Colonisation of Perennial Ryegrass by Endophytic Bacteria

Colonisation of Perennial Ryegrass by Endophytic Bacteria 2. Materials and methods 2.1. Preparation of culture media Nutrient agar (NA) – NA (Oxoid, Basingstoke, Hants, UK) was prepared by weighing out 28g of nutrient agar powder and dissolving in 1L of deionised H2O, by warming on a hot plate. This was autoclaved at 121 °C and 15psi for 15 minutes. The medium was then aseptically dispensed in sterile petri dishes and allowed to solidify. Sucrose glutamate agar (SGA) – SGA was prepared by dissolving 20g of sucrose, 2g of glutamate, 15g of agar bacteriological (Agar No.1) and 1g of K2HPO4 in 1L deionised H2O. This was autoclaved as outline above. The medium was allowed to cool to approximately 60 °C at which 5ml of MgSO4 and 1ml of kanamycin was aseptically added to the medium using a Nalgene Syringe Filters. The medium was then poured into petri dishes and allowed to solidify. Nematode growth medium (NGM) – 1L of NGM was prepared by dissolving 3g of NaCl, 17g of agar, and 2.5g of peptone in 975ml of deionised H2O in a Duran bottle. This was autoclaved as outline above. The media was allowed to cool for 15 minutes at which 1ml of 1M CaCl2, 1ml of 5mg/ml cholesterol in ethanol, 1ml of 1M MgSO4 and 25ml of 1M KPO4 buffer were added aseptically in a Duran bottle. The bottle was swirled to ensure the medium was mixed properly and then aseptically dispensed in sterile petri dishes and allowed to solidify. 2.2. Culturing entomopathogenic nematodes (EPN) 9cm filter paper was placed in sterile petri dish and 1ml of stock nematode suspension was pipetted onto filter paper. Five live Galleria mellonella (wax moths) were placed onto petri dish and sealed with parafilm (Figure 1). The dish were kept in a dark and observed daily for the insect mortality. Once the G. mellonella were dead they were transferred to white traps. Figure 1 Galleria (wax moth) baiting technique. 2.3. Preparation of white traps White traps (White, 1927) were prepared which the lid of a small petri dish (35x10mm) was placed on top of the base and placed in a clear tub, this was then covered with filter paper. 30ml of water was added to the plastic container just to allow absorbing through the filter paper. The dead G. mellonella was placed on the moist filter paper and it was covered and placed in dark for 7 to 14 days (Figure 2). The white traps were observed daily for emergence of infective juveniles (IJs) by using stereoscope (Figure 3). Whenever the water around the stage became densely concentrated with nematodes, the water was decanted into a container and replaced with another 30ml of deionised H2O. The nematodes were stored at 4 °C in 25ml of distilled water until needed for use. Figure 2 – Galleria larvae on white trap. Figure 3 – Galleria larvae under stereoscope (20X). 2.4. Culturing Caenorhabditis elegans (C. elegans) on NGM (Couillault. C, 2002) A stock culture of C. elegans was cultures from a previous stock of C. elegans. In a laminar air flow, the plate was divided into equal sections. A sterilized scalpel was used to cut sections from the stock culture to the centre of a fresh NGM plate (Figure 4). The plates were sealed using a strip of parafilm and stored at room temperature or inside incubator at 21 °C for 3 days. The petri plates were observed regularly using stereoscope (Figure 5). Figure 4 – Culturing C. elegans on NGM (Chunking method). Figure 5 – Observation of C. elegans under stereoscope (20X). 2.5. Culturing of bacterial endophytes The endophytic bacterial strains used in this study were provided by IT Carlow stock collection and have been labelled with gfp (green fluorescent protein). The Pseudomonas strains used were F113, L321 and L228. 1L of Nutrient broth (NB) was prepared and 10ml was pipetted into Mc Cartney bottles and autoclaved. The Pseudomonas strains of L321gfp, L228gfp, and F113gfp were inoculated aseptically using a wire loop and incubated at 30 °C for 24hours in an orbital shaker incubator. The gfp strains in the nutrient broth were then streaked onto fresh non contaminated nutrient agar and SGA in duplicate for each of bacterial strains, using the quadrant streaking method. All plates were covered with parafilm, labelled and incubated at 30 °C for 24hours. A gram stain, catalase test, oxidase test, and observation of morphological characterisation were carried out for the Pseudomonas strains of L321gfp, L228gfp, and F113gfp. 2.6. Quantification of nematodes S. feltiae and C. elegans The stock suspension of nematodes was divided into 50ml samples. 100 µl of the infective juvenile suspension of each sample was pipetted using micropipette onto a counting tray and tally counter was used to count for nematodes under stereoscope (Figure 6). Once they were counted, the sample was discarded and washed with deionised H2O. This was repeated 10 times and the average number of nematodes was calculated. Figure 6 – Counting chamber containing suspension of nematodes. 2.7. Preparation of soil samples 2500g of soil was autoclaved as outlined in section 2.1. The soil was dried in an oven at 55 °C for 24 hours. The soil samples were prepared by weighing out 90g into plastic cups (Figure 7) and temporarily covered with tin foil to prevent any contamination. Figure 7 – Each plastic cup contains 90g of soil. 2.8. Preparation for Sodium alginate beads (Bashan, 2002) The microbeads stock suspension were prepared by dissolving 10g of Sodium alginate in 500ml of deionised H2O , 10g of Calcium Chloride in 500ml of deionised H2O, and 5g of powdered skimmed milk in 50ml of deionised H2O. All components were autoclaved separately and the skimmed milk was autoclaved only for 10 minutes. The working solution was prepared from the stock solutions as follows; 5ml of skimmed milk, 15ml of sodium alginate and 5ml inoculum. The components of the working solution were poured into a sterile petri dish and mixed using a sterile rod. Parafilm was used to plug the spout of 20ml syringe, the alginate mixture and grass seeds were subsequently added. The parafilm was then removed and a sterile rod was used to ensure the coated seeds dropped out individually, into a beaker containing Calcium Chloride on a magnetic stirring plate (Figure 8). The beads were washed at least three times with sterile distilled water and stored in a sealed petri dished until needed for use. Figure 8 – Beaker containing Calcium Chloride on a magnetic stirring plate. 2.9. Isolation of bacterial endophytes from alginate beads (Bashan, Y and Levanony, 1989) In order to isolate and enumerate bacterial endophytes from microbeads seed coating. Six alginate beads containing individual bacterium were dissolved in 10ml of 0.25M Potassium Phosphate buffer in a test tube and incubated at 30 °C for 24 hours. The bead was then shaken on a vortex for 5 minutes to break down the alginate. Using a serial dilution method, 1ml of bead samples containing bacteria was serially diluted in 9ml of sterile ringers from 10-1 to 10-10 (Figure 9); this was carried out onto SGA in triplicate and incubated at 30 °C for 24 hours. Figure 9 – Most probable number (MPN) method/Serial dilution method. 2.10. Isolation of bacterial endophytes from plants (Keogh, E, 2009) Each plant was removed from pots and excess soil was removed. Three samples were taken from each plant (stem, root, and rhizosphere). The stems and roots were surface sterilised with 1% of sodium hydrochloride and washed twice with sterile water. The stems and roots were cut with sterile scalpel and crushed with a pestle and mortar in 5ml of Ringers solution. 100 µl of suspension was added to 900 µl of sterile Ringers in 2.5ml microfuge tubes. The serial dilutions were carried out and the resulting dilutions of 30 µl were then pipetted onto SGA in triplicate and incubated at 30 °C for 24 hours. 3. Results 3.1. Characterisation of bacterial endophytes The classical approach to bacteria identification involves preliminary microscopic examination of the gram-stained preparation for its categorisation which would later form the basis for the selection of biochemical test to be performed to test their identity. Table (Table 1) and figures (Figure 10(a) to (f)) below shows the characterisation for each strain of endophyte. Figure 10 – Characterisation of bacterial endophytes. (a) Culture plate observation for F113. (b) Microscopy examination for F113. (c) Culture plate observation for L228. (d) Microscopy visualisation for L228. (e) Culture plate observation for L321. (f) Microscopy examination for L321. 3.2. Counting of nematodes S. feltiae and C. elegans The number of nematodes was counted per well in four weeks’ time (Table 2) and a chart (Figure 11) was produced comparing the S. feltiae and C. elegans. This was repeated 10 times and the average number of nematodes was calculated. Table 2 – Quantification of nematodes. Figure 11- Comparison between No. of IJ/100 µl with the time of S. feltiae and C. elegans. 3.3. Soil samples inoculation In order to make sure the soil samples free from contamination, the serial dilutions were carried out and the resulting dilutions of 30 µl were then pipetted onto SGA in triplicate and incubated at 30 °C for 24 hours (Figure 12). The results indicated no growth in the soil samples. Figure 12 – No growth in the soil samples. 3.4. Isolation of bacterial endophytes from alginate beads In order to isolate bacterial endophytes from alginate seed coating, the beads were plated onto SGA and incubated at 30 °C for 24 hours. The results indicated that fluorescent which present of green colour pigment (Figure 13). Figure 13 – SGA changed to green colour. 3.5. Colonisation and enumeration of endophytic bacterial within plant tissues Inoculated perennial ryegrass (Lolium perenne) was allowed to grow for 4 weeks (Figure 14) before sampling took place. Total bacterial population of gfp expressing were determined for each of the tissues examined. Endophytic bacteria are considered to be those isolated from the internal tissues of surface sterilised plants. However, it is difficult to determine whether an organism is truly endophytic or merely a survivor of the surface sterilisation process. To ensure that the sterilisation processes were adequate, the sterilised tissues were pressed against the surface of a sterile SGA plate and samples of the third water rinsing were also plated onto SGA plates (Figure 15). Bacterial counts (Figure 16 and 17) on these plates were always between 10-1 to 10-4 CFUs per ml (Table 3 and 4), which was considered to be a good indication that the surface was successfully sterilised. However, under epifluorescent microscopy, the gfp expressing from inoculated plants. Pseudomonas species str ain L321 was detected only in the rhizosphere and the interior root tissues of inoculated plant (Figure 18(a) and (b)). Figure 14 Lolium perenne was allowed to grow. Figure 15 – Bacterial count on SGA plates Table 3 –Plate counts on Pseudomonas strain of L321. Figure 16 – Bacterial counts between S. feltiae and C. elegans. Table 4 – Plate counts on Pseudomonas strain of F113. Figure 17 – Bacterial counts between S. feltiae and C. elegans. Figure 18 – Visualisation under epifluorescent microscope. (a) L321gfp bacteria (400X). (b) L321gfp bacteria (100X). 4. Discussion Bacterial colonisation of the internal tissues of plants has been described in almost all plant species examined so far. Although many of these bacteria are phytopathogenic, a considerable number have also been found that colonise the plant without causing disease. Such bacteria are referred to as bacterial endophytes. Colonisation may take place at the local tissue level or throughout the plant, with bacterial colonies and biofilms residing latently in the intercellular spaces and inside the vascular tissues. This project describes the isolation, identification and colonisation efficiency of perennial ryegrass by gfp labelled bacterial endophytes. Furthermore, this study has shown the successful colonisation of perennial ryegrass by three endophytic bacterial strains under controlled conditions. The Pseudomonas strains, L321 demonstrated efficient colonisation resulting in high population numbers within the plant tissues. This experiment shows that the L321 bacteria endophyte worked successfully with the C. elegans to increase the plant colonisation. In this project, L228 was discarded due to the lawns were very poor and did not fluorescence very well so the experiment carried out only with L321 and F113. During the characterisation of bacteria endophytes, the results were obtained which the genus Pseudomonas appeared in Gram negative bacilli motile by polar flagella. In addition, in catalase test shows positive formed the bubbles when comes into contact with Hydrogen Peroxide. On the other hand, the results show that they are oxidase producing which will be oxidised to deep purple colour. Also, when nematodes had been put on plates and timescale had begun it was noticed that some plates start to dry out which may be due to the media drying out so to overcome this this, the plates have to seal with parafilm to prevent from dry out. Furthermore, other notice when the plates rinsed with water, I noticed crystals formed in the media and this may be due to the tem perature problem. Generally in the experiment there is no physical quantity can be measured with perfect certainty; there are always errors in any measurement. For example, the systematic errors are due to poorly calibrated instrument; observational for example, errors in judgment of an observer when reading the scale of a measuring device to the smallest division. 5. Conclusion In conclusion, this study has shown the successful colonisation of perennial ryegrass by three endophytic bacterial strains under controlled conditions. The Pseudomonas strains, L321 demonstrated efficient colonisation resulting in high population numbers within the plant tissues. Hence, none of the introduced strains showed any signs of pathogenicity towards their host plant and others tested. Many studies have shown that the colonisation levels in field trials are less successful than those in laboratory trials. This is probably due to increased microbial competition and less favourable environmental conditions. Therefore, additional long-term field trials need to be carried out in order to gain a better understanding of the colonisation pattern and population dynamics of endophytic bacteria in the perennial ryegrass. If time permitted future work would include, the carrying out of plant biomass which is a time consuming method that involves drying of cells and to perform by weighi ng the dry and fresh weight of each plant. 6. References Brown, R.H. and Kerry, B.R. (1987). Principles and Practice of Nematode Control in  Crops. Academic Press, Sydney. 447 pp. Evans, D., Trudgill, D.L. and Webster, J.M. (1993). Plant Parasitic Nematodes in  Temperate Agriculture. CAB International, Wallingford. 648 pp.   Luc, M., Sikora, R.A. and Bridge, J. (2005). Plant Parasitic Nematodes in Subtropical  and Tropical Agriculture, 2nd edn. CAB International, Wallingford. 871 pp. Mai, W.F. and Mullin, P.G. (1996). Plant Parasitic Nematodes. A Pictorial Key to  Genera, 5th edn. Comstock, London and Cornell University, Ithaca. 276 pp. Ahmad, F., Ahmad, I., Khan, M.S. 2008. Screening of free-living rhizospheric bacteria for their multiple plant growth promoting activities. Microbiol. Res. 163, 173-181. Barraquio W L, Ladha J K, Watanabe I. Isolation and identification of a N2 fixing Pseudomonas associated with wetland rice. Can J Microbiol. 1983;29:867–873.

Twelve Angry Men and the Jurors of The Kevorkian Trials Essay -- compli

â€Å"I can’t Function as a human being†¦ I want the right to die,† pleaded Sherry Miller, patient of Dr. Jack Kevorkian, the famous â€Å"Death Doctor† (Chermak 107). The entitlement to commit suicide was never a problem, that is until it was challenged by Dr. Jack Kevorkian. Kevorkian was born in Pontiac, Michigan in 1928 (Chermak 101). His infatuation with death started when his mother died from cancer. Kevorkian wanted to find an easier way to help people carry out their death wishes; therefore, his solution was a suicide machine (Chermak 103). In this way, more than 130 were helped by Dr. Kevorkian and his suicide machine (Chua-Eoan). Although Kevorkian was tried five times, he was found innocent four times (Chermak 108-115). His jury had an onerous time coming to their conclusion. Throughout Twelve Angry Men, written by Reginald Rose, Twelve jurors had to decide the fate of one boy convicted of stabbing his father in the chest. In the beginning, eleven of the twelve were positive that the boy was guilty and nothing could change their minds, but Juror Eight disagreed. He showed them the holes in each of the testimonies. After hours of debating, Juror Eight was able to convince most the jury into seeing reasonable doubt in the boys guilt. When the last vote was taken, the tally was eleven to one in favor of innocent. In the end, there are many similarities between the jurors from Twelve Angry Men and the jurors of the Kevorkian trials because they all had a complicated trial, and their rulings were influenced by their personal thoughts and pasts. Throughout Dr. Kevorkian’s career, he challenged the law unlike many. Kevorkian was tried five times, but four out of the five he was found innocent due to the complications with his offences o... ...od, 2007. Print. Chua-Eoan, Howard. â€Å"Jack Kevorkian.† Time 20 June 2011: 16. MAS Ultra - School Edition. Web. 14 Apr. 2014. Cohen, Adam Grace, Julie Tynan, William. "Showdown For Doctor Death." Time 152.23 (1998): 46. MAS Ultra - School Edition. Web. 14 Apr. 2014. Hosenball, Mark. â€Å"The Real Jack KEvorkian.† Newsweek 122.23 (1993): 28. MAS Ultra - School Edition. Web. 14 Apr. 2014. Lessenberry, Jack. "Jury Acquits Kevorkian in Common-Law Case." The New York Times. 14 May 1996. The New York Times. 24 Apr. 2014. Morgenthau, Tom Barrett, Todd. â€Å"Dr. Kevorkian’s Death Wish.† Newsweek 121.0 (1993): 46. MAS Ultra-School Edition. Web. 22 Apr. 2014. Rose, Reginald. Twelve Angry Men. New York: Penguin, 2006. Print. Shapiro, Joseph P.Bowermaster, David. â€Å"Death On Trial. (Cover Story).† U.S. News & World Report 116.16 (1994): 31. MAS Ultra - School Edition. Web. 15 Apr. 2014. 

Child Abuse and Neglect Essay

Almost one in every hundred children gets abused in the United States, and it happens every day. Child abuse is when a child is physically, sexually, emotionally, and mentally abused. Sexual exploitation, neglect, abandonment, and maltreatment are also forms of abuse. Physical abuse includes but not limited to, throwing kicking, burning, or cutting a child. Striking them with a closed fist; shaking them when they are under the age of three, interfering with their breathing, threatening them with a deadly weapon, or doing any other act that is likely to give the child bodily harm and other minor to major injuries to their health, welfare, and safety. Sexual abuse is committing or allowing to be committed to any sexual offense against a child as defined in the criminal code and sexual exploitations when a child is prostituted, being filming in pornographic acts, or being a part of a live action of pornographic acts. Neglect and maltreatment is when the child is not provided with an ade quate amount of food, shelter, clothing, supervision, or healthcare. Abandonment is when a child is deserted by their guardian, left without food, water, shelter, clothing, hygiene, and medical healthcare, or when the guardian is just not doing things they are supposed to do for a child, they relieve themselves from their duties. We are all mandatory reporters. Meaning that any citizen that sees anything suspicious should report, or if you know of a child being abused and don’t report, you are just as guilty. Almost five children die every day as a result of child abuse. Child abuse is no doubt a hidden epidemic in our society today. Ninety percent of child sexual abuse victims know the perpetrators in some way; sixty-eight percent are abused by family members. Child abuse also happens no matter how rich or poor you are, doesn’t matter what color you are, and it doesn’t matter how smart or dumb you are. Thirty percent of women in prison were abused as children, and sixty percent of people in drug rehabilitation centers report being abused or neglected as a child, and are 3.8 times more likely to develop drug addictions. About thirty percent of  abused and neglected as children will continue the cycle with their children as well. Eighty percent of twenty-one year olds that were abused as children met at least one criteria for a psychological disorder. Also abused children are twenty-five percent more likely to experience teen pregnancy. I think child abuse is inclining locally because drugs are becoming more known in younger generations, and their also experimenting sexually earlier, and not being safe about which leads to more teen pregnancies. Some of the reasons child abuse occurs in the first place is because of young parents and drugs/alcohol. So if the rate at which those things or more rapidly happening, and at a younger age, the child abuse rate is bound to incline. It doesn’t help that we are a bad drug city and we have one of the highest count of child abuse cases in our city. One way to help stop and prevent child abuse is to recognize it, look for the signs and report it. Make your children aware of sexual abuse, teach them the proper names, and tell them that if anyone tries to engage with them in a way that their private parts are involved, to tell their â€Å"support person†. Teach young parents how to take care of their children before they leave the hospital. Never discipline children when your anger is out of control, ever leave your children unattended. Listen and believe them. If you see child abuse happening, stop it, don’t let it happen. To help end abuse, first you should learn how to recognize it in the first place. Child abuse will never truly come to an end, but there are ways to make sure it happens less. CPS needs to better investigate into cases. If you’ve ever read â€Å"the child called it† then you would know that if they go to the house and everything checks out, they believe it. I think they should go deeper into the investigation, if that means surveillance outside their house for a week then so be it. CPS also needs to better background check their own foster parents. I saw on the news a while back that CPS supervisor was abusing the kids she was fostering. CPS is supposed to be a safe haven for kids to go, yet they go to further get abused. I myself was in foster care, and my â€Å"parents† weren’t the nicest. It’s hard to honestly say anything about improving â€Å"families† considering we aren’t in there every day life, and we may not even know them. How we can best help families is to  make child abuse awareness more public and known, and if we come across a family that has this problem, then intervene. I couldn’t find much on factors of abuse being â€Å"heredity†, but what I could find is that it is not heredity. What generally happens is the child grows up with the type of abuse they know, and because that’s all they know, that’s how they express their â€Å"love† to their child. Not all abused grow up to do the same to their children, I assume the ones most mentally scarred and got psychological problems from it, grow up to do the same. I don’t think that our society is necessarily feeding the abuse to anybody. I don’t really know who would, that’s a touchy subject in the first place. Although I can infer that all of the drug, drinking, partying, and sexual innuendo that is seen everywhere is what is feeding to a worse environment for our society. Drugs, alcohol, and partying is such a huge and wide topic everywhere you go in our society, because of that, it’s more common. Although no one can honestly end child abuse, it is something that we can make more known so that all the poor children that are dying and suffering everyday can also be loved and saved. Learn the signs, report anything unusual.

Functional or Dysfunctional Families Essays

Functional or Dysfunctional Families Essays Functional or Dysfunctional Families Essay Functional or Dysfunctional Families Essay What Is the first thing thats come Into your mind? Love? Hate? That there Is no such a thing as family? There Is no specific rule that says families should be a certain way. Every single family has problems. Therefore, there is no such thing as a completely functional family. Then, what really is a functional family? Does anybody really know? Or may be- are we Judging something based on a model that has been created by our society? For example: the perfect family Is where the wife stay home ND does her duties and the man of the house brings home the bacon. The perfect children are the ones who never flight; in a perfect family, the mother-in-law is a best friend with the wife. Not Just in the immediate family is there perfection, but also with the cousins, aunts and grandmothers: everybody gets along with everybody else, and nobody talks about anyone behind their backs. In the world there are different types of families, each with their own traditions, customs and values. Each one of them has their own particular ay of thinking. This doesnt necessarily mean this is a dysfunctional family. Actually it would be hard for us to identify what could be a dysfunctional family or not. It Is perfectly normal that females have disagreements and fights because we as humans are not perfect nor have the same way AT twinkling. I en Important thing Is Tort every memoir AT ten family to accept their mistakes and learn to respect each others opinions. : This reminds me of the movie Little Miss Sunshine where the familys elder son decides not speak a word for nine months, the uncle tries to take his own life because of a broken heart, the grandfather is a drug addict and the father does not accept Losers in the family. This is a perfect example of how each and every one of us has our own problems, opinions and ways of dealing with conflict. It shows that at the end no matter what their decisions is, the family should stay together and encourage each other paying no mind to their differences. We have learned that families are not perfect. We should not go by the tankard of functional or dysfunctional because the real issue is not about measuring up to a standard. Since all human beings are complicated, we are bound no to get along all the time. But we must work on doing so, not only in our family circle, but also with all humanity. Family is a very important part of life: what you have forever, even after disappointments, arguments or bog fights. Good families will overcome their emotions and try to help one another. Where this is impossible, friends become the family, but may never be quite the same.

Romeo and Juliet Monologue Essays

Romeo and Juliet Monologue Essays Romeo and Juliet Monologue Paper Romeo and Juliet Monologue Paper Juliet: Hum! Something fears. O Romeo! All I desire is to be with you. I cant stand, seeing you fade away into the dark fog. The wind seems to carry you far away from me but I hope the sun will give you light in the darkness. The more the distance between us the more I dread for you. O Romeo! The moment you leave, your thoughts keep appearing in my mind. Then all I do is dream about what you said all along our meetings. It was on the day my dad gave a party; I wonder how he was there. He looked very nervous. He seemed to be searching for someone; perhaps someone special. Then his keen eyes fell on mine. I couldnt resist the way he stared at me. So I immediately turned away. His hand suddenly touched mine, it was cold and a sudden shiver ran across my body. When I turned, he said If I profane with my unworthiest hand This holy shrine, the gentle sin is this: My lips two blushing pilgrims, ready stand To smooth the rough touch with a tender kiss. I cant forget those words. I really felt as pure and as holy as an angel when he said that. I knew he liked me and wanted to kiss me. I knew it the moment he said O then, dear saint, let lips do what hands do! as we touched each others hands. My cheeks blushed and my heart was too heavy for words as he gave me a loving kiss. The nurse than interrupted, I was so furious, as I hardly met him. I didnt even know his name. After everyone left, me and the nurse were left alone in the massive hall. Thats when I asked her what his name was. She said His name is Romeo, and a Montague. The only son of your great enemy My anger changed to despair as I knew that my only love was my familys greatest enemy. I was shocked. All day I couldnt resist thinking about him. That night, when I was in the balcony opening up my thoughts he suddenly sprang out of the bushes. For a second there, my blood froze. I couldnt say no to him now as he must have heard what I spoke I thought as we spoke Then I feared for him as I told him if they do see thee they will murder thee. I told him again if any of the kinsmen will see him they will kill him. But he didnt care and replied Alack, their lies more peril in thine eye Than twenty of their swords! My heart lightened as he said those words but my fear for him seemed to be deepened. So I said I wouldnt let that happen. Then he replied I have nights cloak to hide me from their eyes. He was exaggerating and dreaming as he said he had nights cloak to hide him from the kinsmen. After that I was quite serious about what was happening. I asked for true commitment for our love. He said Lady, by yonder blessed moon I vow. I was not pleased when he swore on the moon as it constantly rotates round the earth. But then I somehow knew he truly loved me. When he was about to leave he said O, wilt thou leave me so unsatisfied? I was surprised at first as I didnt know what satisfaction he wanted. Then he told me about our marriage and so it was set the next day. It was too quick I thought but I couldnt wait any longer like him. We got married happily, it was a great feeling. Today I didnt want him to go but he had to. I was playing with him until he said come, death, and welcome! Juliet wills it so. How ist, my soul? Lets talk, it is not day. I became serious and didnt want anything to happen to him because of me. So, I rushed him up a bit to get ready for his journey. Then he suddenly said More light and light; more dark and dark our woes. I felt darkness fall over me when he said that. Suddenly, the nurse alerted us as he had to leave quickly as possible because my mother was coming to see me. I became tensed. Then everything went quick and well. Then we gave a farewell kiss and then he departed. When he was walking away an unpleasant thought rushed through my mind. I feared whether we will meet again or not. I was terrorized by the thought. I turned my attention towards my mothers call.

Vaginal Birth is better than Cesarian Section Research Paper

Vaginal Birth is better than Cesarian Section - Research Paper Example   Relevant studies from the United Kingdom echoed findings that babies born from planned vaginal delivery had best outcomes compared to those who had Cesarian sections (Greer, et al. 2009, 675). Of the two approaches, vaginal delivery is cost-effective as it has the least risk for admission to the neonatal unit, least need for supplemental oxygen, least need for hospital and physician services, and least incidence of hypoglycemia and respiratory problems. Guise et al.     explained that Cesarian section is more expensive because of the following reasons: (1) longer stays in the hospital of both mother and child and more intensive observation from physicians; (2) longer delivery and operative time; (3) increased risk for perioperative complications; and (4) increased need for baby’s supplemental oxygen because of respiratory complications (39). As reflected in the studies given, vaginal birth offers the most cost-effective approach of delivery because it does not involve an operation/incision that may take days or weeks to heal. It is cost-effective for the mother because only the necessary medications, treatment and services during delivery are being given. There is no need to avail of any additional supplemental oxygen or treatment for the baby as babies born from vaginal birth have fewer respiratory complications. While discomfort or pain during labor discouraged the mother to have vaginal birth, this was significantly outweighed by a number of risks that may harm the mother and child once they choose the Cesarian section.  

Analysis Of The Book The Cider House Rules Essay

Analysis Of The Book The Cider House Rules - Essay Example   When Homer learns about Larch's secrets- him secretly being an abortionist, driven by the sight of the black-alley horrors- he considers it morally wrong. He befriends Candy Kendall, a couple to Wally Worthington, who need an abortion. He leaves and goes with them to Wally's family's orchard. He develops the love for Candy secretly (Irving 78). They get a child together- Angel. After he knows of Larch’s death, he goes back to St. Clouds as a director.to show his heroism nature; he maintains his dislike for abortions. However, he honors the choice of his parents and continues Dr. Wilbur's legacy.   Even though he has all the training to be an obstetrician and gynecologist and has adequate knowledge on performing abortions, he goes ahead and objects to the latter.   He refuses completely to do them (Irving 34). He is questioned repeatedly throughout this film, on this subject, and he responds to this by declining to perform the abortions with a reason that he is not form ally trained as a doctor, and that they are illegal. He hopes for a day to come when abortions will be legal, free, and safe when he'll no longer be obliged to perform them (Irving 66).  We can describe The Cider House Rules as a coming-of-age story. These tales trail a character from babyhood to adulthood, all the way through his/her moral knowledge, until as a hero; he/she finds a place in life. A true hero, like Homer, is the one who takes decisive action in finding out his/her destiny or fate (Irving 44).