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Ninth arab conference on the peaceful uses of atomic energy, beirut, 3-6 december 2008

Original article Epidemiological Studies on Strongyloides stercoralis at Dilla
District, Ethiopia
Feleke Eriso
Parasitologist, Department of Biology, Dilla University, Dilla, Ethiopia
Corresponding Author

Background and study aim: Some
Results: In the study a total of 710 student
Feleke Eriso children were examined for Strongyloides parthenogenesis or asexual reproduction stercoralis infection out of whom 142 and hermaphroditism (protandrogony) to (in 1st study) were positive, confirming +251916514682 be the only mode of reproduction of the infection rate to be 20% or 198 parasitic female Strongyloides stercoralis positive (in 2nd study) the infection rate in human hosts as parasitic males of it did being 28% by the parasite. Then, the not exist in human hosts. Therefore, the average infection rate was first objective was to work out the developmental stages and sexes of the infection rate of Strongyloides stercoralis parasite were obtained in the study in the population of elementary schools children at Dilla district; secondly, Conclusion: The presence of many
to produce a visible evidence for the prese parasitic males with everted spicules nce of many parasitic males of Strongyloi- observed in fresh stools samples during stercoralis; Diagnosis; Morbidity; des stercoralis as there are parasitic femal this study had been a very strong evidence Mortality; Treatment; -es in fresh stools samples of human for the fact that male and female Immunocompetent hosts; and thirdly, to replace the unfit copulation & fertilization were naturally term by a correct one. taking place among parasitic worms of Strongyloides stercoralis Patients and methods: Stools samples
hosts. Parasitic and free living males of S. were collected from student children of stercoralis have the same morphology elementary schools, and observed under including the curved or coiled posterior parasitology after employing Baermann apparatus technique. article on the internet which stated that the parasitic generation of S. Strongyloides stercoralis is known to stercoralis was known not to have have two life cycles: an internal sexual parasitic males and the parasitic cycle, involving parasitic worms that females used to reproduce only by the constitute the parasitic generation, and asexual method of reproduction [8]. In the parasitic generation, when the interacting among free-living worms filariform larvae are in contact with that represent the free-living generation cutaneous blood vessels and are stated by authors in modern textbooks, carried through the right heart to the journals, and on the internet that the lungs [9]. Then, sexually mature type of reproduction in the parasitic parasitic females settle in the tissues generation of S. stercoralis in human of epithelial mucosa to lay eggs that hosts was only by parthenogenesis of hatch soon and are discharged in the parasitic females in the absence of stools each day [10,11]. When all or parasitic males [4-7]. Due to this metamorphose into parasitic male had been omitted in the figures that demonstrated the life be onset by invading the cycle of the parasitic generation of S. of the ileum or colon, travel to lungs a stercoralis in all modern and relevant nd then return to the intestine to textbooks, journals, and on the mature in the mucosa [12-15]. internet. Not only that, there was an Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article Disseminated strongyloidiasis had been reported Strongyloidiasis [19-21]. Some of the clinical in both of two recipients of kidney allografts presentations of strongyloidiasis can be high- from a single cadaver donor [16]. It was also reported that in a 53-year-old man who had  Cutaneous with larva currens (racing larvae), lung cancer, fulminantly fatal strongyloidiasis pruritic linear or serpiginous, creeping urticarial had developed following postchemotherapy of eruption, dermatologic lesions, and petechiae; immunosuppression, resulting in the death of the  Pulmonary with persistent wheezing, cough, patient within 48 hrs [16]. The development of a and deteriorating respiratory status; and florid strongyloidiasis was observed in a 45-year-old man, following anticancer chemotherapy  Intestinal with vomiting, abdominal pain, when eggs of S. stercoralis were seen in the watery diarrhea and constipation. stools [17]. One scientific study has reported that almost all deaths due to helminths in the United States result from S. stercoralis hyperinfection PATIENTS AND METHODS
mortality rates because the occurrence of The suitable type of study selected to answer the hyperinfection can be as high as 87% [18]. question of this research work was the Cross Sectional Study. The statistical methods pre- Aim of the study planned to be employed in analyzing and The aim of the study has three objectives to work interpreting the results were the expression by percentage and standard deviation. First, to determine the infection rate of S. stercoralis in the population of elementary Eight different elementary schools found at Dilla schools children at Dilla district; district were selected to be the sites of fresh stools sample collection from student children. It  Second, to produce a visible evidence for the presence of many parasitic males of S. geographical points of view, to notify that Dilla stercoralis as there are parasitic females in district is located in Gedio Zone that is found in: fresh stools samples of human hosts; and  Southern Ethiopia,  Thirdly, to replace the unfit morphologic term  The continent of Africa, and by a correct one. Is parthenogenesis or asexual  The northern hemisphere between the tropic of reproduction true in the parasitic generation of cancer and the equator. S. stercoralis in human hosts? Concerning some morphological features of this documenting related information from the parasite, the part of the worm's body that is schools had been carried out from 6/12/2006 to known as the tail is the posterior part of body 10/06/2007 and again repeated in depth from beginning from cloaca in the parasitic males or 10/9/2008 to 25/6/2009. However, the deliberate follow up to be certain about the prevalence and females. Cloaca is the opening through which persistence of the parasite at Dilla district, was spicules are everted at times of copulation & performed by taking fresh stools samples of ten fertilization and it is also the outlet of students from each of the eight different the digestive tract. The 3 stages of human elementary schools every year from 2000 up to strongyloidiasis are Intestinal Strongyloidiasis, the beginning of 2014. Gastropulmonary Strongyloidiasis and Disseminated Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79


Original article Figure (1): Map of administrative regions and zones of Ethiopia. Gedio


Specific sample size:

including the participation of the child for the The sample size taken from the participant necessary information, with the exception of the student children was 710. Each day, Monday column under S. stercoralis, because it had to be through Friday (i.e. every week), fresh stools filled either "–" or "+" for S. stercoralis, by the samples, of ten student children were taken to researcher after examining the fresh stools parasitology laboratory of Dilla University. sample. Writing the name of the student child in When the sample was taken from the student the steps of raw data collection was important to child, he/she gave fresh stools sample in a bottle identify the child for giving treatment if he/she on which his/her I.D. No. was written. In had been found to be positive for the parasite, addition to this, on that very day and moment a because many children could not remember their table that had columns with the headings of Date, I.D. No. Of course, it was decided not to write Name of Child, I.D. No. of child, Class (grade), the name of the student child in the report of the Age in Year, Sex, S. stercoralis, Education of parents, and Job of parents were filled by Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article Diagnostic Examination:
No. of the student child from whom the fresh The diagnostic examination of fresh stools stools sample was taken to be examined. sample of each student child involved the These seven steps were repeated for the fresh following nine steps. stools sample of each of the remaining 9  Baermann funnel apparatus was constructed student children. and the lower opening of the rubber tubing  Using a dropper, a drop of suspension was fitted to the stem of the funnel was closed. taken from the surface of bottom sediment of  Water warmed to 400 C was poured into the the labeled test tube suspension and placed on funnel of the Baermann apparatus and the a clean glass slide and then covered with a cheese cloth, that contained the fresh stools cover slip. The preparation was examined sample of the student child and tied with its under the low power objective of a research peripheral edges to the rim of the funnel, was microscope to confirm the presence or partially immersed in the water warmed to 400 absence of S. stercoralis in the fresh stools C. This was done because if adults as well as sample of the student child. The sample of juveniles of S. stercoralis were present in the each student child was examined in this way. stools, they would be attracted by the warm The column under the heading of S. temperature of water (about 37.5o C as there stercoralis for each student child was marked was dissipation of heat from the initial 40o C "–" indicating the absence or "+" confirming of the added water to the surrounding the presence of S. stercoralis in the fresh materials and equipment) and escape into the stools sample taken. warm water through the pores of the cheese  The suspensions positive for S. stercoralis were fixed and preserved by adding 10%  After staying 1 hour and 30 minutes, the formaldehyde. Each container bottle of closed lower end of the rubber tubing was preservation in 10% formaldehyde was labeled S. opened, releasing the water found in the funnel of stercoralis larvae/other stages including the Baermann apparatus into a 500 ml beaker. The date of collection and kept in a safe place in stools left behind in the cheese cloth was the laboratory of parasitology. Water emergence thrown into the tube of toilet after being semi-concentration technique for detecting treated with a disinfectant (iodine solution) and strongyloides larvae in feces was also used washed away by a current of water. when there were needs to supplement the  The water released and collected in the 500 ml Baermann method [22].
beaker was centrifuged at a speed of 1000 rpm (revolutions of the rotor per minute) for 2 Water emergence semi-concentration technique minutes using a manual centrifuge loaded with for detecting S. stercoralis larvae in feces: 4 centrifuge tubes and anchored to the edge of  A fresh (not more than 2 hours old) formed or semi-formed fecal specimen is required. The  From each centrifuge tube the supernatant was method is as follows: poured off into a waste collecting bucket to be  Using a piece of stick, make a central thrown into the tube of toilet drainage line by depression in the specimen contained in a treating with the disinfectant. vial or bottle. Fill the depression with warm  Using a dropper, about 2 ml of the supernatant water (about 37.50ºC). was added to the sediment of one of the 4  Incubate the specimen in a 35-37.50ºC sediment containing centrifuge tubes and incubator for 1.5 to 3 hours during which time shaked well by closing its mouth with its own the larvae will migrate out of the feces into the fittingly tight lid. The action of shaking was to change the sediment into a transferable  Using a plastic bulb pipette or Pasteur pipette, suspension. The same suspension was transfer some of the water to a slide and cover  Transferred to each of the remaining 3 with a cover glass. Alternatively, transfer all centrifuge tubes one by one where in each the water to a conical tube, centrifuge, and case the centrifuge tube was shaked well and transfer the sediment to a slide. the sediment was changed into suspension.  Examine the preparation, under the low or  Next, the sediment collected in the form of middle power objective lens of a compound suspension from 4 centrifuge tubes was light microscope, for motile larvae of S. poured into a test tube labeled with the I.D. stercoralis. Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article Treatment :
child, infected with S. stercoralis, before he The drug, that was available to treat the student had been given treatment. children infected with S. stercoralis and ordered  Excess water was added to the topsoil of all by the medical doctor assigned to assist the the three petridishes and were incubated at 280 researcher of this study, was albendazole (Avion: C on the same day. Nabros, England) in the study project of  The topsoil of petridish No. 1, 2, and 3 were 6/12/2006 to 10/06/2007. On the other hand, the examined, using Baermann funnel apparatus drug of choice ordered by the medical doctor in technique to check the growth of free-living the study of 10/09/2008 to 25/06/2009 was generation of S. stercoralis, after 11, 30, and ivermectin (Ochoa: Ravenbhel, India). 48 days of initial incubation respectively.  The worms of the free-living generation of S. The dose of albendazole:
stercoralis collected from the three petridishes Each infected child whose age was 9 years and of topsoil using Baermann techniquewere above was advised to take two albendazole fixed and preserved in 10% formaldehyde to tablets at one time after dinner immediately be used for the preparation of permanent before going to bed for night sleep daily for two slides [5].
consecutive days whereas those whose ages were 8 years and below were given 1 bottle (20 ml) Method of Safranin stain preparation :
albendazole oral suspension to take after dinner I.1. Safranin O stock solution: Dissolve 2.5g immediately before going to bed for night sleep safranin O Certistain in 100 ml of 96% daily for three consecutive days. It was notified ethanol. This is a stock solution. that each tablet contained 200 mg albendazole 2. For use: 10 ml of stock solution should be di USP whereas each bottle (20 ml) contained 400 luted with 90 ml of distilled water [23,24]. mg albendazole USP. OR II. 1. Safranin powder……………0.1 g The dose of ivermectin:
2. Distilled water…………….100 ml The prescription was stated as follows in The safranin powder is dissolved in the proportion to individual student child's body distilled water measured above. weight. (Note: in this particular ivermectin 1 tablet is 6 mg in weight). Preparation
permanent
microphotographs :
6 mg tablet of ivermectin
In short, the preparation of S. stercoralis 15-24 kg 0.5 tablet, single dose on empty stomach. permanent slides was effectively done by 25-35 kg 1 tablet, single dose on empty stomach applying the following Yetwin mounting 36-50 kg 1.5 tablets, single dose on empty stomach medium [25].
51-65 kg 2 tablets, single dose on empty stomach 2.5 tablets, single dose on empty stomach Yetwin Mounting Medium:
i 1. 10% bacto-gelatin, granular, aqueous Each student child was advised to take the 2. Glycerin 50.0 ml tablet/s with a glass of water in the morning after 3. 1% chromium potassium sulfate 100.0 ml waking up from bed and begin taking meal at aqueous (Chromium) 4. Phenol (carbolic acid), melted Growth of free-living generation of S. stercoralis ii. Gelatin was dissolved in boiling water (i.e., a in the autoclaved topsoil in petridish incubated at 400 ml beaker, into which 10 g of gelatin & 90 ml of pure water were added, was Topsoil that contained organic substance was immersed in a volume of boiling water in a taken and put into three different petridishes. larger heat-resistant dish) and glycerin was Each of the petridishes was closed with its own added to it. After mixing glycerin and 10% lid and labeled 1, 2, and 3. gelatin solution, 1% chromium potassium sulfate solution and phenol were added to the Next, the petridishes with their contents of mixture of glycerin and 10% gelatin solution. topsoil were autoclaved. The medium was liquefied in 15 minutes at  The topsoil autoclaved in each of the Petri- dishes was inoculated with S. stercoralis from fresh stools sample obtained from a student Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article iii. Thereafter, the S. stercoralis worms were alcoholism; asthma; tuberculosis; malnutrition; transferred from 10% formaldehyde directly chronic pulmonary disease; leprosy; chronic renal into a drop of mounting medium, placed on a failure; impaired bowel motility; immuno- clean slide. The mounting medium with the suppressive therapy for diseases such as worms was covered with a cover slip. rheumatic disease, malignancy or cancer, and organ transplants; and promiscuous defecation. iv. Then, within overnight the gelatin hardened to form a permanent slide of S. stercoralis The difference in the infection rate of S. stercoralis in children due to the difference in the status of environmental sanitation & economic From the permanent slides prepared microphoto- income in the families' residence areas of the graphs of the larvae and other stages of S. children was analyzed by the statistic of standard stercoralis were taken using a digital camera deviation. In this case, the larger the standard from the fields of vision under suitable objective deviation meant the greater the infection rate lenses of the compound light microscope. than the mean rate, manifesting at the epidemic This was the statistical evidence for the fact that The infection rate of S. stercoralis in the the poor status of environmental sanitation and population of student children of elementary poor economic income in the parent families' schools at Dilla district was 20% in the first residence areas of elementary schools children study project (conducted during 6/12/2006 to had been one of the obvious causes for the 10/6/2007), but in the second one (done during increase of infection rate in the student children 10/9/2008 to 25/6/2009), it went up to 28%. Why with S. stercoralis. This sanitation in the was that so? That was so, because a larger residence areas of the children was poor so that amount of sample size was taken & included, in the pathogenic worm-load in the soil would be the second study project than in the first one, high and infect the bare-footed student children from student children who were living in a whose parents were poor and could not buy remote village with poor environmental sanitation and covered with diversity of perennial plants, Both parasitic male and female adults of shrubs of densely planted coffee together with Strongyloides stercoralis other giant trees where the soil was moist developmental stages had been isolated from and warm, and the majority of student children fresh stools samples of the participant student were bare-footed as they used to come from poor parent families. As the result of those environmental conditions the worm-load of S. Growth of free-living generation of S. stercoralis stercoralis in the population of student children in the autoclaved topsoil in petridishes incubated was far higher in this particular remote village at 28ºC, showed the following result. In each of than in any other site school selected for sample the three petridishes that were observed after 11, taking. Due to those environmental and economic 30, and 48 days from the date of initial factors, the infection rate of S. stercoralis grew incubation, adults (males & females) and a large up to 28% in the second study project.With those number of larvae were present. The purpose of practical results in mind, the infection rate of S. growing free-living generation to compare the stercoralis at Dilla district was adjusted to 24%, morphology of free-living males with that of taking the average infection rate of those two parasitic males. Safranin stain is not known at all to stain protozoa or any other parasite here before. When it was tried to stain the worms of S. stercoralis, for the first time, it gave a very good dyeing Several risk factors have been associated with effect. It stained the worms red. human strongyloidiasis, including coinfection Tables 1 & 2, and Fig. 2 are given on following 3 with HIV (Human Immunodeficiency Virus); consecutive pages. HTLT-1 (Human T-cell Lymphotropic Virus type1) infection; diabetes mellitus; chronic Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79


Original article Table (1): The infection rate with Strougyloides stercoralis and the cure rate of the drug albendazole
against human stronglyoidiasis, 6/12/2206 to 10/06/2007 No. of students
No. of students positive
The drug used
No. of students cured
examined
for S. stercoralis
for treatment
by the treatment
≠The percentile quantity in parenthesis adjacent to the value that meant "No. of Students positive for S. stercoralis," represented the infection rate of S. stercoralis in the population of student children whereas the one adjacent to the value that meant "No. of Students cured by the treatment," i.e., †represented the cure rate of the drug albendazole against human strongyloidiasis found at Dilla Table (2): The increase of infection rate with Strougyloides stercoralis due to the poor status of
sanitation and economic income Infection rate of S. stercoralis in children from
Infection rate of S. stercoralis in children
families of better (sanitation and economic)
from families of poor (sanitation and
status-residence areas
economic) status-residence areas
X =12%; S1 =2.2% X or X stands for a sample mean and S1 or S2 represents the standard deviation of a sample. X and S1 are variables for the children from families of better status in sanitation and in economic income whereas X and S2 are for those from families of poor status in sanitation & in economic income.

Figure (2):
Microphotographs of different developmental stages and sexes of Strongyloides
stercoralis isolated from fresh stools samples. (a) parasitic adult male ( stained with Safranin), magn‡. X64 ; (b) parasitic adult female, magn. X64; (c) egg, magn. X640; (d) rhabditiform larva, magn. X640; and (e) filariform larva, magn. X320. Pictures (b), (c), (d) and (e) were colored by a Computer Adobe Phot oshop▪ CS. Each of these five pictures was transformed from its original magnified size to the resolution of 1200 pixels/inch with the quality of 12 (maximum) and large file compatible with A4 page format . ‡magn. stands for the term magnification that gives the value of how many times the actual size of the specimen was magnified. Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article DISCUSSION
strongyloidiasis. On the other hand, the larvae of S. stercoralis, recovered from fresh stools The results obtained in this study project can samples of those infected student children, be defined as a set of achievement scored by were practically observed moving actively way of cross-sectional type of study. Standard in the fields of vision under the objectives of deviation of infection rate, in student children compound light microscopes. With this truth in mind, the student children who were positive sanitation and economic status, was far for S. stercoralis infection and did not less (S1 =2.2%) than in those from poor manifest affectedness with strongyloidiasis environmental sanitation and economic status immunocompetent. (S2 =12.5%). Larger S2 indicated that the infected children, adults and larvae of the observed infection rate went up beyond the parasite were confined to the digestive tract in mean infection rate in the population of which case the children were symptomless student children. Student children from and the S. stercoralis infection they had was families of better economic status did live a asymptomatic intestinal strongyloidiasis. In they used to get water supply lines to wash "gastropulmonary strongyloidiasis" their hands, clothes and bodies at their "disseminated strongyloidiasis" stage in these homes. Families might be in a better economic infected participant student children. position by having enough capital to carry out their own private business work in the central Here it could be understood that the parasite part of the city with better sanitation that was silently hiding in the intestine of each of would be comfortable to be hygienic and buy the infected student children to develop to the shoes for their student children that could not lethal conditions of strongyloidiasis whenever be afforded and done by poor families. the immunity of the student child was broken Parents who had educational skill and down (weakened) by some risk factors. Such government job were economically self- a hidden pathogenic parasite was found out sufficient so that they were able to buy shoes from where it was hiding by carefully for their student children, resulting in reduction employing standard diagnostic procedures in the infection rate with S. stercoralis. The such as Baermann technique and displayed fact that poor environmental sanitation and with all its developmental stages and poor economic income did form one of the sexes. Hence, the parasitic males of S. obvious causes for the increase of infection stercoralis are present together with their rates in the student children was evidenced by parasitic females in the bodies of human hosts the statistic of standard deviation and other and this verified evidence is a spectacularly targetful answer to the major question and objective of this study. When the safranin stain was tried to stain the worms of S. stercoralis, for the first time, it gave a very good dyeing effect. It stained the worms red. Actually, safranin is well known hermaphroditism) were the methods of as the secondary stain (counter stain) applied reproduction for S. stercoralis in human hosts to the fixed preparations of bacteria. If the as the parasitic males did not exist in human bacteria are decolorized with alcohol, they body [6]. S. stercoralis will take up the safranin and appear red parthenogenetic and can produce offspring (gram-negative). If the bacterial cells are not without being fertilized by the male. But the decolorized , the safranin will have no effect fact that parasitic males do exist can be on the already stained preparation, and the demonstrated in experimentally infected dogs bacteria will remain blue or purple (gram- [5]. In other words, this group of thought positive) [23, 24]. stated that the adult female S. stercoralis is parthenogenetic & hermaphroditic in the The student children who were positive for S. mechanism of reproduction. No adult male S. stercoralis infection were not revealing or not stercoralis is known to exist, the adult female manifesting affectedness with the disease, is considered as being parthenogenetic [26]. being active in their daily lives like other Another division of thought had concluded that asexual reproduction was the method for Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article the parasitic females of S. stercoralis to  Applying efficient preventive measures reproduce in human hosts for the very reason and devising effective treatment under that parasitic males did not exist in the body clinical supervision against a pathogenic of humans [8]. However, let us take that both parasite depend on deep and detailed parthenogenesis and asexual reproduction understanding about the biology and life have the same meaning for the method of cycle of the parasite. reproduction. Is there any evidence to  The term curved tail was used by generalize that adult female S. stercoralis is authors for the posterior body part of males parthenogenetic and hermaphroditic? Each of that belong to free living generation these groups of thought did not have any trace found in soil [4]. The term was not
of substantiated and persuasive scientific inclusive and unfit to define the actual proof to be accepted in science. This was so taxonomic morphology of both free-living and parasitic males of S. stercoralis. stercoralis with spicules everted out of their The degree (extent) of being curved in the spicule pouches were practically observed in posterior body part of male S. stercoralis fresh stools samples of participant student is greatly variable among the male worms children. The presence of many parasitic of both free-living and parasitic ones in males of S. stercoralis with everted spicules a similar way in extent. The morphology in fresh stools samples together with parasitic of both parasitic and free living males is females was a very strong evidence for the the same. This was verified by growing fact that there was copulation & fertilization. free-living males in autoclaved topsoil that Everted spicules of males are seen only at was inoculated with fresh stools sample times of mating. obtained from an infected child before giving him treatment and incubated at CONCLUSION
28ºC. When the morphology, including the  The result of this study had identified the variation in the degree of curvature or concepts of both parthenogenesis/asexual coiling of the posterior body part, of these reproduction and protandrogony, in the free-living males was compared with that parasitic generation of S. stercoralis, to be of parasitic males, it was found to be similar in both free living and parasitic evidences, on the reproduction of S. ascertained to be correct to differentiate stercoralis in the parasitic generation in and free-living males human hosts that had been reacted to by their respective this paper could not be denied because they stercoralis was a ventrally "curved or were reported straight forward by authors coiled posterior body part" in the males of in modern textbooks, journals, and on the this very parasite whereas that of the internet. Due to those reports, in all females was straight. modern human parasitology textbooks,  In the life cycle of parasitic generation of journal, and on the internet, the males of S. S. stercoralis both parasitic male & female stercoralis had been excluded (omitted) must be included just like the free-living from the life cycle of its parasitic male & female in their life cycle. generation in human hosts. It was possible for copulation to take place between the  This article is a realistic response to a parasitic males and females to result in chronic global problem that has remained fertilization in the lumen of the human unsolved for generations of man until now host's gut and then the fertilized parasitic and needs world-wide attention of human female could burrow into the intestinal parasitologists. mucosa to lay eggs that would hatch soon. It is just like a domestic cock and a hen Funding:
where it is the hen which goes to a nest The financial support, covering the cost of after mating to lay and incubate eggs and children and technical assistance by assigning a medical doctor for prescription and clinical Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article supervision were given by the Ethiopian REFERENCES
Catholic Church (Dilla Don Bosco). 1. Mitreva M, McCarter JP, Martin J, Dante M, Conflict of interest:
Wylie T, Chiapelli B, et al. Comparative genomics of gene expression in the parasitic I confirm that I don't have any competitive and free-living nematodes Strongyloides conflict of interest with any body. stercoralis and Caenorhabditis elegans. Genome Res [serial on the Internet]. 2004 Ethical approval:
[cited 2012 Oct.24]14: [about 6p.]. Available from:http://genome.cship.org/content/14/2/209. full Ethical permission/clearance to perform the 2. Concha R, Harrington WJR, Rogers AI. research work for the well-being of human strongyloidiasis: subjects was obtained from:- Dilla University, management, and determinants of outcome. J the Office of Gedio-Zone Administration, and Clin Gastroenterol. 2005; 39: 203-11. the Directors of the schools involved in the 3. Hunter CJ, Petrosyan M, Asch M. Dissemination of study. The demand for the continuity of this Strongyloides stercoralis in a patient with study project and participation by the systemic lupus erythematous after initiation of participant student children and their parents albendazole: a case report. J Med Case was unusually high. Reports [serial on the Internet]. 2008 May 14 [cited 2012 Oct. 24];2: [about 5p.]. Available from: http: //www.jmedicalcasereports.com/con Don Bosco, i.e., Dilla Ethiopian Catholic 4. Gilles HM. Soil-transmitted helminths (geohel- minths). In: Cook GC. editor. Manson's tropical Church was very kind and quick to accept the request forwarded by the researcher of this ed., London: WB Saunders; 1996; study for technical and medical assistance. 5. Paniker CKJ. Textbook of medical parasitology. Contribution to and participation in this study 6th ed., New Delhi; Jaypee Brothers Medical project by Don Bosco was immense. It was Publishers 2007; 169-74. this very organization that rescued the student 6. Bogitsh BJ, Carter CE, Oeltmann TN. Human Strongyloides parasitology. 3rd ed., USA; Academic press stercoralis when it understood the potential risk the infection could entail. Then it reacted 7. Dillard KJ, Saari SAM, Anttila M. promptly by covering the financial cost of the Strongyloides stercorali Finnish kennel. Acta Vet Scand [serial on the drug of choice for treatment and assigned a Internet]. 2007 Dec 12 [cited 2012 Oct 24]; 49: medical doctor (Dr. Corazon B. Jaca FMA) [about 5p.].Available from: for prescription and clinical supervision in the http://www.actavetscand.com/content/49/1/37 process of treatment. I am extremely thankful 8. Keiser PB, Nutman TB. Strongyloides stercoralis in to Don Bosco for its extensive financial the immunocompromised population. Clin contribution and participation in favour of the Microbiol Rev [serial on the Internet]. 2004 Jan infected student children. [cited 2012 Oct 24];17(1):[about 3p.]. Available from:http://www.ncbi.nlm.nih.gov>…>Clin I am very grateful to the office of the Microbiol Rev>v.17(1); Jan 2004 President of Dilla University and the main Administrator Office of Gedio Zone for their Hyperinfective strongyloidiasis: S. stercoralis writing letters to the Directors of elementary undergoes autoinfective burst in neonatal schools, requesting them to be cooperative for gerbils. J Parasitol 1999; 85: 286-9. my research activities with their students. 10. Dionisio D, Manneschi LL, Lollo SD, Orsi A, Tani A, Esperti F. Strongyloides stercoralis: I am very strongly thankful to Mr. Tilahun ultrostructural study of newly hatched larvae Kebede, Mr. Mingizem Birhan, and Mr. within human duodenal mucosa. J Clin Pathol Tsegaye Hylu who are computer science 2000; 53: 110-6. teachers & technicians in Dilla University, for 11. Massey HCJ, Nishi M, Chaudhary K, Pakpout their providing me with and installing N, Lok JB. Structure and developmental technical computer tools such as Equation Strongyloides stercoralis. Editor and Adobe Photoshop▪CS that were Int J Parasitol 2003; 33: 1537-44. essential in processing the data. Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79 Original article 12. Garg PK, Perry S, Dorn M, Hardcastle L, 19. Fardet L, Genereau T, Poirot JL, Guidet B, Parsonnet J. Risk of intestinal helminth and Kettaneh A, Cabane L. Severe strongyloidiasis protozoan infection in a refugee population. Am in corticosteroid-treated patients: case series J Trop Med Hyg [serial on the Internet]. 2005 and literature review. J Infect 2007; 54:18-27. Mar 20 [cited 2012 Oct 24]; 83(3): [about 4 p.]. 20. Kerepesi LA, Hess JA, Leon O, Nolan TJ, Available from: http: //www.ajtm.org/content/ Schad GA, Abraham D. Toll-like receptor 4(TLR4) is required for protective immunity to 13. Siddiqui AA, Berk SL. Diagnosis of Strongyloides larval Strongyloides stercoralis in mice. stercoralis. In: Ericsson CD, Steffen R, editors. Microb Infect 2007; 9: 28-34. Clin Infect Dis [serial on the Internet]. 2012 21. Rahif RH, Emara GY, Al-saqur IM. Epidemio- Oct 15 [cited 2012 Oct 24]; 55(8): [about logical study of intestinal nematodes in dogs in 8 p.]. Available from: http: //cid.oxfordjournals.
Basrah(Iraq): prevalence of helminthic ova in org/content/33/7/ 1040.full the fecal deposits. Iraqi J Vet Sci 2002; 16: 71- 14. Siegel MO, Simon GL. Is human immuno- deficiency virus infection a risk factor for 22. Cheesbrough M. District laboratory practice in Strongyloides stercoralis hyperinfection and tropical countries. Part 1. India; Cambridge dissemination? PLoS Negl Trop Dis [serial on University Press 1998; 214-5. the Internet]. 2012 Jul 31 [cited 2012 Oct 24]; 23. Merck KGA. Microscopy: Safranin 0 (C.I. 6(7): [about 4 p.].Available from: http:// www. 50240) Certistain. [serial on the Internet]. 2011; ncbi. nlm.nih.gov>…>PLoS Negl Trop Dis> [about 1 p]. Available from: v.6(7); Jul 2012 15. Ramanathan R, Nutman TB. Strongyloides 24. Boyd PF. Basic medical microbiology. 5th ed. stercoralis USA: Little, Brown and Co. (Inc). 1995; 557. compromised Host. Japan J Parasitol [serial on 25. Beadle GW, Emerson R, Whitaker DM, the Internet]. 2008 May [cited 2012 Oct. 24]; Lawson CA, Lewis RW, Biirmester MA et al. 10(2):[about 4 p.]. Available from: http://www. Full text of "Animal tissue techniques". [serial on the Internet]. USA: W.H. Freeman and Co. 16. Poonam P, Jayshree RS, Acharya RS, Hema S, 1962; [about 1]. Available from: http://archive. Govind B, Suresh TM. Fulminant fatal org/stream/ animaltissuech00hurr Strongyloides stercoralis infection in a 26. Zeibig EA. Clinical parasitology: a practical postchemo-therapy immuno-suppressed cancer approach. London; W.B Saunders Co. 1997; 148. patient. Med Pediatric Oncol 1999; 33: 504-5. 17. Jayshree RS, Hema S, Govind B, Suresh TM. Peer reviewer: Samia Etewa: Professor of
Strongyloides stercoralis Parasitology, Faculty of Medicine, Zagazig the stools during anticancer therapy. Indian J University, Egypt. Gastroenterol 2001; 20: 160- 1. Editor: Tarik Zaher: Professor of Tropical
18. Kim J, Joo HS, Ko HM, Na MS, Hw ASH, Im JC. A case of fatal hyperinfective strongyloidiasis University, Egypt. with discovery of autoinfective filariform larvae in sputum. Korean J Parasitol 2005; 43: 51-5. Eriso, Afro-Egypt J Infect Endem Dis 2014; 4(2): 69-79

Source: http://mis.zu.edu.eg/ajied/Ajied_System_Files/p123.pdf

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‘KSR' has made it easier to prove obviousness Particularly in pharmaceutical cases, this is an increasingly viable patent invalidity defense. By Mark Jansen and Byron ChinIn KSR International Co. v. Teleflex Inc., 550 U.S. 398 (2007), the U.S. Supreme Court heralded more promise in obviousness challenges to patent validity. The Court rejected the U.S. Court of Appeals for the Federal Circuit's "teaching, suggestion and motivation" (TSM) test for patent obviousness as a "rigid preventative rule" and instructed lower courts to take a more "expansive and flexible approach," one applying "common sense" and recognizing that the "combination of familiar elements according to known methods is likely to be obvious when it does no more than yield predictable results."

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