Egg Viability Research Articles

Mutual mate choice and its benefits for both sexes

In mating interactions, it is common in nature for both sexes to choose simultaneously. However, this mutual mate choice and its consequences for progeny has received relatively little study; an approach where both male and female condition is manipulated is thus desirable. We compared both sexes’ preferences in Tenebrio molitor beetles when individual condition varied (healthy vs infected with a fungus), and observed the direct benefits of those preferences. We predicted that: (a) females and males in good condition would prefer high quality mates; (b) preferences would be weaker when the choosing individual is in poor condition (and thus less selective given, for example, time and energetic constrains); and, (c) high quality mates would lay a larger number of total eggs and/or viable eggs than low quality mates. We found that both males and females in good condition were not more likely to choose mates that were also in good condition. However, poor-condition animals were more likely to prefer similar quality animals, while high-condition animals did not necessarily prefer mates of similar condition. Choosing sick males or females had a negative impact on egg number and viability. Our results suggest a non-adaptive mate choice in this species. Possibly, a deteriorated condition may drive individuals to invest more in attracting mates, because their chances of surviving the infection are very low. However, we do not discount the possibility that the fungus is manipulating individuals to increase its transmission during mating.

Ammonium transporter expression in sperm of the disease vector Aedes aegypti mosquito influences male fertility

The ammonium transporter (AMT)/methylammonium permease (MEP)/Rhesus glycoprotein (Rh) family of ammonia (NH3/NH4+) transporters has been identified in organisms from all domains of life. In animals, fundamental roles for AMT and Rh proteins in the specific transport of ammonia across biological membranes to mitigate ammonia toxicity and aid in osmoregulation, acid-base balance, and excretion have been well documented. Here, we observed enriched Amt (AeAmt1) mRNA levels within reproductive organs of the arboviral vector mosquito, Aedes aegypti, prompting us to explore the role of AMTs in reproduction. We show that AeAmt1 is localized to sperm flagella during all stages of spermiogenesis and spermatogenesis in male testes. AeAmt1 expression in sperm flagella persists in spermatozoa that navigate the female reproductive tract following insemination and are stored within the spermathecae, as well as throughout sperm migration along the spermathecal ducts during ovulation to fertilize the descending egg. We demonstrate that RNA interference (RNAi)-mediated AeAmt1 protein knockdown leads to significant reductions (∼40%) of spermatozoa stored in seminal vesicles of males, resulting in decreased egg viability when these males inseminate nonmated females. We suggest that AeAmt1 function in spermatozoa is to protect against ammonia toxicity based on our observations of high NH4+ levels in the densely packed spermathecae of mated females. The presence of AMT proteins, in addition to Rh proteins, across insect taxa may indicate a conserved function for AMTs in sperm viability and reproduction in general.

TÉCNICA PARA IDENTIFICAÇÃO DE OVOS DE Dioctophyme renale EM URINA DE GATOS, EMPREGADA EM SÍLICA

A dioctofimatose é uma doença parasitária causada pelo helminto Dioctophyme renale, tendo como hospedeiros definitivos, cães, gatos, animais silvestres e também o homem, pois este parasito possui potencial zoonótico. Acomete os rins dos seus hospedeiros definitivos, geralmente o direito, mas podem ser encontrados também em outros locais. Embora seja descrito maior ocorrência de casos em cães, também há relatos desta parasitose em gatos. Uma das formas de diagnóstico é através da visualização de ovos do D. renale através do exame de urina. Porém, pelas características comportamentais dos gatos, a coleta de urina nesta espécie é de difícil execução. Portanto, através do estudo de técnicas para pesquisa de ovos deste parasito na urina dos gatos em sílica, presente nas caixas de areia, procura-se facilitar o diagnóstico nestes animais. Para realização deste trabalho, amostras de sílica foram contaminadas com urina contendo ovos de D. renale. As técnicas utilizadas para pesquisa de ovos foram: centrífugo-flutuação em solução hipersaturada glicosada, em duas densidades: 1.230g/ml e 1.275g/ml; flutuação espontânea em solução hipersaturada glicosada, em duas densidades: 1,230g/ml e 1,275g/ml e centrífugo-sedimentação. A avaliação das amostras foi realizada em diferentes tempos, distribuídos da seguinte forma: momento zero, 6 horas, 12 horas, 18 horas, 24 horas, 36 horas, 48 horas, 72 horas, 120 horas, 168 horas, 240 horas e 336 horas após contaminação do material. A técnica de centrífugo-sedimentação foi a que apresentou o melhor resultado, sendo possível identificar ovos morfologicamente viáveis de D. renale durante os 14 dias de análise.

Cladocera resting egg banks in temporary and permanent wetlands

Cladocerans are important filter-feeders transferring energy up the food web to different invertebrate and vertebrate predators. Along the flood period, cladocerans are one of the primary food sources for juvenile fish in floodplain. Resting egg banks allow cladoceran populations to overcome the environmental stress, related to several limnological changes, including complete drying of temporary wetlands. After drought, resting egg banks influence cladoceran community attributes during the cyclic and successional processes driven by episodic flood events. In this study we compared the taxonomic richness of active (from the water column) and dormant (from the sediment) Cladocera assemblages and analyzed the structure of resting egg banks, comparing the diversity, abundance and apparent viability/unviability of the eggs, between six temporary and six permanent wetlands, located along the Ovens River Floodplain, Victoria, Australia. The qualitative analysis shows higher taxonomic richness in active assemblages from temporary (24 taxa) than permanent (13 taxa) wetlands compared to dormant assemblages present in resting egg banks (9 taxa) from temporary and permanent wetlands. However, richness was influenced by taxonomic level of identification, with the majority of resting eggs only being identified to the taxonomic level of family (i.e. Chydoridae). Total taxa richness within egg banks was similar between wetland types, however, on average higher Shannon’s diversity of resting eggs was found within permanent (1.53) than temporary (0.82) wetlands. This is likely to be due to more stable wetlands not providing appropriate cues to trigger dormancy induction or breakage for specific populations, leading to higher values of evenness in permanent than temporary wetlands. Comparing permanent and temporary wetlands, higher abundance of resting eggs (more than four times) consisting of higher abundance of unviable eggs and similar viable egg abundance to permanent wetlands, was found within temporary wetlands, suggesting that the increased resting egg abundance in temporary wetlands is balanced by the losses due to factors such as predation, parasitism or other physical damage, during the terrestrial phase. Despite resistant outer shell structure, this study highlights that the damage to egg integrity is intensified in wetlands that undergo dry phases. Cladoceran resting egg banks represent the potential assemblage to recover after disturbance events such as drying, and information about these is important to ensure appropriate management and conservation of floodplain biodiversity.

Wolbachia's Deleterious Impact on Aedes aegypti Egg Development: The Potential Role of Nutritional Parasitism.

Simple SummaryMosquito-borne viral diseases such as dengue, Zika and chikungunya cause a significant global health burden and are currently increasing in outbreak frequency and geographical reach. Wolbachia pipientis, an endosymbiotic bacterium, offers a solution to this. When Wolbachia is introduced into the main mosquito vector of these viruses, Aedes aegypti, it alters the mosquito’s reproductive biology, as well as reducing the ability of the mosquitoes to transmit viruses. These traits can be leveraged to reduce virus transmission within a community by mass releasing Wolbachia-infected mosquitoes. However, Wolbachia has some negative effects on Aedes aegypti fitness, particularly egg longevity, and the reason behind this remains ambiguous. Insect fitness is very important for the success for Wolbachia-biocontrol strategies as they rely on the released insects being competitive with the wild mosquito population. This review summarises the fitness effects of Wolbachia on Aedes aegypti and investigates the possible contribution of Wolbachia as a nutritional parasite in lowering host fitness. It proposes the next stages of research that can be conducted to address nutritional parasitism to aid in the expansion of Wolbachia-based disease management programs worldwide.The artificial introduction of the endosymbiotic bacterium, Wolbachia pipientis, into Aedes (Ae.) aegypti mosquitoes reduces the ability of mosquitoes to transmit human pathogenic viruses and is now being developed as a biocontrol tool. Successful introgression of Wolbachia-carrying Ae. aegypti into native mosquito populations at field sites in Australia, Indonesia and Malaysia has been associated with reduced disease prevalence in the treated community. In separate field programs, Wolbachia is also being used as a mosquito population suppression tool, where the release of male only Wolbachia-infected Ae. aegypti prevents the native mosquito population from producing viable eggs, subsequently suppressing the wild population. While these technologies show great promise, they require mass rearing of mosquitoes for implementation on a scale that has not previously been done. In addition, Wolbachia induces some negative fitness effects on Ae. aegypti. While these fitness effects differ depending on the Wolbachia strain present, one of the most consistent and significant impacts is the shortened longevity and viability of eggs. This review examines the body of evidence behind Wolbachia’s negative effect on eggs, assesses nutritional parasitism as a key cause and considers how these impacts could be overcome to achieve efficient large-scale rearing of these mosquitoes.

Characterization of field-evolved resistance to pyridalyl in a near-isogenic line of diamondback moth, Plutella xylostella.

Plutella xylostella has developed resistance to a variety of pesticides in the field. Selection, inheritance, a near-isogenic line, cross-resistance and biochemical mechanisms of pyridalyl resistance were characterized in a field-collected resistant population of P. xylostella from China. Compared with a susceptible IVF-S strain, the field-collected FZ population showed ~ 350-fold resistance to pyridalyl. The FZ-PY strain, selected from the FZ population using pyridalyl, developed ~ 640-fold resistance to pyridalyl. Inheritance tests indicated that pyridalyl resistance in the FZ-PY strain was autosomal and incompletely recessive. Through successive backcrossing to IVF-S, a near-isogenic strain (NIL-PY) was established that exhibited 191.21-fold resistance to pyridalyl and no cross-resistance to other tested popular insecticides. No significant effects of synergists and higher activities of metabolic enzymes were observed in NIL-PY compared with IVF-S. Furthermore, the survival rate of NIL-PY larvae, and female oviposition, fecundity and egg viability were markedly reduced in NIL-PY compared with IVF-S. The fitness of NIL-PY was found to be 0.56 compared with IVF-S. Considering that no relevant effects of synergists or oxidative metabolism were observed in NIL-PY, and that pyridalyl resistance results in significant fitness costs compared with IVF-S, further research will be conducted on the mechanism of target-site resistance. © 2020 Society of Chemical Industry.

Features of exogenous development of Trichuris globulosa (Nematoda, Trichuridae)

Parasitic nematodes of the genus Trichuris Röderer, 1761 are hematophagous helminths, capable of parasitizing many different hosts including humans. The domestic and wild ruminants are hosts of several Trichuris species, with Trichuris globulosa (Linstow, 1901) one of the most prevalent parasites found in cattle. The exogenous stages of the helminth’s life cycle develop in the outer environment, and their activity and survival depend on the abiotic factors. Thus the aim of the work was to determine the influence of temperature on the rate and success of development of infectious eggs of T. globulosa in laboratory culture considering their morphological and metric changes. The results of experimental studies showed that the embryogenesis of T. globulosa eggs, cultured in laboratory conditions and obtained from gonads of female nematodes, occurs in six stages regardless of the temperature regime. At the same time, the rate of transition from one stage to another, the term for the formation of infectious eggs and their viability directly depends on the temperature of the external environment. At the optimal temperature for the development of T. globulosa eggs, 25 °C, 76.3% of eggs reached the mobile larva stage. A decrease in temperature to 20 °C and an increase to 30° C led to an increase in the egg mortality of up to 26% and 32%, respectively, and the viability of eggs decreased. Under such temperature conditions, 74% and 68% of eggs of Trichuris reached the mobile larva stage. With an increase in the culture temperature, the time of embryogenesis decreased and amounted to 56 days at 20 °C, 48 days at 25 °C, and 32 days at 30 °C. Depending on the temperature regime, the zygote stage lasted from the release of eggs from the gonads of female nematodes to 12 days, the stage of blastomere formation from 4 to 12 days, the stage of the bean-shaped embryo from 8 to 24 days, the stage of the tadpole embryo from 16 to 36 days, the stage of larval formation lasted from 16 to 48 days, the stage of mobile larva from days 20 to 56. The formation of the infectious T. globulosa egg from a non-infectious one is shown in an experiment to include the metric changes confirmed by metric indicators. Mature eggs with a mobile larva are shorter and wider, with shorter and narrower plugs and a thinner shell than eggs at the zygote stage. The obtained research results on the influence of temperature on the exogenous development of Trichuris globulosa will make it possible to predict the epizootic situation of trichurosis in animal husbandry, as well as take timely measures to arrest the embryonic stages of nematode development in the external environment.

Warming winters threaten peripheral Arctic charr populations of Europe

As the global climate warms, the fate of lacustrine fish is of huge concern, especially given their sensitivity as ectotherms to changes in water temperature. The Arctic charr (Salvelinus alpinus L.) is a salmonid with a Holarctic distribution, with peripheral populations persisting at temperate latitudes, where it is found only in sufficiently cold, deep lakes. Thus, warmer temperatures in these habitats particularly during early life stages could have catastrophic consequences on population dynamics. Here, we combined lake temperature observations, a 1-D hydrodynamic model, and a multi-decadal climate reanalysis to show coherence in warming winter water temperatures in four European charr lakes near the southernmost limit of the species’ distribution. Current maximum and mean winter temperatures are on average ~ 1 °C warmer compared to early the 1980s, and temperatures of 8.5 °C, adverse for high charr egg survival, have frequently been exceeded in recent winters. Simulations of winter lake temperatures toward century-end showed that these warming trends will continue, with further increases of 3–4 °C projected. An additional 324 total accumulated degree-days during winter is projected on average across lakes, which could impair egg quality and viability. We suggest that the perpetuating winter warming trends shown here will imperil the future status of these lakes as charr refugia and generally do not augur well for the fate of coldwater-adapted lake fish in a warming climate.

Changes induced by atrazine in Clarias gariepinus provide insight into alterations in ovarian histoarchitecture and direct effects on oogenesis

Clarias gariepinus juveniles were exposed to environmentally relevant concentrations of 0 (control), 2.5, 25, 250 and 500 μg L−1 atrazine in a quality-controlled 28-day laboratory procedure. Findings revealed a significant decrease in the levels of follicle-stimulating hormone, luteinizing hormone and prolactin relative to control (p < 0.05). Atrazine reduced the levels of testosterone while increasing the concentration of progesterone. Histologically, the control and treatments presented three stages of oocyte maturation: the chromatin nucleolar oocyte stage, early perinucleolar oocyte stage and the vitellogenic oocyte stage. However, in the ovaries of the treatment group with the lowest treatment concentration (2.5 μg L−1), atretic oocytes with broken membranes invaded many of the dead ova and empty spaces. In other treatments (25, 250 and 500 μg L−1), interfollicular spaces, vacuolation in oocyte formation, and dissolution of oocyte walls were observed. Disruption of the yolk vesicle and clumping of the cytoplasm in maturing oocytes was observed only at the highest atrazine concentration (500 μg L−1). Gross alterations in ovarian histoarchitecture and reproductive hormone levels observed in this study showed interference with oogenesis which may result in reduced egg viability and fecundity in fish with ecological implications in water bodies exposed to atrazine even at reduced concentrations.

Effects of novaluron ingestion and topical application on German cockroach (Blattella germanica) development and reproduction.

Insect growth regulators disrupt insect development and reproduction. Chitin synthesis inhibitors (CSIs) allow the insect to grow normally, but because chitin is an essential component of the cuticle, formation of a new cuticle and ecdysis are prevented and the insect dies. CSIs can also kill embryos by disrupting their normal development. We evaluated the potential utility of novaluron in bait formulations against the German cockroach (Blattella germanica L.). The minimum novaluron intake that interfered with molting and reproduction was assessed by exposing nymphs and adult females to novaluron. Results showed that 1 day of feeding on 0.1% novaluron was sufficient to disrupt molting in nymphs and prevent adult females from developing viable oothecae. The long-term effects on gravid females were investigated by feeding females 0.1% novaluron for different 5-day intervals during successive stages of gestation. Results demonstrated that gravid females fed novaluron during any period of gestation were able to produce viable eggs. To determine if ingestion of novaluron affected mating success and fertility of adult males, males were fed novaluron and then allowed to mate with untreated virgin females. Males that fed on novaluron successfully mated, and the females produced viable oothecae. Finally, direct comparisons revealed that novaluron is equally effective by ingestion and topical application. Novaluron caused mortality in nymphs and interfered with ootheca production in adult females, but only before they formed an ootheca. It successfully reduced German cockroach populations in cages and has potential to be incorporated in cockroach baits.

Species Detection within the Echinococcus granulosus sensu lato Complex by Novel Probe-Based Real-Time PCRs.

Infections with eggs of Echinococcus granulosus sensu lato (s.l.) can cause cystic echinococcosis in intermediate host animals and humans. Upon ingestion of viable eggs, oncospheres hatch from the eggs and subsequently develop into fluid-filled larval cysts, most frequently in the liver or the lungs. The slowly growing cysts progressively interfere with organ function. The risk of infection is determined by the host range of the parasite, its pathogenicity and other epidemiologically relevant parameters, which differ significantly among the five species within the E. granulosus s.l. complex. It is therefore essential to diagnose the correct species within E. granulosus s.l. to help understand specific disease epidemiology and to facilitate effective implementation of control measures. For this purpose, simple, fast and cost-effective typing techniques are needed. We developed quantitative real-time polymerase chain reactions (qPCRs) to target polymorphic regions in the mitochondrial genome of E. granulosus s.l. In a single-step typing approach, we distinguished E. granulosus s.l. members in four epidemiologically relevant subgroups. These were E. granulosus sensu stricto, E. equinus, E. ortleppi and the E. canadensis cluster. The technique also allowed identification and differentiation of these species from other Echinococcus or Taenia taxa for samples isolated from cysts or faeces.

Sanitary and parasitological monitoring of environmental objects in the Rostov region

Objective: to study the sanitary condition of various environmental objects of the Rostov region and the degree of their contamination.Materials and methods: in the period from 2015 to 2019, more than 5600 sanitary and parasitological studies of environmental objects of the Rostov region were performed on the basis of the Rostov Research Institute of Microbiology and Parasitology: waste water and its precipitation, soil, water of open surface reservoirs. The work was carried out in accordance with the accepted normative documents.Results: it was found that 33.4 % of all tested samples were positive, and 1.4% of the samples contained viable eggs of various pathogens of helminthiasis and did not meet the requirements of regulatory documents.Conclusions: the results of the study indicate that the environmental objects of the Rostov region are polluted with parasitic pathogens and their unsatisfactory sanitary and parasitological condition.

Herbivory improves the fitness of predatory beetles.

While many predatory arthropods consume non-prey foods from lower trophic levels, little is known about what drives the shift from predator to omnivore. Predatory lady beetles often consume non-prey foods like plant foliage and pollen. One species, Coccinella septempunctata, eats foliage to redress sterol deficits caused by eating sterol-deficient prey. Here we explore how omnivory benefits lady beetle fitness. We reared seven species of lady beetles-from five genera distributed across the tribe Coccinellini-on pea aphids in the presence or absence of fava bean foliage; pea aphids have very low sterol content. Foliage supplements lengthened the development times of four species and decreased survival to adulthood of two species; it had no effect on adult mass. We mated beetles in a 2×2 factorial design (males with or without foliage paired with females with or without foliage). For each species, we observed a profound paternal effect of foliage supplements on fitness. Females mated to foliage-supplemented males laid more eggs and more viable eggs compared to females mated to non-supplemented males. Foliage-supplemented males produced 2.9-4.6 times more sperm compared to non-supplemented males for the three species we examined. We analysed the sterol profile of four beetle species reared on pea aphids-with or without foliage-and compared their sterol profile to field-collected adults. For two laboratory-reared species, sterols were not detected in adult male beetles, and overall levels were generally low (total ng of sterol/beetle range: 3-33ng); the exception being Propylea quatuordecimpunctata females (total ng of sterol/beetle range: 50-157ng). Laboratory-reared lady beetle sterol content was not significantly affected by the presence of foliage. Field-collected beetles had higher levels of sterols compared to laboratory-reared beetles (2,452-145,348ng per beetle); cholesterol and sitosterol were the dominant sterols in both field-collected and laboratory-reared beetles. Our findings indicate that herbivory benefits lady beetle fitness across the Coccinellini, and that this was entirely a paternal effect. Our data provide a rare example of a nutritional constraint impacting fitness in a sex-specific manner. It also shows, more broadly, how a nutritional constraint can drive predators towards omnivory.

Biology and life studies of Tetranychus macfarlanei on okra

Present experiment was carried out on biology and life studies of red spider mite, Tetranychus macfarlanei on okra under laboratory conditions at 30.8-35.10C temperature and 64.5-74% relative humidity at Dr. P. D. K. V., Akola. The total developmental period of T. macfarlanei from egg to adult emergence is recorded less in male (9.0 ± 0.17 days) than female (10.32 ± 0.19 days). T. macfarlanei showed a sexual and arrhenotokous parthenogenesis. Mated female produced a progeny in the ratio 1 male: 4.14 females; whereas, unmated female produced only males. Egg viability was recorded as 93.2 ± 1.17 percent. Adult longevity for male, mated female and unmated female was 11.05 ± 0.99 days, 11.68 ± 1.68 days and 12.45 ± 1.48 days respectively.

Insecticidal and inhibitory effects of Meliaceae and Asteraceae extracts to silverleaf whitefly

ABSTRACT Bemisia tabaci (Hemiptera: Aleyrodidae) is a polyphagous insect that causes direct and indirect damage on tomato. Spraying with synthetic insecticides is one of the main control tools for the whitefly; however, the abusive use of this tool can impact the environment and human beings. In this study we evaluated the bioactivity of extracts from plant species from Asteraceae and Meliaceae families on B. tabaci biotype B in tomato. A preliminary bioassay evaluated the effects of the extracts at 2% (w/v) on infestation and oviposition of this whitefly. Then, the nymphicidal and ovicidal effects of the chloroformic extract of Trichilia pallida - TP-Cl at six concentrations were performed. Finally, the TP-Cl LC50 was estimated. All extracts were repellent to the whitefly, except that ones from T. pallida (ethanolic) and Melia azedarach (hexanic), considered neutral at 24 h. All the extracts caused deterrence of oviposition. All concentrations of TP-Cl revealed nymphicidal activity and the highest ones provided the lowest egg viability averages. The estimated LC50 was 0.0955%. These results provide evidence for the potential of the extracts to be used as alternative control agents of B. tabaci biotype B in tomatoes.

Management of rice moth, Corcyra cephalonica (Stainton) by using non-toxic plant powders in stored groundnut seeds

Laboratory experiment was conducted to study the efficacy of botanical powders viz., neem leaf powder, lantana leaf powder, custard apple leaf powder, eucalyptus leaf powder, marigold flower powder, turmeric rhizome powder, neem seed kernel powder, ginger rhizome powder, and garlic bulb powder @ 2.5 g/100 g kernels in protecting stored groundnuts against rice moth. Neem seed kernel powder was found to be most effective in protecting stored groundnuts against rice moth as it recorded highest oviposition deterrence of 92.16%, maximum reduction of adult emergence of 95.89%, lowest longevity of male (2.85 days) and female (2.89 days) moths, lowest fecundity of 42.80 eggs per female and lowest egg viability of 43.96%. Next effective treatments were neem leaf powder and eucalyptus leaf powder @ 2.5 g/100 g kernels. None of the plant powders had hindered the germination of groundnut seeds during storage periods of up to 120 days.

English

Newcastle disease remains a disease of global concern and serious economic challenge to the poultry industry because of its high mortality rate, in spite of the various intervention programs including vaccination. This research is aimed at studying the antiviral activity of the extract and fractions of Boswellia dalzielii against Newcastle disease virus (NDV) using chicken embryonated eggs. Phytochemical analysis was conducted using standard procedure. The methanol extract of Boswellia dalzielii was subjected to solvent- solvent fractionation using solvents of varing polarity. This process generated four fractions namely hexane fraction, ethyl acetate fraction, n-butanol fraction and aqueous fraction. Nine to eleven day-old viable embryonated chicken eggs (ECE) were used for the antiviral assay; these were divided into seven groups of five eggs each. The methanol extract was also screened for its cytotoxicity, prophylatic, therapeutic and neutralization effects against NDV, while the fractions were screened for their cytotoxicity and neutralization effects. The phytochemical screening of the stem bark extract and fractions of B. dalzielii showed the presence of tannins, flavonoids, saponin, terpenoids and steroids. It was observed from the results that the extract was toxic to the embryo at a concentration above 100 mg/ml. At the end of the bioassay, methanol extract and fractions of B. dalzielii showed antiviral activity against NDV. However, the extract seems to possess the most significant anti-NDV activity than the fractions. The results of the study are very promising and support the use of B. dalzielii in the treatment of viral infections in animals and humans. Key words: Antiviral, Boswellia dalzielii, Newcastle disease virus (NDV), embryonated chicken eggs (ECE).

A comparison of behavioral and reproductive parameters between wild-type, transgenic and mutant zebrafish: Could they all be considered the same “zebrafish” for reglementary assays on endocrine disruption?

Transgenic zebrafish models are efficiently used to study the effects of endocrine disrupting chemicals (EDC); thereby informing on their mechanisms of action. However, given the reported differences between zebrafish strains at the genetical, physiological and behavioral levels; care should be taken before using these transgenic models for EDC testing. In the present study, we undertook a set of experiments in different transgenic and/or mutant zebrafish strains of interest for EDC testing: casper, cyp19a1a-eGFP, cyp19a1a-eGFP-casper, cyp11c1-eGFP, cyp11c1-eGFP-casper. Some behavioral traits, and some biochemical and reproductive physiological endpoints commonly used in EDC testing were assessed and compared to those obtained in WT AB zebrafish to ensure that transgene insertion and/or mutations do not negatively modify basal reproductive physiology or behavior of the fish.Behavioral traits considered as anxiety and sociality have been monitored. Sociality was evaluated by monitoring the time spent near congeners in a shuttle box while anxiety was evaluated using the Novel tank diving test. No critical difference was observed between strains for either sociality or anxiety level. Concerning reproduction, no significant difference in the number of eggs laid per female, in the viability of eggs or in the female circulating VTG concentrations was noted between the 5 transgenic/mutants and the WT AB zebrafish studied.In summary, the transgene insertion and the mutations had no influence on the endpoints measured in basal conditions. These results were a prerequisite to the use of these transgenic/mutant models for EDC testing. Next step will be to determine the sensitivity of these biological models to chemical exposure to accurately validate their use in existing fish assays for EDC testing.

Accelerating the Morphogenetic Cycle of the Viral Vector Aedes aegypti Larvae for Faster Larvicidal Bioassays.

Any bioassay to test new chemically synthesized larvicides or phytolarvicides against Culicidae and more harmful mosquito species, such as Aedes aegypti and Aedes albopictus, which specifically transmit dengue, yellow fever, chikungunya viral fevers as well as Zika virus, or Anopheles gambiae, a vector for malaria and philariasis, requires thousands of well-developed larvae, preferably at the fourth instar stage. The natural morphogenetic cycle of Aedes spp., in the field or in the laboratory, may extend to 19 days at room temperature (e.g., 25°C) from the first permanent contact between viable eggs and water and the last stage of larval growth or metamorphosis into flying adults. Thus, accelerated sequential molting is desirable for swifter bioassays of larvicides. We achieved this goal in Aedes aegypti with very limited strategic and low-cost additions to food, such as coconut water, milk or its casein, yeast extract, and to a lesser extent, glycerol. The naturally rich coconut water was excellent for quickly attaining the population of instar IV larvae, the most advanced one before pupation, saving about a week, for subsequent larvicidal bioassays. Diluted milk, as another food source, allowed an even faster final ecdysis and adults are useful for mosquito taxonomical purpose.

Electro-oxycoagulation Efficiency for the Treatment of Domestic Effluents

Although environment protection efforts worldwide, the chemistry and biological contamination of waters represents an important challenge to be overcome, especially regarding its contamination by domestic wastewater effluents. In this context, this research presented an analysis by using an innovative wastewater treatment system for domestic effluents based on the electro-oxycoagulation approach implanted in wastewater treatment station located at Palma Sola, Santa Catarina, Brazil. We evaluated samples from domestic effluent (before and after treatment), fountain water, and river water collected from the municipality water system, as well as investigated the residual compound yielded by wastewater treatment. In these samples, we performed physicochemical analysis, investigation of viable helminth eggs, and toxicity and microbiological measurements before and after the treatment. Further, the levels of contaminant metals by X-ray fluorescence-based technique were analyzed. Results show an improvement in the quality of treated water, as demonstrated by microbiological, physicochemical, toxicity, and metal analysis of effluent after treatment. After treatment, river water and fountain water levels of metals in accordance with the maximum limits allowed by Brazilian regulatory agencies. Analysis of residual compound indicated that the workers that handle the residue were not contaminated with the identified metals. Thus, the electro-oxycoagulation-based method demonstrated high efficiency for the treatment of domestic effluents and further prevents contamination of the rivers by the released effluent without treatment.