Ovarian Dissection Research Articles

Variability in spruce beetle (Coleoptera: Curculionidae, Scolytinae) adult diapause and evidence for oocyte development prior to winter in a Colorado population.

Diapause regulates seasonal insect life cycles and may be highly variable within and among populations due to genetic and environmental variability. Both types of variation may influence how populations respond plastically or evolutionarily to changing climates. We assessed diapause variability in spruce beetle Dendroctonus rufipennis Kirby (Coleoptera: Curculionidae, Scolytinae), a major forest pest whose life cycle timing is regulated by both prepupal and adult diapauses. Using mating studies and ovary dissections, we tested for variability in adult diapause within and between collection sites in Colorado and Wyoming, USA. Ovary morphology suggested that most females from both sites enter diapause prior to egg formation (oogenesis) when reared at warm temperatures. Though previous studies suggested that adult diapause is obligate, we found that a small proportion of females from both populations terminated diapause without winter chilling in the lab. Moreover, we found that most female beetles sampled at the Colorado field site had mature ovaries relatively early in the fall, suggesting that transient exposure to low temperatures may potentiate pre-winter reproductive development. Adult diapause may act primarily as a block to prevent offspring production late in the season but not necessarily as an overwintering phenotype. Overall, our data do not suggest imminent life cycle shifts mediated by adult diapause, but if the observed variability is heritable, diapause regulation may evolve in response to changing environmental conditions.

The attrition, physical and insecticidal durability of two dual active ingredient nets (Interceptor® G2 and Royal Guard®) in Benin, West Africa: results from a durability study embedded in a cluster randomised controlled trial.

Studies evaluating the attrition, physical and insecticidal durability of dual active ingredient (AI) insecticide-treated nets (ITNs) are essential for makingprogrammatic decisions regarding their deployment. We performed a prospective study embedded in a cluster randomised controlled trial (cRCT) to evaluate the attrition, fabric integrity and insecticidal durability of Interceptor® G2 (alpha-cypermethrin-chlorfenapyr) and Royal Guard® (alpha-cypermethrin-pyriproxyfen), compared to Interceptor® (alpha-cypermethrin) in Benin. A total of 2428 study nets in 1093 randomly selected households in five clusters per arm of the cRCT were monitored for ITN attrition and fabric integrity every 6-12months post-distribution. Householders were further surveyed to investigate non-study net use and their preference for ITN fabric types used in the study nets. A second cohort of 120 nets per ITN type were withdrawn every 12months and assessed for chemical content and insecticidal activity in laboratory bioassays. Alpha-cypermethrin bioefficacy was investigated using the susceptible Anopheles gambiae Kisumu strain, and chlorfenapyr and pyriproxyfen bioefficacy were investigated using the pyrethroid-resistant Anopheles coluzzii Akron strain. Net pieces were tested in WHO cone bioassays and tunnel tests for alpha-cypermethrin and in tunnel tests for chlorfenapyr; pyriproxyfen activity was assessed in cone bioassays as the reduction in fertility of blood-fed survivors using ovary dissection. Bioefficacy was expressed as the proportion of ITNs passing predetermined WHO criteria, namely knock-down ≥ 95% or 24/72h mortality ≥ 80% or reduction in fertility ≥ 50%. Overall ITN survivorship was 52% at 24months and fell to 15% at 36months. Median ITN survival time was lower with Royal Guard® relative to Interceptor® [1.6 vs 2.3years; hazard ratio (HR) 1.49, 95% confidence interval (CI) 1.36-1.66; p < 0.001] and Interceptor® G2 (1.6 vs 2.1years; HR 1.33, 95% CI 1.20-1.47; p < 0.001). Householders overwhelmingly preferred polyester nets over polyethylene nets (96%), and more Royal Guard® nets were replaced with spare polyester nets from previous campaigns. All Royal Guard® nets passed efficacy criteria for alpha-cypermethrin at all time points (100%) while ITN pass rates after 24months had fallen to < 40% for pyriproxyfen and chlorfenapyr. The chemical content analysis showed a higher loss rate of the non-pyrethroid insecticides relative to the pyrethroids in each dual ingredient AI ITN; 74% vs 47% for Royal Guard® and 85% vs 63% for Interceptor® G2 at 36months. The median ITN survival time for Interceptor® G2 (2.1years) and Royal Guard® (1.6years) in Benin is substantially lower than 3years. Royal Guard® nets were discarded more quickly by householders, partly due to their low preference for polyethylene nets. The insecticidal activity of the non-pyrethroid insecticides in both dual AI ITNs was short-lived compared to alpha-cypermethrin. The results corroborate the findings from the cRCT conducted in Benin.

P-795 Robust and simple in vitro maturation of ovarian tissue oocytes

Abstract Study question Using in-vitro gametogenesis results in mice, can we perform simple, robust in-vitro maturation (IVM) of oocytes from human ovarian tissue? Summary answer In vitro oogenesis in mice reveals 3 phases of oocyte maturation, IVD, IVG, and IVM. In vivo IVD and IVG simplifes a more robust IVM. What is known already For fertility preservation in cancer patients, IVM of GV’s obtained from ovary tissue with live birth has been achieved, but the results are variable, and great effort has gone into trying to make the “right” culture media, and most are given booster shots of gonadotropin with questionable results. It is easy to obtain many GV oocytes from tiny follicles using open cortical dissection. But it still has remained a puzzle how to do robust IVM in the best way. Study design, size, duration A total of 146 females underwent ovary tissue cryopreservation since 1977. The most recent 38 consecutive such patients between 2 and 38 years of age underwent in-vitro maturation of oocytes (IVM) derived from their ovarian tissue. This is a prospective non-randomized pilot study based upon our results with in vitro oogenesis. Participants/materials, setting, methods After ovarian cortex dissection, the spent media is examined for cumulous complexes (CC’s), which were placed in culture with widely varying concentrations of FSH and HCG, and in a variety of different media. Based on now established mechanisms of in vitro oogenesis, we postulated that the specific media for IVM and the concentrations of HCG would not impact results. There was no prior hormone administration. Main results and the role of chance The harvested germinal vesicle oocyte developed into mature metaphase II oocytes with simple culture media just as well as with “IVM media”, and with HCG concentration from 10 mIU/l to 1000 mIU/l. Of the 38 cases, 18 had had previous chemotherapy or were pre-pubertal. None of these had CC’s. Of the 20 who were post-pubertal and had not yet undergone chemotherapy, the number of CC’s averaged 20 per patient. Of a total of 402 GV oocytes, 137 became mature MII’s (34.1%), varying between 20% and 56%. The only case with low maturation rate (9%) occurred when the cumulus was stripped immediately instead of at 24 hours. Thus mature oocytes can robustly and easily be obtained from ovary tissue with simple media and no need for ovarian stimulation. Furthermore, ovarian stimulation and ovulation are not necessary for oocyte maturation. Oocytes that have just exited IVD that have been exposed to less than 7 days of gonadotropin (IVG) will not mature. Those oocytes exposed to more than 11 days of IVG will degenerate. Thus about 35% of oocytes at any given time will have already been properly prepared in vivo for IVM. Limitations, reasons for caution This is an early proof of concept study in humans with only 30 participants. However, it is completely consistent with what we have predicted from in vitro oogenesis in the mouse. Wider implications of the findings Ovary tissue freezing along with robust IVM will at once provide cancer patients with two possibilities for fertility preservation, without having to delay cancer treatment. Four months of in vivo IVD (non-hormonal) followed by 11 days of IVG (FSH) have already prepared oocytes for IVM. Trial registration number not applicable

Identifying suitable methods for evaluating the sterilizing effects of pyriproxyfen on adult malaria vectors: a comparison of the oviposition and ovary dissection methods

BackgroundNets containing pyriproxyfen, an insect growth regulator that sterilizes adult mosquitoes, have become available for malaria control. Suitable methods for investigating vector susceptibility to pyriproxyfen and evaluating its efficacy on nets need to be identified. The sterilizing effects of pyriproxyfen on adult malaria vectors can be assessed by measuring oviposition or by dissecting mosquito ovaries to determine damage by pyriproxyfen (ovary dissection).MethodLaboratory bioassays were performed to compare the oviposition and ovary dissection methods for monitoring susceptibility to pyriproxyfen in wild malaria vectors using WHO bottle bioassays and for evaluating its efficacy on nets in cone bioassays. Blood-fed mosquitoes of susceptible and pyrethroid-resistant strains of Anopheles gambiae sensu lato were exposed to pyriproxyfen-treated bottles (100 μg and 200 μg) and to unwashed and washed pieces of a pyriproxyfen long-lasting net in cone bioassays. Survivors were assessed for the sterilizing effects of pyriproxyfen using both methods. The methods were compared in terms of their reliability, sensitivity, specificity, resources (cost and time) required and perceived difficulties by trained laboratory technicians.ResultsThe total number of An. gambiae s.l. mosquitoes assessed for the sterilizing effects of pyriproxyfen were 1745 for the oviposition method and 1698 for the ovary dissection method. Fertility rates of control unexposed mosquitoes were significantly higher with ovary dissection compared to oviposition in both bottle bioassays (99–100% vs. 34–59%, P < 0.05) and cone bioassays (99–100% vs. 18–33%, P < 0.001). Oviposition rates of control unexposed mosquitoes were lower with wild pyrethroid-resistant An. gambiae s.l. Cové, compared to the laboratory-maintained reference susceptible An gambiae sensu stricto Kisumu (18–34% vs. 58–76%, P < 0.05). Sterilization rates of the Kisumu strain in bottle bioassays with the pyriproxyfen diagnostic dose (100 μg) were suboptimal with the oviposition method (90%) but showed full susceptibility with ovary dissection (99%). Wild pyrethroid-resistant Cové mosquitoes were fully susceptible to pyriproxyfen in bottle bioassays using ovary dissection (> 99%), but not with the oviposition method (69%). Both methods showed similar levels of sensitivity (89–98% vs. 89–100%). Specificity was substantially higher with ovary dissection compared to the oviposition method in both bottle bioassays (99–100% vs. 34–48%) and cone tests (100% vs.18–76%). Ovary dissection was also more sensitive for detecting the residual activity of pyriproxyfen in a washed net compared to oviposition. The oviposition method though cheaper, was less reliable and more time-consuming. Laboratory technicians preferred ovary dissection mostly due to its reliability.ConclusionThe ovary dissection method was more accurate, more reliable and more efficient compared to the oviposition method for evaluating the sterilizing effects of pyriproxyfen on adult malaria vectors in susceptibility bioassays and for evaluating the efficacy of pyriproxyfen-treated nets.

Ovary Dissection Is a Sensitive Measure of Sterility in Anopheles gambiae Exposed to the Insect Growth Regulator Pyriproxyfen.

Pyriproxyfen (PPF) is an insect growth regulator used in the co-treatment of long-lasting insecticidal nets for its ability to sterilize female mosquitoes. To evaluate the efficacy of PPF-treated nets on mosquito reproductivity, most studies observe oviposition (egg-laying) rates in the laboratory. This technique has several technical disadvantages. Our study assessed if ovarial dissection could serve as an effective proxy for evaluating sterility in Anopheles gambiae mosquitoes. Blood-fed females were exposed to untreated or PPF-treated nets in cylinder assays and followed over several days to observe oviposition rates or egg development by dissection. For identifying PPF-exposed mosquitoes, both techniques demonstrated high sensitivity (oviposition: 99.1%; dissection: 100.0%), but for identifying non-exposed mosquitoes, specificity was significantly higher in the dissection group (52.5% vs. 18.9%). To assess whether dissection could be applied to nets treated with a pyrethroid or co-treated with a pyrethroid and PPF in tunnel tests, a blinded investigator performed dissections to predict the PPF exposure status across different treatment groups. The exposure status of dissected females was predicted with >90% accuracy. We report that dissection is a sensitive technique to assess sterility in female Anopheles gambiae mosquitoes and can be used as a predictor of PPF exposure.

Vector Surveillance for Lymphatic Filariasis After Mass Drug Administration in an Endemic Area: A Case Study in Bekasi

Introduction: Lymphatic Filariasis (LF) re-transmission in endemic areas that have completed mass drug administration (MDA) should be a concern. Entomological data are required to support the elimination of LF. This research aims to present bionomic and evaluative evidence of Wuchereria bancrofti in Culex quinquefasciatus in Bekasi. Methods: Entomological surveys were carried out in Jatimulya Village, Bekasi, from October to November 2019. Female Cx. quinquefasciatus were caught using Human-baited Double Net traps (HDNs) both indoors and outdoors over a 12-hours (from 6 PM to 6 AM). Female mosquitos were subjected to ovary dissection to determine their longevity. In addition, the Man-Hour Density (MHD), Man-Biting Rate (MBR), Daily Survival Rate (DSR), and estimated longevity were calculated. Wuchereria bancrofti was detected using the Polymerase Chain Reaction (PCR) on dissected mosquitos. Results and Discussion: In total 673 female Cx. quinquefasciatus were collected. Culex quinquefasciatus' peak landing time was demonstrated between 12 and 3 AM. The values of Mosquito Parity Rate (MPR) and DSR are 22.88 and 0.692, respectively, implying that the estimated lifespan of dissected mosquitos ranged up to three days. The PCR analysis has revealed that none of the 48 pooled samples of Cx. quinquefasciatus are tested positive for W. bancrofti. Conclusion: Although this survey has found non-existent microfilaria in the LF vector Cx. quinquefasciatus, routine vector monitoring, and surveillance are still required to ensure the long-term viability of the LF elimination program.

Residual malaria transmission and the role of Anopheles arabiensis and Anopheles melas in central Senegal.

Understanding the behavior and ecology of local malaria vectors is essential for the effectiveness of the commonly used vector-targeted malaria control tools in areas of low malaria transmission. This study was conducted to determine species composition, biting behavior and infectivity of the major Anopheles vectors of Plasmodium falciparum in low transmission settings in central Senegal. Adult mosquitoes were collected using human landing catches during 2 consecutive nights and Pyrethrum Spray Catches in 30-40 randomly selected rooms, from July 2017 to December 2018 in 3 villages. Anopheline mosquitoes were morphologically identified using conventional keys; their reproductive status assessed by ovary dissections, and a sub-sample of Anopheles gambiae s.l. were identified to species level using polymerase chain reaction (PCR). Plasmodium sporozoite infections were detected using real-time quantitative PCR. During this study 3684 Anopheles were collected of which 97% were An. gambiae s.l., 0.6% were Anopheles funestus, and 2.4% were Anopheles pharoensis. Molecular identification of 1,877 An. gambiae s.l. revealed a predominance of Anopheles arabiensis (68.7%), followed by Anopheles melas (28.8%), and Anopheles coluzzii (2.1%). The overall human-biting rate of An. gambiae s.l. was highest in the inland site of Keur Martin with 4.92 bites per person per night, while it was similar in the deltaic site, Diofior (0.51) and the coastal site, Mbine Coly (0.67). Parity rates were similar in An. arabiensis (45%) and An. melas (42%). Sporozoite infections were detected in both An. arabiensis and An. melas with the respective infection rates of 1.39% (N = 8) and 0.41% (N = 1). Results suggest that low residual malaria in central Senegal is transmitted by An. arabiensis and An. melas. Consequently, both vectors will need to be targeted as part of malaria elimination efforts in this area of Senegal.

Effect of age on wingbeat frequency of Aedes aegypti and potential application for age estimation of mosquitoes.

To combat mosquito-borne diseases, a variety of vector control tools have been implemented. Estimating age structure in populations of vector species is important for understanding transmission potential. Age-grading techniques have been used as critical methods for evaluating the efficacy of vector control tools. However, methods like mark-release-recapture and ovarian dissection are laborious and require a high level of training. For decades, scientists have discussed the wide array of acoustic signatures of different mosquito species. These distinguishable wingbeat signatures with spatiotemporal classification allow mosquitoes of the same species to locate one another for mating. In recent years, the use of sensitive acoustic devices like mobile phones have proved effective. Wingbeat signatures can be used to identify mosquito species without the challenge of intensive field collections and morphological and molecular identifications. In this study, laboratory Aedes aegypti (L.) female and male wingbeats were recorded using mobile phones to determine whether sex and age differences with chronological time, and across different physiological stages, can be detected. Our results indicate significantly different wingbeat signatures between male and female Ae. aegypti, and a change of wingbeat frequencies with age and reproduction stage in females.

Dissection, Fixation, and Standard Staining of Adult Drosophila Ovaries.

Studies of the Drosophila ovary have provided significant insight into the molecular and cellular processes that control cell function, tissue organization, and animal development. To characterize mutants with defects in oogenesis or to observe the distribution of gene products involved in egg production, the ovaries must be carefully extracted and prepared for analysis. This chapter describes the manual dissection of ovaries from adult Drosophila females, followed by standard fixation and staining of the isolated tissue. Specifically, this chapter provides procedures for simple DNA and F-actin staining to assess cell and tissue morphology, as well as immunostaining to localize proteins of interest.

Wolbachia inhibits ovarian formation and increases blood feeding rate in female Aedes aegypti.

Wolbachia, a gram-negative endosymbiotic bacterium widespread in arthropods, is well-known for changing the reproduction of its host in ways that increase its rate of spread, but there are also costs to hosts that can reduce this. Here we investigated a novel reproductive alteration of Wolbachia wAlbB on its host Aedes aegypti, using studies on mosquito life history traits, ovarian dissection, as well as gene expression assays. We found that an extended period of the larval stage as well as the egg stage (as previously shown) can increase the proportion of Wolbachia-infected females that become infertile; an effect which was not observed in uninfected females. Infertile females had incomplete ovarian formation and also showed a higher frequency of blood feeding following a prior blood meal, indicating that they do not enter a complete gonotrophic cycle. Treatments leading to infertility also decreased the expression of genes related to reproduction, especially the vitellogenin receptor gene whose product regulates the uptake of vitellogenin (Vg) into ovaries. Our results demonstrate effects associated with the development of infertility in wAlbB-infected Ae. aegypti females with implications for Wolbachia releases. The results also have implications for the evolution of Wolbachia infections in novel hosts.

No association between habitat, autogeny and genetics in Moroccan Culex pipiens populations

BackgroundMosquitoes of the Culex pipiens complex are found across the globe and are the focus of many research studies. Among the temperate species C. pipiens sensu stricto (s.s.), two forms are usually described: molestus and pipiens. These two forms are indistinguishable in terms of morphology but show behavioral and physiological differences that may have consequences for their associated epidemiology. The two forms are well defined in the northern part of the species distribution, where autogeny is strictly associated with the molestus form. However, whether the two remain distinct and show the characteristic differences in behavior is less clear in North Africa, at the southern edge of their range.MethodsThe association between autogeny, as determined by ovarian dissection, and molecular forms, based on the CQ11 microsatellite marker, was studied in six Moroccan populations of C. pipiens.ResultsAn overall low prevalence of autogeny was found at three of the Moroccan regions studied, although it reached 17.5% in the Agadir population. The prevalence of form-specific CQ11 alleles was quite similar across all populations, with the molestus allele being rarer (approx. 15%), except in the Agadir population where it reached 43.3%. We found significant deficits in heterozygotes at the diagnostic CQ11 locus in three populations, but the three other populations showed no significant departure from panmixia, which is in line with the results of a retrospective analysis of the published data. More importantly, we found no association between the autogeny status and CQ11 genotypes, despite the many females analyzed.ConclusionsThere was limited evidence for two discrete forms in Morocco, where individuals carrying pipiens and molestus alleles breed and mate in the same sites and are equally likely to be capable of autogeny. These observations are discussed in the epidemiological context of Morocco, where C. pipiens is the main vector of several arboviruses.Graphical

Durability of three types of dual active ingredient long-lasting insecticidal net compared to a pyrethroid-only LLIN in Tanzania: methodology for a prospective cohort study nested in a cluster randomized controlled trial

BackgroundProgress achieved by long-lasting insecticidal nets (LLINs) against malaria is threatened by widespread selection of pyrethroid resistance among vector populations. LLINs with non-pyrethroid insecticides are urgently needed. This study aims to assess the insecticide and textile durability of three classes of dual-active ingredient (A.I.) LLINs using techniques derived from established WHO LLIN testing methods to set new standards of evaluation.MethodsA WHO Phase 3 active ingredients and textile durability study will be carried out within a cluster randomized controlled trial in 40 clusters in Misungwi district, Tanzania. The following treatments will be evaluated: (1) Interceptor®G2 combining chlorfenapyr and the pyrethroid alpha-cypermethrin, (2) Royal Guard® treated with pyriproxyfen and alpha-cypermethrin, (3) Olyset™ Plus which incorporates a synergist piperonyl butoxide and the pyrethroid permethrin, and (4) a reference standard alpha-cypermethrin only LLIN (Interceptor®). 750 nets will be followed in 5 clusters per intervention arm at 6, 12, 24 and 36 months post distribution for survivorship and hole index assessment. A second cohort of 1950 nets per net type will be identified in 10 clusters, of which 30 LLINs will be withdrawn for bio-efficacy and chemical analysis every 6 months up to 36 months and another 30 collected for experimental hut trials every year. Bio-efficacy will be assessed using cone bioassays and tunnel tests against susceptible and resistant laboratory strains of Anopheles gambiae sensu stricto. Efficacy of field-collected nets will be compared in six experimental huts. The main outcomes will be Anopheles mortality up to 72 h post exposure, blood feeding and egg maturation using ovary dissection to assess impact on fecundity.ConclusionsStudy findings will help develop bio-efficacy and physical durability criteria for partner A.I., in relation to the cRCT epidemiological and entomological outcomes, and refine preferred product characteristics of each class of LLIN. If suitable, the bioassay and hut outcomes will be fitted to transmission models to estimate correlation with cRCT outcomes.Trial registration number: NCT03554616.

The Automatic Classification of Pyriproxyfen-Affected Mosquito Ovaries.

Simple SummaryWith resistance to the pyrethroid family of insecticides increasing, it is more important than ever that tools are available to measure the efficacy of alternatives. Pyriproxyfen (PPF) is an alternative insecticide whose mode of action sterilises adult mosquitoes. Consequently, the efficacy of PPF-based tools can be measured through visual examination of egg development by trained experts. This, however, can be a time-consuming process for which the required expertise can be difficult to train and is not available in many contexts. As such, we propose that an objective machine learning program, which can automatically classify the fertility status of adult mosquito ovaries via a colour image, be developed to improve the speed, accuracy, and consistency of assessment. This study shows that a convolutional neural network, built in Python’s TensorFlow library, can quickly classify images of dissected ovaries into either ‘fertile’ or ‘infertile’ with a high accuracy rate. Such an application would be a practical and accessible tool available to all researchers studying the efficacy of PPF or other insecticides with a similar mode of action.Pyriproxyfen (PPF) may become an alternative insecticide for areas where pyrethroid-resistant vectors are prevalent. The efficacy of PPF can be assessed through the dissection and assessment of vector ovaries. However, this reliance on expertise is subject to limitations. We show here that these limitations can be overcome using a convolutional neural network (CNN) to automate the classification of egg development and thus fertility status. Using TensorFlow, a resnet-50 CNN was pretrained with the ImageNet dataset. This CNN architecture was then retrained using a novel dataset of 524 dissected ovary images from An. gambiae s.l. An. gambiae Akron, and An. funestus s.l., whose fertility status and PPF exposure were known. Data augmentation increased the training set to 6973 images. A test set of 157 images was used to measure accuracy. This CNN model achieved an accuracy score of 94%, and application took a mean time of 38.5 s. Such a CNN can achieve an acceptable level of precision in a quick, robust format and can be distributed in a practical, accessible, and free manner. Furthermore, this approach is useful for measuring the efficacy and durability of PPF treated bednets, and it is applicable to any PPF-treated tool or similarly acting insecticide.

Comparison of surgical outcomes between 3-dimensional and 2-dimensional laparoscopy of ovarian cyst (LOOC): a randomised controlled trial

The purpose of this study was to compare the surgical outcomes and efficacy of 3-dimensional (3D) versus 2-dimensional (2D) imaging systems for the treatment of ovarian cyst. A total of 46 patients undergoing a laparoscopic ovarian cystectomy were randomly assigned to either the 3D or 2D laparoscopy group. The primary outcome measure was the operative blood loss. The secondary outcome measure was visually induced motion sickness (VIMS), task efficacy during laparoscopy, and postoperative complication. There were no differences in baseline demographics between the two groups. The operative blood loss was significantly smaller in the 3D groups (28.7 ± 11.6 mL) than in the 2D groups (46.5 ± 24.4 mL) (p = .012). VIMS score was significantly higher in the 3D groups than the 2D groups (p < .001). 3D laparoscopy was superior to 2D in terms of the task efficacy of ovarian cyst enucleation (p < .001), adhesiolysis or dissection (p < .001), and ovarian suturing (p = .008). None of the patients in both groups developed operative complications. In conclusion, a 3D imaging system showed a more favourable surgical outcome and improved task efficacy than 2D in laparoscopic ovarian cystectomy. However, 3D laparoscopy tends to cause more frequent VIMS in surgeons. Impact statement What is already known on this subject? Several studies examining the possible benefits and drawbacks of a 3D imaging system versus 2D in laparoscopic surgery have brought about conflicting results. However, there have been few studies comparing the surgical outcomes of 3D and 2D laparoscopic ovarian cystectomy. What do the results of this study add? 3D laparoscopy showed favourable surgical outcomes and improved task efficacy than 2D laparoscopy in ovarian cystectomy. What are the implications of these findings for clinical practice and/or further research? More complex procedures, such as suturing and adhesiolysis, might be easier to perform with 3D laparoscopy than with 2D laparoscopy. Therefore, further large studies of 3D gynaecologic laparoscopy with different complexities and for surgeons with different surgical skills are needed.

Acaricidal Properties of Bio-Oil Derived From Slow Pyrolysis of Crambe abyssinica Fruit Against the Cattle Tick Rhipicephalus microplus (Acari: Ixodidae).

Slow pyrolysis is a process for the thermochemical conversion of biomasses into bio-oils that may contain a rich chemical composition with biotechnological potential. Bio-oil produced from crambe fruits was investigated as to their acaricidal effect. Slow pyrolysis of crambe fruits was performed in a batch reactor at 400°C and chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). The bio-oil collected was used in bioassays with larvae and engorged females of the cattle tick Rhipicephalus microplus. Biological assays were performed using the larval packet test (LPT) and adult immersion test. The GC-MS of crambe fruit bio-oil revealed mainly hydrocarbons such as alkanes and alkenes, phenols, and aldehydes. The bio-oil in the LPT exhibited an LC90 of 14.4%. In addition, crambe bio-oil caused female mortality of 91.1% at a concentration of 15% and a high egg-laying inhibition. After ovary dissection of treated females, a significant reduction in gonadosomatic index was observed, indicating that bio-oil interfered in tick oogenesis. Considering these results, it may be concluded that slow pyrolysis of crambe fruit affords a sustainable and eco-friendly product for the control of cattle tick R. microplus.

Planarian Ovary Dissection for Ultrastructural Analysis and Antibody Staining.

Accessibility to germ cells allows the study of germ cell development, meiosis, and recombination. The sexual biotype of the freshwater planarian, Schmidtea mediterranea, is a powerful invertebrate model to study the epigenetic specification of germ cells. Unlike the large number of testis and male germ cells, planarian oocytes are relatively difficult to locate and examine, as there are only two ovaries, each with 5-20 oocytes. Deeper localization within the planarian body and lack of protective epithelial tissues also make it challenging to dissect planarian ovaries directly. This protocol uses a brief fixation step to facilitate the localization and dissection of planarian ovaries for downstream analysis to overcome these difficulties. The dissected ovary is compatible for ultrastructural examination by transmission electron microscopy (TEM) and antibody immunostaining. The dissection technique outlined in this protocol also allows for gene perturbation experiments, in which the ovaries are examined under different RNA interference (RNAi) conditions. Direct access to the intact germ cells in the ovary achieved by this protocol will greatly improve the imaging depth and quality and allow cellular and subcellular interrogation of oocyte biology.

Potential trade-offs between reproduction and migratory flight in Spodoptera frugiperda

Since 2016, the fall armyworm (FAW, Spodoptera frugiperda) has invaded large parts of Africa and Asia, impacting millions of hectares of maize crops and thereby posing a major threat to food security. The rapid geographic spread and outbreak dynamics of S. frugiperda are tied to its unique dispersal ability and long-distance migration capability. Yet, up till present, limited research has been conducted on the physiological determinants of S. frugiperda flight and migration. In this study, we used laboratory experiments to assess whether mating and oviposition affect S. frugiperda flight ability and wingbeat frequency. During 2019–2020, migratory FAW females were trapped in Yunnan (China) and dissected to assess ovarian development. Tethered flight assays showed that gravid S. frugiperda females exhibited strong flight ability at 1–3 days following the onset of oviposition. Flight distance and duration negatively correlated with the number of deposited eggs. Ovarian dissections further showed that over 50% of migrant females were mated and 46–54% had initiated oviposition. Our study shows the complex, yet nuanced effects of reproductive status on flight capacity, with possibly a facultative trade-off between flight and reproduction. These novel insights into S. frugiperda physiology and migration behavior can guide future monitoring and integrated pest management (IPM) programs against this newly-invasive pest in China and abroad.

Species Composition, Parous Rate, and Infection Rate of Anopheles Mosquitoes (Diptera: Culicidae) in Bahir Dar City Administration, Northwest Ethiopia.

Anopheles mosquitoes are the main vectors of malaria. There is little information on the current entomological aspects of Anopheles mosquitoes in Amhara region of northwestern Ethiopia. Therefore, the aim of this study was to assess the prevailing species composition, parous rate, and infection rate of Anopheles mosquitoes in the Bahir Dar city administration. A community-based cross-sectional study was conducted from January through July 2020. For this, six Centers for Disease Control and Prevention light traps (three traps indoor and three traps outdoor) were used to collect adult female Anopheles mosquitoes. The species were morphologically identified, and the parous and infection rates were determined via dissection of ovaries and salivary gland, respectively. A total of 378 adult female Anopheles mosquitoes comprised of three species (Anopheles d'thali, Anopheles rhodesiensis, and Anopheles gambiae complex) were collected and identified at the study sites. Anopheles rhodesiensis was the predominant species accounting for 90% of all collections at the Zenzelima site, followed by An. gambiae complex (6.5%). In contrast, An. gambiae complex was the predominant species at the Tis Abay site, comprising 94% of captures. The overall parous and infection rates were 35 (62.5%) and 1 (2.9%), respectively.

Ameliorative effects of Withania coagulans and Metformin on Ovarian morphology in Polycystic ovarian disease induced rats

Background: Various medicinal herb plants are being used in place of metformin for treatment of polycystic ovarian disease for their less harmful effects. Withania coagulans (WC) is a herb known for its insulin sensitizing and weight reducing properties. The present study was done to determine the influence of aqueous extract of Withania coagulans (aqWC) and metformin on ovarian weight and ovarian folliculogenesis in polycystic ovarian disease-induced rats.&#x0D; Material and Methods: An experimental animal study was carried out at the Physiology Department of Islamic International Medical College, Rawalpindi from April 2016 to March 2017. Forty female Sprague Dawley rats were divided initially into two groups. Group A (Normal Control) and Disease induced group. Standardized laboratory diet was fed to Group A while the disease induced group was given standardized laboratory diet and letrozole solution orally (1mg/kg) for 21 days to induce Polycystic ovary syndrome, which was established by observing estrous cycle of rats. Disease induced group was then split into group B (PCOS control), C (Aqueous Withania coagulans treated) and D (Metformin treated) and observed after 14 days. Groups A and B underwent ovarian dissection after 21 days and groups C and D underwent dissection at the end of experiment (after 35 days). Independent sample t-test was used for the comparison between the control and disease induced group.&#x0D; Results: Group B showed a significant increase in ovarian weight in comparison to group A rats (P &lt; .05). Treatment with Withania coagulans and metformin significantly decreased ovarian weight (P &lt; .05) and increased primary, Graafian, antral follicular count and corpus luteum along with reduction in cystic follicular count in letrozole-induced polycystic ovarian disease rats. Improved folliculogenesis was also observed in the same groups (groups C &amp; D).&#x0D; Conclusions: Withania coagulans can be a substitute for improvement of ovarian follicular development in polycystic ovarian disease.

Apparent Densities of Stomoxys species (Diptera, Muscidae) of different Physiological Ages caught with Vavoua Trap differ With Landscape and trapping period

Stable flies (Stomoxys spp.) are common pests of livestock in the peri-urban rangelands of Vina Division of the Adamawa Plateau. No documented information is available on their diurnal dynamics in relation to physiological age and landscape. The main aim of this study was to determine the trap apparent density (ADT) of Stomoxys and associate it to their diurnal activity rhythm and landscape. Vavoua traps (n=12) were used for Stomoxys collection. Trapping was carried out for seven days i.e. three days consecutively in August and four days consecutively in October, 2016. Three traps were pitched in each of the three biotopes (river, cattle corral and gallery forest) of the four study sites (Galim, Mbidjoro, Velambai and Soukourwo). All female Stomoxys captured underwent ovarian dissection for the determination of their physiological ages (parous or nulliparous). A total of 218 Stomoxys were caught and identified into four species (S. niger niger, S. omega, S. calcitrans, and S. xanthomelas) with an overall ADT of 2.59 flies per trap per day and Stomoxys niger niger (1.13 s.n.n. per trap per day) recorded the highest ADT. Species richness was study site dependent. Higher catches of females 113 (51.83%) were made than that of their male 105 (48.17%) counterparts. Galim recorded the highest Stomoxys apparent density (4.90) as compared to other sites with a statistically significant difference (P&lt;0.05). The proportion of parous 87 (59.30%) was higher than that of their nulliparous 46 (40.71%) counterparts. Parity differed with landscape and capture periods. S. n. niger was the most frequent in all biotopes and Galim recorded the highest ADT. The population of female flies was made up of adults (parous) with a bimodal diurnal activity rhythm whereas males had a unimodal activity rhythm.