1. Introduction
The whitefly
Bemisia tabaci (Gennadius) (Hemiptera, Aleyrodidae), is one of the most economically damaging pest species of crop plants in Africa. It causes physical damage as well as transmitting many plant viruses [
1,
2]. Besides its importance with regard to cassava, the species complex is well known across the world as one of the most important global pests and has been reported on more than 1000 species belonging to over 100 plant families [
3].
Cassava (
Manihot esculenta Crantz) is an important subsistence crop throughout much of sub-Saharan African and is perceived as Africa’s future food security hope, due to its anticipated resilience to the effects of climate change [
4]. However, pests and plant diseases reduce cassava yields substantially, posing a threat to food security throughout the developing world [
5]. Two viral diseases that are spread by whiteflies cause important economic losses to cassava production. These are cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) [
6]. CMD is caused by cassava mosaic begomoviruses (CMBs) [
7] which are transmitted persistently by
B. tabaci [
8]. CMD occurs in all cassava-growing parts of sub-Saharan Africa [
9]. CBSD is caused by cassava brown streak ipomoviruses (CBSIs) [
10], which
B. tabaci transmits in a semi-persistent manner [
11,
12]. Although the distribution of CBSD is confined to coastal East Africa and parts of Central Africa, it is spreading rapidly through the Democratic Republic of Congo and poses a great threat to cassava production zones of global significance in West Africa [
13,
14]. CMD causes losses through reducing plant growth that results from the extensive chlorosis that the viruses cause [
9]. CBSD infection usually causes mild vein-associated chlorosis on lower leaves and can cause stem die-back in severe cases. However, the most common cause of yield loss is the dry, brown, necrotic rot that occurs in the maturing tuberous roots of infected plants [
15]. Overall, these viruses affect more than half of all cassava plants in sub-Saharan Africa and cause annual losses of more than USD 1 billion [
16,
17].
Different methods have been used worldwide to control whitefly, for example, the use of insecticides and biocontrol using natural enemies such as parasitoids. The most widely used conventional insecticides are the neonicotinoids such as imidacloprid. Imidacloprid has been used successfully to manage
B. tabaci and whitefly-borne geminiviruses on tomato in south Florida and elsewhere [
18,
19]. It is relatively effective in controlling
B. tabaci on cassava, when applied either as a soil drench or foliar spray. However, the overuse of neonicotinoids presents a strong risk of cross-resistance between various chemicals of this group and threatens their effectiveness [
20,
21,
22]. Increasing numbers of resistance cases to neonicotinoids (e.g., imidacloprid and thiamethoxam) have been documented [
23,
24]. Recently, it has been reported that flupyradifurone, a new insecticide that belongs to the butenolide class, is effective in reducing levels of virus transmission by
B. tabaci MED species in tomato [
25]. Flupyradifurone, registered name Sivanto (SL 200), belongs to Bayer Crop Science’s own chemical class of butenolides (Bayer AG, Monheim, Germany). It is a systemic insecticide, with flexible modes of application, and is mainly intended for control of sucking pests such as aphids, hoppers and whiteflies. Flupyradifurone’s mode of action is by binding to the insect’s nicotinic acetylcholine receptors (nACHRs), disrupting the nervous system, and thereby causing muscle paralysis and subsequent death of the treated insects [
26]. In practical conditions, flupyradifurone can be considered safe to most beneficial insects including pollinators after having passed tests for its side effects on beneficial arthropods. It is currently approved and registered in Europe by the European commission in accordance with regulation EC 1107/2009, while in East Africa, it is registered in Kenya [
27] and Tanzania [
28].
A study was carried out by [
25] to test flupyradifurone for its efficacy in controlling whiteflies and their transmission of tomato yellow leaf curl virus in comparison to a neonicotinoid insecticide, thiamethoxam. After foliar application at recommended label rates under greenhouse conditions, it suppressed virus transmission by 85%, while levels of suppression after thiamethoxam treatments were just 25% and significantly lower in untreated plots, where 100% of the plants were infected by virus [
25]. A study carried out to evaluate field-collected populations of
B. tabaci MEAM1 response to flupyradifurone in comparison to imidacloprid, thiamethoxam and dinotefuran showed some level of cross-resistance [
29].
Bemisia tabaci is a cryptic species complex with the cassava-colonizing type designated based on mitochondrial DNA cytochrome oxidase I (
COI) sequencing as sub-Saharan Africa (SSA), which consists of five groups (SSA1-5), with SSA1 divided into sub groups (SSA1-SG1 to SG5) [
30,
31]. Recently, six major haplogroups of cassava
B. tabaci were defined (SSA-ECA, SSA-ESA, SSA-CA, SSA-WA, SSA2, SSA4) based on the much more genetically informative SNP genotyping [
32].
Bemisia tabaci cryptic species are known to respond differently to insecticides, with some such as MEAM1 and MED developing resistance to a wide range of current chemistries [
33]. The cryptic species that is predominant in the study area is haplogroup SSA-ESA (mitotype SSA1-SG3) [
31,
32]. In cassava, one recent study reported that imidacloprid applied through cutting dips at planting followed by varying spray intervals was effective at reducing whitefly populations, and significantly reducing cassava yield loss by up to 60% [
34]. In spite of this result, there are currently no effective management strategies being applied for
B. tabaci control on cassava in sub-Saharan Africa. A study was therefore conducted to assess the efficacy of the novel product flupyradifurone with the aim of determining whether or not this novel ‘soft chemistry’ insecticide might have the potential to be incorporated into integrated pest and disease management strategies for whitefly and whitefly-transmitted virus control in cassava. The findings from this study should provide information on the efficacy of flupyradifurone against cassava whiteflies but also, more specifically, on the
B. tabaci haplogroup SSA-ESA.
4. Discussion
The whitefly
Bemisia tabaci is polyphagous pest that causes damage to numerous crops through direct feeding and indirectly through transmission of plant viruses. The major control measure employed against the insect and reduction in virus damage in most crop production systems is the use of chemical pesticides. The occurrence of cryptic species that are known to differ in host plant and chemical response, and rapid insecticide resistance development to existing chemistries necessitates constant evaluation of novel pesticides against this pest [
33]. This study evaluated for the first time the efficacy of flupyradifurone (Sivanto SL 200), a novel chemistry pesticide belonging to the chemical class of butenolides, against cassava
Bemisia tabaci haplogroup SSA-ESA (mitotype SSA1-SG3), which is one of the cryptic species that specializes on cassava [
31,
32]. We employed a range of lab- and field-based approaches to evaluate the effectiveness of flupyradifurone in controlling cassava whiteflies and made comparisons with a current industry standard for sucking pests—the neonicotinoid imidacloprid.
The leaf disc assay and LC
50 tests showed that flupyradifurone caused high levels of mortality compared to untreated controls even at the lowest concentration tested, although this activity was not significantly different to that recorded for imidacloprid. Flupyradifurone caused 70% mortality within 24 h under laboratory conditions (leaf dip assays), rising to 96% after 96 h. Comparable results were obtained for imidacloprid (ranging from 63–92%). This mortality range for flupyradifurone is comparable to what has been reported in other studies that used different cryptic species that were in the approximate ranges of 60–95% [
25,
37]. LC
50 values recorded in our experiments for flupyradifurone (12.7 g a.i/100 L) and imidacloprid (12.6 g a.i/100 L) are slightly higher than those recorded during tests of other whitefly-control products [
38,
39], but this is in part a result of the longer durations used between application and mortality recording for these two studies and different whitefly stages considered.
Screenhouse cutting dip experiments demonstrated high levels of whitefly mortality for plants generated from cuttings soaked for 60 min as opposed to 15 and 30 min. An average of 75% fewer whiteflies settled on flupyradifurone-treated plants compared to the control within 1 day of introduction, and no whiteflies settled on subsequent days for up to 5 days. Imidacloprid had 36% fewer whiteflies settling compared to the control within 1 day and still had whiteflies present on treated plants up to day 5. These findings demonstrate that flupyradifurone is systemically active when administered through soaking for a longer duration than imidacloprid. Field studies testing efficacy through spraying demonstrated large contrasts in whitefly abundances between seasons, and consequently different patterns of effectiveness for application of the two insecticides. There were few whiteflies during the long ‘Masika’ rainy season (March planting), and insecticide treatment effects were therefore not pronounced. This was also accompanied by very low incidences of CMD (<12%), which were observed only in the control treatment of the most susceptible variety Kiroba. Similarly, the highest CBSD incidence of 26% was recorded from the control treatment of the CBSD-susceptible variety Albert. Variety Mkuranga1 did not have any symptoms of CMD or CBSD during the Masika season. Because of low whitefly abundances and similarly low incidences of diseases caused by whitefly-transmitted viruses, there were no measurable yield benefits accrued in the plots treated with either of the two insecticides. The absence of either CMD or CBSD in any treatments of variety Mkuranga1 also highlights the importance of virus resistance as a strategy for managing whitefly-borne viruses.
There were greater abundances of whiteflies during the Vuli season. This included significantly more
B. tabaci adults on Kiroba and Albert in the untreated control than on Mkuranga1 from 2–4 MAP. Similarly, in the cutting dip experiment, abundances of whitefly adults in the control treatment were greater on Albert than those on Mkuranga1 from 1–3 MAP, and there were more nymphs on Albert than on Mkuranga1 at 1 MAP and 6 MAP. These results suggest that Mkuranga1 is less suitable as a host for
B. tabaci than other cassava varieties, an observation that confirms results obtained from a study in coastal Tanzania that did not include any whitefly-control measures, in which Mkuranga1 had the lowest whitefly adult abundance of the seven varieties evaluated [
40]. The performance of Mkuranga1 suggests that resistant or tolerant varieties have an important role to play in management of both vectors and viruses, as noted elsewhere [
34,
41,
42,
43,
44].
Both insecticidal treatments were effective in reducing whitefly populations in each of the spray and cutting experiments. Differences between whitefly abundance in insecticide-treated and control plots were greatest during the earlier assessments (1 MAP and 2 MAP), which fell shortly after spray treatments. The knockdown assessments at 3 WAP and 6 WAP demonstrated the immediate potency of both imidacloprid and flupyradifurone, as whitefly abundances were reduced to close to zero after 24 h, whilst numbers in the control treatment increased. This result contrasts with the pattern observed in the 3 MAP and 4 MAP datasets in which there were generally no significant differences in abundance between control and insecticide treatments. Whiteflies are known to have a high degree of mobility within cassava fields [
45,
46]. Therefore, as insecticidal activity declines over time, it would be expected that short-distance whitefly adult movements between plots would lead to gradual reductions in whitefly abundance differences between plots.
The effectiveness of insecticidal treatments in controlling
B. tabaci whiteflies was reflected in reductions in incidence of whitefly-borne virus disease in both the Vuli spray trial and the cutting dip field experiment. These effects varied greatly, however, depending on the season, variety, and insecticide. Virus incidence reductions caused by insecticidal treatment were most apparent in the Vuli season (for both the spray and cutting dip trials) where whitefly abundances were higher. During Masika, when whitefly populations were low, there was little virus incidence, even in the control treatment. In the cutting dip trial, CBSD disease pressure coupled with high whitefly abundance was such that even insecticide-treated plots had incidences > 90% by 6 MAP. These patterns were also reflected in yield differences. Although there were no statistically significant yield differences between treatments in any of the three field trials, greatest differences in yields (for varieties Albert and Kiroba) occurred in the Vuli spray trial where virus disease incidence differences between treatments were greatest. Contrasts between the high virus disease pressure of the Vuli season and the low pressure in the Masika season were previously described by [
40]. This finding was used as the basis for a recommendation to encourage more farmers in coastal Tanzania to plant during the Masika season. The study by [
40] also highlighted the important and consistent difference in whitefly abundances between the Vuli (high abundance) and Masika (low abundance) seasons.
Flupyradifurone has been shown to be effective in controlling whitefly-transmitted viruses in other crop systems. For example, Tomato yellow leaf curl virus (TYLCV) infection levels were reduced by up to 85% in tomato [
25], Cucurbit yellow stunting disorder virus (CYSDV) transmission and spread were reduced in cantaloupe under greenhouse and field conditions [
47], and reductions in Tomato chlorosis virus (ToCV) transmission in flupyradifurone-treated tomatoes were attributed to reduced phloem activity of the virus [
48].
The superior performance of flupyradifurone in controlling cassava whiteflies when compared to imidacloprid, particularly in the cutting dip experiments, suggest that it has better systemic activity. Although no known studies are reported on the efficacy of flupyradifurone when used to dip planting material for propagation, a study with the neonicotinoids thiamethoxam and acetamiprid showed that this method is effective at controlling
B. tabaci whiteflies on poinsettia [
49]. Whitefly abundances were reduced by 62–88%, although there was no significant difference between foliar spraying and cutting immersion application methods [
49]. A study evaluating efficacy of various insecticides including flupyradifurone using foliar application and root drenching also showed no significant differences in whitefly mortality between the methods [
37]. A striking feature of our evaluation of flupyradifurone for cassava, however, is that a cutting single dip application prior to planting delivered sustained reductions in the abundance of whitefly adults and nymphs up to six months after planting, which in turn resulted in a 60% reduction in CMD incidence for variety Albert and an 81% reduction in CBSD incidence for Mkuranga1. In view of these promising station-based experimental results, there is clear value in testing this whitefly-control approach under on-farm conditions. To obtain the most realistic results, this would need to be conducted by comparing farms fully treated with flupyradifurone with untreated farms. The cutting dipping application of flupyradifurone, if effective under field conditions, could then be widely promoted to reduce cost and to avoid the deleterious effects that direct spraying has on non-target organisms. Although many smallholder cassava farmers in sub-Saharan Africa affected by whitefly-transmitted viruses primarily produce for subsistence purposes, there is an increasing number of commercially oriented producers. Applying effective whitefly control measures such as the systemic application of flupyradifurone or similarly effective insecticides may represent a worthwhile investment as part of a broader IPM strategy also including the purchase of certified planting material of disease tolerant varieties. Although the Masika season has low whitefly numbers, timing of planting may contribute less to control of cassava
B. tabaci, as the majority of farmers plant the crop during the Vuli season. This is attributed to cropping systems where the Masika season is dedicated primarily to cereal crops such as maize.