AÇÃO REGULATÓRIA DO GnRH NO DESENVOLVIMENTO EMBRIONÁRIO PRECOCE E VITRIFICAÇÃO DE EMBRIÕES SUÍNOS PELO MÉTODO DE MICROGOTA (ROLE OF GnRH DURING EARLY EMBRYONIC DEVELOPMENT AND DEVELOPMENT OF SWINE EMBRYOS FOLLOWING VITRIFICATION BY MICRODROPLET METHOD)

The objective of this study was to investigate the role of GnRH on the preimplantation development of mouse embryos in vitro. GnRH-I, GnRH-II, and GnRH agonists: Des-Gly, Des-Trp and histrelin did not improve embryo development. However, treatment with the specific GnRH antagonist SB-75 blocked embryo development at morula stage. The inhibition of embryo development by SB-75 could be rescued by the addition of histrelin. To determine which intracellular signaling cascade is involved following binding of GnRH to the GnRHR, embryos were cultured in the presence of specific PKC (GFX) or PKA (SQ22536) inhibitors. The PKC inhibitor blocked embryo development at a similar stage as SB-75, whereas SQ22536 had an inhibitory effect, diminishing blastocyst formation and hatched rates. There are evidences that GnRH has an essential autocrine effect on mouse embryonic development via GnRHR, probably by activating PKC signaling cascade while the inhibition of the GnRH signaling does not activate apoptotic mechanisms involving caspase-3. In another experiment, development in vitro of embryos from Chinese Meishan (M) and occidental white crossbred (WC) females were investigated after improving the vitrification protocol for pig embryos. Efficient cryopreservation of zona pellucida-intact porcine embryos and studies of the difference among breeds could greatly impact the swine industry. The percentage of embryos surviving 24 h after cryopreservation without lysis or degeneration was higher for M (72%) than WC (44%). However, in vitro development of embryos that survived cryopreservation was not different between M and WC at the expanded (64%) or hatched (22%) blastocyst stages. Developmental rates were significantly higher for control embryos than frozen embryos from both breeds at expanded blastocyst stage, but not at hatched blastocyst stage. Rates of expanded blastocyst formation did not differ between M and WC control embryos (98 and 95%, respectively). With a new procedure to warm vitrified pig embryos, the survival rates may be improved. The optimal stages to vitrify pig embryos using the microdroplet method ranges from late compact morula to early expanded blastocyst. The results suggest that M embryos have a higher capacity to survive the vitrification process than WC embryos. O objetivo do presente estudo foi investigar a importância do GnRH no desenvolvimento embrionário precoce em camundongos. GnRH-I, GnRH-II e os GnRH agonistas: Des-Gly, Des-Trp e histrelina não incrementaram o desenvolvimento embrionário. Entretanto, o tratamento com SB-75, um antagonista específico do GnRH, bloqueou o desenvolvimento embrionário no estádio de mórula. A inibição do desenvolvimento embrionário pelo SB-75 pôde ser revertida com a adição de histrelina. Para determinar a cascata do sinal intracelular desencadeada pela ligação do GnRH com o seu receptor, embriões foram cultivados na presença de inibidores específicos da PKC (GFX) e da PKA (SQ22536). O inibidor da PKC bloqueou o desenvolvimento embrionário em estádio similar ao bloqueio mediado pelo SB- 75, enquanto o SQ22536 teve efeito inibitório diminuindo a formação de blastocisto e taxas de eclosão. Os resultados sugerem que o GnRH tem um efeito autócrino essencial no desenvolvimento embrionário através do GnRHR, provavelmente, ativando a cascata da PKC. Por outro lado, a inibição do sinal do GnRH não ativa mecanismos apoptóticos que involvam caspase-3. Em outro experimento, foi investigado o desenvolvimento in vitro de embriões da raça Meishan (M) e branco cruzado (WC) após vitrificação pelo método microgota. O desenvolvimento de protocolos eficientes para criopreservação de embriões suínos com a zona pelúcida intacta e a avaliação das diferenças entre raças pode ter um significativo impacto na suinocultura. A percentagem de embriões que sobreviveram à criopreservação depois de 24 h foi maior na M (72%) do que na WC (44%). No entanto, o desenvolvimento in vitro dos embriões que sobreviveram à criopreservação não foi diferente entre M e WC nos estádios de blastocisto expandido (64%) ou eclodido (22%). Os índices de desenvolvimento foram significativamente mais altos para os embriões controle do que para os embriões vitrificados nas duas raças no estádio de blastocisto expandido, porém não foram diferentes para o estádio de blastocisto eclodido. A formação de blastocisto expandido não diferiu entre os embriões controle M e WC (98 e 95%, respectivamente). Com o novo procedimento (“hot warm”) para descongelar embriões vitrificados pelo método de microgota, pode-se aumentar dos índices de sobrevivência. Os melhores estádios embrionários para a vitrificação de embriões suínos variam de mórula compacta tardia até blastocisto expandido inicial. Os resultados sugerem que embriões M têm mais capacidade de sobreviver ao processo de vitrificação do que embriões WC.

Never Let a Crisis Go to Waste

The phrase, “never let a crisis go to waste” may take frugality to a whole new level! That’s the sentiment of Lynda Applegate, Professor of Business Administration at the Harvard Business School, in a recent on-line article, Building Businesses in Turbulent Times.

NF99-403 Livestock Waste Management Act (Revised March 2001)

The Livestock Waste Management Act requires all livestock operations with 300 animal units or more to be inspected by the Nebraska Department of Environmental Quality (DEQ) to determine whether livestock wastes contaminate surface or ground water. This NebFact discusses the following parts of the Livestock Waste Management Act: Act (how cited); Terms (defined); Livestock operation, exemption, livestock waste control facility, permit, restriction; Construction permit or operating permit (when required), livestock waste control facilities, classification, restrictions; Section (how construed); Cold water class A streams (designation); Permit (acknowledgment required); Livestock operation (request inspection, when, fees, department, duties); Permits (duration, modification); Permit (application and modification fees, Livestock Waste Management Cash Fund (created, use, investment, report, legislative intent); Applicant (rejection, grounds, application, information required, certification required); Postconstruction inspection requirement; Department (contracts authorized, permit application, notice required); Permit application (approval from Department of Natural Resources and Department of Environmental Quality, powers); Council (rules and regulations); and Enforcement of act (legislative intent).

A Fixed Netting System as a Means of Excluding Birds From a Domestic Waste Landfill

Many bird species are attracted to landfills which take domestic or putrescible waste. These sites provide a reliable, rich source of food which can attract large concentrations of birds. The birds may cause conflicts with human interest with respect to noise, birds carrying litter off site, possible transmission of pathogens in bird droppings and the potential for birdstrikes. In the UK there is an 8 mile safeguarding radius around an airfield, within which any planning applications must pass scrutiny from regulatory bodies to show they will not attract birds into the area and increase the birdstrike risk. Peckfield Landfill site near Leeds, West Yorkshire was chosen for a trial of a netting system designed to exclude birds from domestic waste landfills. The site was assessed for bird numbers before the trial, during the netting trial and after the net had been removed. A ScanCord net was installed for 6 weeks, during which time all household waste was tipped inside the net. Gull numbers decreased on the site from a mean of 1074 per hourly count to 29 per hourly count after two days. The gull numbers increased again after the net had been removed. Bird concentrations in the surroundings were also monitored to assess the effect of the net. Bird numbers in the immediate vicinity of the landfill site were higher than those further away. When the net was installed, the bird concentrations adjacent to the landfill site decreased. Corvids were not affected by the net as they fed on covered waste which was available outside the net throughout the trial. This shows that bird problems on a landfill site are complex, requiring a comprehensive policy of bird control. A supporting bird scaring system and clear operating policy for sites near to airports would be required.