Egg Viability Research Articles

Effect of Insect Growth Regulator Insecticides Novaluron, Teflubenzuron and Lufenuron on the Morphology and Physiology of Euschistus heros

The Neotropical brown stink bug, Euschistus heros (F.) (Heteroptera: Pentatomidae), is an important pest that damages soybean, especially in the central-west region of Brazil. The effect of insect growth regulator insecticides on fourth-instar nymphs and adults of the neotropical brown stink bug was evaluated under laboratory and greenhouse conditions, respectively. In the laboratory, the insecticides (doses in g a.i. ha-1) novaluron (20.0 and 40.0), teflubenzuron (26.2 and 52.5), and lufenuron (20.0 and 40.0), in addition to the control (water), were tested on fourth-instar nymphs of E. heros and their development were followed to adulthood. Mortality, number of adults with and without deformities, and fecundity were determined. In a greenhouse, the effects of the same insecticides on the adult of the stink bug were evaluated by determining the fecundity and viability of the eggs laid. The fourth-instar nymphs of E. heros, when exposed to either dose of the tested insecticides presented mortality and insects with deformations, as well as reduced fecundity of females that reached adulthood. In the trial with adult E. heros, all tested insecticides reduced stink bug fecundity. In the same way, the viability of the produced eggs was reduced in all the chemical treatments, except for novaluron at the lowest dose tested, in which egg viability did not differ from the control treatment. Based on the obtained results, one can infer that the growth-regulator insecticides evaluated, although usually more suitable for the control of caterpillars, can interfere negatively in the development and reproduction of the neotropical brown stink bug, thus constituting a complementary alternative for the management of this pest.

Effects of Tebufenozide on Eggs, Larvae and Adults of Chrysoperla carnea (Neuroptera: Chrysopidae).

Quantifying compatibility among control agents is essential for development of integrated pest management (IPM). Chrysoperla carnea (Siemens) and insect growth regulator insecticides are widely used in IPM of Lepidoptera. C. carnea is a generalist predator naturally present in the Mediterranean agroecosystems and bred in insectariums for commercial purposes. Here, we evaluated lethal and sublethal effects of tebufenozide on C. carnea under laboratory conditions. The treatment of eggs with tebufenozide 24 or 48 h after they were laid did not affect the hatching rate or survival of the neonate larvae. Toxic effects of tebufenozide on topically treated larvae was low; development times of surviving larvae and pupae decreased significantly compared with controls. In choice bioassays, a high percentage of third-instar larvae chose prey (Spodoptera littoralis) treated with tebufenozide in preference to untreated prey. Moreover, second-instar larvae of C. carnea that had previously consumed tebufenozide-treated prey (0.75 mL/L) had significantly reduced larval development time compared with controls, while longevity of surviving adults, fecundity and egg viability were unaffected. Ingestion of tebufenozide by adults of C. carnea at the recommended field dose had no significant effect on female fecundity, egg viability or adult longevity. Tebufenozide exhibited low toxicity towards the developmental stages of C. carnea and is therefore a candidate for inclusion in IPM strategies.

Lack of fitness costs in dsRNA-resistant Leptinotarsa decemlineata ([Coleoptera]: [Chrysomelidae]).

The Colorado potato beetle, Leptinotarsa decemlineata (Say) ([Coleoptera]: [Chrysomelidae]), is the most important defoliator of solanaceous plants worldwide. This insect displays a notorious ability in adapting to biological and synthetic insecticides, although in some cases this adaptation carries relevant fitness costs. Insecticidal gene silencing by RNA interference is a novel mode of action pesticide against L. decemlineata that is activated by ingestion of a double stranded RNA (dsRNA) targeting a vital L. decemlineata gene. We previously reported laboratory selection of a > 11,000-fold resistant strain of L. decemlineata to a dsRNA delivered topically to potato leaves. In this work, we tested the existence of fitness costs in this dsRNA-resistant colony by comparing biological parameters to the parental strain and an additional susceptible reference strain. Biological parameters included length of egg incubation period, number of eggs per clutch, egg viability, larval viability, length of larval and pupal periods, adult emergence, number of eggs laid per day, sex ratio, and adult longevity. Comparisons between the 3 beetle strains detected no fitness costs associated with resistance to dsRNA. This information is important to guide effective insect resistance management plans for dsRNA insecticides against L. decemlineata applied topically to potato leaves.

Selectivity of Bacillus thuringiensis (Bacillales: Bacillaceae) to the polyphagous predator Ceraeochrysa claveri (Navás, 1911) (Neuroptera: Chrysopidae)

AbstractBioinsecticides based on Bacillus thuringiensis (Bt) are widely used to control agricultural pests. However, there are concerns about their impact on non‐target organisms, with for example, the green lacewing Ceraeochrysa claveri, a polyphagous predator with important value in the biological control of insect pests in agroecosystems. This study aimed to analyse the effects of Agree® bioinsecticide (containing Bt serovar aizawai GC‐91) on the post‐embryonic development of C. claveri. For this, neonate larvae of C. claveri were fed with Diatraea saccharalis eggs contaminated with different concentrations of Agree® (0.62, 1.25, 2.50 and 5.00 g L−1) until their pupation. Samples of the midgut of larvae were collected for histological and ultrastructural analysis. The larval and pupal inviability data were collected at the end of the respective developmental stages. After the emergence of adult insects, were considered inviable those with morphological alterations. To evaluate the reproduction, adult insect couples were formed, and the oviposition was collected. No significant differences in the viability of eggs, larvae, pupae and adults of C. claveri contaminated with Agree® were reported. Midgut analyses did not reveal cellular damage in the epithelium, indicating a possible lack of insecticidal proteins receptors. The oviposition and their viability were similar between treated and control groups. Our results demonstrated that Agree® bioinsecticide did not affect the post‐embryonic development of C. claveri. This illustrates that the combined use of Bt and C. claveri for the biological control of arthropod pests can be a sustainable strategy for integrated pest management.

Effects of Fluctuating Thermal Regimes and Pesticides on Egg Hatching of a Natural Enemy Harmonia axyridis (Coleoptera Coccinellidae)

The harlequin ladybird, Harmonia axyridis, is a valuable asset in integrated pest management (IPM); however, issues related to low-temperature storage and transportation have resulted in low hatching rate, while the use of pesticides may lead to non-target effects against this natural enemy during field application. Fluctuating thermal regimes (FTR) have been shown to be beneficial during the low-temperature storage, and the type and concentration of insecticides used are crucial for field application of H. axyridis. Despite this, little research has been conducted on the effects of FTR on the hatching rate of ladybird eggs, and the impact of pesticides on their egg viability remains unclear. To address these gaps, we investigated the effects of different thermal temperatures, recovery frequencies (the number of changes in temperature conditions per unit time), and recovery durations (the duration of the treated temperature condition) on egg hatching under constant low-temperature conditions. We also examined the toxicity and safety of seven commonly used insecticides on egg hatching. Our results indicate that the temperature during FTR application did not significantly affect egg hatching, but the interaction between temperature and recovery frequency can significantly affect egg hatching. Moreover, the recovery frequency and recovery duration had a significant impact on hatching. Under specific conditions, the hatching rate of eggs subjected to FTR was similar to that of eggs stored at 25 °C. Furthermore, we found that matrine (a kind of alkaloid pesticide isolated from Sophora flavescens) had low toxicity to ladybird eggs and is a safe pesticide for use in conjunction with this natural enemy. The study provides valuable information on effectively managing H. axyridis by taking into account both storage temperature and pesticide exposure.

Toxic effects of synthetic and natural indigo dyes on Danio rerio reproduction

The textile industry releases significant amounts of toxic pollutants. In the aquatic environment, the presence of these compounds can impact fauna and flora. The aim of this study was to verify if synthetic (95% purity) and natural indigo blue dye obtained from Indigofera tinctoria plants pose a risk to the aquatic fauna. To investigate ecotoxicological effects of these dyes at concentration of 100 mg.L-1, adult females and males of Danio rerio were exposed in chronic tests. At the end of the twenty-first day, reproduction was performed to evaluate the endpoints: number of eggs, egg viability, larvae survival, and gonadal histology, and the same experiments were repeated after another 21 days in clean water to evaluate the potential of biological recovery. All of the analyzed reproductive parameters were significantly reduced in the organisms exposed to both synthetic and natural indigo blue dyes in relation to the control, and males exposed to the synthetic dye, and females exposed to both natural and synthetic dyes presented gonadal histological alterations, evidencing an important chronic effect of these substances to the reproduction of D. rerio. After the biological recovery period, organisms exposed to both types of dyes showed histological improvements and they recuperated offspring production, but those exposed to the natural dye showed better reproductive performance. Overall, our findings suggested that levels of natural and synthetic indigo dyes permitted by the current legislations must be reviewed, and the substitution of synthetic by natural indigo whenever possible is recommended.

Filial Cannibalism Leads to Chronic Nest Failure of Eastern Hellbender Salamanders (Cryptobranchus alleganiensis).

AbstractIn species that provide parental care, parents will sometimes cannibalize their own young (i.e., filial cannibalism). Here, we quantified the frequency of whole-clutch filial cannibalism in a species of giant salamander (eastern hellbender; Cryptobranchus alleganiensis) that has experienced precipitous population declines with unknown causes. We used underwater artificial nesting shelters deployed across a gradient of upstream forest cover to assess the fates of 182 nests at 10 sites over 8 years. We found strong evidence that nest failure rates increased at sites with low riparian forest cover in the upstream catchment. At several sites, reproductive failure was 100%, mainly due to cannibalism by the caring male. The high incidence of filial cannibalism at degraded sites was not explained by evolutionary hypotheses for filial cannibalism based on poor adult body condition or low reproductive value of small clutches. Instead, larger clutches at degraded sites were most vulnerable to cannibalism. We hypothesize that high frequencies of filial cannibalism of large clutches in areas with low forest cover could be related to changes in water chemistry or siltation that influence parental physiology or that reduce the viability of eggs. Importantly, our results identify chronic nest failure as a possible mechanism contributing to population declines and observed geriatric age structure in this imperiled species.

Natural spawning of the leopard grouper (Mycteroperca rosacea) in captivity in La Paz, Mexico

Natural spawning of the leopard grouper (Mycteroperca rosacea) was studied in captivity from 2004 to 2008 under natural photoperiod conditions. Fish spawned naturally in 2006, but only 90,000 unfertilized eggs were collected. A total of 43 and 27 spawning days were observed in 2007 and 2008, respectively. Spawning season began in March and lasted until May–June and was related to an increase in water temperature from 21.1 to 23.8 °C. The mean (SE) number of eggs released each day was 101,705 (13,876) in 2007 and 40,510 (7,874) in 2008, and the total number of eggs collected in 2007 and 2008 was 4.37 × 106 and 1.07 × 106, respectively. The number of eggs collected per month was not significantly different within each year (P > 0.05), although a significant difference was observed between 2007 and 2008 (P < 0.05). The annual relative number of eggs released was 288,750 eggs·kg–1 in 2007 and 72,919 eggs·kg–1 in 2008, and viable egg production was 184,800 eggs kg–1 in 2007 and 25,375 eggs kg–1 in 2008. The fertilization rate (%) of the eggs collected daily differed significantly throughout the spawning season of 2007 (P < 0.05). The fertilization rate also differed (P < 0.05) between 2007 (61.10%) and 2008 (32.30%). Statistical analysis of the 2 consecutive spawning seasons indicated no correlation between the fertilization rate and fecundity. The effect of the moon phase on spawning was also considered. The fish spawned at dusk before total darkness. Mean (SD) egg diameter was 864.0 ± 22.0 μm.

Mosquito Larvae and Pupae Transport from the Field.

Laboratory study of field-collected mosquitoes can allow researchers to better understand the ways variation within and among mosquito populations shapes burdens of mosquito-borne disease. The Anopheles gambiae complex comprises the most important vectors of malaria, but it can be challenging to keep in the laboratory. For some species of mosquitoes, especially An. gambiae, it is very difficult to bring viable eggs into the laboratory. Instead, it is preferable to collect larvae or pupae and then transport them as carefully as possible back to the laboratory. This simple protocol allows a researcher to start new laboratory colonies from larvae or pupae collected from natural breeding sites or proceed directly to their planned experiments. The use of natural breeding sites provides additional reassurance that the resulting colonies are representative of natural populations.

Automatic method based on deep learning to identify and account Rhipicephalus microplus larval hatching.

Reports of Rhipicephalus microplus resistant populations worldwide have increased extensively, making it difficult to control this ectoparasite. The adult immersion test, commonly used to screen for acaricide resistance, produces the results only after 40 days of the tick collection because it needs the eggs to be laid and larvae to hatch. The present study aims to develop an automatic method, based on deep learning, to predict the hatching of R. microplus larva based on egg morphology. Initially, the time course of embryonic development of tick eggs was performed to discriminate between viable and non-viable eggs. Secondly, using artificial intelligence deep learning techniques, a method was developed to classify and count the eggs. The larval hatching rate of three populations of R. microplus was evaluated for the software validation process. Groups of three and six images of eggs with 12 days of embryonic development were submitted to the software to predict the larval hatching percent automatically. The results obtained by the software were compared with the prediction results of the hatching percentage performed manually by the specialist and with the results of the hatching percentage of larvae obtained in the biological assay. The group with three images of each population submitted to the software for automatic prediction of the larval hatching percent presented mean values of 96.35% ± 3.33 (Piracanjuba population), 95.98% ± 3.5 (Desterro population) and 0.0% ± 0.0 (Barbalha population). For groups with six images, the values were 94.41% ± 3.84 (Piracanjuba population), 95.93% ± 2.36 (Desterro population) and 0.0% ± 0.0 (Barbalha population). Biological assays showed the following hatching percentage values: 98% ± 1.73 (Piracanjuba population); 96% ± 2.1 (Desterro population); and 0.14% ± 0.25 (Barbalha population). There was no statistical difference between the evaluated methods. The automatic method for predicting the hatching percentage of R. microplus larvae was validated and proved to be effective, with considerable reduction in time to obtain results.

Trade-Offs between Temperature and Fitness in Euschistus heros (Fabricius) (Hemiptera: Pentatomidae): Implications for Mass Rearing and Field Management

The brown stink bug, Euschistus heros (Fabricius, 1798) (Hemiptera: Pentatomidae), is one of the most abundant soybean stink bug pests in Brazil. Temperature is a key factor that affects its development and reproduction, and fluctuating temperatures may impact the development and reproduction of E. heros differently from those under constant temperatures. The objective of this study was to evaluate the influence of constant and fluctuating temperature on the biological characteristics of E. heros in three successive generations. Treatments consisted of six constant temperatures (19 °C, 22 °C, 25 °C, 28 °C, 31 °C and 34 °C) and four fluctuating temperatures (25:21 °C, 28:24 °C, 31:27 °C, and 34:30 °C) evaluated for three successive generations. Second-stage nymphs were evaluated daily, and after they reached the adult stage, they were separated by sex, and the individual weight (mg) and pronotum size (mm) were recorded. After pair formation, eggs were collected to evaluate the pre-oviposition period, total number of eggs, and egg viability. The duration of the nymphal stage was reduced with an increase in both constant and fluctuating temperatures; however, at constant temperatures of 19 °C, 31 °C and 34 °C and fluctuating temperatures of 28:24 °C, there was no reproduction in adults. The base temperature and total degree day requirement for nymphal development were 15.5 °C and 197.4 dd, respectively. Pre-oviposition period (d), number of eggs per female, and viability of eggs (%) were affected by temperature across the generations. The multiple decrement life table analysis revealed that mortality was highest during the molting of the second-stage nymphs. These findings have important implications for E. heros' laboratory mass-rearing programs and for its management in fields.

A bacterium against the tiger: further evidence of the potential of noninundative releases of males with manipulated Wolbachia infection in reducing fertility of Aedes albopictus field populations in Italy.

Incompatible insect technique (IIT) is a population suppression approach based on the release of males with manipulated Wolbachia infection inducing egg inviability in wild females. We here present results of multiple field releases of incompatible ARwP males carried out in 2019 in a 2.7-ha green area within urban Rome (Italy) to assess the effect on Aedes albopictus egg viability. Data are compared with results obtained in 2018, when the approach was tested for the first time in Europe. An average of 4674 ARwP males were released weekly for 7 weeks, resulting in a mean ARwP:wild male ratio of 1.1:1 (versus 0.7:1 in 2018). Egg-viability dynamics in ovitraps significantly varied between treated and control sites, with an estimated overall reduction of 35% (versus 15% in 2018). The estimated proportion of females classified as mated with ARwP males was 41.8% and the viability rate of eggs laid by these females (9.5%) was on average significantly lower than that of females only mated with wild males (87.8%); however, high variability in fertility was observed. Values of ARwP male competitiveness were 0.36 and 0.73 based on the overall viability rate of eggs in ovitraps and on female fertility, respectively; thus, well above the conventional 0.2 threshold for an effective suppressive impact in the field. Results further support the potential of IIT as a tool to contribute to Ae. albopictus control in the urban context, stressing the need for larger field trials to evaluate the cost-efficacy of the approach in temperate regions. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Control of Globodera pallida Using Sinapis alba or 4-Hydroxybenzyl Alcohol with the Trap Crop Solanum sisymbriifolium or the Biofumigant Brassica juncea Seed Meal Extract.

The pale cyst nematode Globodera pallida is a highly specialized, economically important pest for potato production. The specialized hatching requirements, ability to adapt, and the loss of effective control strategies such as methyl bromide fumigation increase the challenge to eradicate G. pallida in Idaho. Without a suitable host, this nematode can remain dormant as encysted eggs in soil for up to 20 years. In this study, we first demonstrated that Sinapis alba seed meal extract (SME) or 4-hydroxybenzyl alcohol (HBA), under laboratory and greenhouse conditions, enhances G. pallida egg hatch rate when exposed to potato root diffusate (PRD). This hatch rate enhancement in the presence of PRD is speculated to be due to an increase in egg-shell permeability. We then tested the efficacy of (i) Solanum sisymbriifolium following prior treatment with S. alba SME (0 and 4.48 t/ha) or HBA (0 and 0.12 t/ha) and (ii) Brassica juncea SME (0, 0.14, 0.56, and 1.12 t/ha) following HBA treatment (0 and 4.48 t/ha) on egg viability, hatch rate, and reproduction of G. pallida encysted eggs. S. sisymbriifolium alone reduced the number of encysted eggs compared to the nontreated control by up to 67%, indicating that this trap crop triggered G. pallida eggs to hatch. When combined with S. alba SME or HBA, S. sisymbriifolium significantly reduced egg count, hatch rate, and viability more than S. sisymbriifolium alone. The combination of S. sisymbriifolium with HBA or S. alba SME eliminated G. pallida reproduction on the susceptible potato. All the tested rates of B. juncea SME alone or with HBA reduced egg hatch rate, viability, and reproduction compared to the nontreated control. Combining HBA and B. juncea SME further significantly reduced egg hatch rate, viability, and reproduction than those rates of B. juncea SME alone.

Thymol and eugenol against Rhipicephalus sanguineus sensu lato engorged females: Biological, histopathological and bioinformatic analysis

The aim of this study was to evaluate the reproductive biology and histopathology of the ovaries of engorged females of Rhipicephalus sanguineus sensu lato exposed to combinations of thymol and eugenol, as well as to evaluate in silico the possible interactions of thymol and eugenol in tick cell membranes. To evaluate the tick reproductive biology, the adult immersion test (AIT) was performed, in which the engorged females were immersed in solutions of thymol and eugenol, combined or alone, at concentrations of 2.5 and 5.0 mg/mL. Two control groups (water and 3% DMSO) were also performed. The ticks were kept in a controlled chamber (B.O.D - 27 ± 1 °C and 80% ± 5% RH) to evaluate egg production and viability. To perform the ovaries histopathological evaluation, females were immersed in combination of thymol and eugenol (each at 2.5 and 5.0 mg/mL) and control (water and 3% DMSO) solutions. After immersion, the females were kept in B.O.D (27 ± 1 °C and RH of 80% ± 5%) for four days, they were dissected and the ovaries processed for histological analysis. In addition, an in silico analysis was performed using PASS online® software to predict probability activity (PA) of thymol and eugenol in cell membranes. The treatment with the combination of thymol and eugenol (each at 5.0 mg/mL) caused a reduction (p < 0.01) in oviposition, while the treatments with thymol (5.0 mg/mL) and combination of thymol and eugenol (5.0 mg/mL) reduced (p < 0.05) the egg viability. The treatment with combination of thymol and eugenol (5.0 mg/mL) resulted in a control percentage of 99.9%, while in the other treatments, control percentages below 56% were observed. Oocytes from the females exposed to the combinations of thymol and eugenol (each compound at 2.5 mg/mL) showed histopathological changes, except on oocyte V, while those treated with these compounds alone at 2.5 mg/mL, did not reveal any change. Changes in the shape of the oocyte, presence of vacuoles in the cytoplasm and germinal vesicle, reduction and fusion of yolk granules and rupture of some oocytes were observed. In silico analysis, showed that these compounds can act as membrane permeability inhibitors, membrane permeability agonists, membrane integrity antagonists and apoptosis agonists. We conclude that the combination of thymol and eugenol causes changes in the reproductive biology and morphophysiology of engorged females oocytes. The in silico analysis using thymol and eugenol revealed the possibility of disorganization in the cell membranes, a fact that may explain the histopathological alterations observed.

Single-sex schistosomiasis: a mini review.

Schistosomiasis is a neglected tropical disease caused by dioecious blood flukes of the genus Schistosoma and second to malaria as a parasitic disease with significant socio-economic impacts. Mating is essential for maturation of male and female schistosomes and for females to lay of eggs, which are responsible for the pathogenesis and propagation of the life cycle beyond the mammalian host. Single-sex schistosomes, which do not produce viable eggs without mating, have been overlooked given the symptomatic paucity of the single-sex schistosomiasis and limited diagnostic toolkit. Besides, single-sex schistosomes are less sensitive to praziquantel. Therefore, these issues should be considered to achieve the elimination of this infection disease. The aim of this review is to summarize current progress in research of single-sex schistosomes and host-parasite interactions.

Tyraminergic control of vitellogenin production and release in the blood-feeding insect, Rhodnius prolixus

In insects, the biogenic amine tyramine (TA) has been shown to control several physiological processes. Recently, the involvement of the type 1 tyramine receptor (TAR1) in reproductive processes has been demonstrated in different insects. Here, we investigate the putative role of Rhodnius prolixus TAR1 (RpTAR1) in reproduction in female R. prolixus. RpTAR1 transcript was highly expressed in tissues associated with egg development. Moreover, after a blood meal, which is the stimulus for full egg development, RpTAR1 transcript was upregulated in the ovaries and in the fat body. After RNAi-mediated RpTAR1 knockdown, an ovarian phenotype characterized by the absence or reduction of egg production was observed. Furthermore, protein and Vg accumulation in the fat body was observed, suggesting an impairment in protein release from the fat body into the hemolymph. However, even though fewer eggs were produced and laid, there was no difference in hatching ratio of those laid, in comparison to the controls, indicating that the overall low protein uptake by the ovaries did not influence the viability of individual eggs produced. Interestingly, the eggs from dsTAR1-treated insects appeared more red, indicating a higher content of RHBP compared to the control. A higher colocalization between Vg and Rab11, a marker for the recycling endosome pathway, was observed after dsTAR1 injection, suggesting that a more active lysosome degradation pathway in response to the Vg accumulation may occur. In addition to the Vg accumulation in the fat body, dsTAR1 treatment altered JH pathway. However, it remains to be elucidated whether this event is either directly related to the RpTAR1 downregulation or for a consequence to the Vg accumulation. Lastly, the RpTAR1 action on Vg synthesis and release in the fat body was monitored in the presence or absence of yohimbine, the antagonist of TAR1, in an ex-vivo experiment. Yohimbine antagonises the TAR1 stimulated release of Vg. These results provide critical information concerning the role of TAR1 in Vg synthesis and release in R. prolixus. Furthermore, this work opens the way for further investigation into innovative methods for controlling R. prolixus.

Investing in a nest egg: intraspecific variation in the timing of egg laying across a latitudinal gradient.

Avian reproductive strategies vary widely, and many studies of life-history variation have focused on the incubation and hatching stages of nesting. Birds make proximate decisions regarding reproductive investment during the laying stage, and these decisions likely constrain and tradeoff with other traits and subsequent behaviors. However, we know relatively little about egg-laying stage behaviors given the difficulty of locating and monitoring nest sites from the onset of laying. We used non-invasive continuous video recording to quantify variation in the egg-laying behaviors of burrowing owls (Athene cunicularia) along a 1400-km latitudinal gradient in western North America. Burrowing owls laid eggs disproportionately in the morning hours, and that tendency was strongest among first eggs in a clutch. However, selection appeared to act more strongly on laying intervals (the time between laying of consecutive eggs) than on the diel time of laying, and laying intervals varied widely among and within clutches. Laying intervals declined seasonally and with increasing clutch size but increased with increasing burrow temperature and as a function of laying stage nest attentiveness, which may be a strategy to preserve egg viability. Laying interval was positively correlated with the duration of hatching intervals, suggesting that laying interval duration is one mechanism (along with timing of incubation onset) that generates variation in hatching asynchrony. Our results lend support to two general hypotheses to explain laying schedules; selection favors laying eggs in the morning, but other selective pressures may override that pattern. These conclusions indicate that allocation decisions during laying are an important part of avian life-history strategies which are subject to energetic constraints and tradeoffs with other traits.

Dengue Exposure and Wolbachia wMel Strain Affects the Fertility of Quiescent Eggs of Aedes aegypti.

(1) Background: The deployment of the bacterium Wolbachia to reduce arbovirus transmission is ongoing in several countries worldwide. When Wolbachia-carrying Aedes aegypti are released and established in the field, females may feed on dengue-infected hosts. The effects of simultaneous exposure on life-history traits of Ae. aegypti to Wolbachia wMel strain and dengue-1 virus DENV-1 remain unclear. (2) Methods: We monitored 4 groups (mosquitoes with either DENV-1 or Wolbachia, coinfected with DENV-1 and Wolbachia, as well as negative controls) to estimate Ae. aegypti survival, oviposition success, fecundity, collapsing and fertility of quiescent eggs for 12 weeks. (3) Results: Neither DENV-1 nor Wolbachia had a significant impact on mosquito survival nor on mosquito fecundity, although the last parameter showed a tendency to decrease with ageing. There was a significant decrease in oviposition success in individuals carrying Wolbachia. Wolbachia infection and storage time significantly increased egg collapse parameter on the egg viability assay, while DENV-1 had a slight protective effect on the first four weeks of storage. (4) Conclusions: Despite limitations, our results contribute to better understanding of the tripartite interaction of virus, bacteria and mosquito that may take place in field conditions and aid in guaranteeing the Wolbachia strategy success.

Mesophilic anaerobic digestion of waste activated sludge in an intermittent mixing reactor: Effect of hydraulic retention time and organic loading rate

An anaerobic digester was operated at mesophilic temperature and with intermittent mixing conditions to treat waste activated sludge. The organic loading rate (OLR) was increased by decreasing the hydraulic retention time (HRT), and the effect on process performance, digestate characteristics and inactivation of pathogens was investigated. The removal efficiency of total volatile solids (TVS) was also measured by biogas formation. The HRT varied from 50 to 7 days, corresponding to OLR from 0.38 to 2.31 kgTVS.m−3.d−1. The acidity/alkalinity ratio remained within stable limits (lower than 0.6) at 50-, 25- and 17-day HRT; due to an imbalance between the production and consumption of volatile fatty acids, the ratio increased to 0.7 ± 0.2 at HRT of 9 days and 7 days. The highest TVS removal efficiencies were 16, 12 and 9%, which were obtained at 50-, 25- and 17 day-HRT, respectively. Intermittent mixing provided solids sedimentation greater than 30% for almost all HRT tested. The highest methane yields (0.10–0.05 m3.kgTVSfed−1.d−1) were obtained when the reactor was operated at a higher HRT (50-17 days). At lower HRT, methanogenic reactions were likely limited. Zinc and copper were the main heavy metals found in the digestate, while the most probable number (MPN) of coliform bacteria remained below 106 MPN.g TVS−1. Neither Salmonella nor viable Ascaris eggs were found in the digestate. In general, increasing the OLR by decreasing the HRT to 17 days under intermittent mixing conditions provided an attractive alternative to treat sewage sludge despite some limitations due to biogas and methane yields.

Enhancing the scalability of Wolbachia-based vector-borne disease management: time and temperature limits for storage and transport of Wolbachia-infected Aedes aegypti eggs for field releases

BackgroundIntrogression of the bacterial endosymbiont Wolbachia into Aedes aegypti populations is a biocontrol approach being used to reduce arbovirus transmission. This requires mass release of Wolbachia-infected mosquitoes. While releases have been conducted using a variety of techniques, egg releases, using water-soluble capsules containing mosquito eggs and larval food, offer an attractive method due to its potential to reduce onsite resource requirements. However, optimisation of this approach is required to ensure there is no detrimental impact on mosquito fitness and to promote successful Wolbachia introgression.MethodsWe determined the impact of storage time and temperature on wild-type (WT) and Wolbachia-infected (wMel or wAlbB strains) Ae. aegypti eggs. Eggs were stored inside capsules over 8 weeks at 18 °C or 22 °C and hatch rate, emergence rate and Wolbachia density were determined. We next examined egg quality and Wolbachia density after exposing eggs to 4–40 °C to determine how eggs may be impacted if exposed to extreme temperatures during shipment.ResultsEncapsulating eggs for 8 weeks did not negatively impact egg viability or resulting adult emergence and Wolbachia density compared to controls. When eggs were exposed to temperatures within 4–36 °C for 48 h, their viability and resulting adult Wolbachia density were maintained; however, both were significantly reduced when exposed to 40 °C.ConclusionsWe describe the time and temperature limits for maintaining viability of Wolbachia-infected Ae. aegypti eggs when encapsulated or exposed to extreme temperatures. These findings could improve the efficiency of mass releases by providing transport and storage constraints to ensure only high-quality material is utilised during field releases.Graphical