Alfalfa weevil trial report, Intermountain REC

Ian Grettenberger, with cooperators Rob Wilson and Kevin Goding

Objective: Test a number of treatments for alfalfa weevil control and effects on natural enemies in
alfalfa

Methods
Location: Study was conducted at the Intermountain Research and Extension Center in Tulelake, CA
(41°57'50"N 121°28'08"W / 41.96389, -121.46889)

Plot layout: Plots were arranged in a randomized block design with seven treatments, four replications,
and linear blocks (1 x7 plots) arranged in a 4 x 7 grid.

Alfalfa variety: WL 363 HQ, 4-year-old stand

Crop management: Standard agronomic and weed management practices were followed for
management of this alfalfa stand. The only caveat to normal management was the crop did not receive
additional irrigation due to the water shortage in the Klamath Basin this last year.

Weather at application: 77 °F, 46% RH, wind 2-3 MPH

Application details: All treatments were applied on 2 June, 2021 starting at 10 AM. Applications were
made by foot with a CO2 powered backpack sprayer using a nine-foot handheld spray boom.
Applications were made at 20 GPA and 30 PSI with TeeJet XR8002 nozzles. All treatments included
Rainier-EA nonionic surfactant-activator-spreader at 0.25% v/v. Alfalfa was 18-in tall and pre-bud at
application.

Treatments: Steward and Warrior were both included as standards for weevil management. Pyrethroid
resistance is an issue in some areas of California but does not appear to be an issue in the Tulelake area
to date.
Trt # Product Rate Unit
1 Untreated 0 oz/ac
2 ISM-555 1.03 oz/ac
3 ISM-555 1.54 oz/ac
4 ISM-555 2.05 oz/ac
5 Endigo ZCX 4.5 oz/ac
6 Warrior II 1.92 oz/ac
7 Steward EC 11.3 oz/ac

Insects: The target pest for this trial was alfalfa weevil (Hypera postica). Insect sampling consisted of 10
180° sweeps per plot. Pre-treatment samples were taken at 0 DAT from the just outside of the plot area
(5 locations) to avoid disturbing the plots. Insect samples were taken 0, 7, 10, 14, and 21 DAT. The trial
was initiated after scouting had indicated that alfalfa weevil populations were building in the research
station’s fields. Insect samples were transferred into ethanol in the field and then identified and counted
in the laboratory. Alfalfa weevils and various natural enemies were counted in the lab.

Plants: Phytotoxicity was assessed in each plot at 7 DAT. Damage was visually assessed 7, 10, 14, and 21
DAT in each plot. Canopy cover (percentage) was assessed at two locations in each plot using the
Canopeo app at 14 and 21 DAT. For analysis, values were averaged. Yield was assessed using a plot
harvester at the first cutting timing on 23 June by harvesting ~25 ft of the plots, with a subsample
weighed wet and then dried to correct for percent moisture.

Analyses: All analyses were conducted using R and either linear models (for individual dates/plant
responses), linear mixed models (for insect data and to account for temporal correlation using a random
effect for plot), or Kruskal Wallis H tests (non-parametric, where appropriate). Posthoc comparisons
were made for linear models and within DAT where relevant with a Benjamini-Hochberg procedure to
adjust for multiple comparisons. Posthoc comparisons for Kruskal Wallis H test using Dunn tests. Data
were square root or log transformed as needed. Conducting separate analyses for each DAT for insect
data (linear models, treatment+block only) produced overall very similar results.
Results and Discussion
Throughout, results are presented using both bar and line graphs for the same data. Significant
differences between treatments are indicated only on the bar charts and are indicated using compact
letter displays. See associated text for additional information, although letters associated with the
legend indicate a non-significant treatment*date interaction and differences between treatments across
dates. No letters are used for dates on which analyses were not run because of insufficient data (early
dates for lady beetle larvae) or there were no significant differences.

Alfalfa weevils

Based on a linear mixed model, there was a significant effect of treatment across dates (χ26 = 354.16, P <
0.001), while there was no interaction between date and treatment. The consistent effect of treatment
across evaluation dates showed that all insecticide treatments displayed similar efficacy. All treatments
kept weevil populations below 2/10 sweeps, equating to at least 97% control for all treatments through
10 DAT. Weevil populations were increasing at the time of treatment and then began to decline 14 DAT
through 21 DAT.
Natural enemies

A number of different natural enemies were present in our sweep samples, including lady beetles
(adults and larvae, primarily Coccinella septempunctata), bigeye bugs (adults and nymphs, Geocoris),
damsel bugs (adults and nymphs, Nabidae), minute pirate bugs (primarily adults, Orius sp.), and various
parasitoid wasp species (not identified even to family level).

Lady beetles

Adult lady beetle populations decreased through 10 DAT at which point they began to increase. Dates
were analyzed separately. There was a significant interaction between treatment and date (χ218 = 44.11,
P < 0.001). At 7 DAT, all treatments significantly reduced lady beetle populations relative to the
untreated control. This effect continued through to 10 DAT. At 14 DAT, there was an effect of treatment,
although this was driven by the effect of Endigo, which had fewer lady beetles than all other treatments
(which were equivalent to the untreated). At 21 DAT, the effect of treatment was again significant, with
the most lady beetles in the untreated and variable numbers across other treatments. Endigo still had
the lowest populations.

Lady beetle larvae populations were nonexistent at the beginning of the trial, so analyses were not run
for 7 or 10 DAT. At 14 DAT, populations of larvae began to increase in the untreated plots and to a very
small degree, in the Steward plots. At 21 DAT, all treatments had fewer larvae than the untreated.

For both adults and larvae, aphid populations might explain differences later in the trial (as in a response
of lady beetles to aphid populations), but more for adults than larvae. We did not assess aphid
populations since they were not explicitly part of this trial. They did begin to increase at the last two
sampling dates. Endigo and Warrior plots appeared to have the fewest aphids (at a coarse look). By 21
DAT, at least some aphids were in all plots.
Orius adults

There was significant treatment*date interaction (χ218 = 40.76, P = 0.0016), with significant differences
among treatments for adult Orius populations at 7 and 10 DAT but not 14 and 21 DAT. On both early
dates, the untreated and Steward plots had the most Orius. Meanwhile, the Warrior and Endigo
treatments had the fewest, with the ISM-555 treatments intermediate. The treatments with
intermediate abundances at 7 and 10 DAT did not necessarily separate from other treatments. Patterns
were generally similar between 7 and 10 DAT. While not significant, the trend for both dates was that
ISM-555 treatments did not have as low of populations as the broad-spectrum Warrior and Endigo
treatments. At 10 DAT, several of the ISM-555 treatments had lower numbers than the untreated plots,
which had the most Orius adults.
Parasitoids wasps

Analyzed across dates using a linear mixed model, there was no significant interaction between date and
treatment (P > 0.05), but a significant effect of treatment (χ26 = 14.88, P = 0.021). The untreated plots
had the most wasps, the Warrior plots the fewest, and the remaining treatments intermediate. There
was an “outlier” at 10 DAT for the Endigo treatment wherein one plot had many wasps, which
influenced the results for Endigo.
Overall natural enemies

Effects were mixed on natural enemies and there were only a few taxa that could be analyzed
individually based on their abundance. Adult lady beetle adults were very affected within 7-10 days. At
the 14 day time point, aphid populations may have started to influence adult lady beetle populations vs.
the effect of insecticides alone. The effect of the treatments on lady beetle larvae at 14 DAT likely was a
lagged effect of the insecticides on the adults. There appeared to be some effects of the ISM-555
treatments on Orius, although they did not appear to be as pronounced as those of the broad-spectrum
materials. It appeared there might be some effects on parasitoid wasps for ISM-555, with counts
“intermediate” compared to the broad-spectrum materials and the untreated control (but not
statistically different).

Plants

Phytotoxicity

No phytotoxicity was noted in any of the plots during this trial.

Yield

There was a significant effect of treatment on yield (F6,18 = 4.74, P = 0.0046). The untreated, Steward,
and ISM-555 1.03 oz. treatments had the lowest yields. The two higher rates of ISM-555 (1.54 and 2.05
oz) had the highest yields.
Canopy cover

At 14 DAT, there was no effect of treatment on canopy cover (F 6,18 = 1.44, P = 0.25). At 21 DAT there was
no significant effect of treatment, although it was very close (F 6,18 = 2.24, P = 0.086).

Percent defoliation

For visual assessments of percent defoliation, there were no differences between treatments in the
estimated values for any of the plots through 14 DAT. During this time, all plots were estimated to have
5% defoliation damage. At 21 DAT, the untreated plots did have non-5% damage assessments, which
was statistically different than the other treatments (χ26 = 26.84, P < 0.001).

Relationship between canopy cover and yield

Given the nearly significant differences among treatments for canopy cover at 21DAT and the effect on
yield, we examined the relationship between these two variables using Pearson Correlation. There was a
significant positive correlation between these variables (t26 = 2.87, P = 0.0080). It is possible that weevil
numbers explained this variation, but that does not appear to be the case. Aphid populations may also
possibly explain the coverage patterns, along with the effect on yields, but a quick look at the samples
for a rough aphid assessment suggested this was not the case (ISM-555 plots did not have the fewest
aphids). It also seems unlikely that stand variability across plots would explain the differences/trends we
saw. This would have necessitated the two highest ISM-555 treatments getting randomly assigned to
some of the best-yielding plots.