Terry Gregoire ,John Lukach, Marcia McMullen, Scott Halley, Vern Hofman, Suranjan Panigrahi, Jim Harbour

Fusarium head blight has caused huge losses for wheat and barley growers the past six years. Fungicides have potential to reduce the impact of this disease. The papers following the summary remarks provide details of the experimental work completed at Langdon North Dakota in the summer of 1998. The objective of this work was to evaluate sprayer nozzles, volumes, pressures and nozzle arrangements to improve retention of fungicide on wheat heads during ground application.

Some fifty fungicide trials were conducted in N.D. in 1998. Procedures for this work have and will continue to be in a state of rapid evolution because little information is available on how to target the wheat head when spraying fungicides. A team effort was needed to accomplish this work. The researchers who conducted the spraying technology research at Langdon and at other ND locations included: John Lukach, Langdon Research and Extension Center Terry Gregoire, Northeast District Extension Agronomist, Marcia McMullen and Scott Halley from the NDSU Plant Pathology Dept, Vern Hofman and Suranjan Panigrahi from the NDSU Ag and Bio Sciences Dept and Jim Harbour from the Carrington Research Extension Center. The testing of parameters immediately available to producers were given priority over basic work. Five papers are included in this report.

1997 work at Langdon indicated that nozzles with a small orifice gave the highest dye retention on the head. Spring greenhouse research showed that nozzles oriented front and back at a 30 degree angle from horizontal would give the best coverage of the head. It was decided to minimize use of high water volumes and very small nozzles in 1998 summer research because they are impractical for producers. Small nozzles work well with clean water and red dye but plug easily when using powder fungicides and the flourescent dye used in 1998. Flourescent orange dye (Day Glo blaze orange) was used to allow a camera, coupled with computer digitizing, under ultra violet and low light conditions to measure percent spray coverage. To measure spray coverage four picture of each of 20 to 40 heads per treatment were taken. The pictures were of the front and back of each head under normal low light and under ultraviolet light. The area that fluoresced under ultraviolet was divided by the total head area under normal light to obtain a percentage for each head. The pictures also allow a visual comparison of how much dye stayed on the awns or got onto the glumes. The size of individual trials was severely limited by the need to do laboratory determination of spray coverage on fresh plant material. A major bottle neck was encountered because the anthers on the wheat heads fluoresced the same as the dye. All anthers from all heads had to be removed before pictures could be taken. On some treatments the beards were cut at the top of each kernel to determine the percentage of fungicide on the beards compared to the whole head.

The percent dye coverage of the wheat head was determined on seven dates, six experiments. The coverage for nozzles by spray date is shown in Table 1. The averages are only for nozzles which had the same parameters in that experiment so the comparison was fair. The percent coverage was lower and the variability between dates was higher than was hoped for. These low percent coverage data did not correspond to yield responses or disease reductions, see Appendix Reports. The trials with the higher dye coverage data were generally sprayed on dry heads later in the day when the breeze was stronger. An unproven theory to explain this would be that increased horizontal movement of the spray droplets in the wind allowed increased coverage. The trials which were sprayed at early flower tended to have lower dye coverage. An unproven theory to explain this would be that the flag leaves mixed in with the heads are interfering with head coverage. The larger droplets produced by the Turbo Drop nozzles, 400 microns, tended to have higher coverage than the XR8001 nozzles with 187 micron droplets when spraying at early flower. A analysis of interactions across experiments will be completed to test the validity of this trend. Detailed work will be necessary to prove or disprove these theories.

Table 1. Mean Percent Dye Coverage of the Head by Spray Nozzles, Langdon, 1998

Domain HRS Domain HRS

Munich Durum Munich Durum

Amidon HRS Grandin HRS Munich Durum Russ HRS Russ HRS Russ HRS Russ HRS

25-Jun 7-Jul 9-Jul 13-Jul 18-Jul 22-Jul 24-Jul Nozzle

Nozzle Flowering Early Flower Pre- flower Flowering early flower late flower Flowering Means

Treatments in Average 5 4 4 4 6 6 6

% % % % % % % %

XR8001 * 6.7 1.1 1.5 9.1 2.9 12.0 3.7 5.3

TD01 5.0 1.7 3.0 4.4 3.5 8.3 -- 4.3

XR8002 * -- -- 1.7 2.7 -- -- -- 2.2

TJ8002 -- 1.1 1.4 -- -- -- -- 1.2

XR8002 ** -- -- 0.7 -- -- -- -- 0.7

HC6-70 -- -- -- -- -- -- 2.4 2.4

* - Spray Systems XR8001 and XR8002 flat fan nozzles aimed forward and back at 30 degrees

TD01 - Turbo drop nozzles from Green Leaf Industry aimed forward and back at 30 degrees, 0.1 gpm

TJ8002 - Twin Jet 8002 nozzle sprays forward and back at 60 degrees from horizontal, 0.1 gpm orifices

** - Single XR8002 nozzle spraying straight down

HC6-70 - Hollow cone nozzles aimed forward and back at 30 degrees

Converting a ground sprayer from single nozzles aimed straight down to double nozzles aimed forward and back at 30 degrees from horizontal will greatly increase the fungicide retention on the wheat head. Increasing the volume of spray solution applied per acre increases head coverage even with the improved nozzle system. Large yield increases were documented even with the low head coverage obtained with current spraying methods. Further research on natural or artificial wind to move the spray droplets horizontally or other methods to increase head coverage to a level which will assure good control of Fusarium Head Blight needs to be done.

1. Effect of Fungicides and Sprayer Nozzles on Control of Fusarium Head Blight in Wheat, Langdon, North Dakota, 1998.

Table 1. Effect of Fungicides and Spray Nozzles on Fusarium Head Blight, Langdon, ND, 1998

Head Blight Incidence Head Blight Severity Field Severity Flag Leaf Necrosis Dye Coverage of head Yield Test Weight

% % % % % bu/a lb/bu

Nozzles Averaged over Fungicides

XR8001 68.7 23.3 16.7 24.2 1.2 52.6 57.2

TD01 64.8 20.5 13.9 21.7 1.7 55.6 57.6

TJ8002 62.3 20.6 13.3 17.7 -- 54.8 57.5

lsd 5% ns ns ns ns ns ns ns

Fungicides Averaged over Nozzles

Tilt 4 fl oz/a 71.1 20.7 15.3 25.9 1.3 51.1 56.9

Folicur 4 fl oz/a 58.1 20.3 13.1 15.6 0.9 60.6 57.9

Benlate+Mancozeb 0.5 + 1.0 lb/a 70.5 22.3 16.2 26.0 1.3 52.3 57.7

Quadris+Benlate 12.5 oz + 1.0 lb/a 60.5 21.6 13.7 15.4 2.1 53.9 57.5

Quadris 12.5 fl oz/a 66.1 22.3 14.9 23.1 1.7 53.8 57.3

The nozzle by fungicide interaction was non-significant for all variables.

Table 2. Effect of Fungicides and Spray Nozzles on Fusarium Head Blight, Langdon, ND, 1998

Fungicides Rate Nozzle GPA PSI Head Blight Incidence Head Blight Severity Field Severity Flag Leaf Necrosis Dye Coverage of head Yield Test Weight DON

%%%%%bu/alb/buppm

untreated none -- -- -- 85.8 24.5 41.7 21.8 0.1 40.8 56.1 10.3

Tilt 4 fl oz/a XR8001 18 60 65.8 21.2 24.6 15.1 1.6 49.3 56.8 8.7

Tilt 4 fl oz/a TD01 18 60 80.0 21.4 37.1 17.6 1.0 52.3 56.6 --

Tilt 4 fl oz/a TJ8002 18 60 67.5 19.7 16.1 13.2 na 51.5 57.4 --

Folicur 4 fl oz/a XR8001 18 60 60.0 26.1 19.4 16.8 0.6 60.4 58.0 4.8

Folicur 4 fl oz/a TD01 18 60 54.2 16.2 15.4 9.7 1.1 63.1 58.2 --

Folicur 4 fl oz/a TJ8002 18 60 60.0 18.7 12.1 12.7 0.5 58.2 57.5 --

Benlate+Mancozeb 0.5 + 1.0 lb/a XR8001 18 60 74.2 23.1 38.8 18.5 0.7 50.1 57.5 6.1

Benlate+Mancozeb 0.5 + 1.0 lb/a TD01 18 60 62.2 21.8 18.9 13.9 1.8 56.9 58.0 --

Benlate+Mancozeb 0.5 + 1.0 lb/a TJ8002 18 60 75.0 22.1 20.4 16.3 na 50.0 57.6 --

Quadris+Benlate 12.5 + 1.0 XR8001 18 60 70.0 25.0 14.6 17.5 1.5 50.8 56.8 7.9

Quadris+Benlate 12.5 + 1.0 TD01 18 60 63.3 21.7 18.2 14.5 2.7 53.8 57.8 --

Quadris+Benlate 12.5 + 1.0 TJ8002 18 60 48.3 18.2 13.4 9.1 1.2 57.2 57.8 --

Quadris 12.5 fl oz/a XR8001 18 60 73.3 21.1 23.4 15.6 1.6 52.5 57.2 5.9

Quadris 12.5 fl oz/a TD01 18 60 64.2 21.7 19.2 14.0 1.8 51.8 57.4 --

Quadris 12.5 fl oz/a TJ8002 18 60 60.8 24.2 26.7 15.2 na 57.1 57.2 --

LSD 5% 13.6 ns ns ns 1.0 10.0 1.1 2.1

C.V. % 21.5 28.7 43.1 62.4 57.5 11.3 1.2 16.0

na - data lost to incorrect camera settings

ns - non-significant

Disease control, dye coverage of the head and yield for spray nozzles averaged over fungicides were not significantly different, Table 1. The interaction between nozzles and fungicide treatments was also non-significant. No difference was detected in disease control between fungicides averaged over spray nozzles. The Quadris + Benlate tank mix had significantly higher dye coverage on the head than the Folicur treatments, while other coverage comparisons were non-significant. The mean of Folicur treatments had higher grain yield than all other fungicides. Folicur also had a significantly higher test weight than the Tilt or Quadris treatment means.

The fungicide treatments compared to the untreated had no significant differences in visual fusarium infection of the head or in field severity or foliar disease, Table 2. The foliar disease was not severe in 1998 and the irrigation may have allowed excessive fusarium spread in the head. The fungicide treatments in which FHB Incidence was not significantly lower than the check were Tilt when applied with the TD01 nozzle, Benlate + Mancozeb with either the XR8001 or TJ8002 nozzles and Quadris applied with the XR8001 nozzle. The Folicur and Quadris + Benlate treatments had significant reductions in FHB Incidence with all nozzles. DON was measured only for the XR8001 nozzles. The Folicur, Quadris and Benlate + Mancozeb treatments had the lowest DON while Tilt was not significantly lower than the untreated. The spray coverage of the head data in this experiment are unexplainably low and do not correspond to the differences in yield. The Folicur treatments which had the highest yields had lower percent head coverage. Yield increases in the trial were up to 22 bushels per acre better than the check. The fungicide treatments which were not significantly higher in yield than the untreated were Tilt when applied with the XR8001 nozzle and Benlate + Mancozeb with either the XR8001 or TJ8002 nozzles.

2. Effect of Nozzle Type, Application Volume, and Adjuvants on Fungicide Effectiveness for Control of Fusarium Head Blight in Wheat, Walhalla, North Dakota, 1998.

Effect of Fungicide Application Parameters on Fusarium Head Blight, Wallhalla, ND, 1998

Nozzle Gallons per Acre Adjuvant PSI Head Blight Incidence Head Blight Severity Flag Leaf Necrosis Dye Coverage of head Yield Test Weight

% % % % bu/a lb/bu

untreated -- -- -- 28.0 7.0 65.0 0.2 43.0 53.7

XR8001 4.5 0 60 6.0 1.5 8.8 6.6 47.2 53.8

XR8001 9 0 60 8.0 1.1 18.8 4.4 57.7 53.3

XR8001 18 0 60 8.0 1.1 8.3 11.7 46.7 53.4

TD01 4.5 0 60 9.0 1.9 21.3 2.6 53.8 54.0

TD01 9 0 60 14.0 3.0 26.3 4.5 49.3 53.1

TD01 18 0 60 7.0 1.6 11.3 7.4 54.6 53.1

XR8001 9 CS7 60 3.0 0.6 7.0 5.0 53.8 54.6

XR8001 9 ND72 60 9.0 1.1 6.3 6.1 49.4 55.4

TD01 9 CS7 60 10.0 1.8 7.0 4.8 54.8 54.5

TD01 9 ND72 60 8.0 1.1 8.3 5.6 45.0 53.9

lsd 5% 4.1 1.8 10.6 3.0 8.4 ns

CV % 29.6 64.1 43.0 38.8 11.6 4.8

All fungicide treatments significantly reduced fusarium head blight incidence, severity and foliar disease when compared to the untreated. Increased spray volume increased head coverage for both the XR8001 and TD01 nozzles. The XR8001 nozzles had significantly higher head coverage than the TD01 at 4.5 and 18 gal/a but not at 9 gal/a. Coverage by the TD01 at 4.5 gal/a was not significantly different from the untreated. The addition of adjuvants increased coverage compared to no adjuvant but not significantly. Yield and test weight were not related to differences seen in the disease or coverage data though yields with fungicide tended to be higher. Treatments with CS7 added yielded significantly higher than the untreated while ND72 treatment yields were not different from the untreated.

Treatments were applied to flowering Russ hard red spring wheat July 24th. Application time was early morning with dew on the plants. Air temperature was 68 F, southerly wind 10-15 mph, and the dew point was 50 F during application. Folicur fungicide was applied at 4 oz product per acre. Adjuvants were added at 0.25% v/v. Treatments were applied with a tractor mounted 4 nozzle sprayer using CO2 for pressure. Eight nozzles were spaced 20" apart (2 each at 4 locations on the boom) and oriented forward and back 30 from horizontal. Two nozzle types were used. The hollow cone nozzle was a HC6 70 with a 0.1 gpm orifice at the 40 psi. The second nozzle was a Spray Systems XR 8001. The XR 8001 nozzle produced a MVD of 187 microns. Percent coverage was measured using a spray tracer dye (Day Glo blaze orange). A CCD camera that works under ultra violet low light conditions was used to measure dye on wheat heads. The picture was digitalized and stored in a computer. The picture was analyzed to separate dye coverage from leaf material and converted to percent coverage. Five heads per replication or 20 per treatment were analyzed for dye distribution. Flag leaf disease was visually determined at kernel soft dough stage. A RCB design with 4 replications was used for all treatments and evaluation parameters.

% Disease bu/A lbs/bu Coverage

1 untreated 57.8 53.9 58.7 0.1

2 XR 8001 ND72 50.5 64.6 58.5 4.6

3 XR 8001 CS7 72.0 60.1 57.8 7.2

4 XR 8001 COC 51.8 60.2 58.6 5.4

5 XR 8001 Ammonium Sulfate 46.0 61.5 58.6 0.3

6 XR 8001 28-0-0** 91.5 59.7 56.7 2.7

7 XR 8001 Active-It 45.8 62.4 58.3 1.7

8 HC 6-70 ND72 36.5 63.1 58.0 2.1

9 HC 6-70 CS7 37.5 65.5 58.8 2.9

10 HC 6-70 COC 58.1 60.7 57.9 2.6

11 HC 6-70 Ammonium Sulfate 50.5 56.5 58.6 3.1

12 HC 6-70 28-0-0** 93.5 45.3 57.6 2.1

13 HC 6-70 Active-It 44.3 64.2 58.6 1.4

LSD p>0.05 25.5 9.3 NS 1.7

C.V.% 31.0 11.2 2.2 43.1

**XR8001 nozzles with liquid nitrogen caused increased spike damage compared to HC 6 70 nozzles

Variable Flag Leaf Yield Twt Fungicide %

% Disease bu/A lbs/bu Head Coverage

Nozzle

XR8001 59.6 56.4 58.1 3.7**

HC 6 70 53.2 59.3 58.2 2.4**

Adjuvant

ND72 43.5 63.8 58.2 3.4

CS7 54.8 62.8 58.3 5.0

COC 54.4 60.7 58.2 4.0

Amonium Sulfate 48.3 59.0 58.6 1.7

28-0-0 92.5 37.5 57.1 2.4

Active-It 45.0 63.3 58.4 1.6

Nozzle NS + NS *

Adjuvant 18.4 6.0 NS 1.3

Nozzle x Adjuvant NS * NS *

C.V.% 31.7 10.2 2.0 41.6

*LSD p>0.05

+ LSD p>0.1

** XR8001 nozzles with Liquid Nitrogen caused increased spike damage compared to HC 6 70 nozzles

Flag leaf disease was not reduced by any treatment when compared to the untreated (Table 1). Two treatments using 28-0-0 adjuvant were signficantly higher for % leaf disease than the untreated. Much of this was due to leaf toxicity due to fertilizer salts. Some adjuvants significantly increased yields as compared to the untreated. ND72 adjuvant significantly increased yields at .05 level with 8001 nozzle and at 0.1 level with HC 6-70 nozzle. Latron CS7 and Active-It adjuvants significantly increased yield compared to the untreated at the .05 level when used with a HC 6- 70 nozzle. Using 28-0-0 with either nozzle significantly reduced yields compared to the untreated and any other treatment. Yield increases up to 22 percent were recorded for the best treatment.

Percent head cover was highest when Latron CS7 was used with 8001 nozzles and was significantly higher than any other treatment. COC when used with 8001 nozzle gave significantly higher coverage than any other treatment except Latron CS7. However, better head coverage did not result in significantly different yields or twt compared to other treatment except the 28-0-0 treatments.

Percent head coverage with XR8001 nozzle was signficantly higher than HC 6- 70 nozzles. However, yield was higher (0.1 level) with HC 6-70 nozzles than XR8001 (Table 2).

Treatments were applied to flowering Munich Durum wheat July 10th. Application time was 11:00 to 12:00 a.m. in full sunlight. Air temperature was 81 F, southwest wind 10 mph, and the dew point was 70 during application. Latron CS7 adjuvant were added to folicur treatments at 0.25% v/v. Treatments were applied with a tractor mounted 4 nozzle sprayer using CO2 for pressure. Eight XR 8001 nozzles were spaced 20" apart (2 each at 4 locations on the boom) and oriented forward and back 30 from horizontal. Flag leaf disease was visually

Treatment Rate/A Gallons/A Fusarium Head Blight Flag Leaf Yield Twt

% % % Field % Disease bu/A lbs/bu

Incidence Severity Severity

Untreated 44.2 58.2 28.3 28.8 70.9 59.4

Tilt 4 fl oz 14.2 29.2 63.4 19.0 23.8 66.6 59.1

Folicur 4 fl oz 14.2 30.0 49.2 17.5 28.8 74.6 60.1

Benlate + Mancozeb 8 + 16 fl oz 14.2 20.9 48.6 11.2 43.8 69.2 59.3

Quadris 12.5 fl oz 14.2 17.5 31.9 9.0 23.3 72.9 58.6

Quadris + Benlate 12.5 + 8 fl oz 14.2 27.5 36.2 13.5 30.0 67.7 58.6

Tilt 4 fl oz 23.6 22.5 50.9 11.8 45.0 71.0 60.7

Folicur 4 fl oz 23.6 12.5 15.0 2.5 42.0 69.8 59.6

Benlate + Mancozeb 8 + 16 fl oz 23.6 15.8 37.1 6.9 11.0 69.7 58.0

Quadris + Benlate 12.5 + 8 fl oz 23.6 23.3 55.9 14.9 56.3 58.7 59.7

LSD p>0.05 15.1 25.5 11.7 NS NS 1.3

C.V. % 42.2 40.1 61.2 73.8 10.8 1.5

Treatment Fusarium Head Blight Flag Leaf Yield Twt

% Incidence % Severity % Field % Disease bu/A lbs/bu

Tilt 25.8 57.1 15.4 34.4 68.5 59.9

Folicur 21.3 32.1 10.0 35.4 72.2 59.9

Benlate + Mancozeb 24.6 43.4 11.4 36.9 67.2 59.5

Quadris 16.7 34.5 7.9 17.1 71.3 58.3

Quadris + Benlate 25.4 46.1 14.2 43.1 63.2 59.2

Gallons/A GPA NS NS + NS NS NS

+ significant at p>0.1

All treatments except Tilt and Folicur at 14.2 gpa significantly reduced percent field severity when compared to the check (Table 1). Folicur applied 23.6 gpa significantly reduced percent field severity compared to application at 14.2 gpa. Application volume did not affect performance of other fungicides. Mancozeb-Quadris applied at 23.6 gpa significantly reduced field severity compared to Tilt applied at 14.2 gpa.

Only Tilt at 23.6 gpa significantly increased twt compared to the untreated Mancozeb. Quadris was significantly lower in twt than the untreated. There were significant differences among treatments for twt.

There were no significant differences among treatments for percent flag leaf disease (Table 2). Quadris Benlate treatments were significantly lower for yield than folicur or Quadris alone. Quadris was significantly lower in twt than Tilt, Folicur or Benlate-Mancozeb.

Treatments were applied to flowering Russ hard red spring wheat July 25th. Application time was 10:00 to 11:30 a.m. in full sunlight. Air temperature was 67 F, west wind 10-15 mph, and the dew point was 48 during application. Folicur fungicide was applied at 4 oz product per acre. Latron CS7 adjuvant was added at 0.25% v/v. Treatments were applied with a tractor mounted 4 nozzle sprayer using CO2 for pressure. Eight nozzles were spaced 20" apart (2 each at 4 locations on the boom) and oriented forward and back 30 from horizontal. Three nozzle types were used giving droplet size variation at 60 psi. The turbo drop (TB01) nozzle was a Green Leaf Industry nozzle with a 0.1 gpm orifice. The other nozzles were a Spray Systems XR 8001 and XR 8002. The turbo drop produced MVD droplet size of 400 microns at 60 psi. The XR 8001 nozzle produced a MVD of 187 microns at 60 PSI and the XR 8002 produced a MVD of 240 microns at 60 psi. Percent coverage was measured using a spray tracer dye (Day Glo blaze orange). A CCD camera that works under ultra violet low light conditions was used to measure dye on wheat heads. The picture was digitalized and stored in a computer. The picture was analyzed to separate dye coverage from leaf material and converted to percent coverage. Five heads per replication or 20 per treatment were analyzed for dye distribution. Flag leaf disease was also visually determined at kernel soft dough stage. A RCB design with 4 replications was used for all treatments and evaluation parameters.

Flag Leaf Yield Twt Fungicide %

Trt Nozzle Pressure Adjuvant % Disease bu/A lbs/bu Head Coverage

Untreated none none none 31.3 59.4 60.4 0.1

Folicur XR 8001 60 none 4.0 66.6 61.0 15.8

Folicur XR 8001 60 none 6.3 56.1 59.8 2.9

Folicur TB01 60 none 6.0 62.2 61.0 7.3

Folicur XR 8001 80 none 7.8 64.5 61.4 6.7

Folicur XR 8002 80 none 7.5 61.8 59.5 2.9

Folicur TB01 80 none 9.8 62.7 60.4 3.7

Folicur XR 8001 60 CS 7 5.3 61.5 60.8 12.0

Folicur XR 8002 60 CS 7 5.8 66.2 61.4 2.8

Folicur TB 01 60 CS 7 7.3 67.5 60.3 3.6

Folicur XR 8001 80 CS 7 5.3 66.7 61.9 8.2

Folicur XR 8002 80 CS 7 11.0 63.4 61.4 2.6

Folicur TB 01 80 CS 7 4.3 65.5 62.7 3.1

Folicur XR 8002 20 CS 7 5.7 65.1 60.9 3.6

LSD p>0.05 NS NS NS NS

C.V.% 102.0 9.1 3.2 56.7

Flag Leaf Yield Twt Fungicide %

Nozzle % Disease bu/A lbs/bu Head Coverage

XR 8001 5.6 64.8 61.3 10.7

XR 8002 7.3 62.4 60.5 2.9

TB 01 6.8 64.6 61.1 4.4

LSD p>0.05 NS NS NS 2.3

C.V.% 59.1 9.4 3.1 54.9

Pressure Flag Leaf Yield Twt Fungicide %

(psi) % Disease bu/A lbs/bu Head Coverage

60 5.8 63.3 60.7 7.4

80 7.6 64.1 61.2 4.5

LSD > 0.05 NS NS NS +

C.V. % 58.4 9.4 3.0 83.1

+ Significant at p>0.1

No statistical differences occurred due to treatment for any parameter measured (Table 1). Variability was high in the experiment and removed significance even though there were relatively large reductions in leaf disease and yield increases as high as 14%.

Analysis of interactions did show a significant (.05 level) improvement of head coverage when using the XR 8001 nozzles compared to XR 8002 or TB01 nozzles (Table 2).

Application pressure of 60 psi was signficantly better than 80 psi for percent head coverage at the 0.1 level. However, other measurements were not different (Table 2).

Use of Latron CS 7 adjuvant did not give any significant differences among treatments (Data not shown).