Malaria: Causes, Effects and Cures

Malaria Causes, Effects and CuresSolving a Biological conundrumSummaryThe chapter is slightly the ruleology that is involved in exploring several(prenominal) belowstanding toward solving a biological problem while doing inquiry in quest. With reference of example of malaria as a problem along with diachronic stage involved towards gradual understanding related to its causes leading to the findings to its cure and preventions, an attempt was made to record the betterment that has been involved in addressing the scientific issues in past.A practical approach that is possible to conduct practically at school level to get the students introduced with research at elementary level and reinforced up some interest in students has been presented in the following chapter.Mathematical and statistical tools that ar needed in research cast been introduced to analyze the selective information obtained by victimisation research methodology (hypothetical) to arrive on conclusions about certain aspects of the issues related to malaria to accept, discard or modify the speculation on a scientific query.IntroductionThe homo internal being started putting efforts to explore the world around him right from really beginning. The history is full of examples that show that early kind-hearted being somehow recorded their opinion about different thing. With passage of time, humane beings learnt to evaluate the correctness of their thoughts and opinion on any(prenominal) scientific issue, by setting some experimentations, d defenselessing conclusion (hypothesis) based on results, verification of hypothesis by other experiments and reporting it. Research methodology is latest approach involving these procedures to address a scientific problem.Steps involved in Biological Research ProcedureIdentification of a problemSufferings of mankind that includes diseases, scarcity of viands shelter, utilities etc or related topics be the existing problems from biological origi n.To initiate research in an parklandwealth a problem in a particular heavens is identified. There backside be a wide feed of problems e.g. an approach to find a cure for newly emerging disease, find a more(prenominal) effective dose for a curable disease, increase the shelf life of a product etc.Generation of hypothesisAvailable understanding on an issue that is print in scientific journals is used to draw logical opinion, hypothesis, underlying the biological processes and indicate possibilities that can lead to the trouble of the problem. There are often more than one hypothesis available for a accustomed scientific query. Usually one hypothesis is being testified by prone experiments.(a) ledge life of chadder cheese increases with increase in tot of yeast cells present into it.(b) Quality of spinach deteriorates with increase in rate of transpiration.(c) The rate of step-down in microbial population in a food adjudicate in response of commove treatment at 600C is inversely proportional to its total fundamental content.Laboratory safety ProceduresEvery grammatical case of biological experimentation should comply with necessary safety procedures that take ins the safety of professionals involved in conducting the experiments and other people. That includes use of additional clothing e.g. coats, gloves head cover spectacles etc. All the biological veridical is carried or grown in specially designed containers that ensure no jeopardize for leakage that can be hazards for other people who can come in contact. All the biological material is heated at 121oC under pressure at 15 psi to kill all type of cells before discarding it whereas chemical agents are discarded as mentioned in safety manuals. try DesignExperimentation is a practical approach for investigation a scientific query leading to generation of cards called data. A same question can be investigated by using different experimental approaches. The selection of an experimental desi gn mainly dep lay offs upon time required, extent of precision, availability of resources etc.Each experiment design has got some limitations and it is important to ensure that the selected experiment design can optimize for investigating the required scientific query. Each experiment is design by usually changing a varying.In frame to test a given hypothesis, experiments are set, usually in triplicate and experiments are repeated at least three time to ensure reproducibility of the data. To avoid adding error in the data it is important to set negative andpositive control for a given experiment. Positive control has an ingredient instead of ingredient to be test that should positive result when is added in the reagents and processed as per standard method of experiment in given conditions.On other hand Negative control is set same as positive control but has piss or other solvent added instead of reagent to be tested.It is very important that selection of sample that is used i n the experiment should be random. It is important to ensure that all the experiments should be done at the same conditions. All those factors that can contribute to add errors should be taken care of. info Collection and analytic thinkingObservations can be self- tranquil after completion of experiments or while they are in progress depending on nature of experiments. The data is non invariably a integral values but can be visual observations that can be recorded by photography. The data is usually recorded with allow units in tabular form. This is k directn as raw data. Every data has some error added into it.Mathematic and statistics an integral part of Biological AnalysisThe observations that are compile as numeral value after experiments comprise raw data.Mathematical or statistical methods are used to minimize the effect of errors present into it the raw data. The most widely and commonly statistical method that is used to decline error in data is finding an average i n any given readings.Beside that mathematical or statistical tools are used to deduce a logical ground based on mathematical value to sign, modify or discard any scientific opinion (hypothesis) that is built up on earlier reported scientific findingsUse of ratio proportion and dowerData is analyzed by using mathematical or statistical tools, most commonly that are used include ratio and percentage, for finding an effect of changing a variable on other parameters in a given set of condition.If a mathematical variable a represents excitement of a biological event that needs to be compared with vividness of another similar event represented by numerical variable b then ratios are taken outRatio between intensity of two biological events = a (equating 1) bIn order to represent this comparison on scale of 100, percentage of ratio between variables are taken outThe general formula for percentage is given as below%= Ratio between two numerical variable x100(equation 2)Substitute equa tion 1 in general formula of percentage as shown by equation 2Percent % (ratio between intensity of two biological events) = a x 100 (eq 3) bUsually biological data can comprise of in a range of very small value to very striking and hence it is important to convert it on Log10. The general formula for expressing any numerical variable is shown by equation 4 as followNumber(N) on Log 10 scale= Log10(N) (equation 4)Substitute equation 3 into equation 4Log 10(ratio between intensity of two biological events) =Log10 a x 100(eq 5) bTechnical limitation associated with use of Log10 scaleWhile expressing any data on Log10 the clog arises when it is needed to express integral 0 on Log10 scale (Log10(0)=infinity). In order to cope with this difficulty 1 or any fraction of number that falls within permissible limit of error (10 scale. In later stages the data can easily be plotted on any type of graphs as discussed in later part of this chapter.Significance of error in decision making and predictions on biological dataEvery data collected has certain extent of error present into it depending upon experimentation design, procedures and method of taking observations.This error needs to be evaluated before using the data for testing any hypothesis, decision making or predictions. It is defined as tentative variation on negative and positive scale in a set of observations from existent value.The actual numerical value of a biological effect is (B). An experiment was done to note this biological effect. The experiment was repeated N times e.g. (N1 N2 N3 Nt) to wake observations B as (B1 B2 B3.Bt) respectively.The first stage in calculation the error is to find an averageThe general formula for add up isAverage = center of numerical values of individual observation eq 6Number of times the observation was takenSubstitute the values in equation 6Average =B1+ B2+.Bt eq 7 NtSum of values of observations =B1+ B2+.Btis shown by SBNo of times the experiment was repeated Nt =N Substitute the value in equations in equation 7Average= SB equation 8 NLets value of Average be represented by XSubstitute in equation 7Average = XSecond stage is to find the difference between each numerical value of observation and Average= B1-X, B2-XBt-X,Third stage is to feather each of the difference=B1-X)2,(B2-X)2, (B3-X)2(Bt-X)2Fourth stage is to add the square of differences=(B1-X)2+(B2-X)2+(Bt-X)2Fifth stage is to divide square sum of difference of average of individual numerical variable with number of observations e.g N =S(B1-X)2,( B2-X)2,..(Bt-X)2 NStandard deviation or Error is obtained by taking square root of the quotient obtained by dividing the square sum of difference between the average and the individual numerical variable with number of observations.=S(B1-X)2,( B2-X)2,..(Bt-X)2 NIt is an integral value that is expressed, as on negative as intimately up as positive scale e.g if error calculated is 3 then it would be +3 as well as 3. It represents a range with in which actual value may lie.In olden days, such calculations were done with the assist of calculator but now same work can be done more easily by using different software e.g. excel with computer as a tool.It is often very different to understand the effect underlying in any biological data by merely looking at numerical values. The different types of graphs are used for visual intro of effect by trends available in dataThe most commonly types of graphs that are used for the presentation of data are given in figure 2.The same data for different sample e.g. A, B, C is presented on percentage scale by using horizontal and vertical bars whereas error is shown by error bars .Another set of data for sample D, E, F was presented by line graph.Data psychoanalysis to decide the status of hypothesisAfter the mathematical and statistical treatment of raw data a logical ground built up by comparing certain numerical values or more often represented on graphs to accept, modify or reject an y hypothesis.In order to test the hypothesis The rate of decrease in microbial population in a food sample in response of heat treatment at 600C is inversely proportional to its total organic, 100 cells of Saccharomyces cerevisiae were inoculated in same volume in mediums with concentration as X, 3X and 70X respectively to make last concentration of each medium as 100 cells/ml.(Sample D contains 3X concentration medium, Sample E contains X concentration and Sample F contains 70X concentration). The medium were unbroken at 600C for 1104hours and cells were assayed for viability after 10 hrs, 100, hours, 1000hrs and ten thousand hrs respectively.The viability being dependent value was plotted on a graph Y axis against the time in hours after expressing the value on Log 10 scale. Different data points taken on the graph are sufficiently scattered and can not be joined by a straight line accommodating all the point on it.The trend line can be worn-out manually by accommodating maxim um number of points and leaving as many point above the line as many are in that location below that line. Such a manually drawn line can not be used for any type of scientific predictions. Otherwise highly precise trend line for a given data can be drawn as explained in section given in the end of the chapter that can be used for making decisions on a given hypothesis and for making predictions.The error bars extending on positive as well as negative scale in same magnitude of Y scale is plotted against each observation.The observation that were taken in this experiment are shown as graphs in Figure 2 e.g. Sample D, E and F were presented by using line graphs on Log 10 scale.The trend lines are introduced to show the type of dependency of one parameter on another. As already discussed earlier the biological data is often presented on Log 10 scale to observe the effects in broader prospective and ignore the slight changes especially when carry is made on larger populations of samp les e.g. cells with a wide range of variations.The data show that sample D and E has got similar trend (with an increase in variable on X axis, there is a decrease on Y axis variable, Y axis is dependent on x axis and is inversely proportional to it) whereas in sample F, Y variable is independent of X variable as no change in Y variable is observed with an increase in X variable and this is clear by a straight trend line.The conclusion drawn as evident by three graphs that number of viable yeast cells were found to decrease with passage of time when concentration of organic content in medium was X in sample E or 3X in sample D that is represented by a slopes in respective graphs. But this effect was found to be lost when organic concentration was raised to 70X in sample F and that is.This experiment supports the hypothesis The rate of decrease in microbial population in a food sample in response of heat treatment at 600C is inversely proportional to its total organic content.If re ported literature indicate some other type of experiments done to decelerate the same hypothesis then results are compared and reason of the variations if there are any are discussed with scientific reference and is reported in a scientific journal.Research MethodologyTheory acceptedModification in hypothesisPublication in scientific JournalsDiscarding above hypothesisSupporting above hypothesisResults interpretationDifferent HypothesesData analysisData collectionExperimental design for a given hypothesisLiterature reported in related areaIdentification of Biological ProblemInterpretation of dataThe data that is obtained is interpreted to draw scientific conclusions. The reported literature is used to give history of the conclusion drawn.In the light of conclusion drawn either the hypothesis is accepted, rejected or is modified.If the hypothesis is proven correct with experiments, then it is known as a theory. Scientific articles based on proven hypothesis e.g. theory, disapproved or modified hypothesis are written by scientist involved in research and are sent for publication in scientific journals.Biological Problem as an exampleA school teacher planned an excursion for students and took paramedic staff equipped with sample collection facility to country side area to study the prevalence of any non contiguous disease in human beings and animals. Malaria is an example of non contiguous disease that is spread only by contaminateed female mosquito is malaria and is a matter of great concern.As reported in newspapers and other literature there was a population of 100,000 people living with minimal facilities of life that on a huge natural water system reservoir for tipsiness water. Recent heavy rain resulted in collection of water of stagnant water and much of this water in later stage drained in reservoir.The students prepared a report explaining how did they use biological methodology to study this case.a) Identification of problemPrevalence of certain inc idences of disease symptoms similar to malaria in a given locality was identified as a biological problem. Students wrote the following note after referring the literature about malaria to get basic information about disease. Please refer the picture given below and the literature cited in later part of this chapter.From Mala aria ( bad air) to Malaria-over a period of timeMalaria has been a matter of concerns since very beginning. Understanding about malaria has changed over a period of time. Initially it was considered to be caused by some supernatural power. Supported with the findings of higher incidences near the soggy area, malaria was thought to be caused by bad air that is found near spongelike areas. With the emergence of germ theory malaria by Louis Pasteur (1852-1895) was thought to be caused by any bacteria. It is strongly believed until Charles Louis Alphonse Laveron (1845-1922) during microscopy of Blood from infected patients observed causative agent of malaria in 1 880 and later on reported it to Academy of Medicine in Paris. Albert freeman Africanus King (1841-1940) who was associated with George Washington University on basis of supporting reports presented the idea that mosquitoes are the mean of transmission of malaria (vector) and discarded the opinion that bad air in marshy area has any role in causing malaria. On basis of his data he suggested that proper netting can help to control the incidences of malaria in Washington.Ronald Ross (1857-1932) was a physician, who studied the life motorbike of malarial parasite in mosquites e.g (different stages of parasite infection with a change in morphology) and its transfer to human beings and birds. In 1898 Giovanni Battista Grassi (1854-1925) on basis of understanding about malaria obtained by published literature set an experiment to infect a person who never had malaria (with his permission) in an area of Rome where there was not any case of malaria reported and mosquitoes were not found by exposing the person with Anopheles clavigar for ten night and later on patient developed the symptoms. On basis of his experiment he proved that malaria is spread by mosquitoes in human beings that carries the causative agent Plasmodium.The complete cycle of P. falciparum was observed by Grassi Bignami and Bastianelli in 1899 and the work has been published by Grassi in 1900.The life cycle of Plasmodium has three reproduction stages with different morphologies (shapes). The mosquitoes inject sporozoites in human being s skin that through melody goes to liver where they multiply and change into Merozoites.The second cycle of multiplication does into RBC. Some of the meroziotes after casual through reproductive cycle in RBC converts into gametocytes (male and female gemates) and enters in gut of mosquitoes when they suck livestock from an infected human being where they undergo sexual reproduction to zygote which later on converts Oocyst. Oocyste after under going asexual reproduc tion burst to release newly formed sporozoites that enters in salivary gland of mosquitoesAt the site students made a survey of that area and noted the initial observation with the help of photography. They found that not only human being but also the birds are affected by the diseaseInitial FindingsThey found that the reservoir (a) was associated with dark places where high populations of mosquitoes can be seen (b).Beside that a few sick birds were also found resting on ground. Diseased patients report to an increase in high fever with shivering that stays for some times and then fever become low or even normal with sweating or even without it. These symptoms are repeated with intervals and patient feels weakness.Initial findings support that the disease is malaria.b) On the basis of their initial findings the following hypothesis was built up.The disease may be Malaria and is caused by PlasmodiumExperimentationPlasmodium infect the red blood cells. In order to established that dis eased people are suffering from malaria at least 63 patients having disease were bled to collect blood samples. This type of sample that is under study is called as test sample. Blood were taken from at least 50 healthy who do not have any symptoms of disease. Since these people do not have any symptoms of disease it is very likely that Plasmodium may never be detected in their blood. Such a sample that is known to give a negative test is a called a negative control The blood and water samples were collected in collection tubes specially designed for this purpose.The located slides of infected Red Blood Cell (RBC) were purchased from the market and was taken as positive control.(a)The sample was not only collected from human beings but also from the diseased animals as well. That the blood of each sample was spread on a glass slide, fixed and stained with Giemsia and was observed under microscope.(b)Beside that the blood samples from infected people were inoculated (added) in RPMI- 1640 medium (name of medium used for the growth of Plasmodium that also contain RBC) present in bottle and were incubated to grow causative agent of the disease under laboratory conditions. The sample from these bottles were observed under microscope for presence of Plasmodium after 72 hours of incubation.Furthermore, the surface water from stagnant regions of reservoir was collected in a container and was assayed for presence of larvae of mosquitoes with help of magnifying glass.Laboratory Safety ProcedureAll the containers having biological material was heated at 1210 C at 15 psi for 15 minutes to kill every type of living cells before discarding them. The chemical agents were discarded as described in their respective safety manualsResultMicroscopy results show that the Red Blood Cell of diseased people were found to be infected with Plasmodium.The causative agent of the disease was successfully found to grow in the medium that supports the growth of Plasmodium (Figure 3) and tha t was confirmed by microscopy e.g Plasmodium were found in the sample from medium under microscope and slides observed were found similar as shown in Figure 1 and Figure 2Plasmodium Figure 7 The blood sample after mixing with other reagent (as shown with white arrow indicating towards a tube) is then inoculated into the bottle ( as shown in picture) ( source Nature Protocols dexterity to Nature Publishing Group)The surface water samples that was taken from the stagnant water collected near water reservoir were found of have mosquitoes larvae in large number as is shown in Figure 4.ConclusionThe results that includes, presence of large number of mosquitoes larvae near the site of outbreak, Plasmodium was found in the infected blood of diseased people, that was successfully grown in the medium and conditions specific confirm that the outbreak is of malaria that was caused by Plasmodium. Skill development to solve a Biological ProblemReportA report comprising the incidences of malaria during the years 2002, 2003 and 2004, in three different cities was published in a newspaper. The patient were treated by using drug A and attempt to kill mosquitoes were made by spraying B into environment and adding in water collection. Out of these report related to three cities is given below.In Karachi 530 malarial cases were reported in year of 2002, 534760 in 2003 and 12345668 in 2004 respectively. The mortality reported in these years were 98 in 2002, 120001 in 2003 and 5408889 in 2004.The rain fall recorded in Karachi for year 2002 is 50mm, 2003 is 100 mm and 10,000 mm in 2004.The drug resistance was found in 12 cases in 2002, 60009 cases in 2003 and 9900099 cases in 2004.In 2002, 134 malarial cases were reported in Faisalabad where as in 2003 and 2004 the reported numbers were 1237 and 1379 respectively. The mortality reported in years 2002, 2003 and 2004 were 10, 99, cxv respectively. The annual rainfall reported in these years were 12 mm in 2002, 58 mm in 2003 and 89 mm in 2004.The antimalarial drug drug resistance was found to be in 2 cases in 2002 , 79 cases in 2003 and 91 cases in 2004.In a similar study that was conducted in Gilgit during these three years, it was found that malaria affected 325 people in 2002, 135 people in 2003 and 350 people in 2004.There were 10 people reported to be died of malarial disease in 2002, 8 people in 2003 and 17 people in 2004. The annual rainfall reported in these years were 130, 120, cv mm in 2002, 2003 and 2004 respectively. The resistance against anti malarial drug found in 2002, 2003 and 2004 were 9, 4, 9 respectively.Source( It is an imaginary situation given with data to help student develop research skills)Research MethodologyStep 1 Identification of problem from published literatureAfter reading the above mentioned findings, management of heavy occurrence of malarial disease has been identified as a problem.Step 2 literature search for generating the hypothesis taking malaria as a test caseMalaria is a very common infectious disease that is commonly associated with poverty. It is caused by protozoan parasites Plasmodium species that is transferred to human being blood circulation schema by the vector Anopheles mosquitos bite(1). Literature show that malarial outbreaks can be related with rainfall in that area (2). Malaria is more common in urban area than in cities. However in Africa it is present in both rural and urban areas (3,4)No literature is available about the coitus of malarial incidence with location of the place with height above sea level. The occurrence of malarial outbreak can be related with presence of stagnant water that can support the mosquito survival in populations. Heavy use of anti-malarial drugs and mosquito cidal sprays is reported to produce resistance in the protozoa against commonly used drugs (5).1) cyclooxygenase F (2002). History of Human parasitology. Clin Microbiol Rev 15 (4) 595-612.2) Grover-Kopec E, Kawano M, Klaver R, Blumenthal B, Ceccato P, Connor S. 2005 An online operational rainfall-monitoring resource for epidemic malaria early warning systems in Africa. Malar J 4(1) 6.3) Van Benthem B, Vanwambeke S, Khantikul N, Burghoorn-Maas C, Panart K, Oskam L, Lambin E, Somboon P 2005.Spatial patterns of and risk factors for seropositivity for dengue infection Am J Trop Med Hyg 72 (2) 201-8.4) Keiser J, Utzinger J, Caldas de Castro M, Smith T, Tanner M, Singer B 2004. Urbanization in sub-saharan Africa and implication for malaria control. Am J Trop Med Hyg 71 (2 Suppl) 118-27.5)Rieckmann, K.H.2006 The chequered history of malaria control are new and better tools the ultimate answer? Annals of Tropical Medicine and Parasitology 100(8) 647-6626) en.wikipedia.org/wiki/Malaria(The Scientific literature is presented with citation of references as is shown in above paragraph)Step 3 Deduction of hypothesis with help of published literatureAfter reading the reported literature as mentioned above following hypothesis can be deduced .Incidence of malaria is dependent on amount of rainfall probably through collection of stagnant waterStep 4 Experimental designThe data for the parameters analyzed was collected by using standard methods e.g. microscopy of infected blood samples at different hospitals and was published in a newspaper as a scientific report.Step 5 Presentation of Raw dataThe above mentioned data is presented below in tabular form.KarachiYearMalarial cases reportedMortalityRainfall (mm)No of resistant cases to anti-malarial drug A20025309850122003534760120001100600092004123456685408889100009900099Average/year4293652.66718429963383.333320040FaisalabadYearMalarial cases reportedMort