Developing a novel device, Eggcell, to improve temperature stability during oocyte collection for IVF.

RESEARCH QUESTION: What temperature fluctuations are oocytes exposed to during oocyte retrieval? Can an alternative method of oocyte retrieval be designed to minimize these fluctuations? DESIGN: Mock oocyte retrieval procedures were performed to investigate the change in temperature when the follicular fluid is drained into collection tubes and when the fluid is subsequently poured into dishes to allow identification of the cumulus-oocyte complex (COC). A new device, the Eggcell, has been designed that addresses the problem of these temperature fluctuations. To confirm its safety and demonstrate the clinical applicability of Eggcell, laboratory validation was performed prior to use with human participants (n = 15). RESULTS: Eggcell meets its design specification to provide temperature stability within the physiological range for aspirated follicular fluid. The COC can be successfully retained within the chamber (n = 180) without evidence of loss or damage to the oocytes or compromise of fertilization rate, blastocyst development or clinical outcome. CONCLUSIONS: This study has demonstrated the successful first stages of development of a new medical device. Further studies are needed for comparative evaluation of clinical outcome with standard technology.

An advanced protocol for the purification of whipworm eggs from feces for use as therapeutic agents.

Recent studies have attempted to treat autoimmune diseases using Trichuris suis. whipworm) eggs. Large quantities of eggs can be obtained efficiently by collecting from the feces of the porcine hosts rather than by extracting from the female worm uterus. However, it is difficult to process large amounts of feces using the current methods. In the present study, we propose a method to collect the eggs from bulk feces more efficiently. Collecting the eggs using washing meshes (25 µm sieve) yields 65.7% (56.0-70.7) of eggs (median, min-max) from 100 g feces. Our method, which uses ethyl acetate and simulated gastric fluid, yielded 91.4% (91.4-94.0) of the eggs from 100 g feces into the separated aqueous solution. Egg collection using simulated gastric fluid (SGF) method was also 60 min faster than that using the sieve method. As the SGF used in the experiment is a strongly acidic reagent with a pH of 1-2, embryonation of the eggs was induced by the rapid pH change. As a result, 37.1% (8.0-77.8) of the eggs had embryonated two months after SGF stimulation. Using the developed method, we could process the feces quickly and efficiently. Furthermore, after purification, egg embryonation could be induced without any harmful reagent treatment. This method is expected to be helpful for further research using Trichuris suis eggs.

Methods for quantifying eggs and oviposition rate of Dawestrema cycloancistrium (Monogenea: Dactylogyridae), monogenean parasite of Arapaima gigas (Teleostei: Osteoglossidae).

Dawestrema cycloancistrium is the main ectoparasite causing mortality in fingerlings of pirarucu (Arapaima gigas) in Amazonian aquaculture. Very little is known about the D. cycloancistrium reproductive index and appropriate methods of collecting eggs for investigation. This study aimed to determine the oviposition rate of D. cycloancistrium. To achieve this aim, two egg quantification methods were tested: the estimative method (ME) and the total counting method (MT). Compared with the MT, the ME overestimated the number of eggs counted, which were 2943.5 ± 2840.6 and 1041.5 ± 533 eggs, and the oviposition rate, which was 80.1 ± 58.7 and 31.4 ± 16.4 eggs/parasite/day, for ME and MT, respectively. These results show that for studies quantifying D. cycloancistrium eggs, the total eggs in the sample must be counted, as the estimates made using subsamples are not representative. Using the MT, the oviposition rate for D. cycloancistrium was determined to be 31.4 ± 16.4 eggs per adult parasite per day. The present study demonstrates the egg production capacity of a monogenean species parasite of A. gigas, providing basic biological data for D. cycloancistrium.

A method for collecting eggs of Pseudocapillaria tomentosa (Nematoda: Capillariidae) from zebrafish Danio rerio and efficacy of heat and chlorine for killing the nematode's eggs.

Pseudocapillaria tomentosa is a common pathogen of zebrafish (Danio rerio) in research facilities. We developed a method to collect and concentrate the nematode eggs using a modified sugar centrifugation method and documented their normal development. Embryonating stages with blastomere formation followed by elongation of the embryo prior to larva formation cumulated in developed larvae inside the eggs and hatching after 5-10 day. We then evaluated the efficacy of heat and chlorine to kill them based on a larva development assay. Eggs were exposed to 40, 50, 60 °C for 30 min and 1 h. Chlorine treatment was performed at 100, 250, 500, 1000, 3000 and 6000 ppm for 10 min. Samples exposed to 40 °C for 30 min or 1 h showed incidences of larvated eggs similar to controls. In contrast, no larvation occurred with eggs exposed to either 50 or 60 °C for 30 min or 1 h. Remarkably, in repeated assays, samples exposed to low doses of chlorine (100, 250, 500 and 1000 ppm for 10 min) showed significantly higher incidence of larvation than controls. Eggs treated with 3000 ppm for 10 min did not develop larvae, and no eggs were found after 6000 ppm treatment.

Captive-rearing of Gunnison sage-grouse from egg collection to adulthood to foster proactive conservation and recovery of a conservation-reliant species.

Gunnison sage-grouse (Centrocercus minimus) are distributed across southwestern Colorado and southeastern Utah, United States. Their distribution has decreased over the past century and the species has been listed as threatened by the U.S. Fish and Wildlife Service. Reduced genetic diversity, small population size, and isolation may affect Gunnison sage-grouse population persistence. Population augmentation can be used to counteract or mitigate these issues, but traditional translocation efforts have yielded mixed, and mostly unsuccessful, results. Captive-rearing is a viable, although much debated, conservation approach to bolster wild conservation-reliant species. Although there have been captive-rearing efforts with greater sage-grouse (C. urophasianus), to date, no information exists about captive-rearing methods for Gunnison sage-grouse. Therefore, we investigated techniques for egg collection, artificial incubation, hatch, and captive-rearing of chicks, juveniles, subadults, and adults for Gunnison sage-grouse. In 2009 we established a captive flock that produced viable eggs. From 2009-2011, we collected and artificially incubated 206 Gunnison sage-grouse eggs from 23 wild and 14 captive females. Our hatchability was 90%. Wild-produced eggs were heavier than captive-produced eggs and lost mass similarly during incubation. We produced 148 chicks in captivity and fed them a variety of food sources (e.g. invertebrates to commercial chow). Bacterial infections were the primary cause of chick mortality, but we successfully reduced the overall mortality rate during the course of our study. Conservationists and managers should consider the utility in developing a captive-rearing program or creating a captive population as part of a proactive conservation effort for the conservation-reliant Gunnison sage-grouse.

Evaluation of oviposition traps as an entomological surveillance method for Aedes aegypti (Diptera, Culicidae) / Avaliação das armadilhas de oviposição como método de vigilância entomológica para Aedes aegypti (Diptera, Culicidae)

This study aimed to describe the behavior of oviposition traps for Aedes aegypti over time, to compare it with the larval survey and to investigate the association with climatic variables. It was conducted in São José do Rio Preto city, São Paulo. Daily climatic data and fortnightly measurements for oviposition traps and larval infestation were collected from October 2003 to September 2004. Three different periods were identified in the behavior of oviposition traps' positivity and mean number of eggs: increase, plateau and decrease in values. These measurements followed the variation of climatic data from the first and third periods. High correlation was obtained between the positivity and the mean number of eggs. The oviposition traps showed higher capacity to detect the vector than did larval survey. It was observed that the first (October to December) and third (May to September) periods were considered to be the most suitable to use oviposition traps than larval surveys.

O estudo teve como objetivos descrever o comportamento da armadilha de oviposição para Aedes aegypti ao longo do tempo, comparar com a pesquisa larvária e investigar sua associação com variáveis climáticas. O trabalho foi realizado em São José do Rio Preto, São Paulo. Entre outubro de 2003 a setembro de 2004, as armadilhas e a pesquisa larvária forneceram dados quinzenais e foram obtidos dados climáticos. Três períodos distintos foram identificados no comportamento da positividade das armadilhas e no número médio de ovos: aumento, patamar e decréscimo dos valores. Estas medidas acompanharam as variações climáticas. Alta correlação foi obtida entre a positividade e número de ovos. As armadilhas de oviposição apresentam maior capacidade para detectar o vetor do que a pesquisa larvária. Foi observado que o primeiro (outubro a dezembro) e terceiro (maio a setembro) períodos foram os mais adequados para utilização das armadilhas de oviposição em comparação à pesquisa larvária.