Ovary development and maturation in Nezara viridula (L.) (Hemiptera: Pentatomidae)

Ovary development and maturation of Nezara viridula (L.) were evaluated by examining ovariole morphology and the alterations in the biochemical (protein synthesis related to reproduction) composition of the hemolymph. Quantitative and qualitative protein analyses were performed and ovary structural alterations for the pre-reproductive and reproductive stages were recorded. Total concentration of proteins in female hemolymph gradually increased until the end of the pre-mating stage, remaining unaltered thereafter. Proteins linked to reproduction (vitellogenins) appeared in the hemolymph 10 days after adult emergence and indicated the end of the pre-mating stage. After mating, total protein concentration in the hemolymph was lower compared to virgin females; vitellogenin levels were similar during most of the observation period. Oocyte development and maturation were gradual and age dependent. Ten-day-old females had chorionated oocytes ready for fertilization. Mating did not stimulate oocyte development in N. viridula, but the lack of mating activity appeared to have stimulated oocyte resorption in 17-day-old females.     

Potential use of antibiotic to improve performance of laboratory-reared Nezara viridula (L.) (Heteroptera: Pentatomidae)

The antibiotic streptomycin added to the drinking water at a concentration of 125 mg/ l during nymphal development of Nezara viridula (L.) (Heteroptera: Pentatomidae)accelerated the development in ca. 2 days, increased survivorship, and doubled adult longevity; nymph survivorship and adult body weight were not affected when compared to control insects. Streptomycin has potential in rearing N. viridula, especially in improving quality of field-collected adults, by mitigating the introduction of pathogenic bacteria, and improving the quality of the population.     

Geographical distribution of genetically determined types of Nezara viridula (L.) (Heteroptera: Pentatomidae) in Brazil

Nezara viridula (L.) is a cosmopolitan and polymorphic pentatomid. Several genetically determined types have been described due to body color variation in adults. A survey covering 13 Brazilian states was conducted during 2001 and 2002 to determine the geographical distribution of the main types. Type G (smaragdula--body entirely green), the most common, showed a wide distribution, from south to north (latitude 2 degrees 49' N to 31 degrees 46' S), except in the Central-West Region. Type O (torquata--body green with lateral and median lobes of the head and anterior margin of the pronotum yellow), less abundant than the former, was more frequent at latitudes > 23 degrees 18' S and mean annual temperatures < 20.8 degrees C (Southern Region). Type Y (aurantiaca--body entirely gold or orange), which is rare, was collected only in the Southern Region. N. viridula (smaragdula) was captured in only one place in the Northern Region (Boa Vista, RR, latitude 2 degree 49' N). The abundance of the two most common types, smaragdula and torquata, was not correlated with altitude.     

Development of a dry artificial diet for Nezara viridula (L.) and Euschistus heros (Fabricius) (Heteroptera: Pentatomidae)

Artificial diets prepared with wheat germ, soybean protein, dextrosol, potato starch, sucrose, cellulose, soybean or sunflower oil, and vitamin solution for rearing Nezara viridula (L.) and Euschistus heros (Fabricius) were tested under controlled temperature (25 +/- 1 degrees C), RH (60 +/- 10%), and photophase (14h). Three diets were tested and compared with the natural diet privet [soybean and peanut seeds and privet Ligustrum lucidum Ait. fruit (Oleaceae)]. All three artificial diets allowed full development. The diet containing sunflower oil was the most suitable for N. viridula while E. heros developed better on a diet composed of soybean oil. Data indicated that the artificial diets were inferior to the natural diet. The artificial diets were more adequate for E. heros.     

Phylogeographic relationships of the southern green stink bug Nezara viridula (Hemiptera: Pentatomidae)

Abstract The sequences of 16S rDNA, cytochrome oxidase subunit I (COI) and cytochrome b (Cyt b) genes from nine field collections (seven provinces in China: Guangxi, Hubei, Guangdong, Guizhou, Zhejiang, Hunan, Jiangxi, and also southern and northern part of Iran), plus the sequences of Africa, Europe, Americas and Japan obtained form GenBank were used to re-analyse the genetic variation in the southern green stink bug Nezara viridula (Linnaeus). The phylogeographic re-analysis by using four algorithms (NJ, MP, ML and Bayesian) showed three main lineages. The Iranian haplotypes fell into lineage II formed from Europe and America, rather than in lineage III from Asia; the Chinese haplotypes fell into the Asian clade. Our results suggested that African and non-African gene pools have been isolated since the Miocene era with the molecular clock calibrations for Heteroptera mtDNA, and not since the Pliocene as mentioned previously. The corresponding age of the separation of the eastern and western Asia clades is estimated to be 4.0–1.6 million years ago, coinciding with the Pliocene–Pleistocene epoch and with acute rising events in the Tibet Plateau locates in the western China, which may have formed the barriers observed today.     

Studies on the neurosecretory system and retrocerebral endocrine glands of Nezara viridula Linn. (Heteroptera: Pentatomidae)

The neurosecretory system and retrocerebral endocrine glands of Nezara viridula Linn. have been described on the basis of in situ preparations and histological sections employing the paraldehyde fuchsin (PF) and performic acid-victoria blue (PAVB) techniques. In the brain of N. viridula, there are two medial groups–each consisting of five neurosecretory cells which belong to A-type. The lateral neurosecretory cells are absent. The axons of the two groups of medial neurosecretory cells (MNC) compose the two bundles of neurosecretory pathways (NSP) that decussate in the anterodorsal part of the protocerebrum. The two pathways, after the cross-over, run deep into the protocerebrum and deutocerebrum and emerge as NCC-I from the tritocerebrum. The nervi corporis cardiaci-I (NCC-I) of each side which are heavily loaded with NSM terminate in the aorta wall. Thus, the neurosecretory material (NSM), elaborated in the medial neurosecretory cells of the brain, is stored in the aortic wall and nervi corporis cardiaci-I (NCC-I). The NCC-II are very short nerves that originate from the tritocerebrum and terminate in the corpora cardiaca (CC) of their side. Below the aorta, but dorsal to the oesophagus, lie two oval or spherical corpora cardiaca. A corpus allatum (CA) lies posterior to the corpora cardiaca (CC). The corpora cardiaca do not contain NSM; only the intrinsic secretion of their cells has been occasionally observed which stains orange or green with PF staining method. The corpus allatum sometimes exhibits PF positive granules of cerebral origin. A new connection between the corpus allatum and aorta has been recorded. The suboesophageal ganglion contains two neurosecretory cells of A-type which, in structure and staining behaviour, are similar to the medial neurosecretory cells of the brain. The course and termination of axons of suboesophageal ganglion neurosecretory cells, and the storage organ for the secretion of these cells have been reported. It is suggested that the aortic wall and NCC-I axons function as neurohaemal organ for cerebral and suboesophageal secretions.     

Effect of relative humidity on emergence and on dispersal and regrouping of first instar Nezara viridula (L.) (Hemiptera: Pentatomidae)

Laboratory studies with 1st instar of southern green stink bug, Nezara viridula (L.) indicated that relative humidity (RH) greatly affected nymph emergence and survivorship up to the 2nd instar, reaching the maximum value (approximately 90%) with RH of > 80%. At 60% RH, 60% of the nymphs emerged and survived, while with 0% RH only approximately 15% of eggs hatched, and most nymphs died. Emerged nymphs from egg masses placed in plastic boxes with a gradient of humidity remained on egg shells for ca. one day. After this period, they dispersed and regrouped on top of shells 6.8 +/- 0.67 times, until they abandoned the shells toward the source of humidity, avoiding the water-saturated areas. Duration taken for each rearrangement (dispersal + regroup) increased with time, with a range of approximately 26 min to 44 min. The mean duration of the grouping behavior on egg shells after each rearrangement decreased from approximately 102 min (1st) to 24 min (6th and last grouping). The rearrangement behavior of 1st instars on top of egg shells apparently compensates for the water loss of nymphs.     

Feeding preference and movement of Nezara viridula and Euschistus serous (Hemiptera: Pentatomidae) on individual cotton plants

Experiments were conducted in an environmental growth chamber to determine the movement and feeding preferences of Nezara viridula (L.) and Euschistus serous (Say) on individual cotton plants. Fifth instars were caged by species on a single cotton plant (FM 9063 B2F) containing four discrete boll sizes ranging from 1.1 to 3.0 cm in diameter over a period of 5 d per replication. Two digital video cameras were simultaneously focused on each of the four bolls per plant to visually confirm stink bug resting and movement. During the study, a total of 4,080 h of video footage was recorded and analyzed. Results showed that N. viridula and E. serous did not prefer the exact same boll sizes. In a trial with eight stink bugs per plant, N. viridula spent more time on the three larger boll classes, 1.6-2.0, 2.1-2.5, and 2.6-3.0 cm. In a separate trial with one stink bug per plant, N. viridula spent more time on the larger boll classes while E. serous exhibited the strongest preference for 1.1-1.5 and 2.1-2.5 cm bolls. N. viridula moved more often than E. serous and both species moved more often during photophase compared with scotophase. Regardless of species or number of bugs released, bolls in the smallest boll size class fell off the plant about 3 d after the bugs were released. These results confirm that scouts who are estimating stink bug damage should select bolls in the 2.1-2.5 cm diameter boll size class.     

Possible egg positioning and gluing behavior by ovipositing southern green stink bug, Nezara viridula (L.) (Heteroptera: Pentatomidae)

From March 2002 to January 2004, a colony of southern green stink bug, Nezara viridula (L.), was kept in the laboratory, and its oviposition behavior was observed. During oviposition, soon after the egg is expelled, the female touches the egg mass with the dorsal surface of the last tarsomere; this movement of one leg of the hind pair occurred once. This small component of the oviposition behavior of this pentatomid is little known and not yet fully understood; it may help to position and glue the newly deposited egg to the others.     

Studies on the adult polymorphism in the southern green stink bug,Nezara viridula (hemiptera: Pentatomidae)

Polymorphism in adult colour pattern of Nezara viridula is determined on the genetic basis. The basic colour patterns of adult are classified into four types, i. e. G, O, R and F. No appreciable differences between these types were observed in respect to various physiological traits of nymphs and adults, except that type G seems to be superior in reproductive ability but to be inferior in ability of surviving winters at least to types R and F. Inter-generation changes in percentage frequency of G type were examined from 1959 to 1967 covering more than 30 generations. Percentage frequencies of G types prior to the severe winter of 1962–3 fluctuated greatly around a mean of 87.9%, while they did to a lesser extent than before with a mean of 85.0% after the winter. Unexpected high percentages of G type were recorded frequently in summer generations, viz. 1st and 2nd. On the other hand, the relative frequencies of G and O types decreased after hibernation in contrast to the increases in those of F and R. This sort of changes in genetic composition related to the winter of 1962–3 was observed in several populations segregated from each other. This seasonal alternation of selective activity in the environments is considered to be responsible for retention of the polymorphism. Alternative possible causes, i. e. difference in habitat preference, non-random mating and selective predation by predators among polymorphs, may safely be rejected as irrelevant to the mechanism in maintaining polymorphism. The polymorphism of this insect seems to be in a transient state rather than balanced one contributing little to population regulation, but the persistence of N. viridula in the periphery range may be assisted by retention of the polymorphism.     

Application of neem seed kernel extracts result in mouthpart deformities and subsequent mortality in Nezara viridula (L.) (Hem: Pentatomidae)

Abstract:  The effect of topical application of neem seed kernel aqueous suspension (NSKAS) and neem seed kernel hexane extract (NSKHE) on 2-day-old fourth instar nymphs of Nezara viridula (L.) was studied under laboratory conditions. The applications resulted in loosening of the proboscis, along with separation and deformation of mouthparts, predominantly in the stylets in fifth instar nymphs, adults and in inter-stadial moults. This rendered the insects incapable of feeding and led them to death. Application of NSKAS at 2.5% concentration led to 100% mortality, of which 40% was due to mouthpart deformities. Application of NSKHE at 2.5% concentration led to only 70% mortality, of which 3.33% was due to mouthpart deformities. At 5% concentration, mortality was again 100% for NSKAS and reached close to 100% for the NSKHE application. Dose-dependent mortality was observed for both the applications. However, dose-dependent mouthpart deformities were observed only for NSKAS, the highest being 60% at the 5% concentration. Moreover, for both the applications, more than 50% of the total mortality was observed up to the fifth instar at almost all the concentrations. The LC₅₀ values for NSKAS and NSKHE applications, for mortality up to the fifth instar were 0.36% and 5.22%, and for mortality to the adult stage (10 days old) they were 0.23% and 1.32% respectively. The percentage of bugs showing mouthpart deformities and toxicity was higher for NSKAS than NSKHE, and indicated that NSKAS was more potent than NSKHE.     

Susceptibility of pest Nezara viridula (Heteroptera: Pentatomidae) and parasitoid Trichopoda pennipes (Diptera: Tachinidae) to selected insecticides

Susceptibility of the southern green stink bug, Nezara viridula (L.) (Heteroptera: Pentatomidae), and its endoparasitoid Trichopoda pennipes (F.) (Diptera: Tachinidae) to acetamiprid, cyfluthrin, dicrotophos, indoxacarb, oxamyl, and thiamethoxam was compared in residual and oral toxicity tests. In the residual toxicity test, cyfluthrin, dicrotophos, and oxamyl were highly toxic to N. viridula. Thiamethoxam was moderately toxic to these insects. Each of the four insecticides was highly toxic to T. pennipes after prolonged tarsal contact with dried residues of these chemicals. In the oral toxicity test, where N. viridula fed on food covered with insecticide residues, none of the insecticides were toxic to adults of this stink bug, but acetamiprid, dicrotophos, and thiamethoxam were moderately toxic to the nymphs. In the oral toxicity test, where N. viridula fed on a gel-food containing insecticides, cyfluthrin, dicrotophos, oxamyl, and thiamethoxam were highly toxic to this stink bug. In an oral toxicity test using contaminated sugar water, all of the insecticides were highly toxic to T. pennipes. Because insecticides were as toxic, or more toxic, to T. pennipes than to N. viridula, it is extremely important to conserve this parasitoid by applying these insecticides for control of southern green stink bugs only when the pest reaches economic threshold.     

Assessing Nezara viridula (Hemiptera: Pentatomidae) feeding damage in macadamia nuts by using a biological stain

Damage caused by southern green stink bug, Nezara viridula (L.), to macadamia nuts, Macadamia integrifolia Maiden & Betche, is normally determined after nuts are harvested and processed, which may be many months after damage occurred in the field. We developed a method using ruthenium red dye to stain stink bug feeding probes and indirectly assess feeding activity in macadamia nuts. By using the staining method, feeding probes were easily detected on the husk, shell, and kernel. Husk probing was highly correlated (0.80-0.90) with feeding and damage to the kernel. Failure rate to detect kernel damage from stained husk probes was generally <6%. The staining method was equally effective for immature and mature nuts; therefore, N. viridula feeding activity can be monitored throughout the season to evaluate pest management tactics and forecast outbreak populations.     

Temporal processing of vibratory communication signals at the level of ascending interneurons in Nezara viridula (Hemiptera: Pentatomidae)

During mating, males and females of N. viridula (Heteroptera: Pentatomidae) produce sex- and species-specific calling and courtship substrate-borne vibratory signals, grouped into songs. Recognition and localization of these signals are fundamental for successful mating. The recognition is mainly based on the temporal pattern, i.e. the amplitude modulation, while the frequency spectrum of the signals usually only plays a minor role. We examined the temporal selectivity for vibratory signals in four types of ascending vibratory interneurons in N. viridula. Using intracellular recording and labelling technique, we analyzed the neurons' responses to 30 pulse duration/interval duration (PD/ID) combinations. Two response arrays were created for each neuron type, showing the intensity of the responses either as time-averaged spike counts or as peak instantaneous spike rates. The mean spike rate response arrays showed preference of the neurons for short PDs (below 600 ms) and no selectivity towards interval duration; while the peak spike rate response arrays exhibited either short PD/long ID selectivity or no selectivity at all. The long PD/short ID combinations elicited the weakest responses in all neurons tested. No response arrays showed the receiver preference for either constant period or duty cycle. The vibratory song pattern selectivity matched the PD of N. viridula male vibratory signals, thus pointing to temporal filtering for the conspecific vibratory signals already at level of the ascending interneurons. In some neurons the responses elicited by the vibratory stimuli were followed by distinct, regular oscillations of the membrane potential. The distance between the oscillation peaks matched the temporal structure of the male calling song, indicating a possible resonance based mechanism for signal recognition.     

The effect of vibratory disturbance on sexual behaviour of the southern green stink bug Nezara viridula (Heteroptera, Pentatomidae)

The effect of vibratory disturbance on sexual behaviour and substrate-borne sound communication of the southern green stink bug, Nezara viridula L. was studied. Disturbance signals do not change the time N. viridula males need to locate the source of vibratory signals, but decrease the number of males responding with the calling and courtship song to calling females. Female N. viridula proceed calling during stimulation with disturbance signals but some of them change the song rhythm by skipping one or more signal intervals or emitting the repelling signals. The number of females which change the dominant frequency of the calling song decreases proportionally with increasing differences between the dominant frequency of the disturbance signals and the emitted female calling song. Variation of the song dominant frequency probably serves females to avoid interference by increasing the signal to noise ratio. Signal duration and repetition rate do not change significantly when the female is stimulated with the disturbance signals. This indicates that frequency shift by calling females is the main strategy for reducing interference by competitive signalers in N. viridula vibrational communication.