Observed Termite Activity in Sector 2 in 2009

Termite Report 2009

Prepared by: Tim Myles, Ph.D. Termite Control Officer Community Design and Development Services, Building Services, City of Guelph

COUNTRY CLUB GOLFVIEW R D . GLEN BR OOK D R . ISLIN GTON April, 2010 AVE. FER N D ALE

WOOLWICH ST. D ALEBR OOK PL.

SPEED RIVER W OOD LAW N R D . W GOLFVIEWRD. W OOD LAW N R D . E

W OOD LAW N D EVONSHIRE W IN D SOR ST. CT. C EMETER Y GUELPH JUNCTION RAILWAY

FAIRWAY LANE WINDERMERE INVERNESS DR. INVERNESS

KINGS ETON PL.ETON BALMOR AL D R .

LEY ST.

COUNTRY CLUB GOLF C OU R SE CT.

BERKLEY PL. WINDSOR

SPEED RIVER RIVERVIEW PLACE BALMORALDR.

WOOLWICH ST. WOOLWICH W AVER LEY D R . MARILYN DR.

KEN SIN GTON ST. D ELTA ST.

R IVER SID E PAR K WOLSELEY RD. LAN GSIDST. E VERMONT ST.

BAILEY AVE.

DAKOTA DR. DAKOTA KENSINGTON ST. KENSINGTON

RD N ST. STEVENSON

COLLINGWOODST.

KENSINGTON ST. KENSINGTON

METCALFEST. CLIVE AVE. CLIVE DELHI ST. C ATH C AR T ST. SEN IOR BEATTIE ST BEATTIE LILAC PL.

C EN TR E SPEED RIVER SH AFTESBU R Y AVE.

KATHLEEN ST. KATHLEEN

BAILEY AVE. BAILEY

FREEMAN AVE. FREEMAN

WAVERLY DR. WAVERLY

DUMBARTON ST. DUMBARTON

VICTORIA RD. N RD. VICTORIA

KNIGHTSWOOD BLVD. KNIGHTSWOOD SHERIDAN ST. SHERIDAN FR EEMAN AVE. RIVERVIEW DR. ST. DUMBARTON

RIVERSIDE PARK SU MAC PL. BRIGHTON ST. BRIGHTON

KITCHENER AVE. ST. RENFIELD GEMMEL NELSON RD. LN. AVE. GLAD STON E AVE.

MARLBOROUGH GLADSTONE SPEEDVALE AVE. E

ACORN PL.

CHESTER ST. CT SHERWOOD DR. ALEXANDRA KNIGHTSWOOD MANHATTAN BLVD. OAKWOOD STANLEY ST. C H ESTN U T PL. PHILIP AVE. ANN ST. METCALFEST. BALSAM D R . SPRUCE PL. SHERWOOD DR. BARTON ST. ST. EXHIBITION

PAUL AVE. WALNUT DR. MAC AVE. TAMARACK

HIGHVIEW PL.

GLENWOOD GLENWOOD AVE. KATHLEEN ST. KATHLEEN PL.

PETER AVE.

STULL AVE. DR. PINE

VERNEY ST. RD. MARLBOROUGH EARL ST. EMMA ST. EMMA ST. N ST. STEVENSON H AW TH OR N E

ROBERTSON DR. ROBERTSON PL. ST. ANDREW ST.

SUNNYLEA CRES. LIN D EN PL. CALLANDER DR. CALLANDER

ORCHARD CRES. CLARENCE ST. AVE. DIVISION ST.

PL. TERRY BLVD. TERRY DRUMMOND PL. TER R Y BLVD . CALLANDER DR.

AVONDALE DANWOOD

SUMMIT RENFIELD ST. RENFIELD

GEORGE ST. WOOLWICH ST. WOOLWICH DUFFERIN ST. DUFFERIN W ALN U TDR.

PRINCESS ST. PRINCESS ERAMOSA RD. CLARKE ST. W CLARKE ST. E CRES. SUNNYLEA

KIRSTEN DR. KIRSTEN GREENVIEW ST. JOHN ST. HOMEWOOD PLEASAN T R D . HEALTH CENTRE RD. LINCOLN CRES. PLEASANT WESTMINSTER TOBEY AVE. POWELL ST. W POWELL ST. E ST. DELHI LINCOLN CRES. PIPE ST. METCALFEST. TORRANCE CRES. EXHIBITION SPEED RIVER LAVERNE AVE. PARK C R ES. MEYER TIFFANY ST. W TIFFANY ST. E

DR. VICTORIA RD. VICTORIA SKOV DR. MARCON ST. CAVELL AVE. GUELPH COTE CENTRAL ST. CENTRAL STEVENSON ST. N EXTRA ST. KERR ST. GENERAL

HOSPITAL

EXHIBITION ST. EXHIBITION

KATHLEEN ST. ST. KATHLEEN

PL. CALLANDER DR. CALLANDER TIPPERARY PL. MONT ST. C R ES.

CARDIGAN ST.

SPRING ST. GLENHILL

MEYER D R . WOOLWICH ST. WOOLWICH McTAGUE ST.

ERIN AVE.

DUFFERIN ST. DUFFERIN

VANCOUVER DR. VANCOUVER

OTTAWA CALGARY AVE. CALGARY

DUBLIN ST. DUBLIN WESTOBY

LON D ON R D . W LONDON RD. E ST. HAVELOCK

CARDIGAN ST. CADILLAC DR. LANE ST. LANE DERRY ST. EDWIN ST. PL.

SHIRLEY AVE. SHIRLEY BENNETT AVE. BRUNSWICK BENNETT AVE. AVE. CHARLES ST. ARTHUR ST. N PEARL ST. ST. KING EDMONTON DR. GARTH KIRKLAND ST. R OSED ALE AVE. WOOLWICH ST. NORWICH ST. W ST. QUEEN ERAMOSA RD. GARTH ST. PARK AVE. GREEN NORWICH ST. E VAN C OU VER D R .

GREEN ST. NORWICH ST. E ST. CATHERINE ST. R YAN AVE. ARDMAY

SUFFOLK ST. W LEMON ST. LEMON ST. WILLIAM ST. WILLIAM

KING EDWARD PL WINSTON CRES. WINSTON FRANKLIN AVE FRANKLIN CASSINO AVE. C ASSIN O AVE.

AVE. CASSINO AVE. MITCHELL ST. MITCHELL

LIVERPOOL ST. SUFFOLK ST. E

SPEED RIVER SPEED METCALFE ST. METCALFE

WINSTON CRES. DEL MAR BLVD.

ANTHONY

VICTORIA RD, N N RD, VICTORIA

CT. STUART ST. STUART

KING ST. KING MON TR EAL R D .

OXFORD ST.

DUNKIRK AVE. DUNKIRK LANE ST. LANE

KAR A LEE

LAURINE AVE. LAURINE

QUEEN ST. QUEEN CRESTWOOD PL. CRESTWOOD GLASGOW ST. N PALMER ST. N OR MAN D Y D R . PAISLEY ST. YARMOUTH ST.

DUBLIN ST. N WOOLWICH ST.

ARTHUR ST. N CAMBRIDGE ST. BAKER ST. NORFOLK ST. PAR KH OLM AVE. FRANCHETTO CHAPEL LN. BLVD.

NEW ST. JANE ST JANE COMMERCIAL H EPBU R N AVE.ST JACKSON ST. QUEBEC ST. SPEED RIVER DR. LOUISA CORK ST. W HEFFERNAN ST.

DOUGLAS ST. GRANGE ST. CLARA ST. CLARA

PROSPECT STEVENSON ST. N N ST. STEVENSON

Executive Summary

Guelph’s Termite Management Areas. Guelph has three termite management areas encompassing nearly 900 properties on 50 blocks. Red zone blocks have known termite infestations while blue zone blocks are buffer areas. Each block is assigned a sector number (Fig. 1).

Population Suppression. In 2009 measureable progress was made in area-wide termite population suppression. Total termite trap yield in 2009 was 489,810 compared to 705,261 in 2008, indicating a 30.5% termite population decline. This trend was consistent across most sectors (Fig. 2).

Management Practices. Management practices in 2009 included: monitoring and trapping, installation of new traps in the inner blue zone, reductive trapping, nematode treatments, a yard wood cleanup weekend, notices of required yard wood removal, inspections for sale of properties, issuance of disposal permits, shed treatments, pole treatments, stump and tree removals, and debris clean up along the Guelph Junction Rail line and park margins, and specification for chemical treatments on several properties.

Pattern of Termite Activity. The pattern of termite activity remained similar in 2009 to 2008, with most activity in the Woolwich management area in the sectors north of Tiffany St. (Fig. 3) and in the Emma-Pine area (Fig. 4), while activity in the area south of Tiffany Street and in the Windermere area remained minimal and restricted to relatively few properties (Figs. 3 & 5).

Nematode Treatments. Spring and fall nematode treatments were conducted on 154 properties with the entomopathogenic nematode species, Steinernema carpocapsae.

Yard Wood Cleanup Weekend. A large area-wide yard wood cleanup weekend was held on July 17- 20 with 11 bins of material removed. Five bins were also provided for three smaller cleanups. In total 16 bins and 37.3 tonnes of wood waste were removed in 2009. This was down from a total of 35 bins and 62.25 tonnes removed in 2008, indicating a declining need for large scale clean up weekends. Thus in the future, bins will be provided only for smaller prescribed cleanup projects.

Notices of Required Wood Removal. Notices of required wood removal were a new component of the program this year. Notices were sent to an initial group of 43 property owners, mostly for removal of mulch, stumps, and infested landscaping ties or trees. Compliance has generally been good, although follow up has been required in many instances. A second set of notices will be sent in 2010.

Shed Treatments. Another new component of the program this year was a shed survey and shed treatment program. As a trial run, five infested or at risk sheds were treated in 2009. In 2010, 25 shed treatments are planned. Letters will be sent to selected residents asking for authorization.

Debris Removal and Disposal Permits. Many property owners participated in ongoing yard wood and renovation related debris removal, with 265 disposal permits issued during the 2009 season. Additional tree and stump removals were conducted by both private owners and the operations department. Woody debris was also cleaned up along the Guelph Junction Rail line and margins of Goldie Mill Park and Herb Markle Park.

Chemical Treatments. Structural infestations were discovered on 15 properties. Chemical treatments, at property owner’s expense, were conducted on at least 12 of these, structural renovations or spot treatments on others. Guelph Hydro installed borate rods in utility poles on 10 properties.

Inner Blue Zone Trap Installations. Three traps were installed per property on 86 additional properties of the inner blue zone to improve capacity for early detection in areas bordering known active sectors.

Newly Detected Areas. Termites were newly detected in two areas peripheral to existing red zones. The first of these was a group of five properties on the west side of the Emma Pine Area. Termites were also detected for the first time on the east side of Sector 10. In both cases, this appears to represent improved detection due to better monitoring and awareness within the inner blue zone rather than actual termite expansion within the past year.

Experimental Permits. Applications were submitted for two federal experimental permits to the Pest Management Regulatory Agency in 2009. One permit was for zinc borate as an alternative active for Trap-Treat-Release. A second permit was for Metarhizim anisopliae, a fungal pathogen and agent of green muscardine disease of insects. Several detailed submissions were made related to the permit approval process. In April 2010, a research authorization was approved to use zinc borate. Residents will be asked to sign experimental co-operator letters prior to conducting any experimental treatments.

Reclassification and Constriction of some Termite Management Areas. Continuous inactivity in certain areas allows us to begin reclassifying certain areas and to start constricting termite management areas. Thus, sectors 000, 00, 0, 1, 9, 17, 29, 33, 38, 45, and 46, or portions thereof, formerly blue, have been reclassified as white zones. Parts of sectors 2, 22, and 42, previously classified as red, but those parts never previously infested, have been reclassified as blue zones. Parts of sectors 19, 22, 23, 24, 25, 27, 30, 36, 37, and 42 have been designated as ―continuously inactive‖, defined as being inactive for three years and more than three properties from any known termite activity for three years, indicated by green hatching (Figs. 3 & 5). The number of traps in continuously inactive areas will be reduced to a maximum of five per property, while inner blue zone areas will have a maximum of three per property. The revised termite boundaries and zones for 2010 are shown in Figure 6.

2009 Report. As with previous annual reports, the full 2009 report will be posted on the city’s termite website at www.guelph.ca > quick links > termites by early May.

Plans for 2010. Two half time summer technicians will be hired and start work the second week of May. Traps will be refurbished with new cardboard rolls and lids, missing traps will be replaced, and traps will be removed or reduced in some reclassified zones from May through June. Residents will be asked to sign authorizations letters as research co-operators prior to conducting termite treatments with zinc borate. Trapped termites will then be treated and released via active traps. Later in the season, letters will be sent to selected residents for required wood removal, or to schedule treatments of sheds, fence posts, or retaining walls.

TABLE OF CONTENTS

EXECUTIVE SUMMARY ...... i

LIST OF TABLES ...... iv

LIST OF FIGURES ...... v

LIST OF APPENDICES ...... vii

TABLES ...... 1

FIGURES ...... 12

APPENDICES ...... 48

iii

LIST OF TABLES

Table 1. Extent of Guelph’s Three Termite Management Areas in 2009...... 1

Table 2. Woolwich Termite Management Area ...... 2

Table 3. Windermere Termite Management Area ...... 4

Table 4. Emma-Pine Termite Management Area...... 4

Table 5. Summary of Property and Trap Activity in Red Zone Blocks (1998-2009)...... 5

Table 6. Nematode Treatments (2008-2009)...... 6

Table 7. Guelph Hydro Pole Treatments in 2009 ...... 10

Table 8. Required Wood Removals 2009 ...... 11

LIST OF FIGURES

Figure 1. 2009 Boundaries and sector numbers of Guelph termite management areas ...... 12

Figure 2. Termites trapped by sector and year (1999-2001 & 2007-2009) ...... 13

Figure 3. Areas of detected termite activity in the Woolwich management area ...... 14

Figure 4. Areas of detected termite activity in the Emma-Pine management area ...... 15

Figure 5. Areas of detected termite activity in the Windermere management area ...... 15

Figure 6. Revised 2010 boundaries and zones of termite management areas ...... 16

Figure 7. Trap locations and observed termite activity in sector 2 in 2009 ...... 17

Figure 8. Trap locations and observed termite activity in sector 7 in 2009 ...... 18

Figure 9. Trap locations and observed termite activity in sector 8 in 2009 ...... 19

Figure 10. Trap locations and observed termite activity in sector 10 in 2009 ...... 20

Figure 11. Trap locations and observed termite activity in sector 12 in 2009 ...... 21

Figure 12. Trap locations and observed termite activity in sector 15 in 2009 ...... 22

Figure 13. Trap locations and observed termite activity in sector 16 in 2009 ...... 23

Figure 14. Trap locations and observed termite activity in sector 19 in 2009 ...... 24

Figure 15. Trap locations and observed termite activity in sector 20 in 2009 ...... 25

Figure 16. Trap locations and observed termite activity in sector 21 in 2009 ...... 26

Figure 17. Trap locations and observed termite activity in sector 22 in 2009 ...... 27

Figure 18. Trap locations and observed termite activity in sector 23 in 2009 ...... 28

Figure 19. Trap locations and observed termite activity in sector 24 in 2009 ...... 29

Figure 20. Trap locations and observed termite activity in sector 25 in 2009 ...... 30

Figure 21. Trap locations and observed termite activity in sector 27 in 2009 ...... 31

Figure 22. Trap locations and observed termite activity in sector 30 in 2009 ...... 32

Figure 23. Trap locations and observed termite activity in sector 36 in 2009 ...... 33

Figure 24. Trap locations and observed termite activity in sector 37 in 2009 ...... 34

Figure 25. Trap locations and observed termite activity in sector 41 in 2009 ...... 35

Figure 26. Trap locations and observed termite activity in sector 42 in 2009 ...... 36

Figure 27. Trap locations and observed termite activity in sector 47 in 2009 ...... 37

Figure 28. Inner blue zone areas in 2009 ...... 38

Figure 29. Traps installed in inner blue zone areas of sector 1 in 2009 ...... 39

Figure 30. Traps installed in inner blue zone areas of sector 6 in 2009 ...... 40

Figure 31. Traps installed in inner blue zone areas of sector 9 in 2009 ...... 41

Figure 32. Traps installed in inner blue zone areas of sector 11 in 2009 ...... 42

Figure 33. Traps installed in inner blue zone areas of sector 14 in 2009 ...... 43

Figure 34. Traps installed in inner blue zone areas of sector 31 in 2009 ...... 44

Figure 35. Traps installed in inner blue zone areas of sector 32 in 2009 ...... 45

Figure 36. Traps installed in inner blue zone areas of sector 40 in 2009 ...... 46

Figure 37. Nematode treatment areas in 2008 & 2009...... 47

LIST OF APPENDICES

Media

1) May 19, 2009: City puts bite on termites, Guelph Tribune, p. 5...... 48

2) May 19, 2009: Slow progress beating termites, Guelph Mercury, p. A1...... 49

3) May 26, 2009: Termite control officer gets tough…, Guelph Mercury, p. A4...... 51

4) July 14, 2009: More termite help needed, Guelph Tribune, p. 7...... 52

5) July 17, 2009: Township approved demolition …Wellington Advert., p. A1 ...... 53

6) April 19, 2010: A new weapon in termite control. Guelph Mercury., p. A3 ...... 54

Communications

7) May 8, 2009: Progress Report & Public Meeting, to residents ...... 55

8) June 8, 2009: Spring Nematode Treatment, to residents ...... 56

9) June 9, 2009: Expansion of the Emma-Pine Area, to residents ...... 57

10) June 23, 2009: Installation of Termite Monitoring Traps in Inner Blue Zone ...... 58

11) July 6, 2009: Yard Wood Clean Up Weekend, to residents ...... 59

12) July 8, 2009: Notice of Required Wood Removal ...... 60

13) August 10, 2009: Discovery of a Termite Infestation on Your Block ...... 61

14) September 14, 2009: Update on Management Plan for Sector 10 ...... 63

15) September 21, 2009: Re-Inspection for Required Wood Removal ...... 64

Technical

16) Observations of Native Nematodes…...... 65

17) Sprigtails Associated with Termites…...... 70

18) Parasitic, Phoretic, Predatory and Incidental Mites Associated with Termites ...... 74

19) Molecular Characterization of Two Metarhzium Strains…...... 80

vii

Table 1. Extent of Guelph's Three Termite Management Areas in 2009 (Zones, Blocks and Properties)

Zones Red Blue Red + Blue

Management Areas: blocks properties blocks properties blocks properties

1) Woolwich Area 19 400 19 370 38 770

2) Windermere Area 1 41 4 17 5 58

3) Emma - Pine Area 1 18 4 29 5 47

Totals 21 459 27 416 48 869

Table 2. Woolwich Termite Management Area

Sector Number Traps per Sector Sector Number Properties Name Traps Installed Number Type Properties Installed (SE corner) Installed Property

000 Verney x Woolwich Blue 14 0 0 0.0 00 Earl x Dufferin Blue 18 0 0 0.0 0 Earl x GJR Blue 10 0 0 0.0 1 Division x Woolwich Blue 41 4 12 3.0 2 Clarence x Dufferin Red 22 20 150 7.5 3 Clarence x Speed R. Blue 7 1 3 3.0 4 Avondale x Princess White 16 0 0 0.0 5 Clarke x Princess White 13 0 0 0.0 6 Clarke x Woolwich Blue 26 9 27 3.0 7 Clarke x Dufferin Red 26 25 215 8.6 8 George x Speed R. Red 10 10 86 8.6 9 Powell x Woolwich Blue 22 7 27 3.0 10 Powell x Dufferin Red 20 20 128 6.4 11 John x Speed R. Blue 15 15 45 3.0 12 Pipe x Speed R. Red 12 12 101 8.4 13 Tiffany x Central Blue 22 0 0 0.0 14 Tiffany x Woolwich Blue 22 6 18 3.0 15 Tiffany x Dufferin Red 15 15 124 8.3 16 Tiffany x Speed R. Red 12 12 93 7.8 17 London x Exhibition Blue 1 0 0 0.0 18 Cavell x Central Blue 20 0 0 0.0 19 Extra x Woolwich Red 19 19 117 6.2 20 Kerr x Dufferin Red 15 11 78 7.1 21 Marcon x Speed R. Red 9 8 55 6.9 22 Mont x Woolwich Red 38 38 248 6.5 23 London x Dufferin Red 43 41 245 6.0 24 London x Cardigan Red 35 35 217 6.2 25 McTague x Woolwich Red 40 39 248 6.4

Table 2 (continued). Woolwich Termite Management Area

Number Traps per Sector Sector Name (SE Number Properties Sector Type Traps Installed Number corner) Properties Installed Installed Property

25 McTague x Woolwich Red 40 39 248 6.4 26 London x Dublin Blue 7 0 0 0.0 27 London x Woolwich Red 28 26 160 6.2 28 Suffolk x Park White 32 0 0 0.0 29 K. Edwd x Dublin Blue 43 0 0 0.0 30 Edwin x Woolwich Red 19 19 111 5.8 31 Charles x Woolwich Blue 25 9 27 3.0 32 Norwich x Norfolk Blue 25 6 18 3.0 33 Green x Norfolk Blue 19 0 0 0.0 34 Green x Woolwich White 26 0 0 0.0 35 Liverpool x Norwich White 22 0 0 0.0 36 Norwich x Speed R. Red 10 9 54 6.0 37 Norwich x Cardigan Red 25 24 165 6.9 38 Suffolk x Woolwich Blue 12 0 0 0.0 39 Yarmouth x Norwich White 18 0 0 0.0 40 Woolwich x Cardigan Blue 21 8 24 3.0 41 Eramosa x Speed R. Red 2 1 6 6.0 Red 400 384 2601 6.8 Subtotals Blue 370 65 201 3.1 White 169 0 0 0.0 TOTALS R+B+W 897 449 2802 6.2

Table 3. Windermere Termite Management Area

Number Traps per Sector Sector Number Properties Sector Name Traps Installed Number Type Properties Installed Installed Property

42 Balmoral x Windermere Red 41 41 228 5.6 43 Windsor x Inverness Blue 2 2 6 3.0 44 Balmoral x Inverness Blue 6 6 16 2.7 45 Balmoral x Balmoral Blue 7 6 18 3.0 46 Balmoral x Victoria Blue 2 2 7 3.5 Red 41 41 228 5.6 Subtotals Blue 17 16 47 2.9 TOTALS R + B 58 57 275

Table 4. Emma - Pine Termite Management Area

Number Traps/ Sector Sector Number Properties Sector Name Traps Installed Number Type Properties Installed Installed Property

47 Emma x Pine Red 18 18 201 11.2 48 Metcalfe x Balsam Blue 8 8 29 3.6 49 Emma (south side) Blue 8 8 24 3.0 50 Pine (east side) Blue 8 8 24 3.0 51 Emma x Metcalfe Blue 5 5 15 3.0 Red 18 18 201 11.2 Subtotals Blue 29 29 92 3.2 TOTALS R + B 47 47 293

New

Sup.

2009 2009

New

8 New 8 New

Erad.?

7 7 Sup.

Status

Resurg.

3 3 Erad.?

3 Resurg. 3 Resurg.

395

1,693

4,521

3,513

3,934

1,245

4,231

5,573

1,133

2009

31,406

16,221

11,923

13,120

43,297

489,810

101,020

246,585

279

1,520

6,590

3,542

3,985

1,603

2008

15,742

26,610

23,061

16,753

37,295

705,261

160,183

307,782

100,316

1,333

1,848

2,817

4,186

1,838

8,112

2,069

2007

22,203

606

5,285

4,465

3,211

9,672

2001

34,886

25,059

39,594

20,534

143,312

572

4,097

2000

56,725

46,767

25,532

19,227

22,623

813,774

155,148

132,158

179,156

171,769

Number of Termites Trapped*

4,332

5,526

3,100

1,112

1999

45,985

54,485

54,380

642,791

148,856

121,087

203,918

132

2009

144

2008

2007

2001

163

2000

Number of Active Traps

160

1999

2009

2008

2007◊

2001▫

2000▫

Number of Active Properties

1999▫

TableSummary 5. Propertyof and Trap ZoneBlocksRed in (1998-2009)Activity

101

-- 8 additional new blocks new 2001 8 additional -- after infested

-- 13 blocks known to be infested and managed from 1999-2001 blocks13 from -- managed to and known infested be

1998▪

No.

Sec.

▪ properties with live termites ▪ termitewith or live properties damage termitestraps traps ▫ with active active with or properties live properties ◊

* Inter-year trap yieldstrap not strictly* are Inter-year installedbecause traps late comparable were in the year in 2007

Totals Totals

10 No. Seq.

Table 6. Nematode Treatments 2008-2009

Sector Address Fall 2008 Spring 2009 Fall 2009 1 2 497 Woolwich x x x 2 2 14 Clarence x x x 3 2 22 Clarence x x x 4 2 26 Clarence x x x 5 2 30 Clarence x x x 6 7 11 Clarence x x x 7 7 15 Clarence x x x 8 7 21 Clarence x x x 9 7 23 Clarence x x x 10 7 166 Dufferin x x x 11 7 162 Dufferin x x x 12 7 160 Dufferin x x x 13 7 158 Dufferin x x x 14 7 156 Dufferin x x x 15 7 154 Dufferin x x x 16 7 150 Dufferin x x x 17 7 148 Dufferin x x x 18 7 30 Clarke x x x 19 7 26 Clarke x x x 20 7 20 Clarke x x x 21 7 457 Woolwich x x x 22 7 459 Woolwich x x x 23 7 461 Woolwich x x x 24 7 463 Woolwich x x x 25 7 469 Woolwich x x x 26 7 467 Woolwich x x 27 7 471 Woolwich x x x 28 7 473 Woolwich x x x 29 7 483 Woolwich 30 7 487 Woolwich x x x 31 7 489 Woolwich x x x 32 8 167 Dufferin x x x 33 8 165 Dufferin x x x 34 8 161 Dufferin x x x 35 8 159 Dufferin x x x 36 8 157 Dufferin 37 8 155 Dufferin x x 38 8 153 Dufferin x x x 39 8 151 Dufferin x x x 40 8 12 George x x x

Table 6 (continued). Nematode Treatments 2008-2009

Sector Address Fall 2008 Spring 2009 Fall 2009 41 10 15 Clarke St. E. x 42 10 19 Clarke St. E. x 43 10 23 Clarke St. E. x 44 10 25 Clarke St. E. x 45 10 136 Dufferin St. x 46 10 132 Dufferin St. x 47 10 130 Dufferin St. x 48 10 128 Dufferin St. x 49 10 20 Powell St. E. x 50 10 24 Powell St. E. x 51 10 26 Powell St. E. 52 10 30 Powell St. E. x 53 12 29 John x x 54 12 21 John x x 55 12 15 John x x 56 12 11 John x x 57 12 9 John 58 12 125 Dufferin x x x 59 12 123 Dufferin x x 60 12 2 Pipe x x 61 12 12 Pipe x x 62 12 16 Pipe x x 63 12 20 Pipe x x 64 15 15 Powell x x 65 15 19 Powell x x x 66 15 21 Powell x x x 67 15 112 Dufferin x x x 68 15 110 Dufferin 69 15 32 Tiffany St. E. x x 70 15 28 Tiffany St. E. x x x 71 15 22 Tiffany St. E. x x x 72 15 12 Tiffany St. E. x x x 73 15 6 Tiffany St. E. x x x 74 15 2 Tiffany St. E. x x x 75 15 409 Woolwich x x x 76 15 421 Woolwich x x x 77 16 21 Pipe x x 78 16 19 Pipe x x x 79 16 17 Pipe x x x 80 16 11 Pipe x x x 81 16 115 Dufferin x x x 82 16 113 Dufferin x x x

Table 6 (continued). Nematode Treatments 2008-2009

Sector Address Fall 2008 Spring 2009 Fall 2009 83 16 109 Dufferin x x x 84 16 105 Dufferin x x 85 16 52 Tiffany St. E x x x 86 16 56 Tiffany St. E. x x 87 16 62 Tiffany St. E. x x 88 20 19 Tiffany St. E. x x x 89 20 23 Tiffany St. E. x x x 90 20 27 Tiffany St. E. x x 91 20 22 Kerr St. x 92 20 84 Dufferin St. x 93 20 86 Dufferin St. x 94 20 88 Dufferin St. x 95 20 90 Dufferin St. x 96 20 92 Dufferin St. x 97 20 94 Dufferin St. x 98 21 61 Tiffany St. E. x x 99 21 93 Dufferin St. x 100 21 16 Marcon x 101 21 nr. Herb Markle Park x x x 102 23 17 Kerr St. x x x 103 23 21 Kerr St. x x x 104 23 82 Dufferin x x 105 23 76 Dufferin x x x 106 23 70 Dufferin x x x 107 23 64 Dufferin x x x 108 23 60 Dufferin x x x 109 23 58 Dufferin x x 110 23 54 Dufferin x x 111 23 50 Dufferin x x 112 23 341 Woolwich x x x 113 23 337 Woolwich x x 114 23 329 Woolwich x x 115 23 295 Woolwich x x 116 23 14 London Rd. E. x x 117 24 13 Marcon x x x 118 24 23 Marcon x x x 119 25 15 McTague x x x 120 25 7 McTague x x x 121 25 336 Woolwich x x x 122 25 340 Woolwich x x x 123 25 344 Woolwich x x x 124 25 348 Woolwich x x x 125 25 12 Mont x x

Table 6 (continued). Nematode Treatments 2008-2009

Sector Address Fall 2008 Spring 2009 Fall 2009 126 30 17 Edwin x x 127 30 15 Edwin x x x 128 30 11 Edwin x x x 129 30 14 London Rd. W. x x 130 30 16 London Rd. W. x x 131 30 18 London Rd. W. 132 36 Goldie Mill Park x x x 133 37 25/27 London Rd. E. x x x 134 37 271 Woolwich 135 37 265 Woolwich x x 136 37 259 Woolwich x x x 137 37 255 Woolwich 138 41 nr. Norwich bridge x x x 139 42 5 Windermere Ct. x x x 140 42 7 Windermere Ct. x x x 141 42 9 Windermere Ct. x x x 142 42 11 Windermere Ct. x x x 143 42 15 Windermere Ct. x x x 144 42 17 Windermere Ct. x x x 145 47 126 Emma x x x 146 47 128 Emma x x x 147 47 130 Emma x x x 148 47 132 Emma x x x 149 47 134 Emma x x x 150 47 136 Emma x x x 151 47 6 Pine x x x 152 47 8 Pine x x x 153 47 10 Pine x x x 154 47 12 Pine x x x 155 47 14 Pine x x x 156 47 16 Pine x x x 157 47 251 Metcalfe x x 158 47 253 Metcalfe x x 159 47 255 Metcalfe x x 160 47 257 Metcalfe x x 161 47 259 Metcalfe x x 162 47 271 Metcalfe x x x Total 97 134 154

Table 7. Guelph Hydro Pole Treatments in Termite Management Areas in 2009

Test Date Street Name Street Index St Direction Pole ID Treatment

27-Mar-09 London Road West 47 Boron Rods

14-Apr-09 Dufferin Street 44 Boron Rods

14-Apr-09 Dufferin Street 46 Boron Rods

14-Apr-09 Dufferin Street 48 Boron Rods

14-Apr-09 Earl Street 7 Boron Rods

14-Apr-09 Earl Street 9 Boron Rods

14-Apr-09 Pipe Street 1 Boron Rods

14-Apr-09 Pipe Street 3 Boron Rods

14-Apr-09 Pipe Street 5 Boron Rods

14-Apr-09 Pipe Street 7 Boron Rods

Table 8. Required Wood Removals 2009 Letter sent July 18, 2009 with completion date of Sept 15, 2009 Re-Inspected on Sept 16-21, 2009 # Address No Progress Partially Complete Completed 1 54 Cardigan St. X 2 1 Clarke St. W. X 3 26 Clarke St. E. X 4 15 Clarence St. X 5 23 Clarence St. X 6 2 Dufferin St. X 7 10 Dufferin St. X 8 12 Dufferin St. X 9 116 Dufferin St. X 10 150 Dufferin St. X 11 158 Dufferin St. X 12 166 Dufferin St. X 13 17 Kerr St. X 14 19 London Rd. E. X 15 25 London Rd. E. X 16 251 Metcalfe X 17 253 Metcalfe X 18 257 Metcalfe X 19 259 Metcalfe X 20 263 Metcalfe X 21 20 Norwich St. E. X 22 24 Norwich St. E X 23 40 Norwich St. E X 24 11 Windermere Ct. X 25 265 Woolwich X 26 271 Woolwich X 27 275 Woolwich X 28 279 Woolwich X 29 285 Woolwich X 30 340 Woolwich X 31 341 Woolwich X 32 344 Woolwich X 33 345 Woolwich X 34 348 Woolwich X 35 349 Woolwich X 36 363 Woolwich X 37 365 Woolwich X 38 469 Woolwich X 39 476 Woolwich X 40 489 Woolwich X 41 490 Woolwich X 42 494 Woolwich X 43 497 Woolwich X

COUNTRY CLUB GOLFVIEW R D . GLEN BR OOK D R . ISLIN GTON AVE. FER N D ALE

WOOLWICH ST. D ALEBR OOK PL.

SPEED RIVER W OOD LAW N R D . W GOLFVIEWRD. W OOD LAW N R D . E 43

W OOD LAW N D EVONSHIRE W IN D SOR ST. CT. C EMETER Y GUELPH JUNCTION RAILWAY

WINDERMERE WINDERMERE 46 FAIRWAY LANE 44

42 INVERNESS DR. INVERNESS

KINGS ETON PL.ETON BALMOR AL D R .

LEY ST. 45 COUNTRY CLUB GOLF C OU R SE CT. BERKLEY PL. WINDSOR Windermere Area

SPEED RIVER RIVERVIEW PLACE Sectors 42-BALMORAL46DR. WOOLWICH ST. WOOLWICH W AVER LEY D R . MARILYN DR. (discovered 2000)

KEN SIN GTON ST. D ELTA ST.

R IVER SID E PAR K WOLSELEY RD. LAN GSIDST. E VERMONT ST.

BAILEY AVE.

DAKOTA DR. DAKOTA KENSINGTON ST. KENSINGTON

RD N ST. STEVENSON

COLLINGWOODST.

KENSINGTON ST. KENSINGTON

METCALFEST. CLIVE AVE. CLIVE DELHI ST. C ATH C AR T ST. SEN IOR BEATTIE ST BEATTIE LILAC PL.

C EN TR E SPEED RIVER SH AFTESBU R Y AVE.

KATHLEEN ST. KATHLEEN

BAILEY AVE. BAILEY

FREEMAN AVE. FREEMAN

WAVERLY DR. WAVERLY

DUMBARTON ST. DUMBARTON

VICTORIA RD. N RD. VICTORIA

KNIGHTSWOOD BLVD. KNIGHTSWOOD SHERIDAN ST. SHERIDAN FR EEMAN AVE. RIVERVIEW DR. ST. DUMBARTON

RIVERSIDE PARK SU MAC PL. BRIGHTON ST. BRIGHTON

KITCHENER AVE. ST. RENFIELD GEMMEL NELSON RD. LN. AVE. GLAD STON E AVE.

MARLBOROUGH GLADSTONE SPEEDVALE AVE. E

ACORN PL.

CHESTER ST. CT SHERWOOD DR. ALEXANDRA KNIGHTSWOOD MANHATTAN BLVD. OAKWOOD STANLEY ST. C H ESTN U T PL. PHILIP AVE. ANN ST. METCALFEST. BALSAM D R . SPRUCE PL. SHERWOOD DR. BARTON ST. ST. EXHIBITION

PAUL AVE. WALNUT DR. MAC AVE. TAMARACK

HIGHVIEW PL. 48

GLENWOOD GLENWOOD AVE. KATHLEEN ST. KATHLEEN 000 PL. PETER AVE. 50 STULL AVE. 51 DR. PINE 0 47 VERNEY ST. 00 RD. MARLBOROUGH EARL ST. EMMA ST. EMMA ST. N ST. STEVENSON H AW TH OR N E

ROBERTSON DR. ROBERTSON 49 PL. ST. ANDREW ST. 1 2 3 SUNNYLEA CRES. LIN D EN PL.

Emma-Pine Area DR. CALLANDER ORCHARD CRES. CLARENCE ST. AVE. DIVISION ST. Sectors 47-51 PL. TERRY BLVD. TERRY DRUMMOND 4 8 PL. (discoveredTER R Y BLVD 2007) . CALLANDER DR.

AVONDALE 7 DANWOOD SUMMIT 6 ST. RENFIELD

GEORGE ST. WOOLWICH ST. WOOLWICH 5 ST. DUFFERIN W ALN U TDR.

PRINCESS ST. PRINCESS ERAMOSA RD. SUNNYLEA CRES. SUNNYLEA CLARKE ST. W CLARKE ST. E 11

KIRSTEN DR. KIRSTEN GREENVIEW ST. PLEASAN T R D . 9 JOHN ST. HOMEWOOD 10 HEALTH CENTRE RD. LINCOLN CRES. PLEASANT WESTMINSTER 12 TOBEY AVE. POWELL ST. W POWELL ST. E ST. DELHI LINCOLN CRES. PIPE ST. METCALFEST. TORRANCE CRES. EXHIBITION 13 14 15 16 SPEED RIVER LAVERNE AVE. PARK C R ES. MEYER TIFFANY ST. W TIFFANY ST. E 21

17 18 DR. VICTORIA RD. VICTORIA 19 20 SKOV DR. MARCON ST. CAVELL AVE. GUELPH COTE CENTRAL ST. CENTRAL STEVENSON ST. N GENERAL EXTRA ST. KERR ST.

HOSPITAL EXHIBITION ST. EXHIBITION

KATHLEEN ST. ST. KATHLEEN 22

PL. CALLANDER DR. CALLANDER TIPPERARY PL. MONT ST. C R ES. 24 CARDIGAN ST. 23 SPRING ST. 25 GLENHILL

MEYER D R . WOOLWICH ST. WOOLWICH McTAGUE ST.

ERIN AVE. DUFFERIN ST. DUFFERIN

26 27 DR. VANCOUVER

OTTAWA CALGARY AVE. CALGARY

DUBLIN ST. DUBLIN WESTOBY

LON D ON R D . W LONDON RD. E ST. HAVELOCK CARDIGAN ST. 36 CADILLAC DR. Woolwich Area ST. LANE DERRY ST. EDWIN ST.30 PL.

SHIRLEY AVE. SHIRLEY BENNETT AVE. BRUNSWICK 31 Sectors 1-41 BENNETT AVE. AVE. CHARLES ST. ARTHUR ST. N 37 PEARL ST. ST. KING EDMONTON DR. GARTH KIRKLAND ST. 28 29 (discovered 1960s) R OSED ALE AVE. WOOLWICH ST.

32 ST. QUEEN NORWICH ST. W ERAMOSA RD. GARTH ST. PARK AVE. GREEN NORWICH ST. E 33 VAN C OU VER D R .

GREEN ST. NORWICH ST. E ST. CATHERINE ST. R YAN AVE. ARDMAY SUFFOLK ST. W LEMON ST. LEMON ST. 34 ST. WILLIAM

KING EDWARD PL 40 WINSTON CRES. WINSTON 38 AVE FRANKLIN CASSINO AVE. C ASSIN O AVE.

AVE. CASSINO AVE. 41 ST. MITCHELL LIVERPOOL ST. SUFFOLK ST. E

35 RIVER SPEED METCALFE ST. METCALFE WINSTON CRES. DEL MAR BLVD.

39 ANTHONY

VICTORIA RD, N N RD, VICTORIA

CT. STUART ST. STUART

KING ST. KING MON TR EAL R D .

OXFORD ST.

DUNKIRK AVE. DUNKIRK LANE ST. LANE

KAR A LEE

LAURINE AVE. LAURINE

QUEEN ST. QUEEN CRESTWOOD PL. CRESTWOOD GLASGOW ST. N PALMER ST. N OR MAN D Y D R . PAISLEY ST. YARMOUTH ST.

DUBLIN ST. N WOOLWICH ST.

ARTHUR ST. N CAMBRIDGE ST. BAKER ST. NORFOLK ST. PAR KH OLM AVE. FRANCHETTO CHAPEL LN. BLVD.

NEW ST. JANE ST JANE COMMERCIAL H EPBU R N AVE.ST JACKSON ST. QUEBEC ST. SPEED RIVER DR. LOUISA CORK ST. W HEFFERNAN ST. DOUGLAS ST. GRANGE ST.

CLARA ST. CLARA Figure 1. 2009 boundaries and sector numbersPROSPECT of Guelph termite management areas. N ST. STEVENSON

350,000

300,000

250,000

200,000

Termites Trapped Termites 150,000

100,000

50,000

0 1999 2 7 8 10 2000 12 15 16 2001 19 20 2007 21 22 23 24 2008 25 27 2009 30 36 37 41 42 47 Sectors

Figure 2. Termites trapped by sector and year (1999-2001 & 2007-2009).

PETER AVE. STULL AVE.

VERNEY ST. RD. MARLBOROUGH

EARL ST. EMMA ST. ROBERTSON ROBERTSON DR. ST. ANDREW ST. 2

CLARENCE ST. DIVISION ST. 8

AVONDALE 7 SUMMIT

GEORGE ST.

WOOLWICH ST.

DUFFERIN ST. DUFFERIN PRINCESS ST. CLARKE ST. W CLARKE ST. E

10 JOHN ST. HOMEWOOD 12 HEALTH CENTRE

POWELL ST. W POWELL ST. E DELHI ST. PIPE ST. TORRANCE CRES. 15 EXHIBITION 16 SPEED RIVER PARK TIFFANY ST. W TIFFANY ST. E 21 19 20

CAVELL AVE. MARCON ST. CENTRAL CENTRAL ST.

EXTRA ST. KERR ST. EXHIBITION ST. KATHLEEN ST. 22

TIPPERARY PL. MONT ST. 24

CARDIGAN ST.

25 23 36 SPRING ST. WOOLWICH ST. McTAGUE ST.

27 ST. DUFFERIN DUBLIN DUBLIN ST.

LONDON RD. W LONDON RD. E HAVELOCK ST. CARDIGAN ST. 30 EDWIN ST. DERRY ST.

37 CHARLES ST. ARTHUR ST. N PEARL ST. KING ST. GARTH KIRKLAND ST. WOOLWICH ST. NORWICH ST. W QUEEN ST. GARTH ST. PARK AVE. GREEN NORWICH ST. E GREEN ST. NORWICH ST. E SUFFOLK ST. W

KING EDWARD PL 41 MITCHELL ST. MITCHELL LIVERPOOL ST. SUFFOLK ST. E

Areas inactiveRIVER SPEED for three years and more than three Areas of identified termite activity in 2009. properties from any known activity for three years.

Newly active block in 2009. KING ST.

# Red Zone sector numbers. Maximum extent of infestation. GLASGOW ST. N

Figure 3. Areas of detected termite activity in the Woolwich management area in 2009.

MANHATTAN

METCALFEST. BALSAM DR. PINE DR. PINE

EMMA ST. N ST.STEVENSON

Figure 4. Area of detected termite activity in the Emma-Pine management area in 2009.

TERRY BLVD. TERRY

WINDSOR ST. CT.

WINDERMERE WINDERMERE INVERNESS DR. INVERNESS

BALMORAL DR.

Figure 5. Area of detected termite activity in the Windermere management area in 2009

BALMORAL DR. BALMORAL

TERRACE PARKER PLACE INVERNESS DR. INVERNESS PONDVIEW CRES.

VISTA

COUNTRY CLUB

GOLFVIEW RD. GLENBROOK DR. ISLINGTON

AVE. FERNDALE

BLVD. BIRCHBANK

DALEBROOK PL. WOOLWICH ST.

SPEED RIVER

WOODLAWN RD. W GOLFVIEW RD. W OOD LAW N R D . E

WINDSOR ST. CT. WOODLAWN DEVON SHIRE CEMETERY MONTANA GUELPH JUNCTION RAILWAY

FAIRWAY LANE WINDERMERE INVERNESS DR. INVERNESS

KINGS ETON PL. ETON

BALMORAL DR. LEY ST.

COUNTRY CLUB GOLF COURSE CT.

BERKLEY PL. WINDSOR

SPEED RIVER RIVERVIEW PLACE BALMORAL DR. BALMORAL

WOOLWICH ST. WOOLWICH WAVERLEY DR. MARILYN DR.

ALGONQUIN KENSINGTON ST. DELTA ST.

RIVERSIDE PARK WOLSELEY RD. LANGSIDE ST. VERMONT ST.

BAILEY AVE.

DAKOTA DR. DAKOTA KENSINGTON ST. KENSINGTON

RD N ST. STEVENSON

COLLINGWOOD ST.

KENSINGTON ST. KENSINGTON METCALFE ST. METCALFE

DELHI ST. AVE. CLIVE CATHCART ST. SENIOR BEATTIE ST BEATTIE CENTRE LILAC PL.

SPEED RIVER SHAFTESBURY AVE.

KATHLEEN ST. KATHLEEN

BAILEY AVE. BAILEY

DUMBARTON ST. DUMBARTON DR. WAVERLY

VICTORIA RD. N RD. VICTORIA

KNIGHTSWOOD BLVD. KNIGHTSWOOD SHERIDAN ST. SHERIDAN FREEMAN AVE. RIVERVIEW DR. ST. DUMBARTON SUMAC PL.

RIVERSIDE PARK BRIGHTON ST. BRIGHTON KITCHENER AVE. ST. RENFIELD GEMMEL NELSON RD. LN. AVE.

GLADSTONE AVE. CARMINE PL. CARMINE

MARLBOROUGH GLADSTONE SPEEDVALE AVE. E

ACORN PL.

CHESTER ST. CT SHERWOOD DR. MONTGOMERY ST. ALEXANDRA KNIGHTSWOOD

MANHATTAN BLVD. OAKWOOD STANLEY ST. CHESTNUT PL. PHILIP AVE.

ANN ST. ST. NEWSTEAD

METCALFE ST. METCALFE BALSAM DR. SPRUCE PL.

SHERWOOD DR. BARTON ST. ST. EXHIBITION

PAUL AVE. WALNUT DR. MAC AVE. TAMARACK HIGHVIEW PL.

GLENWOOD GLENWOOD AVE. ERAMOSA RD. KATHLEEN ST. KATHLEEN PL.

PETER AVE.

STULL AVE. DR. PINE HYLAND RD.

VERNEY ST. RD. MARLBOROUGH EARL ST. EMMA ST. EMMA ST. N ST. STEVENSON HAWTHORNE

ROBERTSON DR. ROBERTSON PL. ST. ANDREW ST.

SUNNYLEA CRES. LINDEN PL. CALLANDER DR. CALLANDER

ORCHARD CRES.

CLARENCE ST. AVE. DIVISION ST. PL.

TERRY BLVD. TERRY DRUMMOND PL. DANWOOD TERRY BLVD. CALLANDER DR.

AVONDALE

RENFIELD ST. RENFIELD ELGINFIELD DR. ELGINFIELD GEORGE ST. DR. GLENBURNIE

WALNUT DR.

WOOLWICH ST. WOOLWICH DUFFERIN ST. DUFFERIN

PRINCESS ST. PRINCESS ERAMOSA RD. CLARKE ST. W CLARKE ST. E CRES. SUNNYLEA

KIRSTEN DR. KIRSTEN GREENVIEW ST. PLEASANT RD. JOHN ST. HOMEWOOD HEALTH CENTRE RD. LINCOLN CRES. PLEASANT WESTMINSTER TOBEY AVE. POWELL ST. W POWELL ST. E ST. DELHI LINCOLN CRES.

PIPE ST. ST. METCALFE

EXHIBITION SPEED RIVER LAVERNE AVE. PARK CRES. MEYER CHELTONWOOD AVE. TIFFANY ST. W TIFFANY ST. E

DR. VICTORIA RD. VICTORIA

SKOV DR. MARCON ST.

CAVELL AVE. GUELPH COTE STEVENSON ST. N CENTRAL ST. CENTRAL EXTRA ST. KERR ST. GENERAL

HOSPITAL

EXHIBITION ST. EXHIBITION

KATHLEEN ST. ST. KATHLEEN

PL. CALLANDER DR. CALLANDER TIPPERARY PL. MONT ST. CRES.

CARDIGAN ST.

SPRING ST. GLENHILL

MEYER DR. WOOLWICH ST. WOOLWICH McTAGUE ST.

ERIN AVE.

DUFFERIN ST. DUFFERIN

VANCOUVER DR. VANCOUVER

OTTAWA

CALGARY AVE. CALGARY DUBLIN ST. DUBLIN

LONDON RD. W LONDON RD. E ST. HAVELOCK CADILLAC DR. WESTOBY

CARDIGAN ST. LANE ST. LANE

EDWIN ST. DERRY ST. BENNETT AVE. PL. SHIRLEY AVE. SHIRLEY BRUNSWICK BENNETT AVE. AVE.

CHARLES ST. ARTHUR ST. N

PEARL ST. ST. KING EDMONTON DR.

KIRKLAND ST. ROSEDALE AVE.

WOOLWICH ST. NORWICH ST. W ST. QUEEN ERAMOSA RD.

GARTH ST. PARK AVE.

GREEN NORWICH ST. E GREEN ST. VANCOUVER DR.

NORWICH ST. E ST. CATHERINE ST. RYAN AVE. ARDMAY

SUFFOLK ST. W LEMON ST. LEMON ST. WILLIAM ST.

KING EDWARD PL

WINSTON CRES. WINSTON CASSINO AVE.

FRANKLIN AVE FRANKLIN CASSINO AVE. CASSINO AVE.

AVE. MITCHELL ST. MITCHELL

LIVERPOOL ST. SUFFOLK ST. E

SPEED RIVER SPEED METCALFE ST. METCALFE DEL MAR BLVD.

WINSTON CRES. PALERMO

ANTHONY

VICTORIA RD, N N RD, VICTORIA CT.

STUART ST. STUART MONTREAL RD. KING ST. KING

OXFORD ST.

DUNKIRK AVE. DUNKIRK LANE ST. LANE

KARA LEE

LAURINE AVE. LAURINE

QUEEN ST. QUEEN CRESTWOOD PL. CRESTWOOD GLASGOW ST. N PALMER ST. NORMANDY DR. PAISLEY ST. YARMOUTH ST. WOOLWICH ST. DUBLIN ST. N BAKER ST.

NORFOLK ST. JACKSON ST JACKSON DR. LOUISA Figure 6. Revised 2010 boundaries and zones of termite management areas.

Observed termite activity in Sector 2 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 7. Trap locations and observed termite activity in sector 2 in 2009.

Figure 8. Trap locations and observed trap activity in sector 7 in 2009.

Figure 9. Trap locations and observed trap activity in sector 8 in 2009.

Figure 10. Trap locations and observed trap activity in sector 10 in 2009.

Observed termite activity in Sector 12 in 2009.

PIPE ST. PIPE

JOHN ST. JOHN

12 9

125 2 123

DUFFERIN ST.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 11. Trap locations and observed trap activity in sector 12 in 2009.

Figure 12. Trap locations and observed trap activity in sector 15 in 2009.

Figure 13. Trap locations and observed trap activity in sector 16 in 2009.

Figure 14. Trap locations and observed trap activity in sector 19 in 2009.

Figure 15. Trap locations and observed trap activity in sector 20 in 2009.

Observed termite activity

in Sector 21 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 16. Trap locations and observed trap activity in sector 21 in 2009

Observed termite activity (none)

in Sector 22 in 2009.

Figure 17. Trap locations and observed trap activity in sector 22 in 2009.

Observed termite activity in Sector 23 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 18. Trap locations and observed trap activity in sector 23 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 19. Trap locations and observed trap activity in sector 24 in 2009

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 20. Trap locations and observed trap activity in sector 25 in 2009.

Observed termite activity (none) in Sector 27 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 21. Trap locations and observed trap activity in sector 27 in 2009.

Observed termite activity

in Sector 30 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 22. Trap locations and observed trap activity in sector 30 in 2009.

Observed termite activity in Sector 36 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 23. Trap locations and observed trap activity in sector 36 in 2009.

Observed termite activity in Sector 37 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 24. Trap locations and observed trap activity in sector 37 in 2009

Observed termite activity in Sector 41 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 25. Trap locations and observed trap activity in sector 41 in 2009.

Observed termite activity & nematode

treatments in Sector 42 in 2009.

termite trap active trap termite activity observed in yard survey estimated termite foraging territory

Figure 26. Trap locations and observed trap activity in sector 42 in 2009.

Emma-Pine Termite Management Area

BALSAM DR. 23 275 5 7 9 11

13 15 22

271

267 16 48 15

57 59 DR. PINE

265

13 METCALFE ST.

262 263 50

12 11

260

259

10 9

258

51 257 7

126 8

256 47

255 6

254 5 253

130 136 142

251 128 132 118 118 116 134

EMMA ST.

245 127 129 133 125 135 137 141

143 145 243 termite trap 49 active trap termite activity observed in yard survey estimated termite foraging territory

Figure 27. Trap locations and observed trap activity in sector 47 in 2009.

Figure 28. Inner blue zone boundaries in 2009.

Sector 1 VERNEY ST.

26 24 20 18 14

546

524 528 WOOLWICHST.

512

504 19

500

494 33 29 25 21 17 15

DIVISION ST.

Figure 29. Inner blue zone properties and trap locations in Sector 1 in 2009.

Sector 6

DIVISION ST.

490

34 32 12

30 14

486

480

476

WOOLWICH ST.

PRINCESS ST.

472

468

464

458

25 23

450

11 9

CLARKE ST. W.

Figure 30. Inner blue zone properties and trap locations in Sector 6 in 2009.

POWELLST. W.

CLARKEST. W.

Sector 9 Sector

440 448

424

426

432

436 446

WOOLWICH ST.

Figure 31. Inner blue zone properties and trap locations in Sector 9 in 2009.

DUFFERIN ST.

131 141 143

147

137

GEORGEST.

JOHN JOHN ST.

Sector 11 Sector

30 35

Figure 32. Inner blue zone properties and trap locations in Sector 11 in 2009.

CENTRAL ST

POWELLST. W.

TIFFANYST. W.

SECTOR SECTOR

400

406

410 414

416 2

WOOLWICH ST.

Figure 33. Inner blue zone properties and trap locations in Sector 14 in 2009

EDWINST. CHARLESST.

SECTOR 31 SECTOR

264

258

268 270

WOOLWICH ST.

Figure 34. Inner blue zone properties and trap locations in Sector 31 in 2009.

NORWICHST. W. CHARLESST.

11 12

SECTOR SECTOR

245

250 252

3 1

WOOLWICH ST.

Figure 35. Inner blue zone properties and trap locations in Sector 32 in 2009

40 SECTOR SECTOR

Figure 36. Inner blue zone properties and trap locations in Sector 40 in 2009.

Figure 37. Nematode treatment areas in 2008 & 2009.

Appendix 1.

Appendix 2

Appendix 3

Appendix 4

Appendix 5

Appendix 6

Appendix 7

May 8, 2009

Dear Termite Management Area Resident,

I am writing to provide a progress report on the termite management program and to announce the upcoming annual public meeting. Please find enclosed the Executive Summary of the 2008 Termite Report including a map and graph of termite activity in your area.

There will be two public meetings to review the termite management program, progress to date, and to outline the management plan for the upcoming season. These meetings will be held in the Council Chamber of the new City Hall starting at 7:00 pm, as follows:

Residents south of Tiffany St. and the Windermere Area: Wednesday, May 20, 2009

Residents north of Tiffany St. and the Emma-Pine Area: Thursday, May 21, 2009

I hope to see you at the meeting.

Sincerely,

Tim Myles, Ph.D. Termite Control Officer

Building Services Community Design and Development Services

T 519-837-5615 ext. 2840 City Hall C 519-827-4383 1 Carden St F 519-822-4632 Guelph, ON E [email protected] Canada N1H 3A1

T 519-822-1260 Encl. TTY 519-826-9771

55 guelph.ca

Appendix 8

June 8, 2009

Re: Spring Nematode Treatment

Dear Termite Management Area Resident,

As mentioned in my last mailing, the spring nematode treatment for termite control will be conducted over the next several weeks. A different nematode product, CapsanemTM (Steinernema carpocapsae), will be used for this treatment. An information sheet is enclosed. This nematode species can be used against various soil insect pests including caterpillar larvae, beetle grubs, crickets and termites. This nematode species is safe for humans, pets and plants and can be applied in yards without a permit or need for signage.

Also enclosed is an authorization form. Please sign the upper portion to indicate that you want the nematode treatment or the lower portion to indicate that you do not want the nematode treatment. Please return the signed authorization form by mail, or drop it off at the City Hall drop box (just inside main entrance, or right side of main entrance after hours) as soon as possible.

The nematodes will be applied by watering can using your outdoor faucet or hose. The treatment will be done in the morning from 7:00-11:00 AM or in the evening from 5:00- 9:00 PM. We will leave a sketch showing the treated areas in your mail box. Please water in these areas daily for at least 3 days after the treatment if the weather remains dry, as nematodes require soil moisture.

Many thanks for your assistance and participation.

Sincerely,

Tim Myles, Ph.D. Termite Control Officer

Building Services Community Design and Development Services 1 Carden St.., 3rd Floor, Guelph, ON N1H 3A1 City Hall T 519-837-5615 ext. 2840 1 Carden St Guelph, ON F 519-822-4632 56 Canada E [email protected] N1H 3A1

T 519-822-1260 TTY 519-826-9771

guelph.ca

Appendix 9

June 8, 2009

Dear Resident,

Re: Expansion of the Emma-Pine Termite Management Area

On June 2 termites were discovered in a backyard trap at 255 Metcalfe St. Thus, the termite population has evidently crossed the small lane separating the Metcalfe St. properties from 126 Emma St. Accordingly, we must now reclassify the following five properties: 259, 257, 255, 253, and 251 as Red Zone properties, and the properties opposite on the west side of Metcalfe: 118 Emma, and 254, 256, 258, and 260 Metcalfe as Blue Zone properties. Please see the enclosed revised map of the Emma-Pine Termite Management Area.

Additional traps (10 per property) will be installed in the new Red Zone Properties, and three traps will be installed in each of the new Blue Zone properties. These installations are planned for this coming Thursday and Friday, June 11 and 12. The traps will be checked every two or three weeks through the end of October and will remain in place until the termite population is brought under control.

The Red Zone properties will be included in termite population suppressive treatments, including the upcoming spring nematode treatment, for which information and an authorization form are attached. Please sign and return the authorization form.

A yard wood clean up weekend will be held on July 17-20 and each of you will receive a letter prior to that date indicating yard wood that must be removed and discarded to disposal bins on that weekend.

You will be contacted later in the season regarding further treatments and to arrange indoor inspections. You may find further information about the termite control program at www.guelph.ca > quick links > termites. Please feel free to contact me at any time, if you have any questions about the program.

Sincerely,

Tim Myles, Ph.D. Termite Control Officer City Hall 1 Carden St Guelph, ON Building Services Canada Community Design and Development Services N1H 3A1 rd 57 1 Carden St.., 3 Floor, Guelph, ON N1H 3A1 T 519-822-1260 T 519-837-5615 ext. 2840 TTY 519-826-9771

guelph.ca

Appendix 10

June 23, 2009

Re: Installation of Termite Monitoring Traps in the Inner Blue Zone

I am writing to inform residents of the Inner Blue Zone (see attached map) that we will be installing termite monitoring traps in the yards of all properties over the next few weeks. This is necessary in order to provide better monitoring of areas adjacent to Red Zone Blocks.

Installation of traps will start on June 25. Termite program staff will first knock on your door to identify themselves. If no one is home, we will proceed with trap installation. Traps consist of a roll of corrugated cardboard inside a 4 X 6 inch PVC pipe covered by a plywood lid which is buried about one inch below ground level. Traps will generally be placed in flower beds and yard margins.

Three to six traps will be installed per Inner Blue Zone property. Traps will be marked with a green bamboo stick. A sketch will be left in your mailbox showing the locations of the traps. The traps will be checked by program staff on about a three-week cycle through the active season from May through mid November. Traps may remain in place for several years depending on the progress of the program.

For more information about Guelph’s termite control program please check the City’s web site at www.guelph.ca > quick links > termites. If you have any questions or comments about the termite control program please feel free to contact me at any time (519-827-4383 cell). Thank you for your cooperation and participation.

Sincerely,

Tim Myles, Ph.D. Termite Control Officer

Building Services Community Design and Development Services City Hall 1 Carden St.., 3rd Floor, Guelph, ON N1H 3A1 1 Carden St Guelph, ON Canada T 519-837-5615 ext. 2840 N1H 3A1 F 519-822-4632 E [email protected] T 519-822-1260 58 TTY 519-826-9771

guelph.ca

Appendix 11

July 6, 2009

Termite Management Area Residents

Yard Wood Clean Up Weekend - Friday July 17th to Monday July 20th

As announced at the community meetings, this year’s yard wood clean up weekend is scheduled for the weekend of July 17-20. Disposal bins will be located throughout the Red Zone for residents to use. Adjacent Blue Zone residents are also encouraged to participate. Please see the reverse side of this letter for a map of the planned locations of the bins (actual locations may differ slightly).

The bins will be delivered on the morning of Friday, July 17th and picked up on the morning of Monday, July 20th. The bins will have parking permits, signage, and barriers. If the bin nearest you is full, use another. If all the bins fill, leave excess material on your property or drive, well back from the curb, and we will assist you in disposing of it later.

Yard Wood Management is an important aspect of the City’s termite control program. Dead wood in soil contact provides termites with feeding, breeding, and overwintering habitat. Our primary focus is on removable items such as: wood chip mulch, landscaping ties, edging boards, retaining walls, fire wood, sheds, scrap lumber, old fences, planting barrels, dead trees, and stumps. Wood chip mulch should be raked up and bagged in clear plastic bags in preparation for disposal.

Some dead wood, such as boards or firewood, can safely be retained if stacked properly off the ground on asphalt, patio stones or concrete blocks. Fence posts and utility poles are not the focus at this time. Live trees and branch trimmings are not generally attacked by termites and should not be disposed. Also, please do not dispose any soil, concrete, furniture, building rubble, or other inappropriate material in the bins.

Some Red Zone residents will be provided with a yard map and a specific list of yard wood items which should be removed from your property. This will be left in your mail box by termite program staff prior to July 10. Termite program staff will be on hand on Friday July 17 to assist with removal. Over the weekend, it is hoped that neighbours will assist each other in the disposal task. This is an important opportunity to collectively reduce termite habitat, so please try to remove as much yard wood as possible.

After you have raked up and removed your old wood chip mulch you may want to apply new mulch or ground cover. Approved organic mulches include 100% bark mulches, pine bark nuggets, cocoa beans shells, coco nut husk. Do not use any form of wood chip or straw mulch. Approved non-organic mulches include any kind of gravel, decorative stone, and Rubberific mulch. Approved soil amendments include peat moss, and coconut husk fibre, or any well composted material such as black earth.

If you have excess material to dispose, or can not participate on this weekend, please contact me to make alternative arrangements for disposal. Thank you for your participation in this community project!

City Hall 1 Carden St Guelph, ON N1H 3A1 59 T 519-837-5615, Ext. 2840 Tim Myles, Ph.D. 519-827-4383 (cell)

Termite Control Officer E [email protected]

guelph.ca > quick links > termites

Appendix 12 July 8, 2009

«Owner»

Re: Notice of Required Wood Removal at «Address_for_Wood_Removal»

Under the City’s Termite Control By-law, the Termite Control Officer is empowered to require the removal of yard wood that could promote termite activity. Accordingly you are hereby notified of wood on your property that requires removal.

Items to be Removed (see attached map).

«Items_to_be_removed»

Compliance Date: September 15, 2009

A Yard Wood Clean up Weekend has been scheduled for July 17-20, 2009 as announced in the mailing of July 6. During this weekend bins will be available at various locations for residents to use for disposal of yard wood. Please take advantage of this opportunity to dispose of the above items.

If you are unable to dispose of your yard wood at this time, please haul the material to the Waste Resource Innovation Centre at 80 Dunlop and give the enclosed disposal permit to the weigh scales attendant to waive tipping fees on the first 250 kg. You will be charged at the normal tipping fee rate of $70 per tonne for material in excess of 250 kg.

If you require assistance, additional time, or would like to discuss other possible options, please feel free to contact me.

Thank you for your cooperation.

Tim Myles, Ph.D. Termite Control Officer

Building Services Community Design and Development Services 1 Carden St., 3rd Floor, Guelph, ON N1H 3A1 City Hall T 519-837-5615 ext. 2840 1 Carden St Guelph, ON F 519-822-4632 Canada E [email protected] N1H 3A1

T 519-822-1260 TTY 519-826-9771

60 guelph.ca

Appendix 13 August 10, 2009

Dear Resident:

Re: Discovery of a Termite Infestation on Your Block

This letter is to inform you that a termite infestation was recently discovered on a property on the south side of Clarke St. Accordingly, we must now reclassify the block from a Blue Zone to a Red Zone within the Termite Management Area (see attached map).

As you may know, the City has been monitoring and managing termites in the area for many years. A detailed history of the termite infestation and various management efforts over the years is available on the City’s web site at http://www.guelph.ca > quick links > termites.

Under the City’s Termite Control Bylaw, the Termite Control Officer is empowered to enter and inspect properties to investigate for termites and to prescribe measures aimed at controlling and limiting the spread of termites.

The following management steps will be taken:  Yard inspections  Installation of additional yard traps  Yard wood clean up weekend (September 25-28)  Ongoing monitoring on approximately a 2-3 week cycle to the end of November  Trapped termites will be removed and/or used for colony level treatments  Indoor inspections will be scheduled later in the year

Yard inspections and trap installation: Termite traps will be installed on all properties starting at 8:00 AM this coming Wednesday, August 12 and, if necessary, continuing on August 13 and 14. Termite program staff will first knock on your door, and then proceed to conduct a yard inspection and to install traps. Please leave gates unlocked. Up to ten traps will be installed per property. Traps consist of a 4 X 6‖ plastic pipe with a roll of cardboard and a plywood lid. Traps will be buried about an inch below ground level and marked with a green bamboo stick. A sketch of the locations of the traps will be left in your mailbox along with notification of any discovered termite activity in the yard. Staff will not enter or inspect houses at this time.

Yard wood clean up weekend: A yard wood clean up weekend will be held on September 25-28 and residents will receive a letter prior to that date indicating yard wood that should be prepared for removal to disposal bins on that weekend. If you are unable to prepare the identified wood items for removal by that date, please use the attached disposal permit and take the wood to the Waste Resource Innovation Centre before October 15.

Trees: Live trees are not attacked or killed by termites. Termites may however build City Hall shelter tubes on the bark to reach pruning scars or dead branches. Only trees that have 1 Carden St numerous dead branches or severe heart rot need to be removed. Trees that are taken Guelph, ON Canada down must also have the stump removed. Healthy trees with shelter tubes on the N1H 3A1 exterior bark can be treated chemically. T 519-822-1260 TTY 519-826-9771 61 guelph.ca

Colony level treatments: As part of the program, colony-level treatments are being applied on termite infested properties. Last fall and again this spring, entomopathogenic nematodes were used to treat termite infested properties. In addition, we are expecting soon the approval of experimental permits from the Pest Management Regulatory Agency that will allow us to use two additional experimental control agents: the entomopathogenic fungus, Metarhizium anisopliae, and zinc borate as a topical treatment for Trap-Treat-Release. In any case, you will be notified prior to any of these treatments, provided with further information, and requested to sign an authorization form, before such treatments will be performed on your property.

Short term and long term goals: The goal of the Termite Management Program for this year is to identify the extent of the infestation and to eliminate as much yard wood as possible. We also plan to identify any existing structural infestations and recommend appropriate treatments where necessary to safeguard structures. Our long term goal is to suppress the population by colony level suppressive treatments with baits, transmissible chemicals, or termite-specific biological agents (such as entomopathogenic nematodes and fungi). Using an integrated approach, involving habitat reduction, population treatments and limited use of chemical treatments, the City plans to achieve area-wide control.

Cost sharing: The City of Guelph has taken a proactive approach to termite management with the ultimate goal of eradicating termites from the City. This however, does not mean that the City assumes any responsibility for termite infestation or that it intends to cover all costs associated with termite control. Through the Termite Control Officer the City will monitor termite populations, coordinate control programs, and apply colony level suppression treatments. The City offers a tipping fee waiver for waste disposal up to 500 pounds per trip, and may also from time to time arrange for yard waste disposal bins. The city will also remove any dead city trees or chemically treat any healthy city trees with evidence of shelter tubing. However, costs associated with treatment or removal of dead trees, stumps, poles, sheds, fences, decks, retaining walls, or chemical treatment of structures, or structural renovations on private property must be covered by the property owners.

If you have any questions please feel free to contact either myself or the Chief Building Official, Bruce Poole. Thank you for your cooperation in managing termites in your area and preventing their spread.

Tim Myles, Ph.D. Termite Control Officer Attachment

Building Services Community Design and Development Services City Hall, 1 Carden St., 3rd Floor, Guelph, ON N1H 3A1

T 519-837-5615 ext. 2840 F 519-822-4632 E [email protected]

Appendix 14 September 14, 2009

Re: Update on Termite Management Plan on Your Block (Sector 10)

I am writing to provide an update of the termite situation based on the yard inspections and to inform residents of next steps in the termite management plan for your block.

Update of known activity from yard inspections. Please see the attached map which shows where termite activity was found in the yards. So far, known activity is confined to the east side of the alleyway.

Yard Wood Cleanup Weekend. I want to remind residents of the Yard Wood Clean up Weekend which is scheduled for September 25-28 in your block. A bin will be placed centrally along the alley way as shown in the attached map. The bin will be delivered on Friday morning (Sept. 25) and picked up the following Monday morning (Sept. 28). With this letter, you may also find a sketch of your property showing specific yard wood items that are recommended for disposal in the bin. Wood fence postes and pressure treated wood retaining walls need to be removed, but may require treatment in the future if they become infested.

Nematode Treatment. As mentioned in my last mailing, a nematode treatment for termite control is planned. This will be limited only to those properties on the east side of the alleyway. This will be conducted over the next month. The nematode product, Capsanem TM (Steinernema carpocapsae), will be used for this treatment. An information sheet is enclosed. This nematode species can be used against various soil insect pests including caterpillar larvae, beetle grubs, crickets and termites. This nematode species is safe for humans, pets and plants and can be applied in yards without a permit or need for signage.

If you reside on the east side of the block, you will find enclosed an authorization form for nematode treatment. Please sign the upper portion to indicate that you want the nematode treatment or the lower portion to indicate that you do not want the nematode treatment. Please return the signed authorization form by mail, or drop it off at the City Hall drop box (just inside the main entrance, or on the right side of the main entrance after hours) as soon as possible. The treatment will not be performed on your property unless you sign and return the authorization form.

The nematodes will be applied by watering can using your outdoor faucet or hose. The treatment will be done in the morning from 7:00-11:00 AM. We will leave a sketch showing the treated areas in your mail box. Please water in these areas daily for at least 3 days after the treatment if the weather remains dry, as nematodes require soil moisture.

Indoor Inspections. In addition to the yard inspections and ongoing monitoring of yard traps, I will also be doing indoor inspections of all properties starting October 13. Residents will be called in advance to arrange an inspection time between 7:00 AM and 3:00 PM. These inspections take approximately 15 minutes. In the event that termite activity is discovered indoors, treatment or renovation options will be recommended. If your property has already been inspected within the last 6 months, it will not be re-inspected at this time.

Many thanks for your assistance and participation.

Tim Myles, Ph.D. Termite Control Officer

Building Services City Hall 1 Carden St Community Design and Development Services Guelph, ON 1 Carden St.., 3rd Floor, Guelph, ON N1H 3A1 Canada 63 N1H 3A1 T 519-837-5615 ext. 2840 F 519-822-4632 T 519-822-1260 E [email protected] TTY 519-826-9771

guelph.ca

Appendix 15

October 1, 2009

Re: Re-Inspection for Required Wood Removal

A notice of required wood removal for termite control was sent to you on July 18, 2009 with a designated compliance date of September 15, 2009.

Your property was re-inspected on September 16, 2009. The re-inspection revealed:

 A. Required wood removal has been completed.

 B. Partial required wood removal has been completed.

 C. No progress on required wood removal.

If Box B. or C. above is checked, please contact me at your earliest convenience to let me know your plans for dealing with required wood removal and your estimated date of completion. Let me know if I can assist by providing an additional disposal permit or other assistance. Non-compliance can lead to infractions, so I urge you to communicate with me on this matter by no later than October 30, 2009.

Thank you for your cooperation.

Sincerely,

Tim Myles, Ph.D. Termite Control Officer

Building Services Community Design and Development Services 1 Carden St.., 3rd Floor, Guelph, ON N1H 3A1

T 519-837-5615 ext. 2840 F 519-822-4632 E [email protected]

City Hall 1 Carden St Guelph, ON Canada 64 N1H 3A1

T 519-822-1260 TTY 519-826-9771

guelph.ca

Appendix 16

Observations of Native Nematodes Associated with the Eastern Subterranean Termite, Reticulitermes flavipes, in Ontario.

Timothy G. Myles, Ph.D. Termite Control Officer

City of Guelph, 1 Carden St., Guelph, Ontario, Canada N1H 3A1 [email protected] 519-837-5615 ext. 2840; 519-827-4383 (cell)

Abstract: Four nematode species were observed in cultures that originated from Reticulitermes flavipes collected in Guelph, Ontario. The most rapidly developing species appears to be Rhabditis sp. (possibly R. dolichura). The dauers of this species migrate from water cultures to high points where they nictate in clusters. A more slowly developing species, but one which ultimately is more predominant in cultures is Chroniodiplogaster aerivora. Its daures tend to remain in water cultures rather than migrating out of water cultures. Stomachorhaditis fastidiosus, a very thin and rapid species, is less common, but occasionally seen in some cultures. It may be a predator of the other nematode species. Lastly, many older cultures usually had a very small and slow-moving species living on the surface film of water cultures (Aspidonema sp.). This species is possibly a non-termite specific associate or contaminant.

Key Words: Isoptera, Rhinotermitidae, Nematoda, Bunonematidae, Rhabditidae, Stomachorhabditidae, Diplogasteridae, termite-nematode associations.

In the course maintaining laboratory cultures of the eastern subterranean termite, Reticulitermes flavipes, in Guelph, Ontario, it was observed that sometimes large numbers of termites died shortly after setting up culture boxes. Although some initial mortality is to be expected due to handling injuries, in other cases it seemed that the mortality was excessive. The high mortality was initially attributed to the boxes being either over crowded or over watered. However, when the dead termites in such cultures were examined it was discovered that nematodes were present in the decomposing termites, suggesting that the higher than normal culture box mortality may have resulted from nematode infection of some termite populations with native, termite-associated nematodes. As the nematode cultures continued to be observed, we eventually observed four different nematode species (Figures 1-6).

A literature review revealed that several species of nematodes have previously been reported from North American subterranean termite species. Merrill and Ford (1916) reported Diplogaster aerivora from Reticulitermes sp. (as Leucotermes lucifugus) from Kansas. Banks and Snyder (1920) confirmed this species and another, Rhabditis dolichura Schneider from Reticulitermes flavipes collected in Virginia. Massey (1971) described two new parasitic nematode species from Reticulitermes flavipes collected in Mississippi: Stomachorhabditis fastidiosus (as Termirhabditis fastidiosus) from the termite’s gut, and Rhabpanus ossiculum from the termite’s head. Nguyen and Smart (1994) reported Neosteinernema longicurvicauda as a parasite of Reticulitermes flavipes in Florida. Carta and Osbrink (2005) reported Rhabditis rainai from the Formosan subterranean termite, Coptotermes formosanus, in 65

New Orleans, Louisiana. Kanzaki et al (2008) reported Rhabditis rainai and an unidentified rhabditid and diplogastrid also from Reticulitermes flavipes in Florida.

In addition to these reports of natural associations, some recent studies have also been made on the parasitism and infectivity of North American termites by various nematode species. Yu et al (2006) performed lab bioassays of three species of Steinernema and Heterorhabditis bacteriophora on four species of termites, R. flavipes, R. virginicus, Heterotermes aureus and Gnathamitermes perplexus. Poinar redescribed Diplogaster aerivora and reassigned it to a new genus as Chroniodiplogaster aerivora and studied its parasitism on Reticulitermes tibialis.

Materials and Methods: Attempts were made to isolate, culture, prepare slide mounts and identify the native nematode in our mixed cultures. It was discovered that nematode cultures could be easily maintained by periodically (every week or two) adding more termites to cultures on either wet KimWipes in Petri dishes, or to water cultures in screw top Zip-loc food containers (20 ounce capacity). A standard method of in vivo culture of nematodes utilizes wax moth larvae as hosts. It was confirmed that the nematodes killed, developed and reproduced well on wax moth larvae, as well as butterworm, and meal worm larvae. The nematodes also thrived on in vitro nematode culture media. Nematodes were micro-pipetted from water cultures and mounted in PVA mounting media, and micro-photographed at 200-400X (Figures 1-6).

Results: Table 1. Classification of nematodes naturally associated with or used for subterranean termite control.

Phylum Nematoda Class Chromadorea Subclass Chromadoria Order Rhabditida Suborder Tylenchina Superfamily Strongyloidoidea Family Steinernematidae (Neosteinernema longicurvicauda, Steinernema spp.) Suborder Rhaditina Superfamily Bunonematoidea Family Bunonematidae Subfamily Craspedonematinae (Aspidonema sp.) Superfamily Rhabditoidea Family Rhabditidae Subfamily Rhabditinae (Rhabditis dolichura, Rhabditis rainai, Rhabpanus ossiculum, R. sp.) Subfamily Stomachorhabditinae (Stomachorhabditis fastidiosus) Superfamily Strongyloidea Family Heterorhabitidae (Heterorhabditis bacteriophora) Superfamily Diplogasteroidea Family Diplogasteridae (Chroniodiplogaster aerivora)

Figure 1. Rhabditis sp. (R. dolichura?) adult female.

Figure 2. Rhabditis sp. (R. dolichura?) adult male.

Figure 3. Chroniodiplogaster aerivora. Adult female (top); adult male (bottom).

Figure 4. Chroniodiplogaster aerivora, male spicules.

Figure 5. Stomachorhabditis fastidiosus adult male.

Figure 6. Aspidonema sp.

Acknowledgements: I thank Dr. Robin Giblin-Davis, University of Florida, for helpful discussions and guidance on nematode cultivation, taxonomy and identification.

References:

Banks, N. and T. E. Snyder. 1920. A revision of the Nearctic termites. Bull. Smithsonian Institute. 108: 228 pp.

Carta, L. K. and W. Osbrink. 2005. Rhabditis rainai n. sp. (Nematoda: Rhabditida) associated with the Formosan subterranean termite, Coptotermes formosanus (Isoptera: Rhinotermitidae). Nematology 7 (6): 863-879.

Holovachov, O. and A. M. T. Bongers 2001. Description of Aspidonema scheucherae (Sachs, 1949) Andrassy, 1958 from Turkey (Nematoda: Bunonematidae). Annales Zoologici. 51: 395-401.

Kanzaki, N., R. M. Giblin-Davis, R. H. Scheffrahn, and B.J. Center. 2008?. Poikilolaimus floridensis n. sp. (Rhabditida: Rhabditidae) associated with termites (Kalotermitidae). Nematology . (in press)

Merrill, J. H. and A. L. Ford. 1916. Life history and habits of two new nematodes parasitic on insects. J. Agr. Res. 6: 115-127.

Nguyen, K. B. and G. C. Smart, Jr. 1994. Neosteinernema lonicurvicauda n. gen., n. sp. (Rhabditidae: Steinernematidae), a parasite of the termite Reticulitermes flavipes (Kollar). J. Nematol. 26: 162-174.

Poinar, G. O. Jr. 1972. Nematodes as facultative parasites of insects. Annual Rev. Entomol. 17: 103-122.

Poinar, G. O. Jr. 1990. Redescription of Chroniodiplogaster aerivora (Cobb) gen. n., comb. N. (Rhabditida: Diplogasteridae) from termites. J. Helminthol. Soc. Wash. 57 (1): 26-30.

Simard, L., G. Belair, S. P. Stock, H. Mauleon, and J. Dionne. 2007. Natural occurrence of entomopathogenic nematodes (Rhabditidae: Steinernematidiae) on golf courses in eastern Canada. Nematology 9 (3): 325- 332.

Wang, C. J., E. Powell, and B. M. O’Connor. 2002. Mites and nematodes associated with three subterranean termites species. (Isoptera: Rhinotermitidae). Florida Entomologist. 85 (3): 499-506.

Yu, H., D. H. Gouge, and P. Baker. 2006. Parasitism of subterranean termites (Isoptera: Rhinotermitidae; Termitidae) by entomopathogenic nematodes (Rhabditida: Steinernematidae; Heterorhabditidae). J. Econ. Entomol. 99 (4): 1112-1119.

Appendix 17

Springtails (Collembola: Entomobryidae) Associated with Termites (Isoptera: Rhinotermitidae) in Ontario

Timothy G. Myles, Ph.D. Termite Control Officer

City of Guelph, 59 Carden St., Guelph, Ontario, Canada N1H 3A1 [email protected] 519-837-5615 ext 2840, 519-827-4383 (cell) ______

Abstract: Springtails of the family Entomobryidae have been observed in association with termites in field traps and in laboratory cultures. Three species of entomobryid springtails have been identified in termite cultures from Guelph, Ontario, namely, Sinella (Coecobrya) tenebricosa, Sinella curvisetae, and Isotoma sp. Sinella (C.). tenebricosa has been observed to build up large populations in laboratory cultures that have died from nematode infestation and this species has previously been reported as nematophagous. Although springtails had previously been assumed to be incidental soil organisms, it now appears possible that they play an important role in nematophagy, and as such, may serve to protect termites, at least in some degree, from entomopathogenic nematodes (EPNs) such as Heterorhabditis and Steinernema. Such impacts, however, may be limited because termites and nematodes are likely to occur at deeper soil strata, than springtails. To better understand the potential of EPNs as biocontrol agents of termites, further study of these relationships appears to be warranted.

Introduction: Springtails (Hexapoda: Collembola) are small wingless soil insects that are often found together with termites. However, because of their very small size, about 1-2 mm, springtails are often overlooked or even mistaken as termite larvae. The general occurrence of springtails with termites trapped in the field in cardboard roll traps has been noted over the years. It has also often been observed that springtail numbers may increase considerably in termite laboratory cultures, if kept adequately moist, and that springtail numbers occasionally become very high in such cultures if there is excess moisture and/or after termites have died from nematode infestations in combination with excessive moisture. In lab colonies that have died out from nematode infections, springtails numbers may become very large, probably 10,000 or more and then suddenly collapse. The occurrence of springtails in termite cultures and observations of outbreaks of springtails in nematode infested termite lab cultures suggest an important role in nematophagy. In view of recent efforts to control termites with entomopathogenic nematodes, these ecological relationships of springtails have become of greater interest.

Results: Recent efforts have been made to identify the collembolan species in Guelph termite cultures. Three slender entomobyromorphic springtail species, two white species, and one pinkish species have been noticed in Guelph termite cultures. A fourth entomobryid species, bluish gray in color, possibly Lepidocyrtis cyaneus (a cosmopolitan species according to Zimmerman, 1948) and covered with appressed scales, was previously observed in termite cultures collected in Toronto. In all cases the springtails observed with termite cultures appear to belong to the family Entomobryidae.

The most common, of the species observed in Guelph is white in color, lacks eyes, lacks scales, has a falcate mucro, and is covered with distinctive clavate plumose macrochaetae. This species keyed to Sinella hofti Schaffer 1896, in Mill’s (1934) classic book. However, in Zimmerman (1948) the species keyed to Sinella caeca (Schott) with synonyms listed as Entomobrya caeca Schott 1896, Sinella hofti Schaeffer, 1897, and Sinella tenebricosa Folsom, 1902. The genus Sinella and subgenus Coecobrya have been revised (Chen and Christiansen, 1997, Chen et al, 2005; Greenslade, 1993). The species S. caeca and S. tenebricosa are now both considered valid species placed in the subgenus (Coecobrya). Chen and Christiansen (1997) state that S. (C.) tenebricosa has thin acuminate tenet hairs and that the mucronal base hair is usually long, nearly reaching tip of the apical tooth of the mucro. These features indicate that the species occurring in Guelph matches S. (C.) tenebricosa Folsom 1902. Another common species, very similar but pinkish, with eyes, and body covered with leaf-like scales, as well as feathered macrochaetae, and a bidentate mucro is tentatively identified as Sinella curvisetae. The least common species, slightly smaller and more slender, with trichobothria and no clavate macrochaetae, keyed to Isotoma sp. (possibly I. notabilis, a species reported from Eastern Canada by Sharma and Kevan, 1963).

Table 1. Classification of springtails observed with termites.

Order: Collembola S.O. Arthropleona Family Entomobryidae Subfam. Isotominae (Isotoma sp.) Subfam. Entomobryinae (Sinella (Coecobrya) tenebricosa, S. curvisetae, Lepidocyrtis cyaneus? )

Figure 1A, 1b. Sinella (Coecobrya) tenebricosa Folsom 1902

Figure 2A & 2B. Isotoma sp.

Discussion: The species most commonly found in association with termites in Guelph, Sinella (Caecobrya) tenebricosa is among species that has been documented as nematophagous (Brown, 1954; Gilmore, 1970). The extent to which this effect may depress the efficacy of entomopathogenic nematodes (EPNs) as control agents against termites deserves further investigation.

References

Brown, W. L., Jr. 1954. Collembola feeding on nematodes. Ecol. 35: 421.

Chen, J.-X. and K. Christiansen. 1997. Subgenus Coecobrya of the genus Sinella (Collembola: Entomobryidae) with special reference to the species of China. Ann. Entomol. Soc. Am. 90(1): 1-19.

Chen, J.-X., Leng, Z., and P. Greenslade. 2005. Australian species of Sinella (Sinella) Brook (Collembola: Entomobyidae). Aust. J. Entomol. 44:15-21.

Christiansen, K. The nearctic members of the genus Entomobrya (Collembola). Bull. Mus. Comp. Zool. 118(7).

Christiansen, K. Bionomics of Collembola. Ann. Rev. Entomol. 9: 147-178.

Gilmore, S. K. 1970 Collembola predation on nematodes. Search Agriculture. Cornell Univ. Ag. Exp. Stn. December, Entomology and Limnology 1:1-12..

Greenslade, P. 1992. The identity of Australian specimens recorded as Lepidosinella armata Handschin 1920 Collembola: Entomobryidae) with a key to Australian Sinella and Coecobrya. J. Aust. Entomol. Soc. 31:327-330.

Mills, H. B. 1934. The Collembola of Iowa. Collegiate press, Ames, Iowa.

Nickle, W. R. and Ayre, G. L. Caenorhabditis dolichura (A. Schneider, 1866) Dougherty (Rhabditidae, Nematoda) in the head glands of the ants Camponotus herculeanus (L.) and Acanthomyops claviger (Roger) in Ontario. Proc. Entomol. Soc. Ont. 96: 96-98.

Rockett, G. L. and J. P. Woodring. 1966. Oribatid mites as predators of soil nematodes. Ann. Entomol. Soc. Am. 59: 669-671.

Sharma, G. D. and Kevan, D. K. McE. 1963. Observations on Isotoma notabilis (Collembola: Isotomidae) in eastern Canada. Pedobiol. 3: 34-47.

Zimmerman, E. C. 1948. Insects of Hawaii. Vol. 2, Univ. of Hawaii Press, Honolulu.

Appendix 18

Parasitic, Phoretic, Predatory, and Incidental Mites Associated with the Eastern Subterranean Termite.

Timothy G. Myles, Ph.D. Termite Control Officer

City of Guelph, 59 Carden St., Guelph, Ontario, Canada N1H 3A1 [email protected] 519-837-5615 ext 2840, 519-827-4383 (cell) ______

Abstract: Eleven species of mites have been observed in laboratory cultures and experimental studies of the eastern subterranean termite, Reticulitermes flavipes, in Guelph, Ontario. Five species occur regularly as phoronts, one slime mite and three Acarus occur as non-feeding hypopal stages and one scutacarid species occurs as an adult female phoront. The Histiostoma slime mite and three acarids appear to be involved, along with nematodes, in the decomposition of dead termite corpses, and together with nematodes form a specific bacterial-nematode-mite slime (BNMS) on dead termite bodies. This form of termite corpse decomposition requires damp to wet conditions. Under drier conditions other decomposers, including mainly imperfect fungi (Deuteromycetes) appear to predominate. The of Scutacarus mite appears to be a new species with a unique relationship to the termite, attaching with a high degree of attachment site specificity: in 92% of cases it attaches to the inside surface of the third segment of the left maxillary palpus. In this position it is able to tickle the left mandible and edge of the termite labrum with its long leg setae and thereby induce the termite to frequently clean its mouthparts, thus providing excess saliva for this kleptoparasite. One species of beetle mite appears to be an incidental habitat associate. Although some beetle mites have been reported to have a role in nematophagy. The other five species appear to be loosely associated as soil surface predatory species which may be preying either on nematodes or on other mite species.

Key Words: Isoptera, Rhinotermitidae, Acari, Acaridae, Histiostomatidae, Scutacaridae, Parasitidae, Ascidae, Machrochelidae, termite ecology, termite pathology, termite-mite associations, termite-mite-nematode associations, acarinaria, coevolution of termites and mites.

The phorontomorphic stage is a deutonymphal hypopus which attach via a caudoventral sucker plate to species-specific areas of the termite body. These attachment sites, or ―acarinaria‖ are hypothesized here for the first time as probably specifically adapted isopteran body areas for mite attachment. In all cases, these areas are cuticular surfaces that are protected, smooth, bare areas, free of setae and somewhat protected areas of appropriate size for attachment of mites with sucker plates. Thus the pattern and occurrence of setae on termites may be seen largely or at least partly in termites of a means by which certain mites are excluded or permitted to attach to the termite body. of setae and inaccessible to self grooming, and also relatively inaccessible to allogrooming, and designated as follows: Leg Sites: LT, the bare areas on the inside surface of the tibiae, LFd: bare areas on the distal inside area of the femora LFp; Head Sites: Hd Hg Hv, Thoracic Sites: TPro TMeso TMeta, TS. Bare

areas lacking setae on the inside surfaces of the femora and tibia of the termite legs, and may also be found in lateral depressions on the meso and metanota as well as the posterior-ventral surface of the termite head. Typically, the most commonly observed mites species, is a slime mite of the genus Histiostoma (Acari, Acariformes, Histiostomatidae (=Anoetidae)). This species does not match any known species of Histiostoma from North America and is probably a new, undescribed species, designated here tentatively as Histiostoma sp. RFG (Reticulitermes flavipes, Guelph). The hypopus of this species is the smallest of the observed hypopi and may occupy any of the aforementioned acarinaria but most often attaches to LT and Hv. These mites feed on and vector bacterial slime that develops under wet conditions on dead termite bodies in experimental groups exposed to either entomopathic fungi such as Metarhizium anisopliae or entomopathogenic nematodes such as Heterorhabditis bacteriophora. The populations of the Histiostoma mite can begin to develop explosively within a few weeks in experimental groups or cultures where there is termite mortality. Often surviving termites in such groups become heavily infected with the hypopi, numbering from dozens to hundreds per termite, within about a month to two months after experimental initiation. All live stages: egg, larva, protonymph, deutonymph, tritonymph and adults of both sexes have been observed. They appear to crawl or swim through bacterial slime or pools that develop on dead termites. The mites have feathery palpi which they continuously wave and stroke through bacteria during feeding. If dead termites are placed on a wet paper Kim Wipe or submerged in water nematode cultures the mites are seen developing in abundance shallowly submerged in the slime, evidently able to respire adequately under submersed aquatic necrotic conditions. Alternatively, they can also develop under conditions which are merely damp enough to grow a bacterial slime. A second wave of mites, larger and more slowly developing, of the family Acaridae, follows the initial wave of Histiostomatidae. Three acarid mites belonging to the genera Acarus, tentatively identified as Acarus nr. farris, and Acarus sp. and Caloglyphus nr. oudemansi (Acari: Acariformes, Acaridae). Different acarid species may develop depending on the moisture conditions. Under very moist or wet conditions it is common to find A. farris and C. oudemansi. The hypopi of Acarus spp. are a little larger than that of Histiostoma and do not fit the narrow LT sites, preferring the LF sites. A larger Acarid seems to attach. Under drier conditions an acarid, Acarus sp.?, with two dark spots of the opisthosoma was observed. The largest Acarus and the Caloglyphus hypopi attach to the Hd, Hv sites. In older water culture of the naturally occurring termite associated nematode, Chroniodiplogater aerivora, one the mites, Caloglyphus often predominates. Viewed with ventral illumination through a dissecting microscope one may observe over a period of a week or so the gradual deliquescent of moistened termite corpses. Over time, a sort of nematode and mite slime (NMS) develops in which there is a huge accumulation of all developmental stages of nematodes and mites, and their cast skins (exuvia), and eggs. The large brown eggs of the mite become a conspicuous component of this slimy matrix. As such cultures decline all of these mites transform into hypopi. Many of these hypopi appear to remain as inactive ―latent hypopi‖ remaining unmolted inside the protonymph cuticle. When these hypopi eventually emerge they attach to the acarinaria of the head Hd, Hg, Hv. The even larger hypopi of Caloglyphus oudemansi attach mainly to the. The number of such larger hypopi attached to termites never reaches the very high levels seen with the Histostoma can virtually encrust the termite, but in rare cases may number up to a few dozen per termite. It seems possible that this mite may be preying upon nematodes or their eggs. It is also capable of being apparently able to live and reproduce entirely submerged in the NMS.

Scutacarid mites were observed to develop in old termite cultures and in experimental groups of termites exposed to various species of entomopathogenic nematodes (EPNs). These mites, which I had never previously observed, developed populations levels which became conspicuous enough to notice, e.g. about a tenth to a half of the termite population infected with at least one mite per termite and in rare instances with as many as six mites per termite.

Table 1. Classification of Mites Associated with Reticulitermes flavipes (based on Krantz, 1978).

Class Arachnida Suborder Acari Order Parasitiformes Suborder Gamasida (=Mesostigmata) Cohort Gamasina Supermily Parasitoidea Family Parasitidae (Parasitus americanus, Parasitus sp.) Superfamily Ascoidea Family Ascidae (Proctolaelaps hypadari = P. pygmaeus) Superfamily Eviphidoidea Family Macrochelidae (Macrocheles muscaedomesticae) Order Acariformes Suborder Acaridida Supercohort Acaridides Superfamily Acaroidea Family Acaridae (Acarus farris, Acarus sp. A, Acarus sp. B, Caloglyphus nr. oudemansi) Superfamily Anoetoidea Family Histiostomatidae (Histiostoma sp. (RFG)) Suborder Actinedid Supercohort Promatides Cohort Eleutherengonina Subcohort Heterostigmae Superfamily Pygmephoroidea Family Scutacridae (Scutacrus sp. (RFG)) Suborder Oribatida = Cryptostigmata (beetle mites)

Table 2. Number of Scutacarus sp. Mites per Termite Worker per Group.

No. Mites Group 1 Group 2 Group 3 Group 4 Group 5 Totals 0 mites 13 23 80 83 67 266 1 mite 22 29 56 6 11 124 2 mites 20 25 25 3 1 74 3 mites 10 10 4 1 0 25 4 mites 4 3 1 0 0 8 5 mites 0 1 1 0 0 2 6 mites 0 0 1 0 0 1 termites with mites 56 68 88 10 12 234 total termites 69 90 168 93 79 500 % termites infected 81.16% 75.56% 52.38% 10.75% 15.19% 46.80% total mites 108 121 133 15 13 395 Ave # mites/ infected ter. 1.93 1.78 1.51 1.50 1.08 1.69

Table 3. Attachment Sites of Scutacarus sp. Mites on Worker Termites

Left maxillary palpus 364 92.15% Right maxillary palpus 26 6.58% Left labial palpus 1 0.25% Right antenna 2 0.51% Left antenna 1 0.25% Right meso tarsus 1 0.25%

Caloglyphus nr. oudemansi female C. nr. oudemansi heteromorphic male

Histiostoma sp. female, Histiostoma sp. male Histiostoma sp. hypopus

Macrocheles muscaedomesticae Scutacarus sp.

References

Baker, E. W. and G. W. Wharton, ~1980?. An Introduction to Acarology. MacMillan Co, NY.

Costa-Leonardo, A. M. and H. X. Soares. 1993. Occurrence of phoretic mites on the subterranean termite Heterotermes tenuis (Isoptera, Rhinotermitidae). Sociobiology 23(1): 63-69.

Delfinado, M. D., E. W. Baker, and M. J. Abbatiello. 1976. Terrestrial mites on New York III. The family Scutacaridae (Acarina). J. New York Entomol. Soc. 84: 106-145.

Delfinado, M. D. and E. W. Baker. 1978. Terrestrial mites of New York—VII. Key to the species of Scutacaridae and descriptions of new species. J. New York Entomol. Soc. 86: 87-101.

Ebermann, E. and M. Hall. 2003. First record of sporothecae within the mite family Scutacaridae (Acari, Tarsonemina). Zoo. Anz. 242: 367-375.

Ebermann, E. and J. C. Moser. 2008. Mites (Acari: Scutacaridae) associated with the red imported frie ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), from Louisiana and Tennessee, USA. Internat. J. Acarol. 34(1): 55-69.

Houck, M. A. and B. M. O’Connor. 1991. Ecological and evolutionary significance of phoresy in the Astigmata. Ann. Rev. Entomol. 36: 611-636.

Hughes, R. D. and C. G. Jackson. 1958. A review of the Anoetidae (Acari). Virginia J. Sci. 9, N.S. (1): 5-198.

Hunter, P. E. and C. A. Hunter. 1964. A new Anoetus mite from pitcher plants. Proc. Entomol. Soc. Wash. 66(1): 39-46.

Krantz, G. W. 1978. A Manual of Acarology. 2nd Ed. Oregon State Univ., Corvalis, OR.

McDaniel, B. Year? How to Know the Mites and Ticks. WC Brown Co., Dubuque, Iowa.

Myles, T. G. 2002. Observations on mites (Acari) associated with the eastern subterranean termite, Reticulitermes flavipes (Isoptera: Rhinotermitidae). Sociobiology 39(2): 277-280.

OConnor, B. 2009. Systematics and ecology of North American bee-associated mites: potential threats to native and introduced pollinators. http://insects.ummz.lsa. umich.edu/beemites/Objectives.htm

Phillipsen, W. J. and H. C. Coppel. 1977. Histiostoma formosana sp. n. associated with the Formosan subterranean termite, Coptotermes formosanus Shiraki, (Acarina: Anoetidae--Isoptera: Rhinotermitidae). J. Kans. Entomol. Soc. 50(4): 496-502.

Wang, C. J. E. Powell, and B. M. O’Connor. 2002. Mites and nematodes associated with three subterranean termite species (Isoptera: Rhinotermitidae). Florida Entomologist 85(3): 499-506.

Woodring, J. P. 1973. Four new anoetid mites associated with halictid bees (Acarina: Anoetidae – Hymenoptera: Halictidae). J. Kans. Entomol. Soc. 46(3): 310-327.

Woodring, J. P. and J. C. Moser. 1970. Six new species of Anoetid mites associated with North American Scolytidae. Can. Entomol. 102: 1237-12257.

Woodring, J. P. and J. C. Moser. 1975. Description of Histiostoma conjuncta (new comb.) (Acari: Anoetidae), an associate of Central American bark beetles. Proc. Entomol. Soc. Wash. 77(1):83-86.

Appendix 19

Molecular Characterization of Two Metarhizium Strains Isolated from Termites in Guelph, Ontario.

Timothy G. Myles, Ph.D.

City of Guelph, Building Services, Community Design and Development Services, 1 Carden St., Guelph, Ontario N1H 3A1

and

Richard A. Humber, Ph.D.

USDA-ARS, R.W. Holley Center for Agriculture and Health, Ithaca, New York 14853-2901 ______

Abstract: Two strains of the entomopathogenic fungus, Metarhizium, were isolated from cultures of the eastern subterranean termite, Reticulitermes flavipes, collected in urban residential areas of Guelph, Ontario, Canada. The specificity of Metarhizium species and strains with respect to insect host species is likely to be an important factor in determining efficacy as microbial pest control agents. Therefore it was desirable to undertake a molecular characterization of these strains. The translation elongation factor 1-alpha (TEF-1A) genes were sequenced. Analysis of the sequences indicated that one of the strains is Metarhizium flavoviride var. pemphigi (100% identity) and the other strain is Metarhizium robertsii (99% identity). These are the first records of these species occurring with termites and the first record of M. flavoviride in Canada.

Introduction: Metarhizium anisopliae (Metschnikoff) Sorokin (1879) was originally described by the famous Russian founder of insect pathology, Metschnikoff over 130 years ago, as Entomopththora anisopliae and soon afterwards transferred to the new genus Metarhizium by Sorokin 1883. Thus, Metarhizium, the causative agent of green muscardine disease in insects, is one of the earliest described entomopathogenic fungi and has since become one of the most intensively and well studied. The genus, along with Beauvaria and Bacillus thuringiensis, is one of the leading candidates for the development of microbial pest control agents. Metarhizium has been registered as a myco-insecticide for control of numerous insect pests in many countries around the world (reviewed in Zimmerman, 2007). Several recent studies have investigated its potential as a microbial pest control agent of termites (Zoberi, 1995; Myles, 2002a,b,c; Lenz, 2005; Dong et al, 2007; Chouvenc et al, 2008a,b, 2009a,b). In addition to this applied research, there is much active research on the molecular taxonomy and phylogeny of the genus (Bidochka and Small, 2005; Bischoff et al, 2006, 2009; Driver et al, 2000; Huang et al, 2005a, b; Humber et al, 2005a, b; Pantou et al, 2003; Rombach et al, 1987). Currently, the nucleotide sequence of the translation elongation factor 1-alpha gene is considered to be a good diagnostic marker of the species within the Metarhizium complex (Bischoff et al., 2009). Although, the taxonomy of the genus remains somewhat unsettled at this time. As of 2010, 38 taxa of Metarhizium were listed in the Index Fungorum (Appendix 2).

Materials and Methods: In 2008, two strains of Metarhizium sp. were isolated from lab cultures of the eastern subterranean termite (Reticulitermes flavipes) originally collected in urban residential areas of Guelph, Ontario. One of the strains had green, oblong conidia (asexual spores) and was designated

―Gueph-RF-08a‖ while the other had darker olive-green conidia forming tightly packed clumps, and was designated ―Guelph-RF-08b‖. The conidial chains of Guelph-RF-08a were relatively powdery compared to the much more chunk-like clumps of Guelph-RF-08b. When Guelph-RF-08a was originally isolated on potato dextrose agar, the culture gave rise to conidia in rings around the periphery of the mycelia, but after subculture in the laboratory gave rise to a second growth form which produced conidia uniformly over the mycelium. The ring type was designated Guelph-RF- 08a(1) and the uniform type was designated Guelph-RF-08a(2). Thus, the two strains, 08a and 08b, and the two subtypes of 08a, (1) and (2), may all be distinguished on the basis of gross visual characters.

Initially, on the basis of the green color and oblong shape of the conidia, it was presumed that Guelph- RF-08a was a local strain of Metarhizium anisopliae, a species with green oblong conidia, and on visual appearance, identified as such by myself and by Dr. George Barron, professor of mycology, University of Guelph. The strain was also tentatively identified as M. anisopliae sensu lato by Dr. Richard Humber, curator of the Entomopathogenic Fungus Collection of the United States Agricultural Research Service in Ithaca, New York and accessioned it in the ARSEF collection with Accession No: ARSEF 9358. While Guelph-RF-08b was thought to probably be a different species of Metarhizium.

In order to satisfy the Canadian Pest Management Regulatory Agency’s request for detailed characterization of the strains, for the purposes of a research permit application, samples were submitted to Dr. Michael Bidochka, professor of microbiology, department of biological sciences, at Brock University, St. Catharines, Ontario for molecular characterization. The gene that was sequenced was the translation elongation factor 1-alpha (TEF-1A).

Results: The base sequences for strains Guelph-RF-08a and Guelph-RF-08b were determined as shown in Figures 1 and 2, respectively. The lower case letters had lower signal strength when the sequencing was done and the chromatograms were then checked visually.

cGTAaGTAGCCTTGCTCaaTTCGCGTCttgtaaAAGCCTCTTTACTGACTTGCTCGTCA

TAGGGGTATGTTTTGGAACCTACGCTTTCGAAGTACTCGAAGTTGATTGATAAC

TGACCGATCCTCACAGCCACGTCGACTCCGGCAAGTCTACCACCACTGGTCACT

TGATCTACCAGTGCGGTGGTATCGACAAGCGAACCATTGAGAAGTTCGAGAAG

GTAAGCCAAATTCCCTGTTTTAATGATTCCTGCTTATTTGGGCGATGGGAACAC

TTTTGTTTTCTCACTGCCTGTTGACCATTACCCCTCACTGTCACACAAAATTTTTC

GCGGGGCCTTATCTTGGGCTTTTGGTGGGGCATCGCATACCCCGCCAGCTGTTG

AGGTGTCTTTTGCGTGTCTTGCCTGCTGTTAAGAACCACAACGTGACCATCGCC

TTCAAAACCCAAAAAAAGATTGCAAACTAATTTGCATCTCTGTATAGGAAGCCG

CTGAACTCGGCAAGGGTTCCTTCAAGTACGCATGGGTTCTTGACAAGCTCAAGG

CCGAGCGTGAGCGTGGTATCACCATCGACATTGCCCTCTGGAAGTTCGAGACTC

CCAAGTACTATGTCACTGTCATCGGTATGTCGACTTGCGCAAACTAAACGACAC

GGTTTCGTTTGTCGGATTTGAATGCTAATATCCCTTGCACAGATGCTCCCGGTCA

CCGTGATTTCATCAAGAACAtgat

Figure 1. TEF-1A sequence for Metarhizium strain Guelph-RF-08a(1) and Guelph-RF-08a(2) = ARSEF 9358.

CgtaagatcgCCTGCCTCCaTTTCGAACCTTGTAGAAGCTGTTTACTGACtTGCTTGTC GTAGGGGTATGTTTCGGAGCCTACACTCTTCGCCGTCCCGAGTTTGTGATAACT GACTGGTCCTCaCAGCCACGTCGACTCCGGCAAGTCTACCACCACTGGTCACTT GATCTACCAGTGCGGTGGTATCGACAAGCGTACCATTGAGAAGTTCGAGAAGG TAAGCCAAACCACTCCGATTAATGATCTTTATTGTTTGGCGATGAACATTATTG AGTTTCCCGCTGCCTGTCGGCCATTACCCCTCACTGTGACACGAAAATTTTCGC GGGGCCTTATCTTGGACTTTGGTGGGGCACCATACCCCGCCAGCTGTCGAGGG TGTCTCTGTGTGTCTCTGGCTGTTGAAACCACAATATTGTCGTTGCTTTCAGAG GAAAAAACGTGAAACTAATTTGGATCGCTGTATAGGAAGCCGCTGAACTCGGC AAGGGTTCCTTCAAGTACGCATGGGTTCTTGACAAGCTCAAGGCCGAGCGTGA GCGTGGTATCACCATCGACATTGCCCTCTGGAAGTTCGAGACTCCCAAGTACT ATGTCACCGTCATTGGTATGTCGACTTGCGCAAACTGACCGCATACTTTTCTCC TAAATTGAATGCTAATGCCCCTCCCACAGACGCTCCCGGTCACCGTGACTTTAT CAAGAACATGAt

Figure 2. EF-1A sequence for Metarhizium strain Guelph-RF-08b.

Figure 3. Blast search for sequence similarity for Guelph-RF-08a.

Figure 4. Blast search for sequence similarity for Guelph-RF-08b.

The Basic Local Alignment Search Tool (BLAST) finds regions of local similarity between sequences. The database is the "BLAST" site online. This is the standard for gene to gene comparison. According to the Blast Search (Figure 3), the sequence for RF-08a is most similar to Metarhizium flavoviride var. pemphigi strain ARSEF 7491 (Accession DQ463964.1) (100% identity). The next most similar sequences are of M. frigidum (all strains with 92% identity), M. flavoviride var. flavoviride (90%), M. flavoviride var. minus (88%), and M. robertsii and M. lepiotae (both with 85% identity). Therefore, contrary to the initial identification of strain RF-08a as M. anisopliae, on morphological grounds, the molecular data indicate it to be M. flavoviride var. pemphigi.

The Blast Search for Strain Guelph-RF08b (Figure 4) indicates that the sequence is most similar to a strain ARSEF 8685 of M. brunneum (99% identity). However, labelling of ARSEF 8685 as M. brunneum is thought to be an error, as that sequence is more typical of M. robertsii, and the next three most closely related samples, also at 99% identity, are all strains of M. robertsii. Other strains of M. brunneum expressed 97% identity. Four strains of Metarhizium pingshaense expressed 98% identity. One strain of M. anisopliae anisopliae expressed 97% identity, while all strains of M. guizhouense and M. majus expressed 96% identity. These very close levels of sequence identity show that all these species/strains in the M. anisopliae complex are closely related. Thus, we consider strain Guelph-RF- 08b to be M. robertsii.

Appendix 1 shows the visual alignment of the 733 bases of strain Guelph-RF-08a with the 714 bases of strain Guelph-RF-08b. A difference in sequence lengths implied a number of insertions or deletions whose positions and lengths were located by visual alignment, resulting in an optimally paired sequence of 734 bases. Of the 734 paired bases, 660 base pairs were identical, equal to 77% sequence identity (shown in green), 147 were non-identical (shown in red), and 21 positions were inferred as insertions or deletions (shown in yellow). Two major conserved regions appeared between positions 122-231 and 479-617.

Discussion: A recent taxonomic revisionary study of Metarhizium, based on molecular sequences data has resulted in the splitting of the genus into a number of proposed new cryptic species, which can only be identified on the basis of molecular characters (Bischoff et al, 2009). The genealogical concordance phylogenetic species recognition criterion (GC-PSR) of Taylor et al (2000) was adopted to recognize species based on multigenic molecular sequence analysis. However, not all authorities are in agreement as to what amount of sequence variation should be accepted as constituting species level differentiation and what criteria should be applied to molecular sequence data to delimit valid species. Therefore it is not known if all the proposed species are valid biological species, subspecies, varieties, or merely intra-specific clonal variants. A particular problem is that Metarhizium is an anamorphic genus (asexual phase of a fungi) of the teleomorphic (sexual phase) genus Metacordyceps. Since anamorphs are asexual clones, they are not species in the ordinary sense. In the final analysis, the true measure of a biological species is that it is a sexual population delimited by sexual isolation. Thus until the respective Metacordyceps forms are discovered (and they may not even exist) and fertility crosses of the respective species are conducted, biological species level status must remain provisional. Nevertheless, in real world practice, as with almost all of taxonomy, such crosses have not been conducted and other criteria, morphological or molecular, are almost always applied by taxonomists in respective fields.

A fundamental division exist within the genus Metarhizium between species with white or greenish conidia. Metarhizium album, a pathogen of leafhoppers and planthoppers on rice, is unique and easily distinguished by its white conidia. Those species with greenish conidia, are represented by the type species of the genus M. anisopliae and historically comprise the M. anisopliae lineage or M. anisopliae sensu lato (in the broad sense). Those with greenish conidia are now divided among two sister species complexes, the M. anisopliae and M. flavoviride complexes (Figure 5). As the name flavoviride implies, the type species of the flavoviride complex has distinctly yellowish green or bright green conidia in contrast to anisopliae which have green to olive green, or even brownish conidia. However, as the newer molecular studies have shown, conidia color may not be a consistent and stable character at the species or species complex level. Strains with green conidia may be present in both the anisopliae and flavoviride complexes. Species within the flavoviride complex, as determined by molecular sequencing, may have green conidia which are visually indistinguishable from some species within the color range of the anisopliae complex.

Bidochka has genetically characterized over 500 isolates of Metarhizium from Ontario, but has not before encountered Metarhizium flavoviride nor M. f. var. pemphigi (Bidochka, personal communication). Thus, it appears most likely that this strain is termite-host associated rather than soil- associated in Ontario soils. This is supported by the fact that this strain visually matched isolates previously collected by Myles and Zoberi (then identified as M. anisopliae) which were associated with termites in Toronto (Myles, 2002a,b,c; Zoberi 1995). In contrast, most of Bidochka’s previous isolates were obtained from forest or agricultural soil samples using the wax moth (Galleria sp.) as the bioassay detection organism. Actually, several possibilities may account for the absence of this strain from Bidochka’s previous sampling. It may be that this strain is not biologically detectable with Galleria, or that it tends to be present only in urban areas, or that it is generally rare except in soils inhabited by termites. Perhaps Metarhizium flavoviride var. pemphigi may express some level of termite host specificity, or have been introduced, along with the termites, which themselves are not native to southern Ontario, but were introduced from the United States, and now co-occurs in areas where termites have become established.

Recent studies by Chouvenc et al (2008a,b, 2009a,b) on Metarhizium-termite interactions were conducted with a strain of ―Metarhizium anisopliae anisopliae” from the American Type Culture Collection (ATCC 90448). The ATCC online catalog provides the following information, indicating the provenance of this culture from Austria on codling moth, and originally designated M.a. 43:

ATCC Number: 90448 Organism: Metarhizium anisopliae (Metschnikoff) Sorokin var. anisopliae Designations: M.a. 43 [HRI 275.86]; Isolation: codling moth larvae, Cydia pomonella, Austria

The ARSEF collection holds a sample, ARSEF 1095, also from Austria on codling moth and labelled ―M.a. 43‖:

ARSEF 1095 DW Roberts <-- S Keller (Ma 43). Carpocapsa pomonella [Lepidoptera: Olethreutidae]. Accessioned 16 Feb 1984. Austria. (Strong destruxin producer).

Therefore ARSEF 1095 appears to be identical to the ATCC 90448. ARSEF 1095 has been determined by molecular sequence data to be M. brunneum.

The superior pathogenicity and transmission among termites of Metarhizium reported by Myles (2002c) compared to the results obtained by Chouvenc et al (2008b) are likely due, at least in part, to the use of different Metarhizium species, though both were originally reported as M. anisopliae. Chouvenc’s work was evidently done with M. brunneum and Myles work was evidently done with M. flavoviride var. pemphigi. Alternatively, differences in transmission may have been related to different methods of treatment: dusting with dry conidia by Myles, versus application in Tween solution (Chouvenc, personal communication).

Chouvenc obtained his sample of ATCC 90448 from Rebecca Rosengaus (Chouvenc, personal communication). One may therefore infer that the several research publications dealing with Metarhizium anisopliae by Rosengaus et al (1997, 1998a,b, 1999a,b, 2000, 2003, 2004, 2007) also pertain to M. brunneum. Indeed, misidentification errors are probably systemic in the Metarhizium literature.

The currently available molecular sequence data, suggest that both strains F52 and Guelph-RF-08b belong to the M. anisopliae complex, and these strains, along with M. robertsii, M. brunneum and M. anisopliae anisopliae are all very closely related strains/species within this complex. In contrast, strain Guelph-RF-08a (=ARSEF 9358) clearly belongs to the M. flavoviride complex, which is phylogenetically deeply nested in a different, though sister branch complex of the genus, as shown by the low level of base pair identity (only 77%) between strains Guelph-RF-08a (= M. flavoviride var. pemphigi) and Guelph-RF-08b (probably = M. brunneum or M. robertsii) as illustrated in Appendix 1.

In 2009 ―Metarhizium anisopliae strain F52‖ was granted a conditional registration in Canada for control of root weevils in container-grown ornamentals (PMRA, 2010). According to Richard Humber, strain F52 was sequenced (TEF-1A gene) and proved to be Metarhizium brunneum according to the new system of Bischoff et al, 2009.

The above examples show that even well characterized strains which have traditionally been considered M. anisopliae or M. anisopliae sensu lato, are likely to fall under new species with further refinement and resolution of Metarhizium systematics. From a historical perspective, all species with greenish conidia have been considered M. anisopliae sensu lato. Given that gene sequencing methods have only been widely applied in the last decade, and that a large proportion the Metarhizium species currently considered valid have only been recently described, it is inevitable that most prior research on Metarhizium anisopliae by researchers over the past 130 years may in fact pertain to other species of Metarhizium. This is of considerable concern from a practical perspective, because regulatory approvals of Metarhizium strains for biocontrol purposes will likely to require molecular characterization of species and strain and when the identified strains do not match historical published information reported for M. anisopliae, approvals could be set back with requirements for additional studies on safety and environmental impacts on non-target organisms. Ironically, the more finely Metarhizium systematic is divided the more onerous the requirements for further studies could become for those who wish to develop biocontrol applications. We feel that this would be an unnecessary constraint given the historical safety of the genus as a whole.

Species and strains of Metarhizium are likely to differ slightly with respect to insect host specificity and climatic tolerances but, as specialist, entomopathogenic fungi, it is likely that different species and strains will be essentially similar and innocuous toward non-target organism in the environment. The safety of Metarhizium anisopliae (sensu lato) was recently extensively reviewed by Zimmerman (2007), including effects on non-target organisms, (microorganisms, plants, soil organism, aquatic organisms, predators, parasitoids, honey bees, earthworms, etc.); effects on vertebrates (fish, amphibian, reptiles, and birds), effects on mammals and human health (allergy, pathogenicity/ toxicity), and it was concluded that Metarhizium is safe with minimal risk to vertebrates, humans and the environment. In conformity with this, the American Type Culture Collection offers 76 different strains of Metarhizium for sale commercially including many strains of M. anisopliae and M. flavoviride, all of which carry the least hazardous Biosafety rating BS-1, which are those cultures ―not known to cause disease in healthy adult humans‖ (CDC, 2007).

Acknowledgements: We would like to thank Dr. Michael Bidochka of Brock University for assistance and valuable discussions during this study.

Metarhizium

TRADITIONAL LIMITED MOLECULAR MULTIGENIC (Rombach et al. 1987) (Driver et al. 2000) (Bischoff et al. 2007, 2009)

M. anisopliae M. anisopliae var. anisopliae var. anisopliae M. anisopliae var. majus var. majus M. majus var. acridum M. acridum var. lepidiotum M. lepidiotae M. brunneum M. pingshaense M. guizhouense M. robertsii M. globosum

M. flavoviride M. flavoviride M. flavoviride var. flavoviride var. flavoviride var. flavoviride var. minus var. minus var. minus var. novazealandicum var. novazealandicum var. pemphigi var. pemphigi “Type E” “Type E” M. frigidum

M. album M. album M. album Under construction

Figure 5. Recent history of Metarhizium taxonomy. (Richard Humber, pers. communication).

Bischoff et al, 2009. A multilocus phylogeny of the Metarhizium anisopliae lineage

Multigene phylogram EF-1A phylogram

M. anisopliae complex guizhouense guizhouense majus majus pingshaense brunneum anisopliae pingshaense robertsii robertsii Metarhizium brunneum anisopliae anisopliae lepidiotae lepidiotae lineage acridum acridum sensu lato globosum globosum with greenish conidia flavoviride flavoviride frigidum frigidum

album album

M. flavoviride complex

The topologies of the proposed phylograms differed only slightly in the locations of brunneum and anisopliae.

Figure 6. Phylogeny of Metarhizium (Bischoff et al, 2009).

References

Bidochka, M. J., J. E. Kasperski, and G. A. M. Wild. 1998. Occurrence of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana in soils from temperate and near- northern habitats. Can. J. Bot. 76: 1198-1204.

Bidochka, M. J. and C. L. Small. 2005. Phylogeography of Metarhizium, an insect pathogenic fungus. Pp. 28-50. In: F. E. Vega and M. Blackwell (eds.) Insect-Fungus Associations, Ecology and Evolution. Oxford University Press, New York.

Bischchoff, J. F., S.A. Rehner, and R. A. Humber 2006. Metarhizium frigidum sp. nov.: a cryptic species of M. anisopliae and a member of the M. flavoviride complex. Mycologia 98: 737-745.

Bishchoff, J. F., S. A. Rehner, and R. A. Humber. 2009. A multilocus phylogeny of the Metarhizium anisopliae lineage. Mycologia 101 (4): 512-530.

Centers for Disease Control and Prevention and National Institutes of Health 2007. Biosafety in Microbiological and Biomedical Laboratories, (BMBL) 5th Edition (HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, U.S. Government Printing Office: Washington DC.

Chouvenc, T., N.-Y. Su, and M. L. Elliott. 2008a. Interactions between the subterranean termite Reticulitermes flavipes (Isoptera: Rhinotermitidae) and the entomopathogenic fungus Metarhizium anisopliae in foraging arenas J. Econ. Entomol. 1010: 885-893.

Chouvenc, T., N.-Y. Su, and A. Robert. 2009a. Cellular encapsulation in the eastern subterranean termite, Reticulitermes flavipes (Isoptera), against infection by the entomopathogenic fungus Metarhizium anisopliae. J. Invert. Pathol. 101: 234-241.

Chouvenc, T. N.-Y. Su, and A. Robert. 2009b. Inhibition of Metarhizium anisopliae in the alimentary tract of the eastern subterranean termite Reticulitermes flavipes. J. Invert. Pathol. 101: 130-136.

Chouvenc, T., N.-Y. Su, and A. Robert. 2009c. Susceptibility of seven termite species (Isoptera) to the entomopathogenic fungus Metarhizium anisopliae. Sociobiology. 54 (3): 723-748.

Driver, F., R. J. Milner, and J. W. H. Trueman. 2000. A taxonomic revision of Metarhizium based on a phylogenetic analysis of rDNA sequence data. Mycol. Res. 104: 134-150.

Dong, C., J. Zhang, W. Chen, H. Huang, and Y. Hu. 2007. Characterization of a newly discovered China variety of Metarhizium anisopliae (M. anisopliae var. dcjhyium) for virulence to termites, isoenzyme and phylogenetic analysis. Microbiol. Res. 162: 53-61.

Huang, B., C. Li, R. A. Humber, K. T. Hodge, M. Fan, and Z. Li. 2005a. Molecular evidence for taxonomic status of Metarhizium taii and its teleomoph, Cordyceps taii (Hypocreales, Clavicipitaceae). Mycotaxon 94: 137-147.

Huang, B., R. A. Humber, S. Li, Z. Li, and K. T. Hodge. 2005b. Further notes on the molecular taxonomy of Metarhizium. Mycotaxon. 94:181-187.

Lenz, M. 2005. Biological control in termite management: the potential of nematodes and fungal pathogens. Pp. 47-52. In: C.-Y. Lee and W. H. Robinson (eds.) Proceedings of the Fifth International Conference on Urban Pests. Perniagaan Ph’ng @ P&Y Design Network, Malaysia.

Myles, T. G. 2002a. Alarm, aggregation, and defense by Reticulitermes flavipes in response to a naturally occurring isolate of Metarhizium anisopliae. Sociobiology. 40(2): 234-255.

Myles, T. G. 2002b. Isolation of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes) from Reticulitermes flavipes (Isoptera: Rhinotermitidae) with convenient methods for its culture and collection of conidia. Sociobiology. 40 (2): 257-264.

Myles, T. G. 2002c. Laboratory studies on the transmission of Metarhizium anisopliae in the eastern subterranean termite, Reticuliterms flavipes (Isoptera: Rhinotermitidae), with a method for applying appropriate doses of conidia to trapped termites for release. Sociobiology. 40 (2): 265-276.

Pantou, M.P., A. Mavridou, and M.A. Typas. 2003. IGS sequence variation, group-I introns and the complete nuclear ribosomal DNA of the entomopathogenic fungus Metarhizium: excellent tools for isolate detection and phylogenetic analysis. Fungal Genet. Biol. 38: 159-174.

Pest Management Regulatory Agency, Health Canada. 2010. Evaluation Report: Metarhizium anisopliae strain F52. ERC2010-01.

Rombach, M. C., R. A. Humber, H. C. Evans. 1987. Metarhizium album a fungal pathogen of leaf- and plant-hoppers of rice. Trans. Brit. Mycol. Soc. 37: 37-45.

Rosengaus, R. B. & J. F. A. Traniello 1997. Pathobiology and disease transmission in dampwood termites [Zootermopsis angusticollis (Isoptera: Termopsidae)] infected with the fungus Metarhizium anisopliae (Deuteromycotina: Hypomycetes). Sociobiology 30: 185–195.

Rosengaus, R. B. & J. F. A. Traniello 2001. Disease susceptibility and the adaptive nature of colony demography in the dampwood termite Zootermopsis angusticollis. Behav. Ecol. Sociobiol. 50: 546–556.

Rosengaus, R. B., A. B. Maxmen, L. E. Coates & J. F. A. Traniello 1998a. Disease resistance: a benefit of sociality in the dampwood termite Zootermopsis angusticollis (Isoptera: Termopsidae). Behav. Ecol. Sociobiol. 44: 125–134.

Rosengaus, R. B., M. R. Guldin & J. F. A. Traniello 1998b. Inhibitory effect of termite fecal pellets on fungal spore germination. J. Chem. Ecol. 24: 1697–1706.

Rosengaus, R. B., M. L. Lefebvre, C. Jordan & J. F. A. Traniello 1999a. Pathogen alarm behaviour in a termite: A new form of communication in social insects. Naturwissenschaften 86: 544–548.

Rosengaus, R. B., J. F. A. Traniello, T. Chen, J. J. Brown & R. D. Karp 1999b. Immunity in a social insect. Naturwissenschaften 86: 588–591.

Rosengaus, R. B., M. L. Lefebvre & J. F. A. Traniello 2000. Inhibition of fungal spore germination by Nasutitermes: Evidence for a possible antiseptic role of soldier defensive secretions. J. Chem. Ecol. 26: 21–39.

Rosengaus, R. B., J. E. Moustakas, D.V. II Calleri & J.F.A. Traniello 2003. Nesting ecology and cuticular microbial loads in dampwood (Zootermopsis angusticollis) and drywood termites (Incisitermes minor, I. schwarzi, Cryptotermes cavifrons). J. Insect. Sci. 3: 31– 37.

Rosengaus, R. B., J. F. A. Traniello, M. L. Lefebvre & A. B. Maxmen. 2004. Fungistatic activity of the sternal gland secretion of the dampwood termite Zootermopsis angusticollis, Insect. Soc. 51: 259–264.

Rosengaus, R. B., T. Cornelisse, K. Guschanski & J. F. A. Traniello 2007. Inducible immune proteins in the dampwood termite Zootermopsis angusticollis. Naturwissenschaften. 94: 25–33.

Taylor, J.W., D.J, Jacobson, S. Krolen, T. Kasuga, D.M. Geiser, D.S. Hibbett, and M.C. Fisher. 2000. Phylogenetic species recognition and species concepts in fungi. Fungal Genet. Biol. 31: 21-32.

Zimmerman, G. 2007. Review on safety of the entomopathogenic fungus Metarhizium anisopliae. Biocontrol Science and Technology. 17(9): 879-920.

Zoberi, M. 1995. Metarhizium anisopliae, a fungal pathogen of Reticulitermes flavipes (Isoptera: Rhinotermitidae). Mycologia. 87 (3): 354-359.

Appendix 1. Comparison of sequences of the EF-1A gene for Metarhizium strains G-RF-08A and G-RF-08B.

Base No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 GRF08A c G T A a G T A G C C T T G C T C a a T

GRF08B C g t a a g a t c g C C T G C C T C C a

21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 GRF08A T C G C G T C t t g t a a A A G C C T GRF08B T T T C G A A C C T T G T A G A A G C T

41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 GRF08A C T T T A C T G A C T T G C T C G T C A

GRF08B G T T T A C T G A C t T G C T T G T C G

61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 GRF08A T A G G G G T A T G T T T T G G A A C C

GRF08B T A G G G G T A T G T T T C G G A G C C

81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 GRF08A T A C G C T T T C G A A G T A C T C G A GRF08B T A C A C T C T T C G C C G T C C C G A

101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 GRF08A A G T T G A T T G A T A A C T G A C C G

GRF08B G T T T G T G A T A A C T G A C T G

121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 GRF08A A T C C T C A C A G C C A C G T C G A C

GRF08B G T C C T C a C A G C C A C G T C G A C

141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 GRF08A T C C G G C A A G T C T A C C A C C A C GRF08B T C C G G C A A G T C T A C C A C C A C

161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 GRF08A T G G T C A C T T G A T C T A C C A G T

GRF08B T G G T C A C T T G A T C T A C C A G T

181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200

GRF08A G C G G T G G T A T C G A C A A G C G A

GRF08B G C G G T G G T A T C G A C A A G C G T

201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 GRF08A A C C A T T G A G A A G T T C G A G A A

GRF08B A C C A T T G A G A A G T T C G A G A A

221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 GRF08A G G T A A G C C A A A T T C C C T G T T

GRF08B G G T A A G C C A A A C C A C T C C G A

241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 GRF08A T T A A T G A T T C C T G C T T A T T T

GRF08B T T A A T G A T C T T T A T T G T T T

261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 GRF08A G G G C G A T G G G A A C A C T T T T G

GRF08B G G C G A T G A A C A T T A T T G A G

281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 GRF08A T T T T C T C A C T G C C T G T T G A C

GRF08B T T T C C C G C T G C C T G T C G G C

301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 GRF08A C A T T A C C C C T C A C T G T C A C A

GRF08B C A T T A C C C C T C A C T G T G A C A

321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 GRF08A C A A A A T T T T T C G C G G G G C C T

GRF08B C G A A A A T T T T C G C G G G G C C T

341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 GRF08A T A T C T T G G G C T T T T G G T G G G

GRF08B T A T C T T G G A C T T T G G T G G G

361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 GRF08A G C A T C G C A T A C C C C G C C A G C

GRF08B G C A C C A T A C C C C G C C A G C

381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 GRF08A T G T T G A G G T G T C T T T T G C G T

GRF08B T G T C G A G G G T G T C T C T G T G T

401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 GRF08A G T C T T G C C T G C T G T T A A G A A

GRF08B G T C T C T G G C T G T T G A A A

421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 GRF08A C C A C A A C G T G A C C A T C G C C T GRF08B C C A C A A T A T T G T C G T T G C T T

441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 GRF08A T C A A A A C C C A A A A A A A G A T T

GRF08B T C A G A G G A A A A A A C G T

461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 GRF08A G C A A A C T A A T T T G C A T C T C T

GRF08B G A A A C T A A T T T G G A T C G C T

481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 GRF08A G T A T A G G A A G C C G C T G A A C T GRF08B G T A T A G G A A G C C G C T G A A C T

501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 GRF08A C G G C A A G G G T T C C T T C A A G T

GRF08B C G G C A A G G G T T C C T T C A A G T

521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 GRF08A A C G C A T G G G T T C T T G A C A A G

GRF08B A C G C A T G G G T T C T T G A C A A G

541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 GRF08A C T C A A G G C C G A G C G T G A G C G GRF08B C T C A A G G C C G A G C G T G A G C G

561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580

GRF08A T G G T A T C A C C A T C G A C A T T G

GRF08B T G G T A T C A C C A T C G A C A T T G

581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 GRF08A C C C T C T G G A A G T T C G A G A C T

GRF08B C C C T C T G G A A G T T C G A G A C T

601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 GRF08A C C C A A G T A C T A T G T C A C T G T

GRF08B C C C A A G T A C T A T G T C A C C G T

621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 GRF08A C A T C G G T A T G T C G A C T T G C G

GRF08B C A T T G G T A T G T C G A C T T G C G

641 642 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 GRF08A C A A A C T A A A C G A C A C G G T T T

GRF08B C A A A C T G A C C G C A T A C T T T T

661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 GRF08A C G T T T G T C G G A T T T G A A T G C

GRF08B C T C C T A A A T T G A A T G C T A A T

681 682 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 GRF08A T A A T A T C C C T T G C A C A G A T G

GRF08B G C C C C T C C C A C A G A C G

701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 717 718 719 720 GRF08A C T C C C G G T C A C C G T G A T T T C

GRF08B C T C C C G G T C A C C G T G A C T T T

721 722 723 724 725 726 727 728 729 730 731 732 733 734 GRF08A A T C A A G A A C A t g a t

GRF08B A T C A A G A A C A T G A t

A identical base pairs A

A non-identical base pairs T

A Inferred insertion or deletion

Appendix 2. Described taxa of Metarhizium listed in Index Fungorum. (Names in blue (bold) are names for which there is no taxonomic opinion available, green (italics) are names where a taxonomic opinion has been expressed in Species Fungorum).

1) Metarhizium acridum (Driver & Milner) J.F. Bisch., Rehner & Humber (2009); Anamorphic Metacordyceps 2) Metarhizium album Petch (1931), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 3) Metarhizium anisopliae (Metschn.) Sorokīn (1883), [RSD]; Anamorphic Metacordyceps Metarhizium anisopliae f. anisopliae (Metschn.) Sorokīn (1883), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 4) Metarhizium anisopliae f. major J.R. Johnst. (1915), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 5) Metarhizium anisopliae f. oryctophagum Frieder. (1930), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 6) Metarhizium anisopliae var. acridum Driver & Milner (2000), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 7) Metarhizium anisopliae var. anisopliae (Metschn.) Sorokīn (1883), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 8) Metarhizium anisopliae var. dcjhyium C.J. Dong, Jia M. Zhang, W.G. Chen & Y.Y. Hu (2007), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 9) Metarhizium anisopliae var. frigidum A.C. Rath, C.J. Carr & B.R. Graham (1995), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 10) Metarhizium anisopliae var. lepidiotae Driver & Milner (2000), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 11) Metarhizium anisopliae var. major (J.R. Johnst.) M.C. Tulloch (1976), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 12) Metarhizium brunneum Petch (1935), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 13) Metarhizium chrysorrheae Giard (1888); Anamorphic Metacordyceps 14) Metarhizium cicadinum (Höhn.) Petch (1931); Anamorphic Metacordyceps 15) Metarhizium cylindrosporum Q.T. Chen & H.L. Guo (1986), [RSD]; Anamorphic Metacordyceps 16) Metarhizium flavoviride W. Gams & Rozsypal (1973), [RSD]; Anamorphic Metacordyceps 17) Metarhizium flavoviride var. flavoviride W. Gams & Rozsypal (1986), (= Metarhizium flavoviride), [RSD]; Anamorphic Metacordyceps 18) Metarhizium flavoviride var. minus Rombach, Humber & D.W. Roberts (1986), (= Metarhizium flavoviride), [RSD]; Anamorphic Metacordyceps 19) Metarhizium flavoviride var. novozealandicum Driver & Milner (2000); Anamorphic Metacordyceps 20) Metarhizium flavoviride var. pemphigi Driver & Milner (2000), (= Metarhizium flavoviride), [RSD]; Anamorphic Metacordyceps 21) Metarhizium frigidum J.F. Bisch. & S.A. Rehner (2007), [RSD]; Anamorphic Metacordyceps 22) Metarhizium gigas Sorokīn (1883); Anamorphic Metacordyceps 23) Metarhizium globosum J.F. Bisch., Rehner & Humber (2009); Anamorphic Metacordyceps 24) Metarhizium glutinosum S.A. Pope (1944), (= Myrothecium verrucaria), [RSD]; Anamorphic Hypocreales 25) Metarhizium guizhouense Q.T. Chen & H.L. Guo (1986), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 26) Metarhizium iadini H.L. Guo (1991), (= Metarhizium flavoviride), [RSD]; Anamorphic Metacordyceps 27) Metarhizium lepidiotae (Driver & Milner) J.F. Bisch., Rehner & Humber (2009); Anamorphic Metacordyceps 28) Metarhizium leptophyei Giard (1888); Anamorphic Metacordyceps 29) Metarhizium libelullae Sorokīn (1883); Anamorphic Metacordyceps 30) Metarhizium majus (J.R. Johnst.) J.F. Bisch., Rehner & Humber (2009); Anamorphic Metacordyceps 31) Metarhizium pinghaense Q.T. Chen & H.L. Guo (1986), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 32) Metarhizium polistis Cohn ex Sorokīn (1883); Anamorphic Metacordyceps 33) Metarhizium robertsii J.F. Bisch., Rehner & Humber (2009); Anamorphic Metacordyceps

34) Metarhizium taii Z.Q. Liang & A.Y. Liu (1991), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 35) Metarhizium truncatum (Briard) Petch (1931); Anamorphic Metacordyceps 36) Metarhizium velutinum Borowska, Golonk. & Kotulowa (1970), (= Metarhizium anisopliae), [RSD]; Anamorphic Metacordyceps 37) Metarhizium viridicolumnare (Matsush.) Matsush. (1993); Anamorphic Metacordyceps 38) Metarhizium viridulum (Tzean, L.S. Hsieh, J.L. Chen & W.J. Wu) B. Huang & Z.Z. Li (2004); Anamorphic Metacordyceps

Appendix 3. Taxonomy of Metarhizium with authorship and synonymies (after Bischoff et al, 2009, and Humber, pers. comm.).

I. Names and Synonyms of Accepted Species and Varieties Likely to be Raised to Species.

1) Metarhizium album

―Metarhizium anisopliae‖ complex: 2) M. anisopliae (Metsch.) Sorok., 1883 = Entomophthora anisopliae Metsch (1879) =Isaria anisopliae (Metsch.) 1895 =Penicillium anisopliae (Metsch.)1904 =Isaria destructor Metsch. 1880 =Oospora destructor (Metsch.) 1893 =Isaria anisopliae var. americana Pettit, 1895 =Penicillium cicadinum Holh., 1909 =Metarhizium cicadinum (Hohn.) 1931. = Sporotrichum paranense Marchionatto, 1933 3) M. majus (Johnst.) Bisch, Rehner & Humber, 2009 = Metarhizium anisopliae f. major Johnst., 1915 = Metarhizium anisoplliae f. oryctophagum Friderichs, 1930 = Metarhizium anisopliae var. major Tulloch, 1976 4) M. acridum (Driver & Milner) Bischl, Rehner & Humber = Metarhizium anisopliae var. acridum Driver & Milner, 2000 5) M. lepidiotae (Driver & Milner) Bisch., Rehner & Humber = Metarhizium anisopliae var. lepidiotae Driver & Milner, 2000 6) M. brunneum Petch, 1935 7) M. pingshaense Chen & Guo, 1986 8) M. guizhouense Chen & Guo, 1986 = Metarhizium taii Liang & Liu, 1991 9) M. robertsii Bisch., Rehner & Humber, 2009 10) M. globosum Bisch., Rehner & Humber, 2009

―Metarhzium flavoviride‖ complex 11) M. flavoviride var. flavoviride Gams & Rozsypal, 1973 =Metarhizium iadini H.L. Guo (1991) 12) M. flavoviride var. minus Rombach, Humber & Roberts, 1986 13) M. flavoviride var. novazealandicum Driver & Milner, 2000 14) M. flavoviride var. pemphigi Driver & Milner, 2000 15) M. frigidum Rath, Carr & Graham, 1995

II. Invalidly Named Species (Nomen Nudum)

Metarhizium anisopliae var. dcjhyium C.J. Dong, Jia M. Zhang, W.G. Chen & Y.Y. Hu, 2007

III. Names of Uncertain Status

Metarhizium chrysorrheae Giard, 1888 Metarhizium gigas Sorokīn, 1883 Metarhizium leptophyei Giard, 1888 Metarhizium libelullae Sorokīn, 1883 Metarhizium polistis Cohn ex Sorokīn, 1883 Metarhizium truncatum (Briard) Petch (1931) Metarhizium velutinum Borowska, Golonk. & Kotulowa, 1970 Metarhizium viridicolumnare Matsush., 1993 Metarhizium viridulum (Tzean, L.S. Hsieh, J.L. Chen & W.J. Wu) B. Huang & Z.Z. Li (2004)

IV. Species Moved to Different Genera

Metarhizium glutinosum S.A. Pope (1944), (= Myrothecium verrucaria) Metarhizium cylindrosporum Chen & Guo,1986 = New name: Nomuraea clyindrosporae (Chen and Guo) Tsean et al. (1993)