORDIC Large Mesh NORDIC Large Mesh NORDIC Large Mesh

Apareiodon affinis x x x x x

Astyanax rutilus x x x x

Astyanax sp. x

Bryconamericus iheringii x x x x x

Characidium aff. Zebra x x

Charax stenopterus x

Corydoras paleatus x

Crenicichla minuano x

Crenicichla sp. x

Cyanocharax alburnus x x x x

Cyphocharax voga x x x x

Diapoma terofali x x x

Eigenmannia virescens x

Gymnogeophagus gymnogenys x

Gymnogeophagus sp. x x

Hoplias lacerdae x

Hoplias malabaricus x x

Hypostomus commersoni x

Iheringichthys labrosus x x x x x x

Loricariichthys anus x

Mojarra (no ID) x

Odontesthes humensis x x x x

Oligosarcus jenynsii x x x x x

Oligosarcus oligolepis x x x x x

Pachyurus bonariensis x x x

Parapimelodus valenciennis x

Pimelodus maculatus x x

Platanichthys platana x x

Rhamdia quelen x

Rhinelepis strigosa x x x x x x

Trachelyopterus lucenai x

Trachelyopterus teaguei x x

Vieja (no ID) x

- Identifes species with conservation status.

bold - identifies a species that is threatened in Uraguay

Ref US of Intake DS of Intake

Day Night Day Day Night

15

Environmental IntelligenceFish Community Island Channels (Preliminary Results)

NORDIC Large Mesh

Ref

0

20

40

60

80

100

120

140

160

180

CP

UE

(#/h

ou

r)

*

NORDIC Large Mesh

Ref

0

500

1000

1500

2000

2500

3000

3500

4000

4500

BP

UE

(g

/ho

ur)

*

NORDIC Large Mesh

Ref

0

2

4

6

8

10

12

14R

ich

ness

*

Zone

1

Zone

3

A

Area

# of Net Sets 2 2

Net Type NORDIC Large Mesh

Apareiodon affinis x

Astyanax rutilus x

Bryconamericus iheringii x

Cyanocharax alburnus x

Cyphocharax voga x

Iheringichthys labrosus x

Mojarra (no ID) x

Odontesthes humensis x

Oligosarcus jenynsii x

Oligosarcus oligolepis x

Pachyurus bonariensis x

Platanichthys platana x

- Identifes species with conservation status.

Ref

bold - identifies a species that is threatened in Uraguay(A)

(A) (A)


16

Environmental IntelligenceSummary of Fish Sampling Program (Preliminary Results)

• Main Channel – CPUE, BPUE, Richness and Species similar between areas.

• Lagoons - CPUE, BPUE, Richness and Species was similar between lagoons D and E.

Lagoon B has similar richness and species composition to D and E but has lower CPUE

and BPUE than D and E – this may be due to variability or size (I.e. lagoon B is 1/4 the

size of lagoon D and 1/8 the size of lagoon E.

• Island Channel – yet to be determined.

• Fish sampling program to be completed in March.

• Once the fish sampling program has been completed a full report will be issued.

17

Environmental IntelligenceFurther Discussion

Gracias!

ANEXO II »SIC 14 - ECOSISTEMAS ACUÁTICOS II

Fish Habitat Assessment

UPM Pulp Mill Fish Habitat Assessment for a Proposed Pulp Mill along the Río Negro at Paso de los Toros, Durazno Department, Uruguay

Report prepared for: Blanvira S.A. Avenida Italia 7519, Piso 2 Montevideo, Uruguay Report prepared by: ECOMETRIX INCORPORATED www.ecometrix.ca Mississauga, ON Ref. 18-2534 1 March 2019

http://www.ecometrix.ca/

UPM Pulp Mill Fish Habitat Assessment for a Proposed Pulp Mill along the Río Negro at Paso de los Toros, Durazno Department, Uruguay Michael White, Ph.D Project Manager

______________________________________ Nicholas Edmunds, M.Sc. Biologist and GIS Analyst Brian Fraser, M.Sc. Project Principal and Reviewer

FISH HABITAT ASSESSMENT Executive Summary

Ref. 18-25 1 March 2019 i

EXECUTIVE SUMMARY

UPM retained EcoMetrix Incorporated (EcoMetrix) in partnership with Laboratorio Tecnológico del Uruguay (LATU) to characterize aquatic habitats in an area of the Rio Negro near Paso de los Toros, Durazno Department, Uruguay. The characterization was completed to support efforts related to the environmental approvals processes for the proposed UPM pulp mill development.

Field surveys were undertaken in February 2019 within a study area comprising an approximate 10 km reach of the Rio Negro downstream of Paso de los Toros. The study area was divided into three zones; Zone 1 - in the immediate vicinity of the proposed mill effluent outfall and water intake; Zone 2 - downstream of Zone 1 to the maximum extent of the hydrodynamic mixing zone as determined by predictive modeling; and, Zone 3 - upstream of the proposed mill site. Shoreline and vegetation features, bottom substrate types and river bathymetry were mapped to better understand the spatial distribution of fish habitat and potential fish habitat use within the study area.

Within the study area no uncommon features were identified pertaining to fish habitat. While variability in habitat exists, the river provides similar fish habitat among each of the study zones.

The following features were identified as potentially important areas of fish habitat:

• Lagoons – Lagoons are backwater areas that occur outside the margins of the main river channel. These areas provide macrophyte complexity and are lentic or quiescent zones that provide refuge from the high velocity flows often seen in the main river channel. The lagoons provide nursery and rearing habitat for young-of-the-year, juvenile and other small fish.

• Island channels – Island channels occur between the river shoreline and islands that occur in the main river channel. These areas provide similar macrophyte habitat complexity as that found in lagoons with some overbank cover. The gentler flows and cover provided in the island channels provide nursery and rearing habitat for young-of-the-year, juvenile and other small fish.

• River Margins (Riparian and Littoral Zones) – River margins of the main channel are vegetated and also provide areas of refuge for young-of-the-year, juvenile and other small fish in particular. In general, the riparian and littoral zones of the main channel were limited in spatial extent due to local land uses and relatively steep banks compared to lagoon and island channel habitats.

The area occupied by the effluent mixing zone is not expected to have an effect on fish habitat as this substrate type is common throughout the main channel of the study area.

FISH HABITAT ASSESSMENT Table of Contents

Ref. 18-25 1 March 2019 ii

TABLE OF CONTENTS Page 1.0 INTRODUCTION ...................................................................................................1.1

1.1 Terms of Reference ...............................................................................................1.2

1.2 Objectives ..............................................................................................................1.2

1.3 Report Format .......................................................................................................1.3

2.0 WORK SCOPE ......................................................................................................2.1

2.1 Study Area .............................................................................................................2.1

2.2 Characterization of Shoreline Habitats and Vegetation ..........................................2.3

2.2.1 Field Procedure ........................................................................................2.3

2.2.2 ArcGIS .....................................................................................................2.3

2.3 Characterization of Bottom Substrates ...................................................................2.3

2.3.1 Field Procedure ........................................................................................2.3

2.3.2 Arc GIS.....................................................................................................2.4

2.4 Characterization of Bathymetry ..............................................................................2.4

3.0 SHORELINE HABITAT AND VEGETATION .........................................................3.1

4.0 BOTTOM SUBSTRATES ......................................................................................4.1

5.0 Bathymetry ...........................................................................................................5.1

6.0 KEY RESULTS .....................................................................................................6.1

Appendix A Terms of Reference ............................................................................... A.1

Appendix B Shoreline Habitat Field Sheets ............................................................. B.2

Appendix C Petite Ponar Sampling Field Sheets ..................................................... C.3

Appendix D Photo Documentation ........................................................................... D.4

FISH HABITAT ASSESSMENT Table of Contents

Ref. 18-25 1 March 2019 iii

LIST OF TABLES Table 3-1: Percent length of shoreline coverage for each aquatic vegetation type within the study area located on the Rio Negro. ................................................................................3.2 Table 3-2: Percent area coverage for each aquatic vegetation type within the study area located on the Rio Negro...................................................................................................3.2 Table 4-1: Percent area coverage for each substrate type within the study area located on the Rio Negro. ...................................................................................................................4.2

LIST OF FIGURES Figure 1-1: Site Location – Proposed UPM Mill Site .......................................................1.2 Figure 2-1: Location Map of Study Area located on the Rio Negro. ...................................2.2 Figure 3-1: Vegetation types found within the study area of the Rio Negro. ......................3.3 Figure 3-2: Aquatic vegetation map of the full study area located on the Rio Negro. .........3.4 Figure 3-3: Aquatic vegetation map of Zone 1 located on the Rio Negro. ..........................3.5 Figure 3-4: Aquatic vegetation map of Zone 2 located on the Rio Negro. ..........................3.6 Figure 3-5: Aquatic vegetation map of Zone 3 located on the Rio Negro. ..........................3.7 Figure 4-1: Substrate types found within the study area of the Rio Negro. ........................4.3 Figure 4-2: Substrate map of the full study area located on the Rio Negro. .......................4.4 Figure 4-3: Substrate map of Zone 1 located on the Rio Negro. ........................................4.5 Figure 4-4: Substrate map of Zone 2 located on the Rio Negro. ........................................4.6 Figure 4-5: Substrate map of Zone 3 located on the Rio Negro. ........................................4.7 Figure 5-1 Bathymetry of the full study area located on the Rio Negro. .............................5.2

FISH HABITAT ASSESSMENT Introduction

Ref. 18-25 1 March 2019 1.1

1.0 INTRODUCTION UPM is considering alternatives for long-term development in Uruguay. This involves the construction of a state-of-the-art pulp mill near Paso de los Toros, Durazno Department, Uruguay, as shown in Figure 1-1. The mill will have a production capacity of 2,100,000 air dry tonnes per year (ADt/yr).

UPM set a design criteria for the project to include compliance with current Uruguayan legislation, as well as compliance with international standards and recommendations for modern mills as stated in the EU Best Available Techniques (BAT).

To move forward the project requires an environmental authorization (Autorización Ambiental Previa, or AAP). To obtain the AAP, the project has to be communicated to the Ministerio de Vivienda, Ordenamiento Territorial y Medio Ambiente (MVOTMA) through a Project Communication, an Environmental Location Viability (VAL) has to be obtained and, following project classification by MVOTMA, an Environmental Impact Study (EIS) will be completed.

UPM retained EcoMetrix Incorporated (EcoMetrix) in partnership with Laboratorio Tecnológico del Uruguay (LATU) to characterize aquatic habitats in an area of the Rio Negro near the proposed mill site. The characterization was completed to support efforts related to the environmental approvals processes for the mill.

This report presents the findings of the field survey described in the Terms of Reference (ToR) as it relates to fish habitat. In particular, shoreline habitat and bottom substrates were mapped to better understand the spatial distribution of fish habitat within the relevant section of the Rio Negro.


Ref. 18-25 1 March 2019 1.2

Figure 1-1: Site Location – Proposed UPM Mill Site

1.1 Terms of Reference

A Terms of Reference (ToR) for an aquatic habitat survey was developed by EcoMetrix in partnership with LATU (see Appendix A). The ToR was submitted to and reviewed by DINAMA and recommendations were provided.

The revised aquatic habitat survey work scope that is described in detail in Section 2.0 was executed in February 2019. Briefly, aquatic habitats were characterized within a study area comprising an approximate 9 km reach of the Rio Negro downstream of Paso de los Toros. The study area was divided into three zones --- Zone 1 – in the immediate vicinity of the proposed mill effluent outfall and water intake; Zone 2 – downstream of Zone 1 to the maximum extent of the hydrodynamic mixing zone as determined by predictive modeling; and, Zone 3 – upstream of the proposed mill site. Shoreline and vegetation features and bottom substrate types were mapped in each zone.

Minor deviations from the planned sampling program were made due to conditions encountered in the field. These deviations are highlighted in Appendix A. The deviations did not negatively affect the habitat characterization program, nor the interpretation of the habitat information collected during the program.

1.2 Objectives

The primary objective of this assessment was to characterize aquatic habitats to develop a


Ref. 18-25 1 March 2019 1.3

greater understanding of the spatial distribution of fish habitat and potential fish habitat use within the study area. The results of this assessment support the ongoing assessment of the potential effects (if any) that may be associated with the development and operations of the proposed pulp mill.

1.3 Report Format

Following this introductory section, the remainder of this report is organized as follows:

• Section 2.0 describes the work scope and detailed methodologies that were employed to execute the habitat characterization program;

• Section 3.0 describes the results of program with regards to shoreline habitats;

• Section 4.0 describes the results of program with regards to bottom substrates;

• Section 5.0 describes the results of program with regards to bathymetry; and,

• Section 6.0 provides and overall summary and interpretation of the habitat characterization survey program.

Raw data and photographic records collected during the field sampling program are provided in Appendix B, Appendix C and Appendix D.

.

FISH HABITAT ASSESSMENT Work Scope

Ref. 18-25 1 March 2019 2.1

2.0 WORK SCOPE The work scope is described below in terms of the three main components, shoreline habitats and vegetation, bottom substrates and bathymetry. The field survey for this study was conducted between the 4th and 9th of February, 2019.

2.1 Study Area

The study area is inclusive of a length of the river that extends from approximately 4 km upstream of the proposed water intake and effluent outfall location to 5 km downstream, covering a total river length of approximately 9 km. This area has been sectioned into three survey zones, Zone 1, Zone 2 and Zone 3 (Figure 2-1).

Zone 1 is the area in the vicinity of the proposed water intake and effluent outfall structures. Zone 1 encompasses a length of river extending from approximately 1 km upstream of the proposed water intake and effluent outfall structures to approximately 2 km downstream and includes the main river channel, as well as backwater areas (lagoons and island side channels). Survey efforts were most intense in this zone relative to the other zones, as potential direct project-related effects, such as those related to construction of the water intake and effluent outfall and entrainment of aquatic biota during operations, could be manifested. The approximate location and area of the 2% effluent mixing zone1 is depicted in this zone (Figure 2-1).

Zone 2 extends along the river beyond Zone 1, encompassing a length of river of approximately 3 km. The aquatic survey program was completed in the main river channel, as well as backwater areas (lagoons and island side channels). Survey components undertaken in Zone 2 included three ponar samples per transect as oppose to five in Zone 1, but were otherwise identical.

Zone 3 is the area upstream of Zone 1 and is therefore upstream of the location of the proposed pulp mill. Under the current proposed mill configuration, this area would have no direct or indirect interaction will the mill and can serve as a reference area. Zone 3 encompasses a 3 km stretch of river immediately upstream of Zone 1. The aquatic survey program was completed in the main river channel, as well as backwater areas (lagoons and island side channels). Survey components undertaken in Zone 3 were the same as those in Zone 2 in terms of both scope and effort.

1 Response to the 28th December 2018 letter from MVOTMA, mixing zone - UPDATED, 7th March 2019.


Ref. 18-25 1 March 2019 2.2

Figure 2-1: Location Map of Study Area located on the Rio Negro.


Ref. 18-25 1 March 2019 2.3

2.2 Characterization of Shoreline Habitats and Vegetation

To identify areas that may be important as fish habitat shoreline vegetation and habitat features were mapped.

2.2.1 Field Procedure

Habitat mapping consisted of the boat operator slowly driving the shoreline while an observer recorded habitat features on waterproof field sheets that were preprinted with the outline of the shoreline. Each lagoon had its own outline map (8.5 by 11 inch) and the main channel was comprised of multiple outline maps, each defining a separate portion of the main channel.

Mapped shoreline habitat included the following:

• Shoreline vegetation type and location;

• Macrophyte type and location;

• General habitat features (boulders, logs, other); and,

• Photo documentation of different habitat types.

2.2.2 ArcGIS

All information was input into Arc GIS and tables for each zone were generated for percent vegetation type (length and area) within the main channel (including island side channels) and lagoon habitats.

2.3 Characterization of Bottom Substrates

Petite ponar grabs of the substrate were taken at transects spaced at 200 meter intervals in all zones. In Zone 1, five petite ponars were taken along each transect and in Zones 2 and 3, three petite ponars were taken in each transect. Island channels were identified as a prominent feature, and single ponar grab were taken in the center of each island channel at 200 meter intervals in each zone. Petite ponar sampling was also conducted in lagoons. In large and small lagoons, 10 and 5 petite ponar grabs were taken, respectively.

2.3.1 Field Procedure

At each location the following was conducted:

• Boat was anchored;

• Technician 1 lowered petite ponar, retrieved sample, and placed sample in bin;


Ref. 18-25 1 March 2019 2.4

• Technician 1 then deployed a velocity meter at 1 meter depth;

• On a custom field sheet (Appendix B), Technician 2 recorded water velocity, recorded GPS coordinates, recorded sample depth, took photo(s) of the substrate, recorded photo numbers, recorded sediment character, and recorded any vegetation if present.

2.3.2 Arc GIS

Substrate information was input into Arc GIS and the natural neighbor interpolation function was used to create a substrate map of the study area. Tables for each zone were generated for percent substrate type (area) within the main channel (including island side channels) and lagoon habitats.

2.4 Characterization of Bathymetry

The depth data that was acquired at each petite ponar sampling station (Section 2.3) was used to generate a general bathymetric map of the study area. The depth data was input into Arc GIS and the natural neighbor interpolation function was used to create depth contours of the river bathymetry.

FISH HABITAT ASSESSMENT Shoreline Habitat

Ref. 18-25 1 March 2019 3.1

3.0 SHORELINE HABITAT AND VEGETATION A total of seven aquatic vegetation types were identified in the study area: flooded shrub (FS); emergent grass (EG); emergent macrophyte (EM); floating macrophyte (FM); emergent reed (ER); submergent macrophyte low (SL); and, submergent macrophyte high (SH) (Figure 3-1).

The seven aquatic vegetation types would often occur in various combinations as can be seen in the vegetation maps (Figure 3-2 to Figure 3-5). The vegetation maps presented in this report are representative of the length of shoreline covered (Figure 3-2 to Figure 3-5) and not actual area coverage. However, macrophyte area was recorded and the results are presented in tabular format.

Within the main channel, submergent low macrophytes and submerged grass covered more than 80% of the shoreline in all zones (Table 3-1). Similarly, with the exception of 57% emergent grass cover in Zone 1 lagoons, submergent low macrophytes and emergent grass also covered more than 80% of the shoreline within lagoons.

Flooded shrub (Figure 3-1,a) was found at slightly lower coverage (~70%) but was also similar between zones, as well as, main channel and lagoon habitats (Table 3-1).

Emergent macrophytes covered 55% of Zone 1 lagoon shorelines and 30% of Zone 2 main channel and lagoon shorelines, in all other areas they covered 5% or less of the shoreline (Table 3-1).

The highest percentage of floating macrophyte was found in Zone 2 lagoons (45%) with all other areas covering 6% or less of the shoreline.

Emergent reed covered 29% of the lagoon shorelines in Zone 2, and 9% and 11% of Zone 1 lagoon and main channel shorelines, respectively.

Submergent high macrophyte (Figure 3-1,g) was rarely encountered in the study area (Table 3-1). Tabulation of percent macrophyte surface area coverage (Table 3-2) presents analogous findings to that of percent linear coverage.

While the submergent low and emergent grass provides marginal habitat for fish, as they are sparsely distributed, the other vegetation types provide excellent habitat for forage fish as they provide structural complexity within the water column and occur with sufficient density to provide refuge from predators.

In areas where only emergent grass and submergent low macrophyte exist, it is apparent that the lack of other aquatic vegetation is largely due to cattle having access to the shoreline.


Ref. 18-25 1 March 2019 3.2

Due to the small area of the mixing zone within the central portion of the main channel (Figure 3-3), the proposed pulp mill is not expected to have an effect on the shoreline vegetation or the fish habitat it provides.

Table 3-1: Percent length of shoreline coverage for each aquatic vegetation type within the study area located on the Rio Negro.

Table 3-2: Percent area coverage for each aquatic vegetation type within the study area located on the Rio Negro.

Zone 1 Zone 2 Zone 3 Zone 1 Zone 2 Zone 3 Zone 1 Zone 2 Zone 3EM 24% 31% 5% 5% 31% 5% 55% 29% 0%EG 70% 90% 96% 78% 87% 95% 57% 100% 100%FM 3% 16% 0% 5% 6% 0% 0% 45% 0%FS 60% 70% 63% 78% 66% 63% 30% 81% 64%SH 0% 0% 0% 0% 0% 0% 0% 0% 0%SL 95% 98% 99% 92% 97% 99% 100% 100% 100%ER 3% 15% 0% 0% 11% 0% 9% 29% 0%

Total Main Channel LagoonHabitat

Zone 1 Zone 2 Zone 3 Zone 1 Zone 2 Zone 3 Zone 1 Zone 2 Zone 3EM 0.12% 1.23% 0.09% 0.01% 1.02% 0.10% 3.47% 2.60% 0.00%EG 0.29% 3.20% 2.90% 0.16% 2.39% 2.61% 4.43% 8.46% 8.64%FM 0.03% 0.60% 0.00% 0.03% 0.21% 0.00% 0.00% 3.16% 0.00%FS 0.25% 2.86% 1.94% 0.10% 2.26% 1.72% 4.62% 6.79% 6.32%SH 0.05% 0.06% 0.00% 0.00% 0.00% 0.00% 1.44% 0.46% 0.00%SL 0.36% 3.76% 2.94% 0.17% 3.04% 2.66% 6.14% 8.46% 8.64%ER 0.05% 2.02% 0.32% 0.00% 1.10% 0.33% 1.47% 7.98% 0.00%

Total Main Channel LagoonHabitat


Ref. 18-25 1 March 2019 3.3

Figure 3-1: Vegetation types found within the study area of the Rio Negro.

Panel a) flooded shrub (FS), b) emergent grass (EG), c) emergent macrophyte (EM), d) floating macrophyte (FM), e) emergent reed (ER), f) submergent macrophyte low (SL), and g) submergent macrophyte high (SH).


Ref. 18-25 1 March 2019 3.4

Figure 3-2: Aquatic vegetation map of the full study area located on the Rio Negro.


Ref. 18-25 1 March 2019 3.5

Figure 3-3: Aquatic vegetation map of Zone 1 located on the Rio Negro.


Ref. 18-25 1 March 2019 3.6



Ref. 18-25 1 March 2019 3.7


FISH HABITAT ASSESSMENT Bottom Substrate

Ref. 18-25 1 March 2019 4.1

4.0 BOTTOM SUBSTRATES A total of six substrate types were identified in the study area: clay; silt/mud; sand; gravel; cobble; and, bedrock (Figure 4-1). Across the whole study area, substrate was sampled using a petite ponar at 251 sampling locations (Figure 4-2 to Figure 4-5, blue circles depict ponar sampling locations). No aquatic vegetation was found in any of the substrate samples.

Main Channel Substrate (including side channels)

All six substrate types were found in the main channel section of each zone (Table 4-1). However, clay and silt/mud substrate were uncommon and covered approximately 2% and 7% of the river bottom, respectively. Zone 1 and Zone 2 had very similar percent composition of substrates with Zone 1 being dominated by gravel (40%) followed by sand (35%) and Zone 2 being dominated by sand (50%) followed by gravel (23%)(Table 4-1). Zone 3 had similar proportions of sand, gravel, cobble and bedrock of approximately 20% each.

It is noted that a commercial sand dredging barge was seen operating along the west shore of the island chain located in Zone 2.

Lagoon Substrate

Clay and bedrock substrate types were not present in any lagoons (Table 4-1). Lagoons in Zones 2 and 3 were dominated by sand with values of 81% and 91%, respectively; whereas, lagoons in Zone 1 were dominated by silt/mud (50%) followed by sand (44%). The greater percentage of silt/mud in Zone 1 lagoons may be attributed to their larger size which may allow tributary sediments to settle in the lagoon before entering the main channel.

Water velocity

Although water velocity is largely regulated by hydroelectric dam activity, it is noted that flow was not present in the central portions of the lagoons. The flow through the island channels was approximately 75% less than that of the midsection of the main channel. Water velocity at each ponar station is presented in Appendix B.

The lentic and shallow nature of the lagoons provides nursery habitat for young of year fish, while the island channel section provides forage habitat for fish before they are able to adequately navigate the stronger flows of the main channel section.

The proposed location of the effluent diffuser and associated area of the mixing zone lies over a small area of gravel substrate (Figure 4-3). This substrate is plentiful throughout the study area of the Rio Negro (Figure 4-2), as such, the proposed pulp mill is not expected to effect fish habitat within the study area of the Rio Negro.


Ref. 18-25 1 March 2019 4.2

Table 4-1: Percent area coverage for each substrate type within the study area located on the Rio Negro.

Zone 1 Zone 2 Zone 3 Zone 1 Zone 2 Zone 3 Zone 1 Zone 2 Zone 3Clay 2% 1% 2% 2% 1% 3% 0% 0% 0%Silt/mud 19% 8% 7% 7% 7% 7% 50% 16% 4%Sand 37% 53% 25% 35% 50% 22% 44% 81% 91%Gravel 31% 21% 25% 40% 23% 26% 7% 3% 5%Cobble 9% 12% 20% 12% 14% 20% 0% 0% 0%Bedrock 3% 5% 21% 4% 5% 21% 0% 0% 0%

Habitat Total Main Channel Lagoon


Ref. 18-25 1 March 2019 4.3

Figure 4-1: Substrate types found within the study area of the Rio Negro.

Panel a) clay, b) silt/mud, c) sand, d) gravel, e) cobble, and f) bedrock.


Ref. 18-25 1 March 2019 4.4

Figure 4-2: Substrate map of the full study area located on the Rio Negro.

Blue circles identify ponar sampling locations.


Ref. 18-25 1 March 2019 4.5

Figure 4-3: Substrate map of Zone 1 located on the Rio Negro.



Ref. 18-25 1 March 2019 4.6




Ref. 18-25 1 March 2019 4.7




Ref. 18-25 1 March 2019 5.1

5.0 Bathymetry A general bathymetric map was also created using the depth data from each ponar sampling station (Figure 4-6). This map indicates that the mid channel depth of Zone 3 is relatively deeper than that of Zones 1 and 2 each having average depths of approximately 10, 8 and 7 meters, respectively. Similarly, the slope of the submerged bank of the main channel is steeper in Zone 3 compared to the other two zones. The bathymetric map also indicates that lagoons are roughly 2 to 3 meters deep and that the narrow island channels are approximately 3 to 4 meters deep.


Ref. 18-25 1 March 2019 5.2

Figure 5-1 Bathymetry of the full study area located on the Rio Negro.

FISH HABITAT ASSESSMENT References

Ref. 18-25 1 March 2019 6.1

6.0 KEY RESULTS Within the study area no uncommon features were identified pertaining to fish habitat. While variability in habitat exists, the river provides similar fish habitat among each of the study zones.

Lagoons are important areas of fish habitat. Lagoons are backwater areas that occur outside the margins of the main river channel. These areas provide macrophyte complexity and are lentic or quiescent zones that provide refuge from the high velocity flows often seen in the main river channel. The lagoons provide nursery and rearing habitat for young-of-the-year, juvenile and other small fish.

Island channels occur between the river shoreline and islands that occur in the main river channel. These areas provide similar macrophyte habitat complexity as that found in lagoons with some overbank cover. The gentler flows and cover provided in the island channels provide nursery and rearing habitat for young-of-the-year, juvenile and other small fish.

Areas along the river margins of the main channel are vegetated and also provide areas of refuge for young-of-the-year, juvenile and other small fish in particular. In general, the riparian and littoral zones of the main channel were limited in spatial extent due to local land uses and relatively steep banks compared to lagoon and island channel habitats.

The area occupied by the effluent mixing zone is not expected to have an effect on fish habitat as this substrate type is common throughout the main channel of the study area.

FISH HABITAT ASSESSMENT Appendix A

Appendix A Terms of Reference

6800 Campobello Road, Mississauga, Ontario, Canada L5N 2L8 Tel: (905) 794-2325 Fax: (905) 794-2338 Toll-Free: 1-800-361-2325 www.ecometrix.ca

MEMO

To: Gervasio Gonzalez

From: Brian Fraser, Michael

White, Rob Eakins, Nicholas Edmunds, Paul Patrick, Bruce Rodgers

Ref: Terms of Reference – Rio Negro Aquatic Survey (February 2019)

Date: 28 January 2019

The following provides a Terms of Reference (ToR) for the implementation of an aquatic field survey to support efforts related to environmental impacts assessment and licensing processes for the proposed UPM pulp mill development on the Rio Negro near Paso de les Toros, Duranzo Department, Uruguay. The overall objective of the field survey is to collect detailed fish and aquatic habitat information within an area that encompasses the proposed water intake and effluent outfall structures and extends downstream to capture the full extent of the effluent mixing zone, as defined by predictive modelling completed by Dr. Piedra-Cueva. This ToR has been developed in consideration of discussions between UPM and DINAMA regarding recommendations for additional information that will aid in DINAMA’s decision-making process to advance the proposed project to the next phase. Study Area The proposed study area is shown in Figure 1. The study area is inclusive of a length of the river that extends from approximately 1 km upstream of the proposed water intake and effluent outfall location to the edge of the effluent mixing zone, a total river length of approximately 6.5 km. For the purpose of the study two sampling zones have been defined – Zone 1 and Zone 2. Zone 1 is the area in the vicinity of the proposed water intake and effluent outfall structures. Zone 1 encompasses a length of river extending from about 1 km upstream of the proposed water intake and effluent outfall structures to approximately 2 km downstream and includes the main river channel, as well as backwater areas (lagoons). Survey efforts will be relatively intensive in this area, as potential direct project-related effects, such as those related to construction of the water intake and effluent outfall and entrainment of aquatic biota during operations, could be manifested. In addition, effluent concentrations in this initial, near-field area of the mixing zone will be relatively high and therefore represent the upper-bound condition for effluent exposure of aquatic biota.

25 January 2019 Gervasio Gonzalez Page 2 of 14

Reference: Terms of Reference – Rio Negro Aquatic Survey (February 2019)

Zone 2 extends along the river beyond Zone 1 to the full extent of the mixing zone, encompassing a length of river of approximately 3.5 km. The aquatic survey program will be completed in the main river channel, as well as backwater areas (lagoons), as appropriate. Survey components undertaken in Zone 2 will be the same as in Zone 1, though sampling will less intensive. The primary interaction between the project and aquatic biota in this area in via the effluent exposure pathway, with Zone 2 representing an area where lower effluent concentrations (on a % v/v in river water basis) will be found as effluent continues to be diluted through hydrodynamic mixing process.

Figure 1. Aquatic Assessment Study Area



Schedule and Level of Effort The field work is scheduled to begin on-site on February 4th, 2019. It is expected that the work scope described below can be accomplished over a five- to seven-day period assuming that three field crews (two to three people each) can be mobilized to the site with sufficient resources (e.g., boats, gill nets, depth sounders, etc.). It is envisioned that two field crews would complete the fish community assessment (one crew in Zone 1 and one crew in Zone 2) and one field crew would be responsible for the habitat aquatic mapping over the entirety of the study area. Aquatic Habitat Characterization Aquatic habitat will be surveyed to allow a detailed habitat map to be produced. Aquatic habitat characterization consists of two parts, one involves petite Ponar sampling to confirm bottom substrate type and the other involves shoreline habitat mapping to document shoreline features, in particular those that may provide unique or preferred aquatic habitat for resident aquatic species. Bottom Substrates In Zones 1 and 2, petite ponar grabs of the substrate will be taken at transects that will be spaced at 100 meter intervals. In Zone 1, five petite ponars will be taken along each transect and in Zone 2, three ponars will be taken in each transect (Figure 2). Island channels have also been identified as a prominent feature, and single ponar grab will be taken in the center of each island channel at 100 meter intervals. Ponar sampling will also occur in lagoons. Dependent upon the size of the lagoon, 5 to 10 petite ponar grabs will be taken in each lagoon. It is estimated that all ponar sampling will take 4 days (assuming a 10-hour work day); 2 day for main channel zone one, 1 day for main channel zone two, and 1 day for lagoons. An additional day will be needed to conduct the visual shoreline habitat assessment. Therefore, it is anticipated the field program to conduct the aquatic habitat characterization component will require 5 (10-hour) work days. The following sampling routine for petite Ponar sampling is proposed:

• Have GPS coordinates preloaded into GPS unit, identifying sample transects and sampling locations.

• Follow coordinates to the first transect. • Sample each transect from one shoreline to the opposite shoreline • Once at sample location boat captain will hold boat in location (depending on

current and wind condition boat may need to be anchored). • Technician 1 lowers petite ponar, retrieves sample, and places sample in bin. • If a successful sample is taken, technician 1 then lowers a velocity meter.



• If a successful sample is taken, technician 2 records water velocity, records GPS coordinates (if different than those provided), records sample depth, takes 2 photos (1 of minimally disturbed ponar the other after the sediment has been stirred with a spoon), records photo numbers, records sediment character, and records any vegetation that is present.

• After technician 1 has recorded GPS, water velocity, and depth the boat captain can proceed to the next sample station while technician 1 records the remaining information descripted in the previous bullet.

• In lagoons, habitat mapping should occur with ponar sampling, in the main channel habitat mapping should occur on a separate day. The rational for this is to increase efficiency of the overall field program.

A field sampling form is attached for reference. Shoreline Habitat Habitat mapping consists of the boat operator slowing driving the shoreline while an observer records habitat on waterproof field sheets that have been preprinted with the outline of the shoreline. Each lagoon will have its own outline map (8.5 by 11 inch) and the main channel will be comprised of multiple outline maps, each defining a separate portion of the main channel. At a minimum the following will be mapped:

• North arrow; • Shoreline vegetation type and location; • Macrophyte type and location; • Habitat features (boulders, logs, other); and, • Noticeable current location.

An example of a habitat map that has been marked up in the field during surveying (Figure 3) and a final report-ready map (Figure 4) are provided for reference.



Figure 2: Hypothetical river cross section with ponar sampling locations.

- Facing upstream ponars are sampled from left to right.- Shoreline ponars are sampled between 1.0 and 1.5 meters depth- A centre ponar is taken in the middle of the channel- For zone one, ponars 3 and 4 are to be taken midway between the centre and

shoreline ponars.

T1P1Depth (m)

1

2

3

4

0

T1P2 T1P3 T1P4 T1P5



Figure 3: Sample field survey map



Figure 4: Sample report ready habitat map.

Depth (m)

11

10

9

8

7

6

5

4

3

2

1

0

Habitat Legend N

Description Typical Shoreline* Shallow Sand Beach Boulders Along Shore Logs Bog (shrubs directly over water) Submergent / Emergent Macrophytes Submerged Sand Bar

Inlet / Outlet Profile Location

100m

*Rocky shore with overhanging shrubs, quickly transitioning to sandy bottom, and shallow sandy beaches with patchy emergent vegetation

Key Features

Survey Date: 16-17 September 2016

Surface Area: 137.94 ha

Volume: 6,411,000 m3

Mean Depth: 4.6 m

Maximum Depth: 11.2 m

Lake Elevation: 514.25 m

Secchi Depth: 1.5 m

0 5 10 15 20 0

2

(m)

4

Dep

th

6

8

10

12

Temperature (°C) Dissolved Oxygen (mg/L)

Field pH (units)

Conductivity (µS/cm)

Fish Species Observed Northern Pike White Sucker Lake Whitefish Spottail Shiner Ninespine Stickleback Walleye Yellow Perch

Aquatic Plants Observed Burreed Pondweeds Sedges Muskgrass Water Lobelia Yellow Pond Lily

Wheeler River Project

Lake LA-9 Bathymetry and Aquatic Habitat Map

EcoMetrix Figure 3-7 I N C O R P O R A T E D


Fish Community Assessment Sampling of the resident fish community of the Rio Negro should be undertaken within Zones 1 and 2, in littoral and mid-channel habitats (Figure 5). Littoral habitats include nearshore areas with depths < 2 m, whereas mid-channel habitats are characterized by deeper water (>2 m). In Zone 1, nets should be set in the immediate vicinity of the proposed intake and outfall (mid channel). Fishing should also take place within the lagoons and inlets that occur upstream, adjacent to, and downstream of the proposed intake location, along the south shore (littoral and mid channel), as well as within the side channels along the north shore (littoral). In addition, nets should be set within a similar habitats upstream of Zone 1, and within Zone 2. Fishing within each location should consist of both daytime and nighttime sets, if possible. Exact replicates are not required, but a minimum of three nets per area/habitat is preferred, especially within Zone 1. Fishing methods should include Nordic nets standardized according to EESTI Standard for fish sampling (EESTI, 2015), that were used previously in the Bonete and Baygorria sampling areas. Nets were 30m long, by 1.5m wide and were composed of 12, 2.5m panels of varying mesh sizes as follows 5, 6.25, 8, 10, 12.5, 15.5, 19.5, 24, 29, 35, 43 and 55 mm. In addition, larger mesh experimental gillnets, 23m long by 1.8m high with mesh sizes of 76, 101 and 127 mm should also be employed. All net set and fish collection information will be recorded on Fish Collection Forms (see attached). Recorded information should include waterbody, station no., location, coordinates, datum, gear type, net size, mesh size(s), set date/time, set lift date/time, depth of set, substrate and habitat characteristics. All fish collected should be identified to species, enumerated, measured (length/weight) and released if alive. Representative photographs of each species should be taken of live/fresh specimens. Vouchers should be collected if specimens need to be retained to confirm species identity.



Figure 5: Fish community assessment netting locations.



Equipment, Materials and Supplies Based on the survey components described above, the following list of equipment, materials and supplies that we believe will be needed to complete the field program is provided for consideration. EcoMetrix can potentially provide any of the items in the instance that LATU does not have (nor cannot acquire given the limited lead time) the full complement of equipment, materials and supplies that has been suggested.

• Aquatic Habitat Assessment Equipment List • 1 petite Ponar; • 3 plastic tubs, to put contents of petite ponar in for photo documentation and

substrate characterization; • 1 stainless steel spoon; • 1 digital camera; • 1 GPS; • 1 depth sounder; • 1 velocity meter; • river maps; and, • field forms.

• Fish Community

• Large-mesh experimental gill nets (minimum 6); • Nordic nets (minimum 6); • Net tubs, floats, anchors, ropes; • Depth sounder; • GPS; • Camera; • 25L pails (for fish); • Fish collection forms; • Fish measuring board(s); and, • Weigh scales.

Information Management At the end of each field day any records (field forms, written documentation, maps, field notes, etc…) should be backed up (e.g., scanned or photocopied). Photographs should be downloaded from cameras (or phones) onto a computer or onto a secure file storage server. GPS coordinates that have not been documented on field forms should be downloaded onto a computer or onto a secure file storage server.



All project-related information and documents provided by UPM and generated on behalf of UPM during the execution of the field survey and associated reporting will be treated as confidential and will not be shared with third-parties without prior approval from UPM.

All project-related documents will be handled in keeping with the requirements of the EcoMetrix Quality Management Program as detailed in Section 4.2 of the Quality Management Plan, and associated SOPs. Reporting EcoMetrix will initiate preparation of the aquatic survey report immediately once the in-field sampling has been completed and data have been provided. The report will include the following information:

• A clear, concise statement of the study objective, including the intended use of the data;

• A summary of site- and/or project-related information that is relevant to establish the context for the study;

• A detailed description of the methods by which the field sampling program was implemented, by which the samples generated by the field sampling were analyzed and by which the data generated by the field sampling program and sample analyses were summarized and analyzed;

• A robust presentation of the results and analyses of the study, including graphs, figures, tables and photographs, as appropriate;

• A complete and scientifically defensible interpretation of the results and analysis of the study, supported by existing scientific literature, as appropriate; and,

• A summary of the key findings and conclusions of the study, pertaining to the overall study objectives, as well as recommendations as appropriate.

Survey-component specific aspects of the report are provided as follows:

• Aquatic Habitat – Aquatic habitat information will be summarized, for reporting purposes, pictorially using maps in combination with the photographs. The mapping will cover the entire study area and will document the habitat features described above, and will in particular identify and demarcate “sensitive” habitat areas as can be deduced from the survey data.

• Fish Community - All fish that are collected will be identified, enumerated, weighed and measured and assigned to an age class (young-of-the-year, juvenile, adult). Fish collection data will be summarized by gear type to provide estimates of number of fish collected per unit effort (e.g., per hour of netting). Fish species numbers will be reported by age classes present. The length and weight data can be used to determine “condition”, an index that provides a measure of overall fish health. If



sufficient numbers of fish are collected age class, length and weight distributions will be determined. Fish data will be used to assess the spatial distribution of fish within the study area and to identify any patterns that may be noted with respect to distribution given habitat conditions.

• Fish Species with Conservation Status – Special attention will be afforded to consideration of fish species that have conservation status in terms of presence, numbers, distribution and habitat availability.

All references cited in the report will be listed in full. Raw data, field sheets, photographic records and statistical analysis results will be provided as appendix material. As required by the EcoMetrix Quality Management Program the report will be reviewed by internally by senior staff prior to submission to UPM. Briefly, at a minimum, the review will consider report completeness (outputs compared to the stated project objectives/ requirements), technical content (transparency, citation of sources, reasonable results, appropriate methods and conclusions) and clarity of message. Any issues identified in the report will be corrected prior to its issue. In addition, some or all calculations may be verified, which involves a comprehensive check of input data against cited sources, and output results against independent calculations using the cited methodology. The draft report will be provided to UPM for review prior to finalization. EcoMetrix will provide dispositions to any review comments received from UPM, as appropriate, and in sufficient detail to allow UPM to fully understand the disposition and achieve resolution. Following acceptance of the dispositions, a revised report will be provided to UPM. The final report will be provided in electronic format. All information will be provided in a format acceptable to UPM. Health and Safety EcoMetrix will work with LATU to develop and Project Safety Plan (PSP), or alternatively work under the LATU safety plan, as may be appropriate. The intent of the PSP is to establish safe work procedures for project personnel to follow, thereby reducing the risk of exposure to hazards associated with project-related work activities. The details of the PSP, or other equivalent health and safety procedures can be worked out with LATU prior to initiating the field work. Quality Management EcoMetrix was certified to the ISO 9001:2015 standard by NSF International in 2018. We ensure the integrity of our by following our corporate Quality Management Plan (QMP) and the associated quality documentation, including SOPs. Standard operating procedures



for field sampling and report preparation ensure that each phase of the project runs smoothly and according to established principles of quality. Our quality management documentation ensures that specific QA/QC measures are incorporated into every our work. Examples of some general QA/QC measures that are relevant to this current work scope include:

• Field Sampling o all personnel involved in the field sampling will have appropriate training,

experience with field equipment and objectives and will execute activities according to SOPs;

o all safety measures will be identified, understood and adhered to; o contamination during chemical sampling will be checked with trip blanks; o detailed field notes will be maintained in a bound notebook or custom field

sheets; and o chain-of-custody forms and appropriate shipping and storage procedures will

be used.

• Analysis and Reporting o conduct screening exercises data reviews to identify transcription errors,

outliers and other suspicious data points; o provide raw data in an electronic database format and appendices to reports

which summarize the data; o document the methods (specific statistical tests) and software (as

appropriate) used for analysis; o check for editorial, grammatical and spelling errors and data entry errors; o check for consistency of format and the completeness of each section; o check for data handling and reporting (entry checks, missing values,

methods, QC); o ensure that pertinent information has been reported in detail (including field

notes, accurate site locations); o ensure that any changes in protocol, study design, or other components of

the study have been reported; and o ensure that all QA/QC documentation is complete and included in the report.

We will work with LATU on issues related to quality management and quality program documentation prior to initiating the field work. Closure We trust this ToR suits your needs at this time. We are available to discuss any aspect of the ToR with you and the team at LATU at your convenience and move forward with planning and logistics.



References EESTI. 2015. Water quality – Sampling of fish with multi-mesh gillnets (EVS-EN

14757:2015). https://www.evs.ee/products/evs-en-14757-2015

https://www.evs.ee/products/evs-en-14757-2015

Study Area:Weather:Investigators:

Transect#

Ponar#

UTMZone UTM North UTM East Depth

(m)Velocity

(m/s)

DominantSubstrate

Type1

SecondSubstrate

Type1

VegetationType(s)2 Vegetation Name(s)

OrganicDebris3

PhotoNumbers

P1

P2

P3

P4

P5Notes:

P1

P2

P3

P4

P5Notes:

P1

P2

P3

P4

P5Notes:

1 Substrate Types - 1 Clay, 2 Silt/Mud, 3 Sand, 4 Gravel, 5 Cobble, 6 Bedrock, 7 None2 Vegetation Types - 1 None, 2 Floating, 3 Emergent, 4 Subsuface low, 5 Subsurface high3 Organic Debirs - 1 None, 2 Some, 3 Lots

Client:Project No.:Date & Time:

Ponar Habitat Sampling(Petite Ponar)

6800 Campobello RoadMississauga, Ontario

www.ecometrix.ca(905) 794-2325

Signature: ________________________ .

Client: __________________________ ___ of ___ Project No. ______________________ Investigators: ____________________

ECOMETRIX INCORPORATED FISH COLLECTION FORM

Waterbody: ______________________ Station No.: __________ Location: ___________________ Coordinates: __________________ N ____________________ W Datum: _______________ Gear Type: Gillnet ( ) Trapnet ( ) Other ( ): _______________ Net Size: (l x w x h): ____________________ Mesh Size (inch): _____________ Net Set Date: _________________________ Time: ______________ Net Removed Date: _________________________ Time: ______________ Total Fishing Time: _______________ (hrs) Depth of Set: _______________ (m) Substrate Characteristics: __________________________________________________________

Fish Species

Length (cm)

Weight

(g)

Notes (sex, stomach contents, parasites, etc.)

Client: __________________________ ___ of ___ Project No. ______________________ Investigators: ____________________

ECOMETRIX INCORPORATED FISH COLLECTION FORM

Waterbody: ______________________ Station No.: __________ Location: ___________________ Coordinates: __________________ N ____________________ W Datum: _______________ Gear Type: Gillnet ( ) Trapnet ( ) Other ( ): _______________ Net Size: (l x w x h): ____________________ Mesh Size (inch): _____________ Net Set Date: _________________________ Time: ______________ Net Removed Date: _________________________ Time: ______________ Total Fishing Time: _______________ (hrs) Depth of Set: _______________ (m) Substrate Characteristics: __________________________________________________________

Fish Species

Length (cm)

Weight

(g)

Notes (sex, stomach contents, parasites, etc.)

Fish Habitat Study Deviations from the Terms of Reference

The following is a list of study components that were changed from that which is stated in the Terms of Reference. Rational for the change is also provided.

1. Transect spacing of ponar sampling was changed from 100 meters between transects to 200 meters between transects. Upon completion of the first four transects it became evident that the spatial distribution of substrate could be adequately mapped using a greater distance between transects.

2. Water velocity measurements were taken at 1 meter below the water surface. Water velocity measurement at 60% depth was not possible as the rod holding the probe would break due to the strong current. Therefore, a choice was made to take velocity measurements at a consistent depth of 1 m below the surface of the water.

These were the only two changes that were made concerning the sampling plan defined in the Terms of Reference.

FISH HABITAT ASSESSMENT Appendix B

Appendix B Shoreline Habitat Field Sheets

FISH HABITAT ASSESSMENT Appendix C

Appendix C Petite Ponar Sampling Field Sheets

FISH HABITAT ASSESSMENT Appendix D

Appendix D Photo Documentation

(Photo numbers are referenced in Appendix C)

Photo numbers range from 1 to 307; however, some photos numbers are not present as they were either bedrock samples or not taken.

Note, photo numbers 72 to 84 were used twice in error. Therefore, the zone to which they

pertain to is identified in this appendix.

a

a

a

b

b

bNote: a – denotes pictures taken from Zone 3 lagoon 2, and b – denotes pictures taken from Zone 3 main channel.

a

a

a

b

b

b

a b

Note: a – denotes pictures taken from Zone 3 lagoon 2, and b – denotes pictures taken from Zone 3 main channel.

a

a

a

b

b

b

a b


a

a

b

b

a b


ANEXO III »SIC 14 - ECOSISTEMAS ACUÁTICOS II

Potential for EDC Effects in Modern Mill Effluents


MEMO

To: Gervasio Gonzalez, UPM

From: Don Hart, Ph.D.; Brian Fraser,

M.Sc.

Ref: Potential for EDC Effects in

Modern Mill Effluents

Date: 19 March, 2019

Copies to: Bruce Rodgers, EcoMetrix

This memo addresses the potential for effects from endocrine disrupting compounds

(EDCs) in association with modern pulp mill effluents. A presentation on this subject was

given to staff with the Ministerio de Vivienda Ordenamiento Territorial y Medio Ambiente

(MVOTMA) on February 21st, 2019, including the information provided herein.

The presentation and this memo were prepared in response to concerns raised by

MVOTMA related to potential for EDC effects from the proposed UPM mill on the Rio

Negro. The concerns have been fueled in part by a thesis (Miguez-Carames, 2013) that

suggested EDC effects related to the Fray Bentos mill on the Rio Uruguay. Our critique of

that work has been presented in a separate memo (EcoMetrix, 2019). In short, the thesis

does not present credible evidence for EDC effects from the Fray Bentos mill. The

concerns were based also on a published paper about EDC effects observed in zebra fish

exposed to effluent from a Brazilian pulp mill (Castro et al., 2018). Our critique (EcoMetrix,

2019) explains that the Brazilian mill is not a modern mill using best available technology

(BAT) and is thus not in any way comparable to the Fray Bentos mill or the proposed UPM

mill on the Rio Negro.

This memo presents an overview of EDC sources, effect endpoints and organisms

potentially affected by EDC exposure. It also describes work that has been undertaken in

Canada toward prediction and mitigation of EDC effects in pulp mill effluents, and explains

that modern mills using BAT are not expected to produce EDC effects.

EDC Sources and Chemicals

There are many sources of EDC chemicals released to aquatic environments, and a wide

variety of chemicals with endocrine effects (Environment Canada, 2000). In municipal

effluents EDC chemicals include natural hormones from human waste, synthetic steroids

from contraceptives, and alkyl phenols. In agricultural runoff many pesticides are EDCs. In

industrial effluents, EDC chemicals include alkyl phenols and phthalates. In pulp mill

19 March, 2019

Gervasio Gonzalez, UPM

Page 2 of 8

Reference: Potential for EDC Effects in Modern Mill Effluents

effluents, EDC chemicals include phytosterols, phenolics, resin acids and metabolites, and

in mills using free chlorine, chlorophenols and polychlorinated dioxins/furans (Hewitt et al.,

2008; Cotrim et al., 2016). In addition, most harbours contain organotin compounds which

are EDCs, used as antifouling agents on the hulls of vessels.

Hewitt et al. (2008) note that the elimination of free chlorine use at pulp mills, and the

biotreatment of mill effluent, largely eliminate EDC effects from pulp mill effluents. Studies

show that mills with these mitigations typically show no significant effects, or effects only

with exposure to 100% effluent. Modern BAT mills, including the Fray Bentos mill and the

proposed UPM mill, employ these mitigations.

EDC Effect Endpoints

Substances that produce endocrine effects either mimic or antagonize animal hormones. In

so doing, EDCs can disrupt the normal developmental processes that are controlled by the

natural hormones. Typical responses to EDC exposure include disruption of gonadal

development, resulting in masculinization of females, feminization of males, altered sex

ratios in the population, delayed sexual maturation, reduced (or increased) egg production

and/or reduced egg viability. Egg production has been shown to be a sensitive endpoint as

compared to other reproductive endpoints in fish bioassays (Parrott, 2005).

Reproductive effects, particularly those involving reduced production or viability of eggs, are

considered to be of primary importance to population success, and are therefore the most

studied endpoints of EDC effect. However, other biological systems can be affected also.

For example, stress responses, mediated by adrenocorticotropic hormone, and growth

responses, mediated by thyroid hormones, can also be affected (Matthiessen et al., 2015).

Effect endpoints should not be confused with exposure endpoints. For example, levels of a

precursor egg protein (vitellogenin) may be measured as an indication of response to

estrogenic substances, and gene transcription activity associated with production of egg

protein may also be measured. However, such endpoints have little ecological relevance

because they are not predictive of effects on spawning or egg production or egg viability.

By themselves, they only indicate EDC exposure.

Organisms Affected by EDCs

Aquatic organisms are of primary concern for EDCs found in liquid effluents, because these

organisms are most exposed to an aquatic release. Fish are the most studied aquatic

organisms, because they are vertebrates with well understood endocrine systems, and

because standardized test systems exist for evaluating reproductive effects. Fish are a

19 March, 2019


Page 3 of 8


good surrogate for other aquatic organisms, such as amphibians, since all vertebrates have

similar hormonal systems.

The fish test system has been widely used in developing our current understanding of EDC

effects from exposure to mill effluents in Canada (Parrott, 2005; Martel et al., 2012, 2017).

Prediction/Mitigation of EDC Effects in Mill Effluents

Many chemicals in mill effluent have potential to cause reproductive effect at some

concentration. Not all are known, nor are all concentration-response relatinships known.

Nevertheless, we have effluent indicators of EDC activity, based on reproductive effect

studies over a wide range of mills. As noted by Hewitt et al. (2008), after free chlorine has

been eliminated, wood extractives are the main causative agents. Collectively, these are

reflected in the organic content of the effluent. Biological oxygen demand (BOD5) is a

good indicator of organic content.

Investigation of Cause studies were undertaken in Canada as part of the Environmental

Effects Monitoring (EEM) program (Martel et al., 2012, 2017). The objective was to find

chemical indicators in mill effluent that correlate with reproductive effect in fishes, and to

devise mitigation strategies to remove such effects. A standard egg production bioassay

with fathead minnows was used to measure effluent effect. The egg production endpoint in

this short-term test correlates well with the same endpoint in a full lifecycle test; it is a

sensitive and ecologically meaningfull indicator of reproductive effect (Parrott, 2005).

The Canadian studies involved 81 effluents from 20 mills, under normal and upset

conditions. The Canadian mills are older mills. While there have been process upgrades

since the 1990’s in many cases, they are not BAT mills. The study involved 8 bleached

kraft mills, 6 mechanical pulp mills, 4 ecycled fibre mills and 2 sulphite mills. Results for the

bleached kraft mills are shown below (Figure 1).

All the bleached kraft effluents with BOD5 greater than 25 mg/L produced significant

impairment of egg production. Only 3 effluents with BOD5 less that 25 mg/L produced

significant impairment of egg production. In two of them, from mill K3, the effect was

attributed to a polymer flocculant in the treatment plant. When this additive was changed,

the reproductive effect went away.

The study concluded that, for most mills, reducing the loading of organics in final effluent is

the best strategy for mitigating effluent effects on reproduction of fishes. Suggestions to

accomplish this were use of BAT to reduce organic losses from the mill, and optimization of

biotreatment.

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Figure 1: Egg Production (% of Control) vs BOD5 (mg/L) for Bleached Kraft Mills

Since the Fray Bentos mill is a BAT mill with average BOD5 of 9 mg/L (2012-2016), and 95th

percentile 17.7 mg/L, we would expect no reproductive effect from exposure to its effluent.

Effluent Concentration for EDC Effects

The concentration of effluent needed to cause reproductive effects depends entirely on the

effluent quality, and especially on the organic content. For modern mills using BAT, under

normal operation, we expect no effect, even at 100% effluent (Hewitt et al., 2008).

Munkittrick et al. (1998) noted that there is potential for effect at effluent concentrations as

low as 1%. The authors cited work by Kovacs et al. (1995) on a problem mill. Kovacs

found an effect threshold concentration (IC25) of 1.7% effluent for fathead minnow

spawning and egg production. The mill had only 45% ClO2 substitution, its foul condensate

was not steam stripped, and it ran town sewage through its aerated lagoon. The final

effluent contained chlorinated phenolics at 110 ug/L (51 to 218 ug/L) and AOX at 14 mg/L

(10 to 16 mg/L).

19 March, 2019


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After mill upgrade, Kovacs et al. (1996) reported that there was no reproductive effect at the

highest effluent concentration tested (20%). The final effluent contained chlorinated

phenolics at 5.6 ug/L (2 to 10 ug/L) and AOX at 2.8 mg/L (1.6 to 3.7 mg/L).

In comparison, the Fray Bentos mill effluent (2012-2016) had chlorinated phenolics at 0.49

ug/L and AOX at 1.6 mg/L. Given its better quality, we would expect no reproductive effect

from fish exposure at 20% effluent or higher concentrations.

Martel et al. (2011) studied another bleached kraft mill before and after implementation of

biotreatment. Before biotreatment, with BOD5 at 53 mg/L, fathead minnow egg production

was significantly reduced at an effluent concentration of 65% (Figure 2). After biotreatment,

with BOD5 at 19 mg/L, fathead minnow egg production was not significantly reduced, even

at 100% effluent concentration (Figure 3).

Figure 2: Egg Production Before Biotreatment (BOD5 53 mg/L) (Martel et al., 2011)

19 March, 2019


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Figure 3: Egg Production After Biotreatment (BOD5 19 mg/L) (Martel et al., 2011)

As discussed above, for the Fray Bentos mill, with average BOD5 of 9 mg/L (2012-2016),

and 95th percentile 17.7 mg/L, we would expect no reproductive effect from exposure to the

mill effluent.

History of EDC Concern at Mills in Canada and Europe

Studies of EDC effects in fish began at older mills in Sweden, and then in Canada, in

1988/89 (Munkittrick et al., 1998). Pulp and paper effluent regulations, established in

Canada in 1992, required environmental effects monitoring to look for reproductive and

other effects in fish in the receiving environment near the mills. Throughout the 1990’s

there was a push to reduce/eliminate use of free chlorine in mills, and to start/improve

biotreatment. In Europe, BAT standards were implemented, and new mills are expected to

meet that standard. As a result, fish reproductive effects from mills have been greatly

reduced or eliminated. Investigation of Cause studies in Canada point to mitigation of such

effects by reducing organic losses from the mill, consistent with BAT, and by optimizing

biotreatment (Martel et al. 2012, 2017).

The Fray Bentos mill, and the proposed UPM mill, are modern BAT mills, and as such, they

have good effluent quality and are not expected to produce fish reproductive effects.

19 March, 2019


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Overall Characterization of Risks from EDCs from the Proposed Pulp Mill Project at

Paso de los Toros

Based on consideration of our current understanding of EDCs that is based on design

information for the proposed mill and operating experience at modern BAT mills, published

research, literature and professional experience no significant effects as the result of EDCs

are expected as the result of the project.

References Cited

Cotrim, G., C.S. Fanning, G.O. da Roacha, and V. Hatje. 2016. Endocrine disruptors: strategies for determination and occurance in marine environments. J. Integrated Coastal Zone Management 16(3): 299-326.

EcoMetrix Incorporated (EcoMetrix). 2019. Memo to Gervasio Gonzalez - Review of EDC References Provided by DINAMA. Dated 19 March 2019.

Environment Canada. 2000. Endocrine Disrupting Substances in the Environment. Cited in: C. Labelle, 2000, Endocrine Disruptors Update, PRB-00-01E. Science and Technology Division.

Hewitt, L.M., T.G. Kovacs, M.G. Dube, D.L. MacLatchy, P.H. Martel, M.E. McMaster, M.G. Paice, J.L. Parrott, M.R. van den Heuvel and G.J. Van Der Kraak. 2008. Altered reproduction in fish exposed to pulp and paper mill effluents: Roles of individual compounds and mill operating conditions. Environ. Toxicol. And Chem. 27(3): 682-697.

Kovacs, T.G., J.S. Gibbons, L.A. Tremblay, B.I. O’Connor, P.H. Martel and R.H. Voss. 1995. The effects of a secondary treated bleached kraft mill effluent on aquatic organisms as assessed by short-term and long-term laboratory tests. Ecotoxicol. and Environ. Safety 31: 7-22.

Kovacs, T.G., J.S. Gibbons, P.H. Martel, and R.H. Voss. 1996. Improved effluent quality at a bleached kraft mill as determined by laboratory biotests. J. Toxicol. and Environ. Health Part A 49(5): 533-561.

Martel, P.H., T.G. Kovacs, B.L. O’Connor, S. Semeniuk, L.M. Hewitt, D.L. MacLatchy, M.E. McMaster, J.L. Parrott, M.R. van den Heuvel and G.J. Van Der Kraak. 2011. Effluent monitoring at a bleached kraft mill: Directions for best management practices for eliminating effects on fish reproduction. J. Environ. Science and Health Part A 46: 833-843.

Martel, P., T. Kovacs, B. O’Connor, M. Hewitt, M. McMaster, J. Parrott, D. MacLatchy, G. Van Der Kraak and M. van den Heuvel. 2012. Towards Cost Effective Solutions for

19 March, 2019


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Elimination of Reproductive Effects in Fish Exposed to Pulp and Paper Mill Effluents. Cycle 6 Investigation of Cause Project. Environment Canada.

Martel, P.H., B.I. O’Connor, T.G. Kovacs, M.R. van den Heuvel, J.L. Parrott, M.E. McMaster, D.L. MacLatchy, G.J. Van Der Kraak and L.M. Hewitt. 2017. The relationship between organic loading and effects on fish reproduction for pulp mill effluents across Canada. Environ. Science and Technol. 51: 3499-3507.

Matthiessesn, P., J.R. Wheeler and L. Weltje. 2018. A review of evidence for endocrine disrupting effects of current-use chemicals on wildlife populations. Critical Reviews in Toxicology 48(3):195-216.

Munkittrick, K.R., M.E. McMaster, L.H. McCarthy, M.R. Servos, and G.J. VanDer Kraak. 1998. Overview of recent studies on the potential of pulp mill effluents to alter reproductive parameters in fish. J. Toxicol. Environ. Health, Part B, 1:347-371.

ANEXO IV »SIC 14 - ECOSISTEMAS ACUÁTICOS II

Review of EDC References Provided by DINAMA


MEMO

To: Gervasio Gonzalez, UPM

From: Don Hart, Ph.D.; Brian Fraser,

M.Sc.

Ref: Review of EDC References

Provided by DINAMA

Date: 19 March, 2019

Copies to: Bruce Rodgers, EcoMetrix

In support of UPM’s ongoing consideration of construction of a state-of-the-art, best

available technology (BAT) pulp mill near Paso de los Toros, Uruguay, EcoMetrix

incorporated (EcoMetrix) has undertaken various tasks, including assessing potential water

quality related effects that may be associated with mill operations on the Rio Negro. The

assessment has considered the potential effects of nutrients (in terms of both enrichment

and toxicity), dissolved oxygen, pH, resins acids, dioxins and furans and endocrine

disrupting compounds (EDCs). A presentation was given to staff with the Ministerio de

Vivienda Ordenamiento Territorial y Medio Ambiente (MVOTMA) on February 21st, 2019,

that summarized this assessment. The presentation included review of EDCs (history of

concern, sources, mode of action, assessment endpoints) and EcoMetrix’ experience with

EDCs as gained through our long-term involvement with the pulp and paper industry, and

more specifically the Environmental Effects Monitoring (EEM) program, in Canada on which

much of the important early EDC research was completed .

It is our understanding that MVOTMA staff have requested a review of specific publications

regarding endocrine disrupting compounds (EDCs) and their implications as to potential for

EDC effects in the Rio Negro in the vicinity of the proposed mill site. Specifically, it has

been requested that the following documents should be reviewed:

Miguez-Carames, D. (2013). Integrated risk assessment of endocrine disruptors

in the Uruguay River. Ph.D. Thesis. Cranfield University. Cranfield Water

Science Institute.

Castro, A.J.G., et al. (2018). Exposure to a Brazilian pulp mill effluent impacts

the testis and liver in the zebrafish. Comp. Biochem. and Physiol., Part C (206-

207):41-47.

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Reference: Review of EDC References Provided by DINAMA

Carnikian-Fernandes, A. (undated). Evaluacion de los efectos de efluentes

industriales y urbanos sobre aspectos de la reproduccion en el pez Pimephales

promelas. M.Sc. Tesis. Universidad de la Republica Oriental del Uruguay.

Keel-Morgan, K. (2012). Disruptores Endocrinos. Efectos en peces Pimephales

promelas. M.Sc. Tesis. Instituto Pasteur Montevideo. Facultad de Ciencias.

This memo has been prepared in response to the above-referenced information request. A

review of each document is provided below. A summary that considers these documents in

light of other literature on EDC effects as the result of pulp mill effluent exposure is also

provided for reference.

It is noted that we were familiar with two of the four documents directly - Miguez-Carmes,

2013; Castro et al., 2018 - as well as the other two indirectly as they were referenced in the

Miguez-Carmes PhD thesis, but did not include them in our EDC review. In the former

document, our interpretation of the results differed than those presented by the author;

whereas, in the latter document the results were not relevant as the mill in question was not

based on BAT.

Review of Miguez-Carames (2013)

This thesis is presented as an assessment of risk from endocrine disruptors in the Rio

Uruguay. It includes a review of literature, a screening assessment of EDC hazards in the

river, an Exposure Assessment (based on monitoring of both EDCs and biological effects in

the river), a Risk Estimation (calculation of hazard quotients) and a Risk Characterization

(interpretation of risks related to particular industrial/municipal sources).

Our review is focused on evidence and interpretations relevant to potential for EDC effects

from the pulp mill at Fray Bentos, in consideration of the both the in-field and laboratory

data that are presented. In this context, our overall opinion of the thesis is that it is

speculative in nature and that it greatly overstates the potential risks from EDCs on aquatic

biota that may be mill related. The evidence linking the mill discharge to effects does not

hold up to scrutiny.

The review below focuses on two main aspects of the thesis. First, the review considers

the information or evidence provided in the thesis concerning effluent mixing the likelihood

of effluent-exposure for aquatic biota. Secondly, the review considers the analysis of the

reproductive endpoints derived from field surveys and laboratory testing.

19 March, 2019


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1.0 Effluent Dilution and Effluent Exposure

Upstream Reference Stations

Table 4-1, page 63, of the thesis summarizes the monitoring stations. The only station

identified as reference is R1 located approximately 30 km upstream near Nuevo Berlin. Two

other stations should also be listed as reference stations, R2 and R4.

Station R2 is located approximately 1.5 km upstream near the international bridge. It is not

identified as a reference station even though it is located upstream from the discharge.

Station R2 is identified as a station to “evaluate reverse flow”.

The flow in the river can be influenced by wind driven seiche within the Atlantic Ocean and

Río de la Plata. Such seiche events can cause the flow within the Río Uruguay to

temporarily increase or decrease. Under rare occasions (a few times per year or less), the

flow can even reverse direction and travel upstream for a few hours. These flow reversals

have been observed during extreme low flow conditions, but are not expected to occur

when the flow at the Salto Grande Dam is greater than 1,000 m3/s (EcoMetrix, 2006).

The thesis did not present all of the data used for the assessment, so we cannot determine

which data were recorded during a flow reversal event or not. But given the short duration

and extreme nature of such events, it is unlikely that the data recorded at station R2

occurred during a flow reversal. As such, station R2 is most likely a reference station and

not an exposure station.

The thesis concludes that the “probability of finding contamination from the water route is

negligible for Yaguareté Bay (R4)”. The dilutions reported at Yaguareté Bay range from

3,000,000:1 at an extreme low flow of 500 m3/s to 28,000:1 at average flows of 6,000 m3/s.

Environment Canada defines areas that exceed 1,000:1 dilution as reference areas and

considers them representative of reference conditions unaffected by the wastewater

discharge. As such, the monitoring stations within Yaguareté Bay (R4, S4 and F4) can all

be considered reference stations.

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Predicted Dilution

Figure 4-4 of the thesis shows the calculated dilutions from the thesis at Yaguareté Bay

(R4), Ubici Beach (R5), the drinking water intake (R6) and Las Cañas (R8). For each

station, the predicted dilutions are 3,000,000:1, 890:1, 400:1 and 600:1, respectively, at 500

m3/s, and 28,000:1, 2,400:1, 7,500:1 and 11,000:1, respectively, at 6,000 m3/s.

A flow of 500 m3/s is expected to occur in the Río Uruguay at Fray Bentos once every 5 to

20 years based on a 7Q1 of 950 m3/s; 7Q5 of 640 m3/s; and 7Q20 of 400 m3/s (EcoMetrix,

2006). The average flow in the Río Uruguay at the Salto Grande dam is approx. 6,230

m33/s.

19 March, 2019


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Figure 7-12 shows identical results to those shown in Figure 4-4, except Figure 7-12 is as

% effluent and Figure 4-4 is as dilution.

19 March, 2019


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Misrepresentation of the Effluent Plume

Figure 7-3 presents the iso-concentration maps at 500 m3/s and 1,000 m3/s at Yaguareté

Bay (R4), Ubici Beach (R5), and further downstream. The scale on this figure is illegible

given the poor clarity of the figure. Regardless of the scale, the figure provides a gross

misrepresentation of the plume delineation from the mill discharge.

The colours depicted at Yaguareté Bay (R4) and Ubici Beach (R5) are bright orange and

yellow, which usually implies a high concentration of effluent at levels that may cause harm

to the environment.

Figure 4-4 and Figure 7-12 declare that at Yaguareté Bay (R4) the effluent is diluted

3,000,000:1 (0.00003% effluent) under the two flow conditions (500 and 1,000 m3/s). This

implies no exposure to effluent; hence the colour at Yaguareté Bay (R4) should be blue

similar to the reference station at the international bridge (R2).

At Ubici Beach (R5), the effluent is diluted 890:1 and 610:1 (0.11% and 0.16%) under the

two flow conditions. Such high dilutions would not be expected to harm the environment.

Given the high dilutions predicted in Figure 4-4 (and percent effluent predicted in Figure 7-

12), the iso-concentration map shown in Figure 7-3 should more accurately be represented

19 March, 2019


Page 7 of 33


by slight differences in blue with green limited to the mixing zone limited to the immediate

vicinity of the discharge.

Figure 1 presents the delineation of the effluent plume for the mill discharge based on field

measurements conducted during low, typical and high flow conditions (EcoMetrix, 2009).

The plume delineation shown in Figure 1 based on field measurements differs significantly

from that presented in Figure 7-3.

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Figure 1: Delineation of effluent plume under low (742 m3/s), typical (3,784 m

3/s) and high

(8,192 m3/s) flow conditions from field measurements (EcoMetrix, 2009)

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Conductivity Along the Río Uruguay

Figure 7-5 shows the measured conductivity along the river from Nuevo Berlin (R1) to Las

Cañas (R8) for periods pre- and post- start-up of the mill.

It is apparent that the conductivity within the Río Uruguay has decreased post- mill start-up

as compared to the period pre- mill start-up. This decrease extends along the entire length

of the river, including the reference stations at Nuevo Berlin (R1) and international bridge

(R2).

Furthermore, for the period post- mill start-up, the conductivity increases along the length of

the river from approx. 58 µS/cm at Nuevo Berlin (R1), to approx. 60 µS/cm at the

international bridge (R2), to approx. 62 µS/cm at Fray Bentos (R3), to approx. 64 µS/cm at

Las Cañas (R8). Generally, an increasing spatial trend indicates a non-point source rather

than a point source. A non-point source implies a continuous input of waters with elevated

conductivity along the entire length of the river, such as from agricultural runoff. A point-

source input generally shows as an abrupt increase in conductivity at the point of inflow

followed by a gradual decrease in conductivity with distance downstream due to dispersion.

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Wind and Currents

On page 140, the thesis reads “Readings of wind direction and wind velocity related to the

sampling moment were used to evaluate if there could be influence of Yaguareté stream

(C1) on R4 conductivity as it drained into the bay”. Wind speed and direction affect wave

generation not necessarily currents. Waves may have been observed on the surface of the

water during sampling but these surface waves in no way affect the advective transport of

waters from Yaguareté stream within the bay.

Table 7-2 presents four surveys within Yaguareté Bay (R4) with specific reference to wind.

On these four dates, the flows at the Grande Salto Dam were 7,089 m3/s, 2,270 m3/s, 5,755

m3/s and 9,349 m3/s. Under such high flows, the currents within Yaguareté Bay are most

likely dominated by the flow within the Río Uruguay and less so by local winds.

It is unclear whether or not data were discarded due to wind speed and direction.

Measured Conductivity, Phenols and AOX Data Presented in Thesis

Table 7-2, Table 7-3, Table 7-4, Figure 7-15 and 7-16 present the measured data used to

support the thesis. A few other summary tables and figures were provided but not a detailed

account of all available data. The sections below reference the data presented in these

tables and figures.

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Yaguareté Bay (R4)

As previously stated, the thesis correctly concludes that the “probability of finding

contamination from the water route is negligible for Yaguareté Bay (R4)”. This conclusion is

based on the predicted dilutions reported at Yaguareté Bay ranging from 3,000,000:1 at an

extreme low flow of 500 m3/s to 28,000:1 at average flows of 6,000 m3/s. As such, the

monitoring stations within Yaguareté Bay (R4, S4 and F4) can all be considered reference

stations.

From Table 7-2, the conductivity within Yaguareté Bay (R4) ranges from 71 to 96 µS/cm

base on four surveys. In comparison, the reference conductivity at the international bridge

(R2) ranges from 72 to 99 µS/cm based on three surveys. The conductivity in Yaguareté

Bay (R4) exceeds the reference conductivity by 5 and 19 µS/cm based on two surveys, and

is lower than the reference conductivity by 22 µS/cm for one survey.

According to Figure 4-4, the dilution of mill effluent at Yaguareté Bay (R4) ranged from

600,000:1 to 28,000:1 (0.00017% to 0.0036%). Given this level of dilution, the potential

change in conductivity in Yaguareté Bay (R4) attributed to the mill discharge would be less

than 0.1 µS/cm, far less than the differences measured. The mill discharge cannot be

attributed to the observed increase in conductivity on the two survey dates.

The conductivity in Yaguareté stream (C1) ranged from 195 to 594 µS/cm based on three

surveys. These values translate into dilutions ranging from 6:1 and 79:1. It is far more likely

that the water quality within Yaguareté stream (C1) influences water quality in Yaguareté

Bay (R4) than mill effluent.

From Figure 7-15, the average concentration of phenols in Yaguareté Bay (R4) is approx.

2.1 µg/L compared with a reference concentration at Nuevo Berlin (R1) and the

international bridge (R2) of approx. 0.6 µg/L. The difference of 1.5 µg/L cannot be attributed

to the mill discharge given the reported dilution, but it can be attributed to the discharge

from Yaguareté stream (C1) since the concentrations compare.

From Figure 7-16, the concentration of AOX in Yaguareté Bay (R4) was non-detectable

based on what seems to be one measurement. In comparison, the reference concentration

at Nuevo Berlin (R1) and the international bridge (R2) was approx. 2.5 µg/L. The data do

not support an interpretation of potential sources. The mill discharge is not expected to

affect the concentration of AOX in Yaguareté Bay (R4) given the reported dilution.

Yaguareté stream (C1) is expected to affect the concentration of AOX in Yaguareté Bay

(R4) given the elevated concentration of AOX of approx. 12.5 µg/L and the limited dilution

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of the Yaguareté stream (C1) discharge within Yaguareté Bay (R4) estimated from

measured phenols and conductivity.

The limited data provided in the thesis for conductivity, phenols and AOX implicates

Yaguareté stream (C1) as the most likely cause of water quality effects within Yaguareté

Bay (R4). The reported dilution of the mill effluent within Yaguareté Bay (R4) identifies it as

the least likely cause of water quality effects within Yaguareté Bay (R4), as concluded by

the thesis. This also implies all effects to sediment quality and biota to be attributed to

Yaguareté stream (C1) and not the mill discharge.

Ubici Beach (R5)

From Table 7-2, the measured conductivities at Ubici beach (R5) ranged from 76 to 130

µS/cm based on four surveys. In comparison, the reference conductivities at the

international bridge (R2) ranged from 72 to 99 µS/cm based on three surveys. The

conductivity in Yaguareté Bay (R4) exceeds the reference conductivity by 5 to 31 µS/cm.

According to Figure 4-4, the dilution of mill effluent at Ubici beach (R5) ranged from 640:1

to 2,400:1 (0.16% to 0.04%). Given this level of dilution, the potential change in conductivity

at Ubici beach (R5) attributed to the mill discharge would range from 1 to 5 µS/cm,

insufficient to account for the observed increase in conductivity at Ubici beach (R5).

The discharge from Yaguareté stream (C1) could account for the observed increase in

conductivity at Ubici beach (R5). The measured conductivity in Yaguareté stream (C1)

ranged from 195 to 594 µS/cm based on three surveys, resulting in dilutions at Ubici beach

(R5) ranging from 16:1 to 39:1. These are realistic dilutions to expect for a shoreline

discharge from a creek into a river.

From Figure 7-15, the average concentration of phenols at Ubici beach (R5) was approx.

4.5 µg/L compared with a reference concentration at Nuevo Berlin (R1) and the

international bridge (R2) of approx. 0.6 µg/L. The difference of 3.9 µg/L cannot be attributed

to the mill discharge given the reported dilution. The mill discharge would only account for a

0.01 µg/L change in phenols at Ubici beach (R5), not the observed 3.9 µg/L change. The

difference can also not be attributed to the discharge from Yaguareté stream (C1) since the

concentration in the stream is approx. 1.8 µg/L, lower than the reported concentration at

Ubici beach (R5). Another source of phenols is required to explain the reported

concentration of phenols at Ubici beach (R5).

Figure 2 shows Google Earth images of the Ubici Beach, Yaguareté Bay, and the adjacent

watershed. The images reveal three potential sources.

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1. The municipal landfill for the city of Fray Bentos is located south of Ubici Beach

(towards the lower portion of Figure 2). The municipal landfill was apparently

constructed without impervious barriers, so all leachate generated within the landfill

reports to the local watershed. This leachate would then presumably enter

Yaguareté Bay via Yaguareté stream or the smaller creek towards the west. The

quality of the leachate has not been reported.

2. Sewage from urban areas within the city of Fray Bentos do not have collection or

treatment systems. The sewage from these areas reports to the local watershed,

which presumably enters Yaguareté Bay via Yaguareté stream or the smaller creek

to towards the west. The quality of the leachate has not been reported.

3. Two factories, meat packing and juice packaging, are located towards the south

east of Ubici Beach. The Google Earth image shows a wastewater treatment

system, which presumably collects and treats the wastewaters from these two

factors. The treated wastewaters are discharged to Yaguareté Bay via the small

creek within approximately 700 m from Ubici Beach. The quality of the discharge

has not been reported.

From Figure 7-16, the concentration of AOX at Ubici beach (R5) was non-detectable based

on what seems to be one measurement. In comparison, the reference concentration at

Nuevo Berlin (R1) and the international bridge (R2) was approx. 2.5 µg/L. The limited AOX

data do not support an interpretation of potential sources.

The limited data provided in the thesis for conductivity, phenols and AOX does not provide

conclusive evidence of the source of water quality effects at Ubici beach (R5). The mill

discharge cannot be attributed to the observed effects given the significant level of dilution.

Yaguareté stream (C1) as the most likely cause of water quality effects within Yaguareté

Bay (R4). The reported dilution of the mill effluent within Yaguareté Bay (R4) identifies it as

the least likely cause of water quality effects within Yaguareté Bay (R4), as concluded by

the thesis. This also implies all effects to sediment quality and biota to be attributed to

Yaguareté stream (C1) and not the mill discharge.

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Figure 2: Google Earth images of Ubici Beach, Yaguareté Bay, and the adjacent watershed

showing potential sources to the waterfront

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Las Cañas (R8)

The conductivity at Las Cañas (R8) was 74.2 µS/cm as a median from Table 7-3 and 69.6

µS/cm as a mean from Table 7-4. In comparison, the reference conductivity at the

international bridge (R2) was 67 µS/cm as a median from Table 7-3 and 64.0 µS/cm as a

mean from Table 7-4. The conductivity at Las Cañas (R8) exceeds the reference

conductivity by 7.2 and 5.6 µS/cm based on the two pairs of values reported in the

respective tables.

According to Figure 4-4, the dilution of mill effluent at Las Cañas (R8) ranged from 4,100:1

to 11,000:1 (0.02% to 0.01%). Given this level of dilution, the potential change in

conductivity at Las Cañas (R8) attributed to the mill discharge would range from 0.3 to 0.8

µS/cm, insufficient to account for the observed increase in conductivity at Las Cañas (R8).

From Figure 7-15, the average concentration of phenols at Las Cañas (R8) was approx. 0.8

µg/L compared with a reference concentration at Nuevo Berlin (R1) and the international

bridge (R2) of approx. 0.6 µg/L. The difference of 0.2 µg/L cannot be attributed to the mill

discharge given the reported dilution. The mill discharge would only account for a 0.001

µg/L change in phenols at Las Cañas (R8). Another source of phenols is required to explain

the reported concentration of phenols at Las Cañas (R8). Possible sources include urban

runoff from Fray Bentos

From Figure 7-16, the concentration of AOX at Las Cañas (R8) appears to be similar, if not

less than, the reference concentration at Nuevo Berlin (R1) and the international bridge

(R2). There is no indication from the measured data that the mill effluent affects the

concentration of AOX at Las Cañas (R8).

The limited data provided in the thesis for conductivity, phenols and AOX does not provide

conclusive evidence of the source of water quality effects at Las Cañas (R8). The mill

discharge cannot be attributed to the observed effects given the significant level of dilution.

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2.0 Reproductive Endpoints Derived from Field Surveys and Laboratory Testing

Effluent Testing for Biological Effects

Most relevant to the question of pulp mill effects, the fathead minnow (Pimephales

promelas) was exposed to 100% pulp mill effluent. In the 21-day egg production test, it was

noted that egg production was reduced (Table B-4, shown below). From the information

provided, it is not clear that a statistically significant reduction in egg production occurred.

Note: C2 stream receives municipal wastewater; E2 is -oestradiol (positive control)

The test protocol requires a minimum of 4 replicate exposures per treatment, and statistical analysis of the data according to specifications (US EPA, 2002a). The thesis does not identify the number of replicate exposures, nor present any statistical analysis of the data. It is stated that there were 2 males 4 females per tank, and that “n=36” (pp B-401-402). We infer from this that there may have been 36 fish and 6 tanks per sample tested, but it is unclear how they were allocated between the treatments and the controls. In the absence of statistical analysis, it is not clear that an effluent effect on egg production has been demonstrated. Statistical methods were described by Keel-Morgan (2012) and are reviewed below. They did not follow the test protocol, and seem to be inappropriate.

What is clear is that total egg production can differ by a factor of two among the various

controls, similar to the magnitude of difference between the control and the effluent

exposed treatment. Effluent exposed fish produced eggs in numbers similar to most of the

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control groups. The control associated with the effluent test had unusually high egg

production, while the fish exposed to mill effluent were similar to the other controls.

In the 14-day reproduction test with fathead minnows, effluent exposed fish were examined for effects on gonadal development (gonadosomatic index, GSI), development of secondary sexual characteristics (nuptial tubercles), condition factor, and induction of egg protein (vitellogenin). In all these cases, no significant effect of pulp mill effluent was observed (Tables B-5, B-6; see un-numbered table from thesis). In addition, histological examination of gonads revealed no effect of pulp mill effluent.

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Un-numbered Table from Thesis: VTG concentrations in liver homogenate of exposed and

control P. promelas

*Significant differences (concentration units were not stated)

Field Testing for Biological Effects

In field studies of fish health, it was noted that the gonadosomatic index (GSI) for Stage 2 testes of the banded tetra (Astyanax fasciatus) was reduced in August 2010 in fish collected at Ubici Beach (F3) as compared to fish from Yaguarate Bay (F2) (Figure 7-38, Figure 7-42). This was interpreted as a pulp mill effect. However, since neither location is appreciably exposed to mill effluent, the interpretation is not defensible. For low to average flow (2,000 m3/s to 6,000 m3/s) the dilution of mill effluent is 640:1 to 2,400:1 at Ubici Beach (R5) and 28,000:1 to 600,000:1 at Yaguarate Bay (R4) (Figure 4-4, presented above) (mill effluent of 0.16% to 0.04% at R5; 0.0036% to 0.00017% at R4). A more likely cause of the observed GSI response at Ubici Beach (F3) would be exposure to agricultural runoff from Stream C1, or exposure to industrial wastewater effluents, both of which discharge upstream of F3.

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Moreover, since fish exposed experimentally to 100% mill effluent did not show a GSI response, it seems unlikely that mill effluent is the cause of the GSI response at F3.

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Figure 7-42 Fish sampling locations for GSI of male A. fasciatus

In testing of river sediment elutriate toxicity to fathead minnows, the embryo-larval stage was exposed to elutriate over a 7-day period. It was noted that elutriate from the S2 river transect, downstream of the Fray Bentos mill, produced 3.3% spinal cord malformations. No malformations were observed at Nuevo Berlin (S1), upstream of the mill, or at Las Canas (S3), downstream. This was suggested as evidence of endocrine effect from the mill.

However, the embryo-larval mortality was 21.5% at Nuevo Berlin, 7.5% at Fray Bentos, and 0% at Las Canas, indicating greater reproductive effect upstream of the mill.

The test protocol (US EPA, 2002b) calls for five effluent concentrations and a control, each with four replicates. The test acceptability is defined in terms of control results. No control results were presented in the thesis, so the validity of the test cannot be determined. Nor could we find any indication of test replication. The protocol requires statistical analysis of the concentration-response curve to estimate ECx, NOEC and LOEC concentrations. No such analysis was presented. In the absence of these required elements, it cannot be determined whether there was any statistically significant effect on embryo-larval development or survival, at S2 or any other location. The 3.3% frequency of malformations is small, and unlikely to be significant, either statistically or ecologically.

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The concentration-response curve shown for the Fray Bentos elutriate exposure is very unusual, due to high mortality at the lowest elutriate concentration (Figure 7-27). This suggests some problem with the experimental technique in the study.

Figure 7-27 Concentration-response curve for fish embryo-larval lethality and developmental effects with exposure to sediment elutriate (S2)

Conclusions Regarding Endocrine Effect from the Fray Bentos Mill

Based on the monitoring results presented in the thesis, there is very little evidence to support the suggestion of endocrine effect due to Fray Bentos mill effluent. While gene transcription biomarkers of exposure to EDCs were found in fish exposed to mill effluent,

and certain EDCs (-sitosterol, endosulfan, alkylphenols) were measured in fish throughout the river, biomarkers of exposure should not be construed as an effect, and the similarity of EDC levels in fish throughout the river argues against a significant mill contribution.

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Overall, based on our review the evidence cited, the evidence for actual reproductive or developmental effects in the river due to mill effluent is weak.

Exposure Assessment and Risk Estimation

The thesis presents calculated hazard quotients (HQ) for selected EDCs based on estimated exposures and doses, compared to reference doses (RfD) for people, and to toxicity reference values (TRV) for selected ecological receptors. In general, the calculations as presented are not transparent, or reproducible. The HQ presentation seldom points to the exposure concentrations used in the calculation, or to the human exposure factors used (e.g. body weights, inhalation rates, dermal contact rates). No example calculations are provided.

The thesis concludes that there is a low risk of endocrine disruption in humans through fish and water ingestion, and a low to moderate risk to freshwater biota. However, we find that the risk estimations are poorly explained, with obscure linkage to the underlying data.

Nevertheless, we have been able to reproduce a few of the HQ calculations. Table 7-24 presents EDC concentrations in edible fish tissues (Pimelodus maculatus, from the Fray Bentos market) and the resulting human doses from fish consumption. Table 8-1 presents the associated HQ values (HQ = dose / RfD). Based on comparison of the two tables, three of the seven HQ values in Table 8-1 appear to be incorrect.

We have verified the doses shown in Table 7-24. Using the RfDs in Table 8-1, we calculate HQs for 4 Nonylphenol (1.3x10-5), Endosulfan sulfate (3.7x10-3) and PCBs (4.4x10-3). These all differ from the HQs shown in Table 8-1.

In the case of 4-Nonylphenol, we can reproduce the value shown in Table 8-1 by adding the HQ from drinking water (predicted concentration at R6 (0.36 ug/L), dose 7.7x10-6 mg/kg/d, HQ 1.5x10-3) and then rounding. In the other two cases we cannot explain the discrepancy.

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Notwithstanding the errors, from the risk estimates we have been able to review, we would agree that risks to people from EDCs in the Rio Uruguay are low.

Risk estimates were presented in Section 8.3.2 for benthic invertebrates based on maximum measured glyphosate in sediments of Yaguarete Stream (S5), and in Section 8.3.3 for fish (A. fasciatus), based on maximum measurements of endosulfan in fish collected at F1, F2, F3, and F4. The HQ for benthic invertebrates was 0.002 based on glyphosate in sediment at S5. The HQs for A. fasciatus were 0.6, 0.9, 0.2 and 1.9 at F1, F2, F3, and F4, respectively, suggesting endosulfan residues of concern throughout the river, and above the TRV for fish lethality at Las Canas (F4).

Based on these ecological risk estimates, we would agree with a finding of low to moderate risk to freshwater biota in the river, but the HQ values point to an agricultural runoff issue rather than a pulp mill effluent issue.

Conclusions Regarding Risk to People and Freshwater Biota

The risk estimations in the thesis are poorly explained, and difficult to check based on the information provided. The exposure data and assumptions used in the risk calculations are generally unclear. Where we have been able to check, there seem to be errors in the HQ calculations. None of the HQ calculations suggest significant risks to people or freshwater biota that are attributable to pulp mill effluent. The results do support low to moderate risk to freshwater biota, related to agricultural runoff.

Review of Castro et al. (2018)

This paper describes a study of male zebrafish exposure to a pulp and paper mill effluent in Brazil. The mill uses a pine feedstock, and a Kraft bleaching process with the sequence O, C1, E1, D1, E2. The mill has secondary treatment in an aerated lagoon with a 7-day retention time.

The mill is using free chlorine, and is therefore not a BAT mill. It should not be considered representative of a modern mill in Uruguay, using a Eucalyptus feedstock. The mill effluent that was tested had high levels of total phenols (3,800 ug/L) and resin acids (9,300 ug/L). Although not specifically reported, due to the use of free chlorine, we also expect high levels of chlorophenols.

The male zebrafish were exposed to 4% mill effluent over a 14-day period, which included the period of gonadal maturation. The gross morphology and histology of the developing testis was studied to detect any effects of effluent exposure. The GSI was not affected; however, histological differences were observed between control and exposed fish. The number of cysts containing spermatids was reduced by effluent exposure, although the

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number of spermatozoa produced was not affected. There were also histological effects in liver cells.

Biochemical effects were noted in the testes, including reduced levels of lactate and reduced activity of lactate dehydrogenase. This may have been related to the histological changes, since lactate is involved in normal development of germ cells.

Conclusions Regarding Effects of Effluent from the Proposed Mill

We can draw no conclusions from this study about potential effects of effluent from the proposed pulp mill on the Rio Negro. The proposed mill is a modern mill which meets BAT standards. The mill studied in this paper was not in any way similar.

Review of Carnikian-Fernandes (undated)

This thesis presents information on the bioassay testing with fathead minnows that contributed to the Ph.D. thesis of Miguez-Carames (2013), reviewed above. It presents additional detail that was not stated in the Ph.D. thesis, and thus facilitates our review.

Our review of this thesis is focused on evidence and interpretations relevant to potential for EDC effects from the pulp mill at Fray Bentos, and particularly on aspects of that work that we can understand better by review of this thesis.

An important line of evidence in the thesis by Miguez-Carames (2013) was the bioassay testing of effluent from the Fray Bentos pulp mill. That testing is also presented by Carnikian-Fernandes (undated), but with more methodological detail. Specifically, in the 21-day egg production bioassay, Carnikian-Fernandes clarifies that there were 3 control tanks and 3 effluent exposure tanks. Thus, n=3 for each treatment (control, effluent). This is below the minimum of 4 replicates required by the test protocol (US EPA. 2002a).

No statistical analysis is presented to support the claim that the lower egg production in the effluent treatment as compared to the control is statistically significant. As noted above, it is not clear that an effluent effect on egg production has been demonstrated. Statistical methods were described by Keel-Morgan (2012) and are reviewed below. They did not follow the test protocol, and seem to be inappropriate; thus, we do not believe that a mill effluent effect on fish egg production was demonstrated.

Carnikian-Fernandes (undated) indicates that histological examination of the gonads of exposed fathead minnows did not show any statistically significant changes as compared to controls, either for males or females. It is difficult to believe that egg production has been impaired when ovarian histology is normal.

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Review of Keel-Morgan (2012)

This thesis presents information on the bioassay testing with fathead minnows that contributed to the Ph.D. thesis of Miguez-Carames (2013) and the M.Sc. thesis of Carnikian (undated), both reviewed above. This thesis presents detail on the bioassay testing that was not stated in the other two theses, and thus facilitates our review of the work.

Our review of this thesis is focused on evidence and interpretations relevant to potential for EDC effects from the pulp mill at Fray Bentos, and particularly on aspects of that work that we can understand better by review of this thesis.

An important line of evidence in the thesis by Miguez-Carames (2013) was the bioassay testing of effluent from the Fray Bentos pulp mill. That testing is also presented by Keel-Morgan (2012), but with more methodological detail. Specifically, in the 21-day egg production bioassay, the experimental design and statistical analysis are better described by Keel-Morgan (2012).

The experimental design for the egg production bioassay is clarified. For each effluent sample there were 3 control tanks and 3 effluent exposure tanks, each containing 2 males and 4 females (36 fish in total). Thus, n=3 for each treatment (control, effluent). This is below the minimum of 4 replicates required by the test protocol (US EPA. 2002a).

The statistical analysis of the egg production data consisted of a Kolmogorov-Smirnov (K-S) test for equality of two probability distributions. The plot of total egg production against day, for each treatment, was considered as a cumulative probability distribution, and the distributions were compared between control and effluent exposed fish. This approach is referenced to Thorpe et al. (2007); however, it does not follow Thorpe et al. (2007). Thorpe et al. (2007) used the K-S test to compare the temporal pattern of egg production in pre-exposure vs exposure periods, for the same group of fish.

The K-S approach is not in accordance with the statistical analysis section of the test protocol (US EPA. 2002a). In particular, it does not consider the variability among replicates, and does not test whether the difference in egg production between treatments is greater than expected based on the variability within treatments (between replicates). This is particularly important because individual fish are variable, and differences can arise by chance, simply due to the selection of fish for control and treatment groups. A number of appropriate statistical approaches for evaluating treatment effects on egg production are described in the test protocol. The K-S test is not one of them.

Based on the departure from test protocol, and the inappropriate statistical analysis, we do not believe that an effect of pulp mill effluent exposure on fathead minnow egg production was demonstrated.

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Keel-Morgan (2012) indicates that gene transcription biomarkers of exposure to EDCs were found in the fish exposed to mill effluent. This is hardly surprising, given the detection of EDCs in fish throughout the Rio Uruguay (Miguez-Carames, 2013). Biomarkers of exposure do not constitute evidence of endocrine effect.

Summary - Consideration of Other Literature on EDC Effects in Mill Effluent

The author interpretation of results in the three theses mentioned above, i.e. that evidence is provided for EDC effects from a BAT mill on the Rio Uruguay, is in marked contrast to other literature on prediction/mitigation of such effects. Much of the literature on EDC effects from mill effluent has involved studies of Canadian mills. The Canadian mills are not modern mills. Mill effluent effects on fish reproduction have been demonstrated at some of these older mills. Systematic investigations have been undertaken to identify mill effluent characteristics that correlate with these effects, and mitigative measures that are likely to eliminate these effects (Martel et al., 2012; Kovacs et al., 2013; Martel et al. 2017).

The fathead minnow egg production bioassay test has been used as a standard test in these investigations. Martel et al. (2012, 2017) tested 81 effluents, from 20 mills, including 8 bleached kraft mills, 6 mechanical mills, 4 recycled fibre mills and 2 sulphite mills. For the bleached kraft mills (Figure 3), they found that effluents with more than ~25 mg/L BOD5 caused significantly reduced egg production, while those with less than 25 mg/L BOD5 did not, with a few exceptions that were attributed to additives. Investigation of two exceptions showed that a polymer flocculant in the treatment plant was likely the causative agent. Apart from this, the BOD5 correlation implicates organic losses, likely wood extractives, as causative agents when effects occur.

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Figure 3: Egg Production vs BOD5 in Fathead Minnows Exposed to Bleached Kraft Mill Effluent (Martel et al., 2012).

Common wood extractives include resin acids, plant sterols, phenolics, and in mills using free chlorine, chlorophenolics (Hewitt et al., 2008). The strongest reproductive effects that have been reported for mill effluents are in mills that still use free chlorine (e.g. Kovacs et al., 1995; Castro et al., 2018). Kovacs et al. (1996) reported elimination of reproductive effects at one such mill following mill upgrade, which included elimination of free chlorine. As most mills move away from free chlorine, and adopt secondary treatment, mitigation is focused on control of organic losses and optimization of biotreatment. Martel et al. (2011) report elimination of reproductive effects at a mill following implementation of biotreatment. Hewitt et al. (2008) note that, while biotreatment may not eliminate all causative agents, continuous exposure to 100% effluent is typically required to maintain effects, if any.

In consideration of the body of literature on fish reproductive effects from bleached kraft mill effluent, it is clear that such effects are not expected from effluent exposure at a BAT mill with BOD5 below 25 mg/L. Given this fact, and the questions raised in our review of the three theses mentioned above, we believe that the theses have not demonstrated reproductive effect from the BAT mill effluent that they studied. At most, they provide biomarker evidence that fish exposed to mill effluent were also exposed to EDC compounds. This is not evidence of reproductive effect.

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References Cited

EcoMetrix, 2006. Cumulative Impact Study, Uruguay Pulp Mills. Annex D: Water Quality. A report prepared for the International Finance Corporation, World Bank Group. September 2006.

EcoMetrix, 2009. Orion Pulp Mill, Uruguay. Independent Performance Monitoring as required by the International Finance Corporation. Phase 3: Environmental Performance Review, 2008 Monitoring Year. A report prepared for Botnia S.A. for the International Finance Corporation, World Bank Group. March 2009.

Hewitt, L.M., T.G. Kovacs, M.G. Dube, D.L. MacLatchy, P.H. Martel, M.E. McMaster, M.G. Paice, J.L. Parrott, M.R. van den Heuvel and G.J. Van Der Kraak. 2008. Altered reproduction in fish exposed to pulp and paper mill effluents: Roles of individual compounds and mill operating conditions. Environ. Toxicol. And Chem. 27(3): 682-697.

Kovacs, T.G., J.S. Gibbons, L.A. Tremblay, B.I. O’Connor, P.H. Martel and R.H. Voss. 1995. The effects of a secondary treated bleached kraft mill effluent on aquatic organisms as assessed by short-term and long-term laboratory tests. Ecotoxicol. and Environ. Safety 31: 7-22.

Kovacs, T.G., J.S. Gibbons, P.H. Martel, and R.H. Voss. 1996. Improved effluent quality at a bleached kraft mill as determined by laboratory biotests. J. Toxicol. and Environ. Health Part A 49(5): 533-561.

Kovacs, T.G., P.H. Martel, B.I. O’Connor, L.M. Hewitt, J.L. Parrott, M.E. McMaster, D.L. MacLatchy, G.J. Van Der Kraak and M.R, van den Heuvel. 2013. A survey of Canadian mechanical pulp and paper mill effluents: Insights concerning the potential to affect fish reproduction. J. Environ. Science and Health Part A. 48: 1178-1189.

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US EPA. 2002b. Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms. Fourth Ed. EPA-821-R-02-013. U.S. Environmental Protection Agency. Washington, DC.

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INFORME TÉCNICO · 2019-03-25 · 1 INFORME TÉCNICO RESPUESTA A SIC 14 – ECOSISTEMAS ACUÁTICOS II CUECAR S.A. Y BLANVIRA S.A. Exp. 2018/14000/011210 El presente informe tiene