Host range evaluation of the leaf-feeding beetle,Zygogramma bicolorata and the stem-boring weevil,Listronotus setosipennis demonstrates their suitability for biological control of the invasive weed,Parthenium hysterophorus in Ethiopia

The invasive weed, parthenium (Parthenium hysterophorus L.) (Asteraceae: tribe Heliantheae), damages agriculture, adversely impacts biodiversity and is hazardous to human and animal health in Ethiopia. The host range of two natural enemies, a leaf-feeding beetle, Zygogramma bicolorata (Coleoptera: Chrysomelidae) and a stem-boring weevil, Listronotus setosipennis (Coleoptera: Curculionidae) was evaluated for biological control of the weed in Ethiopia. The specificity of Z. bicolorata and L. setosipennis was assessed against 29 and 31 non-target plant species, respectively. The host range of Z. bicolorata and L. setosipennis was first assessed using no-choice tests to examine their oviposition and feeding response on non-target plants. Although oviposition by Z. bicolorata occurred on six non-target species in four Asteraceae species in no-choice tests, it was significantly lower than on parthenium and no larvae developed. Zygogramma bicolorata nibbled the leaves of one of the five niger seed (Guizotia abyssinica L. – an oil seed crop closely related to parthenium) cultivars tested, but feeding and oviposition were significantly less than on parthenium. Furthermore, choice tests indicated that Z. bicolorata did not oviposit nor feed on G. abyssinica when parthenium was present. In no-choice tests, L. setosipennis did not oviposit on any of the non-target species assessed. Mean oviposition on parthenium was 39.0?±?3.4 eggs per plant whereas no eggs were laid on any of the 31 species tested. Based on these and other host range tests, permission was obtained to field release Z. bicolorata and L. setosipennis in Ethiopia.     

Stage specific feeding attributes and mobilization of nutrients in Mexican beetle,Zygogramma bicolorata Pallister on Parthenium hysterophorus L.

Parthenium hysterophorus L. (Asteraceae) is a toxic weed of agricultural farms, pastures and wastelands with a pan-tropical distribution. The weed causes a reduction in crop production of agricultural fields and severe health problems in humans. The Mexican beetle, Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae) is among the most promising candidates for the weed biocontrol. However, no previous study has evaluated assimilation of nutrients in this beetle. In the present study, feeding attributes and assimilation of nutrients by larvae of Z. bicolorata were assessed on the weed. Results revealed that the first larval instars of Z. bicolorata exhibited lowest consumption rates, and accumulated minimum concentrations of glucose, proteins and triglycerides in their body. They showed compensatory feeding, and displayed highest food utilization efficiencies and developmental rates. In contrast, the fourth larval instars exhibited higher food consumption rates and conversion efficiencies, but displayed lowest developmental rates. Accumulation of food reserves was also recorded maximum for the fourth larval instars. Overall, the findings may aid current mass-rearing efforts for Z. bicolorata in laboratories, but field trials are still needed to strengthen the present findings.     

Host range and risk assessment of Zygogramma bicolorata,a defoliating agent released in South Africa for the biological control of Parthenium hysterophorus

Parthenium hysterophorus (parthenium) is a weed of international importance and is spreading rapidly in sub-Saharan Africa. Consequently, it has been targeted for biocontrol in South Africa since 2003. Based on precedents elsewhere in the world, the defoliating beetle Zygogramma bicolorata was prioritised as a candidate agent. Although no-choice tests, involving some 48 test plant species, indicated a significant preference for parthenium, significantly reduced feeding and oviposition was recorded on some species. Multiple-choice tests resolved many of these non-target results; however, Helianthus annuus (sunflower) was still selected for oviposition and feeding. Of the 12 sunflower cultivars tested, four were selected for oviposition, while two were selected for oviposition and feeding. These six cultivars were then subjected to larval development trials, together with three native and two weed species (in the Asteraceae). These trials showed high levels of complete development on parthenium, significantly reduced development on sunflower cultivars, and partial development on only one of the weed species. Finally, a risk assessment was conducted on the six sunflower cultivars to quantify Z. bicolorata feeding and reproductive performance. Feeding risk calculations revealed these cultivars to have an extremely low risk (<0.2%) of supporting Z. bicolorata feeding and development. Similarly, reproductive risk calculations showed a very low risk (<0.16%) of supporting viable Z. bicolorata populations. These data are supported by findings from both the native (Mexico) and introduced ranges (Australia, India) of Z. bicolorata, where it has never been recorded as a pest of sunflower. These considerations were accepted by the regulatory authorities and in August 2013, Z. bicolorata became the second insect agent to be released in South Africa for the biocontrol of parthenium.     

Impact of defoliation by the biocontrol agentZygogramma bicolorataon the weed Partheniumhysterophorus in Australia

The leaf-feeding beetle Zygogrammabicolorata Pallister was introduced from Mexico intoAustralia in 1980 as a biocontrol agent for the weedParthenium hysterophorus L. (Asteraceae). Z. bicolorata became abundant in 1990, and since 1992there has been regular outbreaks resulting in thedefoliation of the weed in central Queensland. In thisstudy we evaluated the impact of defoliation by Z. bicolorata on P. hysterophorus from 1996 to1998. Z. bicolorata caused 91–100% defoliationresulting in reductions in weed density by 32–93%,plant height by 18–65%, plant biomass by 55–89%,flower production by 75–100%, soil seed-bank by13–86% and seedling emergence in the following seasonby 73–90%. At sites with continued outbreaks ofZ. bicolorata, it is expected that the existing soilseed-bank will be minimised, resulting in reduceddensity of parthenium in 6 to 7 years.     

Activity enhancement of Zygogramma bicolorata,a biocontrol agent of Parthenium hysterophorus,by temperature regulated diapause aversion

Diapause is a unique strategy of dormancy in insects to avoid unfavourable conditions. The exotic beetle Zygogramma bicolorata, is an effective biological control agent of Parthenium hysterophorus in India, Adults diapause in soil during December to May. As a result, there is delay in its effectiveness on the plant that reaches to flowering and seed production by the time the beetle is able to build up its population after emerging from diapause. Therefore, a study was conducted to explore possibilities of diapause aversion by temperature regulation. Results indicated that exposure of newly emerged adults to heat treatment of 35°C and to low temperature of 10°C could reduce diapause in Z. bicolorata. The low temperature can also be used as a medium for the storage of the mass reared beetles for a long time without having negative effect on their longevity and fecundity.     

Temperature and altitude modulate feeding attributes of Mexican beetle,Zygogramma bicolorata Pallister on Parthenium hysterophorus

Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae) is an effective biocontrol agent of Parthenium hysterophorus L. which is an alien invasive herbaceous weed with a pan-tropical distribution. The present study aimed to assess the effects of temperature and altitude on feeding attributes (consumption rate, conversion efficiency and growth rate) of adults from the wild populations of Z. bicolorata inhabiting India and Nepal. Results revealed that adults inhabiting areas of low temperature (24°C ‒ 25°C) and high altitude (415 m ‒1400 m) were large and had higher food consumption rates. In contrast, those inhabiting areas of high temperature (34°C ‒ 36°C) and low altitude (81 m ‒ 229 m) were smaller and had higher food utilization efficiencies. In all the eco-climatic regions, females were larger than males and had higher feeding attributes than their counterparts. Temperature between 27°C and 30°C was found optimal for Z. bicolorata adults to convert and utilize the food biomass to body mass. Above the optimal temperature the feeding attributes decreased. Present results suggest that there exists a possibility for decrease in body size, and thereby weed biocontrol efficiency of Z. bicolorata adults with an increase in temperature due to global climate change.     

Evaluation of Darwin's fecundity advantage hypothesis in Parthenium beetle,Zygogramma bicolorata

In the Parthenium beetle, Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae), variation in body size exists between and within the sexes. The females are larger than the males. Darwin (1874) proposed the fecundity advantage hypothesis, that is, large‐sized females produce more progeny, with subsequent studies supporting, as well as, refuting the hypothesis. Thus, in order to evaluate whether this hypothesis stands in Z. bicolorata we performed experiments to investigate the role of body size in influencing: (i) assortative mating; (ii) reproductive attributes; and (iii) growth, development and survival of offspring. It is the first attempt in this beetle. We found that size influenced assortative mating, reproductive output and offspring fitness. Larger males and females were preferred as mates over smaller ones. The pairs, having larger adults as mates, had higher fecundity, while the egg viability was influenced by the male size only. The offspring of larger parents had fast development and higher survival, indicating thereby possible better nutrient allotment by the female and supply of accessory gland proteins by the male in addition to better quality of genes.     

Rearing the Mexican beetle Zygogramma bicolorata (Chrysomelidae: Coleoptera) on a semi-synthetic diet

A semi-synthetic diet utilising parthenium leaf powder was evaluated for rearing the Mexican beetle Zygogramma bicolorata Pallister under in vitro conditions at 26±1°C, 65% RH. Significant differences were observed in the biological parameters of the beetle viz., the grub survival (54.3%), pupation (47.2%), fecundity (54.6/day) and total developmental period (37.7 days), when reared on the semi-synthetic diet compared to those when reared on natural parthenium leaves (91.5, 85.5, 74.8 and 29.8, respectively). The feasibility of utilising the semi-synthetic diet for mass production of the beetle is discussed.     

Post introductory risk assessment studies on Zygogramma bicolorata (Coleoptera: Chrysomelidae), a classical biological control agent of Parthenium hysterophorus (Asteraceae) in India

Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae), a biological control agent of Parthenium hysterophorus L. (Asteraceae), was feeding on leaves of Helianthus annuus in India, raising concerns of its host range. Based on the age grading technique, it was shown that the majority of Z. bicolorata on sunflower plants at any time were reproductively immature. This establishes that Z. bicolorata does not pose any risk to sunflower in India.     

Performance of a semiochemical‐baited autoinoculation device treated with Metarhizium anisopliae for control of Frankliniella occidentalis on French bean in field cages

The performance of an autoinoculation device was evaluated in field cage experiments for control of Western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), in French bean, Phaseolus vulgaris L. var. Samantha (Fabaceae). Treatments consisted of a fungus‐treated device with and without semiochemical (Lurem‐TR), and a fungus‐free device as control; the fungus used was Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae). The overall mean number of conidia acquired by a single insect was higher in the fungus‐treated semiochemical‐baited device than in the device without semiochemical: 5.0 ± 0.6 × 10⁴ vs. 2.2 ± 0.4 × 10⁴ conidia per insect. The overall thrips mortality was also higher in the semiochemical‐baited device than in the device without the semiochemical: 59.3 ± 3.9 vs. 41.7 ± 3.5%. Conidial viability was not affected in the M. anisopliae‐treated device without semiochemicals, 7 days after treatment, whereas it was considerably decreased – from 81.0 ± 1.3 to 6.5 ± 1.1%, 2 and 7 days post‐inoculation – in the M. anisopliae‐treated semiochemical‐baited device. Thrips mortality was positively correlated with M. anisopliae conidial viability, and conidial viability was negatively correlated with conidial acquisition. This suggests that the semiochemical volatiles reduced the conidial viability, which in turn resulted in significant reduction in thrips mortality, despite the highest conidial acquisition in the semiochemical‐baited fungus‐treated device. Thrips density per plant was significantly reduced in both fungus‐treated treatments (with semiochemical: 8.7 ± 1.7 adults per plant; without semiochemical: 6.6 ± 1.4 adults per plant) compared with the fungus‐free control (19.8 ± 2.6 adults per plant). These results demonstrate the potential for an autoinoculation device strategy for the control of thrips, particularly in screenhouses.     

Molecular phylogenetic analysis supports the synonymy of Prodontria modesta (Broun) and Prodontria bicolorata Given (Coleoptera: Scarabaeidae: Melolonthinae)

The validity of the synonymy of Prodontria modesta (Broun) and the nationally endangered Prodontria bicolorata Given, both flightless with very restricted distributions in Central Otago, New Zealand, was investigated using molecular phylogenetic analyses of a section of the mitochondrial gene region coding for cytochrome oxidase subunit 1 (COI) and nuclear rDNA internal transcribed spacer region (ITS1). Maximum likelihood analysis of the COI data found no separate, monophyletic lineages linked to colour form among six populations of the beetle and the genetic variation observed was less than that recorded within many other beetle species, including other melolonthines. The synonymisation of P. bicolorata with P. modesta should be considered valid and conservation efforts should focus on protecting habitats and populations, rather than on preserving colour forms.     

Review of the biology and ecology of a ragweed leaf beetle,Ophraella communa (Coleoptera: Chrysomelidae), which is a biological control agent of an invasive common ragweed,Ambrosia artemisiifolia (Asterales: Asteraceae)

A ragweed leaf beetle, Ophraella communa (Coleoptera: Chrysomelidae), has been highlighted as a potential biological control agent of Ambrosia artemisiifolia. O. communa and A. artemisiifolia are native in North America and alien species in East Asia and Europe. As an invasive weed, A. artemisiifolia causes severe economic losses as reducing agricultural production as well as producing severe allergenic pollen. As an herbivore insect, O. communa has strong host preference on A. artemisiifolia. All the developmental stages of O. communa can be found on A. artemisiifolia and it attacks a single plant in repeated and extended manners. With few individuals on A. artemisiifolia, O. communa can completely defoliate before pollen production. Therefore, O. communa had been focused as a biological control of this invasive weed, but its introduction was denied because of possible damage on an important crop, Helianthus annuus. O. communa was accidentally introduced in East Asia and Europe in 1990s and 2010s, respectively. Fortunately, O. communa population was well established to suppress A. artemisiifolia in the introduced areas. Following detailed field surveys and host specificity tests of O. communa were conducted and proved a strong potential of O. communa as a biological control agent of A. artemisiifolia. Moreover, O. communa has been investigated in physiological and evolutionary studies. In this study, the potential of O. communa as a biological control agent and a study organism are reviewed.     

Effects of Bacillus thuringiensis δ-endotoxins Cry1Ab and Cry1Ac on the coccinellid beetle,Cheilomenes sexmaculatus (Coleoptera,Coccinellidae) under direct and indirect exposure conditions

Genes encoding cry1Ab and cry1Ac δ-endotoxins from the bacterium, Bacillus thuringiensis (Bt) that have been incorporated in several crops to enhance their resistance to insect pests may possibly influence the activity and abundance of natural enemies of insect pests. The ladybird beetle, Cheilomenes sexmaculatus (L.) might ingest Bt toxins expressed by genetically modified plants by feeding on aphids, early instar larvae of lepidopterans, and other soft bodied insects feeding on transgenic plants. Therefore, we studied the effects of Cry1Ab and Cry1Ac Bt toxins on C. sexmaculatus under direct and indirect exposure conditions. For direct exposure, the neonate C. sexmaculatus larvae were fed either pure 2M sucrose (control) or sucrose solution containing Cry1Ab or Cry1Ac (0.1%), and on alternate days with aphids till pupation. Direct exposure of C. sexmaculatus larvae to Bt toxins resulted in reduced larval survival and adult emergence as compared to the controls, which might be due to long-term direct exposure. However, there were no adverse effects of the Bt toxins on C. sexmaculatus when the larvae were reared on Aphis craccivora Koch fed on different concentrations of Cry1Ab or Cry1Ac in the artificial diet. A significant and positive correlation was observed between the presence of Bt toxins in aphids, and coccinellid larvae and adults (r=0.53** to 0.86**). The results suggested that a direct exposure to Bt toxins expressed in transgenic plants or predation on H. armigera on Bt-transgenic plants will have little effect on the activity and abundance of the ladybird, C. sexmaculatus.     

Benefits of polyandry in Parthenium beetle,Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae)

The influence of polyandry on the reproductive performance of females and on offspring fitness in Zygogramma bicolorata Pallister was investigated using four experimental treatments, viz. (A) monandrous, limited mating, (B) monandrous, unlimited mating, (C) polyandrous, no-choice limited mating, and (D) polyandrous, mate choice unlimited mating. Polyandrous females had higher reproductive performance than monandrous ones. Monandrous females subjected to unlimited matings had higher egg viability than those subjected to limited matings, but fecundity did not differ significantly. In polyandrous females, the freedom to choose mates did not affect reproductive performance. However, offspring of polyandrous females allowed mate choice developed fastest and had the highest survival at 25, 27, and 30 °C. Thus, polyandry in Z. bicolorata appears to provide both direct (material) and indirect (genetic) benefits resulting in better reproductive performance and increased adaptability of the offspring to counter environmental stresses. The present study not only adds to the knowledge of reproductive biology of Z. bicolorata but it could also be of economic value as it may help in the mass rearing of Z. bicolorata and in the management of Parthenium hysterophorus.     

Host specificity of Metriona elatior, a potential biological control agent of tropical soda apple, Solanum viarum, in the USA

The leaf beetle Metriona elatior from Brazil-Argentina was screened in the Florida (USA)State quarantine facility as a potential biological control agent of tropical soda apple, Solanum viarum, a recently arrived weed species. Multiple-choice host-specificity tests were conducted in small cages (60 cm × 60 cm × 60 cm) using 95 plant species in 29 families. Adults fed heavily on the main target weed (S. viarum), and on turkey berry,Solanum torvum (noxious weed of Asiatic origin); fed moderately on red soda apple, Solanum capsicoides (weed of South American origin), and eggplant, Solanum melongena (economic crop); and fed lightly on aquatic soda apple, Solanum tampicense (weed of Mexican-Caribbean-Central American origin), and onsilverleaf nightshade, Solanum elaeagnifolium(native weed widely distributed). M.elatior adults laid 84 to 97% of their egg masses onS. viarum, and 3 to 16% on S. melongena. Non-choice host-specificity tests were also conducted in quarantine in which M. elatior adults and neonate larvae were exposed to 17 and 19 plant species, respectively. Tests with the neonates indicate that this insect was able to complete its development on S. viarum, S. torvum, S. melongena, and S. capsicoides. Although some adult feeding and oviposition occurred on S.melongena in quarantine on potted plants in small cages, no feeding or oviposition by M. elatiorwas observed in field experiments conducted in Brazil. Surveys in unsprayed S. melongena fields in Argentina and Brazil indicated that M. elatioris not a pest of S. melongena in South America. The evidence obtained from the South-American field surveys, Brazil open-field experiments, and Florida quarantine host specificity tests indicate that M. elatior causes significant feeding damage toS. viarum, and does not represent a threat to S. melongena crops in the USA. Therefore an application for permission to releaseM. elatior against S. viarum in the USA was submitted in October 1998. This revised version was published online in July 2006 with corrections to the Cover Date.     

Sympatric occurrence of three leaf beetle species of Macroplea Samouelle, 1819 (Coleoptera,Chrysomelidae, Donaciinae) in Finland with a key to species in Northern Europe

Macroplea Samouelle, 1819 is the only known fully aquatic leaf beetle genus with three European species that have earlier been classified by their assumed water salinity preferences. We studied the inter- and intraspecific variation of the specimens living in Northern Europe using both molecular (cytochrome c oxidase subunit I, COI) and morphological evidence. The variation in the COI sequences between M. mutica (Fabricius, 1792) and M. pubipennis (Reuter, 1875) was 8.4%–9%, M. mutica and M. appendiculata (Panzer, 1794) – 3.9%–4.9%, and M. appendiculata and M. pubipennis – 8.8%–9.2%. All three species were sampled together in the Bothnian Sea on the same water plants, showing that neither salinity nor plant species bear a decisive importance in their occurrence in the region. Phylogenetic results suggest the existence of two currently unknown Macroplea species that are evolutionarily close to M. appendiculata. A key to the Nordic species is provided.     

Slow and fast development in Parthenium beetle and its effect on reproductive attributes

The present study was designed to investigate the existence of slow and fast developing individuals and to evaluate their effect on reproductive attributes of Parthenium beetle, Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae); a biocontrol agent of Parthenium weed. Significantly different rates of development were found in a cohort of the beetle. Under constant rearing conditions, the emergence ratio of slow and fast developers was determined to be 0.47 and 0.53, respectively. Males developed faster than females, and significant differences in development were found among all immature stages and total developmental period. Slow developing females laid higher numbers of eggs with a higher percentage egg viability than fast developing females. This is the first attempt of such a study for the Parthenium beetle, and provides great insight into Parthenium beetle development for evolutionary biologists, improving our understanding of their ecological thresholds. The information could help in the selection of fast developing lines and thus in the mass multiplication of this bioagent.     

Electrophysiological and behavioural responses of the pollen beetle, Meligethes aeneus, to volatiles from a non-host plant, lavender, Lavandula angustifolia (Lamiaceae)

A semiochemical based push-pull strategy for control of oilseed rape pests is being developed at Rothamsted Research. This strategy uses insect and plant derived semiochemicals to manipulate pests and their natural enemies. An important element within this strategy is an understanding of the importance of non-host plant cues for pest insects and how such signals could be used to manipulate their behaviour. Previous studies using a range of non-host plants have shown that, for the pollen beetle Meligethes aeneus (Coleoptera: Nitidulidae), the essential oil of lavender, Lavandula angustifolia (Lamiaceae), was the most repellent. The aim of this study was to identify the active components in L. angustifolia oil, and to investigate the behaviour of M. aeneus to these chemicals, to establish the most effective use of repellent stimuli to disrupt colonisation of oilseed rape crops. Coupled gas chromatography-electroantennography (GC-EAG) and gas chromatography-mass spectrometry (GC-MS) resulted in the identification of seven active compounds which were tested for behavioural activity using a 4-way olfactometer. Repellent responses were observed with (±)-linalool and (±)-linalyl acetate. The use of these chemicals within a push-pull pest control strategy is discussed. Handling Editor: Yvan Rahbe.     

Maternal cannibalistic preferences affect offspring preferences and development in Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae): A transgenerational investigation

Cannibalism has been widely reported across taxa. However, the heritability and expression of cannibalistic traits have been least explored. The variation in the expression of cannibalism is likely to exist amongst the population affecting the propensity of cannibalism. Thus, to know whether the mother has any role in the transgenerational transmission of this trait in a ladybird beetle, Menochilus sexmaculatus, we studied the interaction between maternal and offspring prey preferences and its effect on, development duration and body weight of offspring over generations. An insignificant effect of maternal dietary history on offspring prey preference was observed across generations except for the non-cannibalistic adults who significantly preferred aphids over eggs. The long-term detrimental effect of cannibalism was found in cannibals with increased developmental duration and decreased body weight of offspring over generations. In conclusion, the results show that maternal diet did not affect the offspring preferences in M. sexmaculatus but cannibalism had a profound generational effect on the cannibalistic propensity, development and body weight of offspring across generations shows that larval dietary history and nutritional composition of prey contribute to the expression of cannibalistic behaviour across generations.