Individual Pupae Research Articles

Energetic costs of raising brood in honey bee colonies are high, but heater bees are cheap

Little is known about the energetic costs to insects of raising young. Honey bees collectively raise young, or brood, through a series of complex behaviors that appear to accelerate and synchronize the timing of brood maturation. These include maintaining the brood nest at warmer and consistent temperatures (33–36 °C) and the exceptional activity of heater bees. Heater bees are a part of the larger group of nurse bees that care for brood by rapidly contracting thoracic muscles to generate high body temperatures, from 42 to 47 °C. Heater bees move among brood cells and display this behavior to regulate the temperatures of individual larvae and pupae. We constructed three sets of experimental hives to explore the energy costs of raising brood in general and the cost of heater bees specifically. One set was designed to estimate the numerical allocation of individuals to the heater bee task. The second set was designed to contain only brood, which eliminated foraging and allowed us to quantify stored honey use when rearing juveniles at 10 and 30 °C. The final set was used to measure the respiration rates and energy expenditure of individual bees displaying resting, walking, heating, and agitated behavior. By integrating honey used by brood-only experimental colonies with whole-colony measurements of honey storage in the literature, we estimated that raising brood costs colonies half of their annual energy budgets stored as honey, or approximately 43.7 ± 0.9 kg·yr−1. We estimated that roughly 2 % of individuals in a colony perform as heater bees. Respiration rates of heater bees (19 mW) were more than those of resting bees (8 mW) but similar to those of walking bees (20 mW) and about half of those that were agitated (46 mW). The energetic cost of heating was more than an order of magnitude lower than the reported values for the energetic cost of flying. By integrating data from our three experimental hives, we estimate that the annual cost of raising brood is relatively high. However, heater bee behavior and physiology may require only about 7 % of the annual honey stored by a colony.

Analysis of the bacterial communities associated with pupae and winged or wingless adults of Lipoptena fortisetosa collected from cervids in Italy.

The hippoboscid Lipoptena fortisetosa Maa, 1965 is a hematophagous ectoparasite of cervids that can bite humans. This fly is expanding its geographical range and is of concern for animal and human health since it can potentially harbour harmful microorganisms. This study was aimed at characterizing the bacterial communities of L. fortisetosa in its different life-cycle stages. Pupae and wingless adults were collected from cervids hunted in Tuscan-Emilian Apennines (central Italy) and pooled into groups of 10 by life stage (30 individual pupae; 1420 individual wingless adults). Winged flies were caught by sweep netting and separated into five pools of 10 insects. After DNA extraction, the bacterial content of each pool was analysed using 16 S metabarcoding. Results revealed that the composition and relative abundance of different taxa greatly differed in the three analysed groups. Wingless adults showed a high abundance of Bartonella (33.07%), which is almost absent in winged flies and pupae. Among the detected pathogens, four genera of concern for human health were found: Bartonella, Moraxella, Mycobacterium and Rickettsia. Interestingly reads similar to Bartonella bovis, Moraxella osloensis and Arsenophonus lipopteni Operational Taxonomic Unit (OTUs)were detected. These findings suggest the possible role of L. fortisetosa as a reservoir of pathogenic microorganisms, confirming the need for further investigation to ascertain its vectorial capacity.

Kinome Analysis of Honeybee (Apis mellifera L.) Dark-Eyed Pupae Identifies Biomarkers and Mechanisms of Tolerance to Varroa Mite Infestation

The mite Varroa destructor is a serious threat to honeybee populations. Selective breeding for Varroa mite tolerance could be accelerated by biomarkers within individual bees that could be applied to evaluate a colony phenotype. Previously, we demonstrated differences in kinase-mediated signaling between bees from colonies of extreme phenotypes of mite susceptibility. We expand these findings by defining a panel of 19 phosphorylation events that differ significantly between individual pupae from multiple colonies with distinct Varroa mite tolerant phenotypes. The predictive capacity of these biomarkers was evaluated by analyzing uninfested pupae from eight colonies representing a spectrum of mite tolerance. The pool of biomarkers effectively discriminated individual pupae on the basis of colony susceptibility to mite infestation. Kinome analysis of uninfested pupae from mite tolerant colonies highlighted an increased innate immune response capacity. The implication that differences in innate immunity contribute to mite susceptibility is supported by the observation that induction of innate immune signaling responses to infestation is compromised in pupae of the susceptible colonies. Collectively, biomarkers within individual pupae that are predictive of the susceptibility of colonies to mite infestation could provide a molecular tool for selective breeding of tolerant colonies.

Behavioral Genetics of the Interactions between Apis mellifera and Varroa destructor.

The western honeybee Apis mellifera exhibits a diverse set of adaptations in response to infestations by its most virulent disease-causing agent, the ectoparasitic mite Varroa destructor. In this study, we investigated the effect of honeybee pupae genotype on the expression of four host and parasite traits that are associated with the reproductive phase of the mite in the brood of its host. We first phenotyped cells containing bee pupae to assess their infestation status, their infestation level, the reproductive status of the mites, and the recapping of cells by adult workers. We then genotyped individual pupae with five microsatellites markers to compare these phenotypes across full sister groups. We found that the four phenotypes varied significantly in time but did not across the subfamilies within the colonies. These findings show that V. destructor mites do not differentially infest or reproduce on some particular honeybee patrilines, and that workers do not target preferentially specific pupae genotypes when performing recapping. These findings bring new insights that can help designing sustainable mite control strategies through breeding and provide new insights into the interactions between A. mellifera and V. destructor.

Iris Yellow Spot Virus Prolongs the Adult Lifespan of Its Primary Vector, Onion Thrips (Thrips tabaci) (Thysanoptera: Thripidae).

Iris yellow spot virus (IYSV) from the genus Tospovirus, family Peribunyaviridae, reduces yield in several crops, especially Allium spp. IYSV is primarily transmitted by onion thrips (Thrips tabaci), but little is known about how IYSV impacts the biology of its principal vector. In a controlled experiment, the effect of IYSV on the lifespan and fecundity of onion thrips was examined. Larvae were reared on IYSV-infected onions until pupation. Individual pupae were confined until adults eclosed, and the lifespan and total progeny produced per adult were monitored daily. Thrips were tested for the virus in reverse-transcriptase polymerase chain reaction using specific primers to confirm the presence of IYSV. Results indicated that 114 and 35 out of 149 eclosing adults tested positive (viruliferous) and negative (nonviruliferous) for IYSV, respectively. The viruliferous adults lived 1.1–6.1 d longer (average of 3.6 d) than nonviruliferous adults. Fecundity of viruliferous and nonviruliferous onion thrips was similar with 2.0 ± 0.1 and 2.3 ± 0.3 offspring produced per female per day, respectively. Fecundity for both viruliferous and nonviruliferous thrips also was significantly positively correlated with lifespan. These findings suggest that the longer lifespan of viruliferous onion thrips adults may allow this primary vector of IYSV to infect more plants, thereby exacerbating IYSV epidemics.

Impact of the parasitic wasp Nasonia spp. (Walker, 1836) Hymenoptera: Pteromalidae) on populations of bird blowflies Protocalliphora spp. Hough, 1899 (Diptera: Calliphoridae)

In this work we focused on the prevalence of infection of Nasonia spp in pupae of Protocalliphora spp collected from nests n of different avian hosts from different sites in Slovakia Our research was focused on the impact of this parasitoid wasp on bird blowflies under natural conditions We studied its occurrence which was of all nests where bird blowflies were present The consequent mortality of host populations was with the number of wasps localized in individual pupa estimated on almost The aim of this work was to show that parasitoid wasp Nasonia spp significantly affects populations of Protocalliphora spp under natural conditions and that they have important role in regulation of population of bird blowflies

Prevalence of common honey bee pathogens at selected apiaries in Kenya, 2013/2014

AbstractThe present study was part of a larger surveillance effort to identify the determinants of African honey bee health, and, particularly, to detect honey bee pathogens across Kenya, where 160 colonies were examined from 32 apiaries (five colonies/apiary). From each colony, 20 individual foragers, nurse bees, worker pupae, and drone pupae were sampled separately. These were organized as 30 foragers, 32 nurse bees, 28 worker pupae, and 10 drone pupae pools. Nucleic acid was extracted from the pooled homogenates and tested using a panel of 18 different (RT-)PCR methods targeted at detectingPaenibacillus larvae,Melissococcus plutonius,Ascophaera apis,Aspergillusspp.,Nosema ceranae, N. apis,Deformed wing virus(DWV),Varroa destructor virus 1(VDV 1),Acute bee paralysis virus(ABPV),Sacbrood virus(SBV),Israeli acute paralysis virus(IAPV),Black queen cell virus(BQCV),Chronic bee paralysis virus(CBPV), andKashmir bee virus. All amplified bands were sequenced and compared to the GenBank database. VDV 1 was the most abundant virus at 50% prevalence in the 100 bee pools. It was closely followed by DWV at 44%. The others were BQCV (36%), SBV (14%), IAPV (9%), ABPV (8%), andN. ceranae(5%). The pathogens co-existed within apiaries. VDV 1 was present in 66% of the apiaries, DWV in 69%, BQCV in 69%, SBV in 28%, IAPV in 22%, ABPV in 19%, andN. ceranaein 13%. The study concludes that these pathogens should be incorporated in honey bee disease surveillance activities in the region.

Characterization of the Copy Number and Variants of Deformed Wing Virus (DWV) in the Pairs of Honey Bee Pupa and Infesting Varroa destructor or Tropilaelaps mercedesae

Recent honey bee colony losses, particularly during the winter, have been shown to be associated with the presence of both ectoparasitic mites and Deformed Wing Virus (DWV). Whilst the role of Varroa destructor mites as a viral vector is well established, the role of Tropilaelaps mercedesae mites in viral transmission has not been fully investigated. In this study, we tested the effects that V. destructor and T. mercedesae infestation have on fluctuation of the DWV copy number and alteration of the virus variants in honey bees by characterizing individual pupae and their infesting mites. We observed that both mite species were associated with increased viral copy number in honey bee pupae. We found a positive correlation between DWV copy number in pupae and copy number in infesting mites, and the same DWV type A variant was present in either low or high copy number in both honey bee pupae and infesting V. destructor. These data also suggest that variant diversity is similar between honey bee pupae and the mites that infest them. These results support a previously proposed hypothesis that DWV suppresses the honey bee immune system when virus copy number reaches a specific threshold, promoting greater replication.

Effects of X-ray irradiation on development and reproduction of the oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae)

All stages of the life cycle of Helicoverpa assulta were irradiated with X-rays to determine the inhibitory dose for development and reproduction to serve as a quarantine treatment. The 100-Gy dose was effective for irradiation of eggs and larvae, and the 200-Gy dose was effective for pupae and mixed-sex adults. When either adult males or females were irradiated, however, 500 Gy was required to prevent the F1 eggs from hatching, and thus single-sex adults required much higher doses of X-ray irradiation. To gather confirmatory data applicable to phytosanitary quarantine regulations, pupae—the immature stage most resistant to X-ray irradiation—were placed inside paprika in boxes for exportation and were irradiated with 300 Gy as a small-scale confirmatory test. The dose given to 1,007 individual pupae resulted in 12.62 % survival, and 1.79 % of pupae emerged as normal adults; however, these adults produced only a few eggs that did not hatch, suggesting that a minimum dose of 300 Gy of X-ray irradiation will provide quarantine security for immature H. assulta in paprika exports.

Pupicidal Activity of Ethanol and Water Extracts of <i>Phytolacca dodecandra </i> on <i> Anopheles gambiae</i> (Diptera Culicidae) Pupae

Pupae are an integral part of Anopheles gambiae life history and a tool that effectively targets them is likely to impact on malaria vector density. In this study we evaluated pupicidal activities of crude ethanol and water extracts of Phytolacca dodecandra (L’ Herit) on An. gambiae (Diptera: Culicidae) under laboratory condition. Individual early pupae of An. gambiae mosquitoes were exposed to concentrations of 40, 20, 10, 5, 2.5 mgs/100 ml of mature green fruits and leaves of the shoot and midsection of P. dodecandra in containers measuring 6 cm top × 5.7 cm bottom × 3.5 cm height. P. dodecandra was sourced from Eldoret (highland) and Nyando (lowland). Extraction of P. dodecandra bioactives was done using 80% ethanol and distilled water. Extracts of Neem leaves and untreated rain water were used as positive and negative controls respectively. WHO, 2005 mortality threshold of >80% was used to determine the effectiveness of the extracts as pupicide 24 hours post exposure. Mortality of exposed pupae was dose dependent. Concentrations of 20 mgs/100 mls and higher and 40 mgs/ 100mls of mature green fruits from Nyando and Eldoret met the mortality threshold respectively. Extracts of P. dodecandra from Nyando was more potent than that from Eldoret as was water compared to ethanol extracts. Positive (Neem) control killed < 80% of exposed pupae. It was concluded that crude extracts of P. dodecandra are potent as pupicide against An. gambiae mosquitoes and is a likely alternative to synthetic insecticides against An. gambiae in future.

Foraging Activity ofPalmistichus elaeisis(Hymenoptera: Eulophidae) at Various Densities on Pupae of the Eucalyptus DefoliatorThyrinteina arnobia(Lepidoptera: Geometridae)

Parasitoids are the main component of biological control of Lepidoptera defoliators in forested areas, and the densities of host insects can affect the efficiency of these natural enemies. The aim of this work was to study the biology and parasitism of Palmistichus elaeisis Delvare & LaSalle (Hymenoptera: Eulophidae) at various densities of females on individual pupae of Thyrinteina arnobia Stoll (Lepidoptera: Geometridae), the major Lepidoptera pest in eucalyptus (Myrtaceae) plantations in Brazil. Thyrinteina arnobia pupae were exposed to P. elaeisis females at various parasitoid-to-host ratios: (1:1, 3:1, 6:1, 9:1, 12:1, 15:1, 18:1, and 21:1), with 12 replications. The parasitism (81.8%) and emergence (100%) rates of P. elaeisis on T. arnobia pupae were highest with 15 ovipositing females per host. The life cycle (egg to adult) of P. elaeisis was shortest (19.8 ± 0.5 d) at a ratio of 21:1. The largest production of P. elaeisis per T. arnobia pupa was obtained at the ratios of 15:1 (298.4 ± 5.0) and 18:1 (287.4 ± 4.3). The female sex ratio of parasitoid offspring was highest (0.97 ± 0.01) with a parasitoid-to-host ratio of 12:1. The density of 15 P. elaeisis females per T. arnobia pupa was the most appropriate one for providing high parasitism (81.8%), emergence (100%), and progeny (298.4 ± 5.0) of this parasitoid. ResumoParasitoides sao os principais componentes de controle biologico de lagartas desfolhadoras em areas florestais e o numero de femeas por hospedeiro pode afetar a eficiencia desses inimigos naturais. O objetivo deste trabalho foi estudar a biologia e o parasitismo de Palmistichus elaeisis Delvare e LaSalle (Hymenoptera: Eulophidae) em diferentes densidades de femeas desse parasitoide por pupa de Thyrinteina arnobia Stoll (Lepidoptera: Geometridae), principal lepidoptero-praga em plantacoes de eucalipto no Brasil. Pupas de T. arnobia foram expostas ao parasitismo por femeas P. elaeisis nas seguinte proporcoes de parasitoides/hospedeiro: 1:1, 3:1, 6:1, 9:1, 12:1, 15:1, 18:1 e 21 :1, com 12 repeticoes. O parasitismo (81,8%) e emergencia (100%) de P. elaeisis em pupas de T. arnobia foram maiores com 15:1 parasitoides/hospedeiro. O ciclo de vida (ovo-adulto) de P. elaeisis foi mais curto (19,8 ± 0,5 dias) em uma proporcao de 21:1. A maior producao de P. elaeisis por pupa de T. arnobia foram obtidas nas densidades de 15:1 (298,4 ± 5,0) e 18:1 (287,4 ± 4,3), respectivamente. A razao sexual do parasitoide foi maior (0,97 ± 0,01), com 12:1. A densidade de 15 P. elaeisis femeas por pupa de T. arnobia foi o mais adequado por apresentar maior parasitismo (81,8%), emergencia (100%) e progenie (298,4 ± 5,0) do parasitoide. View this article in BioOne

Bumblebee pupae contain high levels of aluminium.

The causes of declines in bees and other pollinators remains an on-going debate. While recent attention has focussed upon pesticides, other environmental pollutants have largely been ignored. Aluminium is the most significant environmental contaminant of recent times and we speculated that it could be a factor in pollinator decline. Herein we have measured the content of aluminium in bumblebee pupae taken from naturally foraging colonies in the UK. Individual pupae were acid-digested in a microwave oven and their aluminium content determined using transversely heated graphite furnace atomic absorption spectrometry. Pupae were heavily contaminated with aluminium giving values between 13.4 and 193.4 μg/g dry wt. and a mean (SD) value of 51.0 (33.0) μg/g dry wt. for the 72 pupae tested. Mean aluminium content was shown to be a significant negative predictor of average pupal weight in colonies. While no other statistically significant relationships were found relating aluminium to bee or colony health, the actual content of aluminium in pupae are extremely high and demonstrate significant exposure to aluminium. Bees rely heavily on cognitive function and aluminium is a known neurotoxin with links, for example, to Alzheimer’s disease in humans. The significant contamination of bumblebee pupae by aluminium raises the intriguing spectre of cognitive dysfunction playing a role in their population decline.

Susceptibility of different life stages of Tribolium confusum to pyrethrin aerosol: effects of a flour food source on insecticidal efficacy

Accumulated grain dust and flour residues in flour mills can potentially decrease the efficacy of contact surface treatments and aerosol insecticides used for control of adult and immature stages of stored product insects. A study was conducted to evaluate the effects of flour residues on the efficacy of piperonyl butoxide-synergized pyrethrin aerosol against different life stages of the confused flour beetle, Tribolium confusum Jacquelin du Val. Twenty individual adults, pupae, or larvae of T. confusum, confined in Petri dishes containing 0, 0.1, 1, 5, or 10 g of wheat flour, were exposed separately inside an empty shed to the label rate of the aerosol. The formulation contained 1 % active ingredient pyrethrin and the label application rate was 1.04 ml/m3. After 2 h of exposure, all dishes were removed from the shed and placed in an incubator set at 27 °C and 60 % RH. Mortality of T. confusum adults decreased with increasing amount of flour. Recovery of moribund adults 15 days after exposure was greater in the 5 and 10 g flour dishes (15 and 46 %) compared to the 0.1 or 1 g flour dishes (0.7 and 5 %). As the flour amounts increased, more larvae and pupae were able to emerge as adults. Results show accumulated flour residues during aerosol application can compromise aerosol efficacy, and also suggest the flour impaired penetration ability of the aerosol. These results also emphasize the importance of sanitation and cleaning to remove spillage and extraneous material prior to an aerosol application.

How Varroa Parasitism Affects the Immunological and Nutritional Status of the Honey Bee, Apis mellifera.

We investigated the effect of the parasitic mite Varroadestructor on the immunological and nutritional condition of honey bees, Apis mellifera, from the perspective of the individual bee and the colony. Pupae, newly-emerged adults and foraging adults were sampled from honey bee colonies at one site in S. Texas, USA. Varroa‑infested bees displayed elevated titer of Deformed Wing Virus (DWV), suggestive of depressed capacity to limit viral replication. Expression of genes coding three anti-microbial peptides (defensin1, abaecin, hymenoptaecin) was either not significantly different between Varroa-infested and uninfested bees or was significantly elevated in Varroa-infested bees, varying with sampling date and bee developmental age. The effect of Varroa on nutritional indices of the bees was complex, with protein, triglyceride, glycogen and sugar levels strongly influenced by life-stage of the bee and individual colony. Protein content was depressed and free amino acid content elevated in Varroa-infested pupae, suggesting that protein synthesis, and consequently growth, may be limited in these insects. No simple relationship between the values of nutritional and immune-related indices was observed, and colony-scale effects were indicated by the reduced weight of pupae in colonies with high Varroa abundance, irrespective of whether the individual pupa bore Varroa.

Effect of host‐cocoon mass on adult size in the secondary hyperparasitoid wasp, Pteromalus semotus (Hymenoptera: Pteromalidae)

Abstract Parasitoids have long proven to be model organisms in studying resource‐related constraints on immature development. Here we examine the relationship between host cocoon (= pupal) size in the gregarious endoparasitoid wasp, Cotesia glomerata, and development time and adult size in the solitary idiobiont hyperparasitoid, Pteromalus semotus. Little is known about the biology or ecology of this ecto‐hyperparasitoid species, although it is one of the major secondary hyperparasitoids of C. glomerata. The size of the adult wasp covaried with the size of the host cocoon at parasitism. Moreover, female wasps were larger than male wasps for a given cocoon size. Adult wasps have remarkably long life‐spans, 3 months on average. Longevity did not significantly differ with sex. We also examined how larvae of P. semotus exclude other potential competitors. P. semotus is protandrous, with females taking significantly longer to complete their development than males. In experiments where several eggs of P. semotus were placed on individual pupae of C. glomerata, newly hatched hyperparasitoid larvae moved rapidly over the surface of the host and destroyed the eggs of any conspecifics by biting them before they would initiate feeding on host tissues. Our results are discussed in relation to those with other studies with solitary ichneumonid idiobiont hyperparasitoids of C. glomerata.

Recombinants between Deformed wing virus and Varroa destructor virus-1 may prevail in Varroa destructor-infested honeybee colonies

We have used high-throughput Illumina sequencing to identify novel recombinants between Deformed wing virus (DWV) and Varroa destructor virus-1 (VDV-1), which accumulate to higher levels than DWV in both honeybees and Varroa destructor mites. The recombinants, VDV-1(VVD) and VDV-1(DVD), exhibit crossovers between the 5'-UTR and the regions encoding the structural (capsid) and non-structural viral proteins. This implies that the genomes are modular and that each region may evolve independently, as demonstrated in human enteroviruses. Individual honeybee pupae were infected with a mixture of observed recombinants and DWV. A strong correlation was observed between VDV-1(DVD) levels in honeybee pupae and associated mites, suggesting that this recombinant, with a DWV-derived 5'-UTR and non-structural protein region flanking a VDV-1-derived capsid-encoding region, is better adapted to transmission between V. destructor and honeybees than the parental DWV or a recombinant bearing the VDV-1-derived 5'-UTR (VDV-1(VVD)).

Host size, superparasitism and sex ratio in mass-reared Diachasmimorpha longicaudata, a fruit fly parasitoid

We analyzed the relationship among host size, superparasitism and sex-ratio in mass reared Diachasmimorpha longicaudata (Hymenoptera: Braconidae). Individual host pupae of Anastrepha ludens (Diptera: Tephritidae) were measured (length and width), and the number of oviposition scars per pupa was used as a reliable indicator of superparasitism. The probability of an emerging parasitoid being a female was positively associated with the number of oviposition scars on the host cuticle, but not with the host size. The number of scars per host pupae from which females emerged was slightly but significantly higher than in those pupae giving raise to males. In D. longicaudata, the influence of host size on sex allocation decisions of individual females seems to be overridden by the level of superparasitism, which itself was positively correlated with pupa length. This suggests that larger pupae could experience a higher number of ovipositions than their smaller counterparts, and that a high level of superparasitism may conduct to a female biased sex ratio. We discuss the relevance of these findings which could provide new elements (e.g., the manipulation of superparasitism) for optimizing the mass rearing of this parasitoid.

Effects on microhymenopteran progeny of different host exposure periods (Chrysomya megacephala, Calliphoridae) to the parasitoid wasp Nasonia vitripennis (Hymenoptera: Pteromalidae)

To test the assumption that exposure of the host to parasitoid for long periods could provoke superparasitism, the aim of this work was to test the consequences on the immature development time, productivity of parasitoids per pupa, sex ratio and rate of parasitism of Nasonia vitripennis bred in Chrysomya megacephala pupae. Each individual pupa was placed in a glass tube with one parasitoid female for 24, 48, 72 and 96 h period of exposure, under controlled laboratory conditions. Twenty replicates of each period were performed. ANOVA with a 5% significance level was applied. The 72 h exposure caused the immature development time to increase. The mean parasitoids emergence per pupa did not vary significantly with the time of exposure. There were a significantly higher number of females than males and a trend in sex ratio deviation towards the females in all of the treatments. An increase in the number of unviable pupae rates were observed with increased exposure time.

Rapid Identification of Aedes albopictus, Aedes scutellaris, and Aedes aegypti Life Stages Using Real-time Polymerase Chain Reaction Assays

In 2005, a widespread infestation of Aedes albopictus was discovered in the Torres Strait, the region between northern Australia and New Guinea. To contain this species, an eradication program was implemented in 2006. However, the progress of this program is impeded by the difficulty of morphologically separating Ae. albopictus larvae from the endemic species Aedes scutellaris. In this study, three real-time TaqMan polymerase chain reaction assays that target the ribosomal internal transcribed spacer 1 region were developed to rapidly identify Aedes aegypti, Ae. albopictus, and Ae. scutellaris from northern Australia. Individual eggs, larvae, pupae, and adults, as well as the species composition of mixed pools were accurately identified. The assay method was validated using 703 field-collected specimens from the Torres Strait.

SSR based linkage and mapping analysis of C, a yellow cocoon gene in the silkworm, Bombyx mori

AbstractThe yellow color of the cocoon of the silkworm Bombyx mori is controlled by three genes, Y (Yellow haemolymph), I (Yellow inhibitor) and C (Outer‐layer yellow cocoon), which are located on linkage groups 2, 9 and 12, respectively. Taking advantage of a lack of crossing over in females, reciprocal backcrossed F1 (BC1) progeny were used for linkage analysis and mapping of the C gene using silkworm strains C108 and KY, which spin white and yellow cocoons, respectively. DNA was extracted from individual pupae and analyzed for simple sequence repeat (SSR) markers. The C gene was found to be linked to seven SSR markers. All the yellow cocoon individuals from a female heterozygous backcross (BC1 F) showed a heterozygous profile for SSR markers on linkage group 12, whereas individuals with light yellow cocoons showed the homozygous profile of the strain C108. Using a reciprocal heterozygous male backcross (BC1 M), we constructed a linkage map of 36.4 cM with the C gene located at the distal end, and the closest SSR marker at a distance of 13.9 cM.