Search results for: Newcastle vaccine
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 250

Search results for: Newcastle vaccine

40 Molecular Detection of Acute Virus Infection in Children Hospitalized with Diarrhea in North India during 2014-2016

Authors: Ali Ilter Akdag, Pratima Ray

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Background:This acute gastroenteritis viruses such as rotavirus, astrovirus, and adenovirus are mainly responsible for diarrhea in children below < 5 years old. Molecular detection of these viruses is crucially important to the understand development of the effective cure. This study aimed to determine the prevalence of common these viruses in children < 5 years old presented with diarrhea from Lala Lajpat Rai Memorial Medical College (LLRM) centre (Meerut) North India, India Methods: Total 312 fecal samples were collected from diarrheal children duration 3 years: in year 2014 (n = 118), 2015 (n = 128) and 2016 (n = 66) ,< 5 years of age who presented with acute diarrhea at the Lala Lajpat Rai Memorial Medical College (LLRM) centre(Meerut) North India, India. All samples were the first detection by EIA/RT-PCR for rotaviruses, adenovirus and astrovirus. Results: In 312 samples from children with acute diarrhea in sample viral agent was found, rotavirus A was the most frequent virus identified (57 cases; 18.2%), followed by Astrovirus in 28 cases (8.9%), adenovirus in 21 cases (6.7%). Mixed infections were found in 14 cases, all of which presented with acute diarrhea (14/312; 4.48%). Conclusions: These viruses are a major cause of diarrhea in children <5 years old in North India. Rotavirus A is the most common etiological agent, follow by astrovirus. This surveillance is important to vaccine development of the entire population. There is variation detection of virus year wise due to differences in the season of sampling, method of sampling, hygiene condition, socioeconomic level of the entire people, enrolment criteria, and virus detection methods. It was found Astrovirus higher then Rotavirus in 2015, but overall three years study Rotavirus A is mainly responsible for causing severe diarrhea in children <5 years old in North India. It emphasizes the required for cost-effective diagnostic assays for Rotaviruses which would help to determine the disease burden.

Keywords: adenovirus, Astrovirus, hospitalized children, Rotavirus

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39 The Comparison between bFGF and Small Molecules in Derivation of Chicken Primordial Germ Cells and Embryonic Germ Cells

Authors: Maryam Farzaneh, Seyyedeh Nafiseh Hassani, Hossein Baharvand

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Objective: Chicken gonadal tissue has a two population such primordial germ cells (PGCs) and stromal cells (somatic cells). PGCs and embryonic germ cells (EGCs) that is a pluripotent type of PGCs in long-term culture are suitable sources for the production of chicken pluripotent stem cell lines, transgenic birds, vaccine and recombinant protein production. In general, the effect of growth factors such bFGF and mouse LIF on derivation of PGCs in vitro are important and in this study we could see the unique effect of small molecules such PD032 and SB43 as a chemical, in comparison to growth factors. Materials and Methods: After incubation of fertilized chicken egg up to 6 days and isolation of primary gonadal tissues and culture of mixed cells like PGCs and stromal cells. PGCs proliferate in the present of fetal calf serum (FCS) and small molecules and in another group bFGF, that these factors are important for PGCs culture and derivation. Somatic cells produce a multilayer feeder under the PGCs in primary culture and PGCs make a small cluster under these cells. Results: In present of small molecules and high volume of FCS (15%), the present of EGCs as a pluripotent stem cells were clear four weeks, that they had a positive immune-staining and periodic acid-Schiff staining (PAS), but in present of growth factors like bFGF without any chemicals, the present of PGCs were clear but after 7 until 10 days, there were disappear. Conclusion: Until now we have seen many researches about derivation and maintenance of chicken PGCs, in the hope of understanding the mechanisms that occur during germline development and production of a therapeutic product by transgenic birds. There are still many unknowns in this area and this project will try to have efficient conditions for identification of suitable culture medium for long-term culture of PGCs in vitro without serum and feeder cells.

Keywords: chicken gonadal primordial germ cells, pluripotent stem cells, growth factors, small molecules, transgenic birds

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38 Functional Characterization of Rv1019, a Putative TetR Family Transcriptional Regulator of Mycobacterium Tuberculosis H37Rv

Authors: Akhil Raj Pushparajan, Ranjit Ramachandran, Jijimole Gopi Reji, Ajay Kumar Ramakrishnan

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Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is one of the leading causes of death by an infectious disease. In spite of the availability of effective drugs and a vaccine, TB is a major health concern and was declared a global emergency by the World Health Organization (WHO). The success of intracellular pathogens like Mtb depends on its ability to overcome the challenging environment in the host. Gene regulation controlled by transcriptional regulators (TRs) plays a crucial role for the bacteria to adapt to the host environment. In vitro studies on gene regulatory mechanisms during dormancy and reactivation have provided insights into the adaptations employed by Mtb to survive in the host. Here we present our efforts to functionally characterize Rv1019, a putative TR of Mtb H37Rv which was found to be present at significantly varying levels during dormancy and reactivation in vitro. The expression of this protein in the dormancy-reactivation model was validated by qRT-PCR and western blot. By DNA- protein interaction studies and reporter assays we found that under normal laboratory conditions of growth this protein behaves as an auto-repressor and tetracycline was found to abrogate this repression by interfering with its ability to bind DNA. Further, by cDNA analysis, we found that this TR is co-transcribed with its downstream genes Rv1020 (mfd) and Rv1021 (mazG) which are involved in DNA damage response in Mtb. Constitutive expression of this regulator in the surrogate host M. smegmatis showed downregulation of the orthologues of downstream genes suggested that Rv1019 could negatively regulate these genes. Our finds also show that M. smegmatis expressing Rv1019 is sensitive to DNA damage suggests the role of this protein in regulating DNA damage response induced by oxidative stress. Because of its role in regulating DNA damage response which may help in the persistence of Mtb, Rv1019 could be used as a prospective target for therapeutic intervention to fight TB.

Keywords: auto-repressor, DNA repair, mycobacterium smegmatis, mycobacterium tuberculosis, tuberculosis

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37 Longitudinal Profile of Antibody Response to SARS-CoV-2 in Patients with Covid-19 in a Setting from Sub–Saharan Africa: A Prospective Longitudinal Study

Authors: Teklay Gebrecherkos

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Background: Serological testing for SARS-CoV-2 plays an important role in epidemiological studies, in aiding the diagnosis of COVID-19 and assess vaccine responses. Little is known about the dynamics of SARS-CoV-2 serology in African settings. Here, we aimed to characterize the longitudinal antibody response profile to SARS-CoV-2 in Ethiopia. Methods: In this prospective study, a total of 102 PCR-confirmed COVID-19 patients were enrolled. We obtained 802 plasma samples collected serially. SARS-CoV-2 antibodies were determined using four lateral flow immune assays (LFIAs) and an electrochemiluminescent immunoassay. We determined longitudinal antibody response to SARS-CoV-2 as well as seroconversion dynamics. Results: Serological positivity rate ranged between 12%-91%, depending on timing after symptom onset. There was no difference in the positivity rate between severe and non-severe COVID-19 cases. The specificity ranged between 90%-97%. Agreement between different assays ranged between 84%-92%. The estimated positive predictive value (PPV) for IgM or IgG in a scenario with seroprevalence at 5% varies from 33% to 58%. Nonetheless, when the population seroprevalence increases to 25% and 50%, there is a corresponding increase in the estimated PPVs. The estimated negative-predictive value (NPV) in a low seroprevalence scenario (5%) is high (>99%). However, the estimated NPV in a high seroprevalence scenario (50%) for IgM or IgG is reduced significantly from 80% to 85%. Overall, 28/102 (27.5%) seroconverted by one or more assays tested within a median time of 11 (IQR: 9–15) days post symptom onset. The median seroconversion time among symptomatic cases tended to be shorter when compared to asymptomatic patients [9 (IQR: 6–11) vs. 15 (IQR: 13–21) days; p = 0.002]. Overall, seroconversion reached 100% 5.5 weeks after the onset of symptoms. Notably, of the remaining 74 COVID-19 patients included in the cohort, 64 (62.8%) were positive for antibodies at the time of enrollment, and 10 (9.8%) patients failed to mount a detectable antibody response by any of the assays tested during follow-up. Conclusions: Longitudinal assessment of antibody response in African COVID-19 patients revealed heterogeneous responses. This underscores the need for a comprehensive evaluation of serum assays before implementation. Factors associated with failure to seroconvert need further research.

Keywords: COVID-19, antibody, rapid diagnostic tests, ethiopia

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36 Raising Antibodies against Epoxyscillirosidine, the Toxic Principle Contained in Moraea pallida Bak. in Rabbits

Authors: Hamza I. Isa, Gezina C. H. Ferreira, Jan E. Crafford, Christoffel J. Botha

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Moraea pallida Bak. (yellow tulip) poisoning is the most important plant-induced cardiac glycoside toxicosis in South Africa. Cardiac glycoside poisonings collectively account for about 33 and 10 % mortalities due to plants, in large and small stock respectively, in South Africa. The toxic principle is 1α, 2α-epoxyscillirosidine, a bufadienolide. The aim of the study was to investigate the potential to develop a vaccine against epoxyscillirosidine. Epoxyscillirosidine and the related bufadienolides proscillaridin and bufalin, which are commercially available, were conjugated to the carrier proteins [Hen ovalbumin (OVA), bovine serum albumin (BSA) and keyhole limpet haemocyanin (KLH)], rendering them immunogenic. Adult male New Zealand White rabbits were immunized. In Trials 1 and 2, rabbits (n=6) were, each assigned to two groups. Experimental animals (n=3; n=4) were vaccinated with epoxyscillirosidine-OVA conjugate, while the control (n=3; n=2) were vaccinated with OVA, using Freund’s complete and incomplete and Montanide adjuvants, for Trials 1 and 2, respectively. In Trial 3, rabbits (n=15), randomly allocated to 5 equal groups (I, II, III, IV and V), were vaccinated with proscillaridin-BSA, bufalin-BSA, epoxyscillirosidine-KLH, epoxyscillirosidine-BSA conjugates, and BSA respectively, using Montanide as adjuvant. Vaccination was on Days 0, 21 and 42. Additional vaccinations were done on Day 56 and 63 for Trial 1. Vaccination was by intradermal injection of 0.4 ml of the immunogen (4 mg/ml [Trial 1] and 8 mg/ml for Trials 2 and Trial 3, respectively). Blood was collected pre-vaccination and at 3 week intervals following each vaccination. Antibody response was determined using an indirect ELISA. There was poor immune response associated with the dose (0.4 mg per rabbit) and adjuvant used in Trial 1. Antibodies were synthesized against the conjugate administered in Trial 2. For Trail 3, antibodies against the immunogens were successfully raised in rabbits with epoxyscillirosidine-KLH inducing the highest immune response. The antibodies raised against proscillaridin and bufalin cross-reacted with epoxyscillirosidine when used as antigen in the ELISA. The study successfully demonstrated the synthesis of antibodies against the bufadienolide conjugates administered. The cross-reactivity of proscillaridin and bufalin with epoxyscillirosidine could potentially be utilized as alternative to epoxyscillirosidine in future studies to prevent yellow tulp poisoning by vaccination.

Keywords: antibodies , bufadienolides, cross-reactivity, epoxyscillirosidine, Moraea pallida, poisoning

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35 Risk Factors for Severe Typhoid Fever in Children: A French Retrospective Study about 78 Cases from 2000-2017 in Six Parisian Hospitals

Authors: Jonathan Soliman, Thomas Cavasino, Virginie Pommelet, Lahouari Amor, Pierre Mornand, Simon Escoda, Nina Droz, Soraya Matczak, Julie Toubiana, François Angoulvant, Etienne Carbonnelle, Albert Faye, Loic de Pontual, Luu-Ly Pham

Abstract:

Background: Typhoid and paratyphoid fever are systemic infections caused by Salmonella enterica serovar Typhi or paratyphi (A, B, C). Children traveling to tropical areas are at risk to contract these diseases which can be complicated. Methods: Clinical, biological and bacteriological data were collected from 78 pediatric cases reported between 2000 and 2017 in six Parisian hospitals. Children aged 0 to 18 years old, with a diagnosis of typhoid or paratyphoid fever confirmed by bacteriological exams, were included. Epidemiologic, clinical, biological features and presence of multidrug-resistant (MDR) bacteria or intermediate susceptibility to ciprofloxacin (nalidixic acid resistant) were examined by univariate analysis and by logistic regression analysis to identify risk factors of severe typhoid in children. Results: 84,6% of the children were imported cases of typhoid fever (n=66/78) and 15,4% were autochthonous cases (n=12/78). 89,7% were caused by S.typhi (n=70/78) and 12,8% by S.paratyphi (n=10/78) including 2 co-infections. 19,2% were intrafamilial cases (n=15/78). Median age at diagnosis was 6,4 years-old [6 months-17,9 years]. 28,2% of the cases were complicated forms (n=22/78): digestive (n=8; 10,3%), neurological (n=7; 9%), pulmonary complications (n=4; 5,1%) and hemophagocytic syndrome (n=4; 5,1%). Only 5% of the children had prior immunization with typhoid non-conjugated vaccine (n=4/78). 28% of the cases (n=22/78) were caused by resistant bacteria. Thrombocytopenia and diagnosis delay was significantly associated with severe infection (p= 0.029 and p=0,01). Complicated forms were more common with MDR (p=0,1) and not statistically associated with a young age or sex in this study. Conclusions: Typhoid and paratyphoid fever are not rare in children back from tropical areas. This multicentric pediatric study seems to show that thrombocytopenia, diagnosis delay, and multidrug resistant bacteria are associated with severe typhoid fever and complicated forms in children.

Keywords: antimicrobial resistance, children, Salmonella enterica typhi and paratyphi, severe typhoid

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34 Molecular Characterisation and Expression of Glutathione S-Transferase of Fasciola Gigantica

Authors: J. Adeppa, S. Samanta, O. K. Raina

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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

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33 Chikungunya Virus Infection among Patients with Febrile Illness Attending University of Maiduguri Teaching Hospital, Nigeria

Authors: Abdul-Dahiru El-Yuguda, Saka Saheed Baba, Tawa Monilade Adisa, Mustapha Bala Abubakar

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Background: Chikungunya (CHIK) virus, a previously anecdotally described arbovirus, is now assuming a worldwide public health burden. The CHIK virus infection is characterized by potentially life threatening and debilitating arthritis in addition to the high fever, arthralgia, myalgia, headache and rash. Method: Three hundred and seventy (370) serum samples were collected from outpatients with febrile illness attending University of Maiduguri Teaching Hospital, Nigeria, and was used to detect for Chikungunya (CHIK) virus IgG and IgM antibodies using the Enzyme Linked Immunosorbent Assays (ELISAs). Result: Out of the 370 sera tested, 39 (10.5%) were positive for presence of CHIK virus antibodies. A total of 24 (6.5%) tested positive for CHIK virus IgM only while none (0.0%) was positive for presence of CHIK virus IgG only and 15 (4.1%) of the serum samples were positive for both IgG and IgM antibodies. A significant difference (p<0.0001) was observed in the distribution of CHIK virus antibodies in relation to gender. The males had prevalence of 8.5% IgM antibodies as against 4.6% observed in females. On the other hand 4.6% of the females were positive for concurrent CHIK virus IgG and IgM antibodies when compared to a prevalence of 3.4% observed in males. Only the age groups ≤ 60 years and the undisclosed age group were positive for presence of CHIK virus IgG and/or IgM antibodies. No significant difference (p>0.05) was observed in the seasonal prevalence of CHIK virus antibodies among the study subjects Analysis of the prevalence of CHIK virus antibodies in relation to clinical presentation (as observed by Clinicians) of the patients revealed that headache and fever were the most frequently encountered ailments. Conclusion: The CHIK virus IgM and concurrent IgM and IgG antibody prevalence rates of 6.5% and 4.1% observed in this study indicates a current infection and the lack of IgG antibody alone observed shows that the infection is not endemic but sporadic. Recommendation: Further studies should be carried to establish the seasonal prevalence of CHIK virus infection vis-à-vis vector dynamics in the study area. A comprehensive study need to be carried out on the molecular characterization of the CHIK virus circulating in Nigeria with a view to developing CHIK virus vaccine.

Keywords: Chikungunya virus, IgM and IgG antibodies, febrile patients, enzyme linked immunosorbent assay

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32 An Ecofriendly Approach for the Management of Aedes aegypti L (Diptera: Culicidae) by Ocimum sanctum

Authors: Mohd Shazad, Kamal Kumar Gupta

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Aedes aegypti (Diptera: Culicidae), commonly known as tiger mosquito is the vector of dengue fever, yellow fever, chikungunya and zika virus. In the absence of any effective vaccine against these diseases, control the mosquito population is the only promising mean to prevent the diseases. Currently used chemical insecticides cause environmental contamination, high mammalian toxicity and hazards to non-target organisms, insecticide resistance and vector resurgence. Present research work aimed to explore the potentials of phytochemicals present in the Ocimum sanctum in management of mosquito population. The leaves of Ocimum were extracted with ethanol by ‘cold extraction method’. 0-24h old fourth instar larvae of Aedes aegypti were treated with the extract of concentrations 50ppm, 100ppm, 200ppm and 400ppm for 24h. Survival, growth and development of the treated larvae were evaluated. The adults emerged from the treated larvae were used for the reproductive fitness studies. Our results indicate 77.2% mortality in the larvae exposed to 400 ppm. At lower doses, although there was no significant reduction in the survival after 24h however, it decreased during subsequent days of observations. In control experiments, no mortality was observed. It was also observed that the larvae survived after treatment showed severe growth and developmental abnormalities. There was significant increase in larval duration. In control, fourth instar moulted into pupa after 3 days while larvae treated with 400 ppm extract were moulted after 4.6 days. Larva-pupa intermediates and the pupa-adult intermediates were observed in many cases. The adults emerged from the treated larvae showed impaired mating and oviposition behaviour. The females exhibited longer preoviposition period, reduced oviposition rate and decreased egg output. GCMS analysis of the ethanol extract revealed presence of JH mimics and intermediates of JH biosynthetic pathway. Potentials of Ocimum sanctum in integrated vector management programme of Aedes aegypti were discussed.

Keywords: Aedes aegypti, Ocimum sanctum, oviposition, survival

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31 Identification and Characterization of Polysaccharide Biosynthesis Protein (CAPD) of Enterococcus faecium

Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc

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Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.

Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation

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30 In Silico Screening, Identification and Validation of Cryptosporidium hominis Hypothetical Protein and Virtual Screening of Inhibitors as Therapeutics

Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

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Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

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29 A Retrospective Study on the Spectrum of Infection and Emerging Antimicrobial Resistance in Type 2 Diabetes Mellitus

Authors: Pampita Chakraborty, Sukumar Mukherjee

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People with diabetes mellitus are more susceptible to developing infections, as high blood sugar levels can weaken the patient's immune system defences. People with diabetes are more adversely affected when they get an infection than someone without the disease, because you have weakened immune defences in diabetes. People who have minimally elevated blood sugar levels experience worse outcomes with infections. Diabetic patients in hospitals do not necessarily have a higher mortality rate due to infections, but they do face longer hospitalisation and recovery times. A study was done in a tertiary care unit in eastern India. Patients with type 2 diabetes mellitus infection were recruited in the study. A total of 520 cases of Type 2 Diabetes Mellitus were recorded out of which 200 infectious cases was included in the study. All subjects underwent detailed history & clinical examination. Microbiological samples were collected from respective site of the infection for microbial culture and antibiotic sensitivity test. Out of the 200 infectious cases urinary tract infection(UTI) was found in majority of the cases followed by diabetic foot ulcer (DFU), respiratory tract infection(RTI) and sepsis. It was observed that Escherichia coli was the most commonest pathogen isolated from UTI cases and Staphylococcus aureus was predominant in foot ulcers followed by other organisms. Klebsiella pneumonia was the major organism isolated from RTI and Enterobacter aerogenes was commonly observed in patients with sepsis. Isolated bacteria showed differential sensitivity pattern against commonly used antibiotics. The majority of the isolates were resistant to several antibiotics that are usually prescribed on an empirical basis. These observations are important, especially for patient management and the development of antibiotic treatment guidelines. It is recommended that diabetic patients receive pneumococcal and influenza vaccine annually to reduce morbidity and mortality. Appropriate usage of antibiotics based on local antibiogram pattern can certainly help the clinician in reducing the burden of infections.

Keywords: antimicrobial resistance, diabetic foot ulcer, respiratory tract infection, urinary tract infection

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28 Interrelationship of Socio-Demographic Factors, Health Belief Dimensions and Compliance to Measles Vaccination among Filipino Mothers

Authors: Beryl Rene R. Lopez, Lesley Anne M. Lipat, Rhogene Barbette C. Lirio, Laurice Joy H. Llanes, Karl Philippe M. Llapitan, Einstein James R. Lopez, Socorro S. GuanHing

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Background: Measles remain as one of the most common childhood diseases despite the availability of the vaccine that is safe and cost-effective. Because of morbidity and mortality associated with the recent measles outbreak in the Philippines, there is an increasing concern from the health care professionals. Objective: The purpose of this study is to determine the relationship between the compliance of Filipino mothers to measles vaccination and their health beliefs when grouped according to the given socio-demographic factors using a researcher-made questionnaire. Research Methodology: This research utilized the descriptive-correlational research design. With the use of purposive sampling technique, the study involved 200 Filipino mothers aged 18 years old and above excluding those who are healthcare professionals with children aged 2-3 years old with either urban or rural as their settlements. Pre-testing was done prior to the actual data gathering. A questionnaire composed of 26 items involving socio-demographic, compliance, and health beliefs was distributed to the sample population. Statistical analysis was done with the use of Exploratory Factor Analysis (EFA) for the first research question and Structural Equation Model (SEM) for the second research question. Results: Four dimensions were generated with the use of EFA namely: Vulnerability-Oriented Beliefs (VOB), Knowledge-Oriented Beliefs (KOB), Accessibility-Oriented Beliefs (AOB), and Outcomes-Oriented Beliefs (OOB). These were then correlated with the mothers’ socio-demographic factors (age, educational attainment, the area of residence, the number of children, and family income) and their compliance to the measles vaccination schedule. Results showed significant and direct relationships between area of residence and compliance, family income and compliance, KOB and compliance, education and KOB, KOB and VOB, KOB and OOB, AOB and KOB, AOB and OOB, AOB and VOB, and lastly, OOB and VOB. Conclusion: The Knowledge – Oriented Belief dimension greatly influence compliance to measles vaccination. Other determinants of compliance like the area of residence, educational attainment, and family income significantly increase the Filipino mothers’ likelihood of compliance to measles vaccination, which have implications to health education.

Keywords: socio-demographic, health beliefs, compliance, measles vaccination

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27 Change of Epidemiological Characteristics and Disease Burden of Varicella Due to Implementation of Mass Immunization Program in Taiwan from 2000 to 2012

Authors: En-Tzu Wang, Ting-Ann Wang, Yi-Hui Shen, Yu-Min Chou, Chi-Tai Fang, Chin-Hui Yang

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Background and purpose: A mass varicella immunization program was established to provide free 1-dose vaccination for all 1-year-old children throughout Taiwan since 2004. The epidemiological characteristics and disease burden of varicella from 2000 to 2012 was investigated and the results will be essential to refine the national immunization policy. Method: We included patients (n = 17,838–164,245) with ICD-9-CM codes 052 (chickenpox) from the 2000 to 2012 National Health Insurance Database. The age, period, and cohort-specific incidence of varicella were calculated. The hospital admission rate, medical costs and indirect costs from the societal perspective of varicella including travel costs to the medical facility, registration fee, productivity losses of the patients and caregivers were also estimated. Result: There were 979,252 patients for medical treatment due to varicella from 2000 to 2012 in Taiwan. The implementation of a routine childhood varicella vaccination program has resulted in 87% decline in morbidity (881.49 to 115.17 per 100,000). The average age of patients increased from 7.9 years to 16.3 years. The overall varicella-related hospital admission rate was 15.5 per 1000 patients, and peaked in the groups of infants younger than 1 year, adults aged from 20 to 39 years and elders over 70 years. Among patients admitted to hospital, 33.5% of them had one or more complications. Patients with underlying diseases had higher admission rate (241.6 per 1,000) and longer duration of hospital stay (6.61 days vs. 4.76 days). The annual varicella-related medical expense declined after 2002 and the proportion of medical costs for admission has increased to 42%. The annual indirect costs from the societal perspective of varicella were 5.29 to 9.63 times higher than varicella-related medical costs. Every one dollar invested in the varicella immunization program, 2.97 dollars of medical and social costs were saved on average. Conclusion: The dramatic decline in morbidity, hospitalization, medical and social costs of varicella can be directly attributed to the implementation of the mass immunization program. Two-dose vaccination is recommended for both children with underlying diseases and susceptible adults to prevent serious complications and hospitalizations.

Keywords: disease burden, epidemiology, medical and social costs, varicella, varicella vaccine

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26 Breast Cancer Cellular Immunotherapies

Authors: Zahra Shokrolahi, Mohammad Reza Atashzar

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The goals of treating patients with breast cancer are to cure the disease, prolong survival, and improve quality of life. Immune cells in the tumor microenvironment have an important role in regulating tumor progression. The term of cellular immunotherapy refers to the administration of living cells to a patient; this type of immunotherapy can be active, such as a dendritic cell (DC) vaccine, in that the cells can stimulate an anti-tumour response in the patient, or the therapy can be passive, whereby the cells have intrinsic anti-tumour activity; this is known as adoptive cell transfer (ACT) and includes the use of autologous or allogeneic lymphocytes that may, or may not, be modified. The most important breast cancer cellular immunotherapies involving the use of T cells and natural killer (NK) cells in adoptive cell transfer, as well as dendritic cells vaccines. T cell-based therapies including tumour-infiltrating lymphocytes (TILs), engineered TCR-T cells, chimeric antigen receptor (CAR T cell), Gamma-delta (γδ) T cells, natural killer T (NKT) cells. NK cell-based therapies including lymphokine-activated killers (LAK), cytokine-induced killer (CIK) cells, CAR-NK cells. Adoptive cell therapy has some advantages and disadvantages some. TILs cell strictly directed against tumor-specific antigens but are inactive against tumor changes due to immunoediting. CIK cell have MHC-independent cytotoxic effect and also need concurrent high dose IL-2 administration. CAR T cell are MHC-independent; overcome tumor MHC molecule downregulation; potent in recognizing any cell surface antigen (protein, carbohydrate or glycolipid); applicable to a broad range of patients and T cell populations; production of large numbers of tumor-specific cells in a moderately short period of time. Meanwhile CAR T cells capable of targeting only cell surface antigens; lethal toxicity due to cytokine storm reported. Here we present the most popular cancer cellular immunotherapy approaches and discuss their clinical relevance referring to data acquired from clinical trials .To date, clinical experience and efficacy suggest that combining more than one immunotherapy interventions, in conjunction with other treatment options like chemotherapy, radiotherapy and targeted or epigenetic therapy, should guide the way to cancer cure.

Keywords: breast cancer , cell therapy , CAR T cell , CIK cells

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25 Human Rabies Survivors in India: Epidemiological, Immunological and Virological Studies

Authors: Madhusudana S. N., Reeta Mani, Ashwini S. Satishchandra P., Netravati, Udhani V., Fiaz A., Karande S.

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Rabies is an acute encephalitis which is considered 100% fatal despite occasional reports of survivors. However, in recent times more cases of human rabies survivors are being reported. In the last 5 years, there are six laboratories confirmed human rabies survivors in India alone. All cases were children below 15 years and all contracted the disease by dog bites. All of them also had received the full or partial course of rabies vaccination and 4 out of 6 had also received rabies immunoglobulin. All cases were treated in intensive care units in hospitals at Bangalore, Mumbai, Chandigarh, Lucknow and Goa. We report here the results of immunological and virological studies conducted at our laboratory on these patients. The clinical samples that were obtained from these patients were Serum, CSF, nuchal skin biopsy and saliva. Serum and CSF samples were subjected to standard RFFIT for estimation of rabies neutralizing antibodies. Skin biopsy, CSF and saliva were processed by TaqMan real-time PCR for detection of viral RNA. CSF, saliva and skin homogenates were also processed for virus isolation by inoculation of suckling mice. The PBMCs isolated from fresh blood was subjected to ELISPOT assay to determine the type of immune response (Th1/Th2). Both CSF and serum were also investigated for selected cytokines by Luminex assay. The level of antibodies to virus G protein and N protein were determined by ELISA. All survivors had very high titers of RVNA in serum and CSF 100 fold higher than non-survivors and vaccine controls. A five-fold rise in titer could be demonstrated in 4 out of 6 patients. All survivors had a significant increase in antibodies to G protein in both CSF and serum when compared to non-survivors. There was a profound and robust Th1 response in all survivors indicating that interferon gamma could play an important factor in virus clearance. We could isolate viral RNA in only one patient four years after he had developed symptoms. The partial N gene sequencing revealed 99% homology to species I strain prevalent in India. Levels of selected cytokines in CSF and serum did not reveal any difference between survivors and non-survivors. To conclude, survival from rabies is mediated by virus-specific immune responses of the host and clearance of rabies virus from CNS may involve the participation of both Th2 and Th1 immune responses.

Keywords: rabies, rabies treatment, rabies survivors, immune reponse in rabies encephalitis

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24 Exploiting Charges on Medicinal Synthetic Aluminum Magnesium Silicate's {Al₄ (SiO₄)₃ + 3Mg₂SiO₄ → 2Al₂Mg₃ (SiO₄)₃} Nanoparticles in Treating Viral Diseases, Tumors, Antimicrobial Resistant Infections

Authors: M. C. O. Ezeibe, F. I. O. Ezeibe

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Reasons viral diseases (including AI, HIV/AIDS, and COVID-19), tumors (including Cancers and Prostrate enlargement), and antimicrobial-resistant infections (AMR) are difficult to cure are features of the pathogens which normal cells do not have or need (biomedical markers) have not been identified; medicines that can counter the markers have not been invented; strategies and mechanisms for their treatments have not been developed. When cells become abnormal, they acquire negative electrical charges, and viruses are either positively charged or negatively charged, while normal cells remain neutral (without electrical charges). So, opposite charges' electrostatic attraction is a treatment mechanism for viral diseases and tumors. Medicines that have positive electrical charges would mop abnormal (infected and tumor) cells and DNA viruses (negatively charged), while negatively charged medicines would mop RNA viruses (positively charged). Molecules of Aluminum-magnesium silicate [AMS: Al₂Mg₃ (SiO₄)₃], an approved medicine and pharmaceutical stabilizing agent, consist of nanoparticles which have both positive electrically charged ends and negative electrically charged ends. The very small size (0.96 nm) of the nanoparticles allows them to reach all cells in every organ. By stabilizing antimicrobials, AMS reduces the rate at which the body metabolizes them so that they remain at high concentrations for extended periods. When drugs remain at high concentrations for longer periods, their efficacies improve. Again, nanoparticles enhance the delivery of medicines to effect targets. Both remaining at high concentrations for longer periods and better delivery to effect targets improve efficacy and make lower doses achieve desired effects so that side effects of medicines are reduced to allow the immunity of patients to be enhanced. Silicates also enhance the immune responses of treated patients. Improving antimicrobial efficacies and enhancing patients` immunity terminate infections so that none remains that could develop resistance. Some countries do not have natural deposits of AMS, but they may have Aluminum silicate (AS: Al₄ (SiO₄)₃) and Magnesium silicate (MS: Mg₂SiO₄), which are also approved medicines. So, AS and MS were used to formulate an AMS-brand, named Medicinal synthetic AMS {Al₄ (SiO₄)₃ + 3Mg₂SiO₄ → 2Al₂Mg₃ (SiO₄)₃}. To overcome the challenge of AMS, AS, and MS being un-absorbable, Dextrose monohydrate is incorporated in MSAMS-formulations for the simple sugar to convey the electrically charged nanoparticles into blood circulation by the principle of active transport so that MSAMS-antimicrobial formulations function systemically. In vitro, MSAMS reduced (P≤0.05) titers of viruses, including Avian influenza virus and HIV. When used to treat virus-infected animals, it cured Newcastle disease and Infectious bursa disease of chickens, Parvovirus disease of dogs, and Peste des petits ruminants disease of sheep and goats. A number of HIV/AIDS patients treated with it have been reported to become HIV-negative (antibody and antigen). COVID-19 patients are also reported to recover and test virus negative when treated with MSAMS. PSA titers of prostate cancer/enlargement patients normalize (≤4) following treatment with MSAMS. MSAMS has also potentiated ampicillin trihydrate, sulfadimidin, cotrimoxazole, piparazine citrate and chloroquine phosphate to achieve ≥ 95 % infection-load reductions (AMR-prevention). At 75 % of doses of ampicillin, cotrimoxazole, and streptomycin, supporting MSAMS-formulations' treatments with antioxidants led to the termination of even already resistant infections.

Keywords: electrical charges, viruses, abnormal cells, aluminum-magnesium silicate

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23 Genotyping of Rotaviruses in Pediatric Patients with Gastroenteritis by Using Real-Time Reverse Transcription Polymerase Chain Reaction

Authors: Recep Kesli, Cengiz Demir, Riza Durmaz, Zekiye Bakkaloglu, Aysegul Bukulmez

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Objective: Acute diarrhea disease in children is a major cause of morbidity worldwide and is a leading cause of mortality, and it is the most common agent responsible for acute gastroenteritis in developing countries. With hospitalized children suffering from acute enteric disease up to 50% of the analyzed specimen were positive for rotavirus. Further molecular surveillance could provide a sound basis for improving the response to epidemic gastroenteritis and could provide data needed for the introduction of vaccination programmes in the country. The aim of this study was to investigate the prevalence of viral etiology of the gastroenteritis in children aged 0-6 years with acute gastroenteritis and to determine predominant genotypes of rotaviruses in the province of Afyonkarahisar, Turkey. Methods: An epidemiological study on rotavirus was carried out during 2016. Fecal samples obtained from the 144 rotavirus positive children with 0-6 years of ages and applied to the Pediatric Diseases Outpatient of ANS Research and Practice Hospital, Afyon Kocatepe University with the complaint of diarrhea. Bacterial agents causing gastroenteritis were excluded by using bacteriological culture methods and finally, no growth observed. Rotavirus antigen was examined by both the immunochromatographic (One Step Rotavirus and Adenovirus Combo Test, China) and ELISA (Premier Rotaclone, USA) methods in stool samples. Rotavirus RNA was detected by using one step real-time reverse transcription-polymerase chain reaction (RT-PCR). G and P genotypes were determined using RT-PCR with consensus primers of VP7 and VP4 genes, followed by semi nested type-specific multiplex PCR. Results: Of the total 144 rotavirus antigen-positive samples with RT-PCR, 4 (2,8%) were rejected, 95 (66%) were examined, and 45 (31,2%) have not been examined for PCR yet. Ninety-one (95,8%) of the 95 examined samples were found to be rotavirus positive with RT-PCR. Rotavirus subgenotyping distributions in G, P and G/P genotype groups were determined as; G1:45%, G2:27%, G3:13%, G9:13%, G4:1% and G12:1% for G genotype, and P[4]:33%, P[8]:66%, P[10]:1% for P genotype, and G1P[8]:%37, G2P[4]:%21, G3P[8]:%10, G4P[8]:%1, G9P[8]:%8, G2P[8]:%3 for G/P genotype . Not common genotype combination were %20 in G/P genotype. Conclusions: This study subscribes to the global agreement of the molecular epidemiology of rotavirus which will be useful in guiding the alternative and application of rotavirus vaccines or effective control and interception. Determining the diversity and rates of rotavirus genotypes will definitely provide guidelines for developing the most suitable vaccine.

Keywords: gastroenteritis, genotyping, rotavirus, RT-PCR

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22 Burden of Dengue in Northern India

Authors: Ashutosh Biswas, Poonam Coushic, Kalpana Baruah, Paras Singla, A. C. Dhariwal, Pawana Murthy

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Burden of Dengue in Northern India Ashutosh Biswas, Poonam Coushic, Kalpana Baruah, Paras Singla, AC Dhariwal, Pawana Murthy. All India Institute of Medical Sciences, NVBDCP,WHO New Delhi, India Aim: This study was conducted to estimate the burden of dengue in capital region of India. Methodology:Seropositivity of Dengue for IgM Ab, NS1 Ag and IgG Ab were performed among the blood donors’ samples from blood bank, those who were coming to donate blood for the requirement of blood for the admitted patients in hospital. Blood samplles were collected through out the year to estimate seroprevalance of dengue with or without outbreak season. All the subjects were asymptomatic at the time of blood donation. Results: A total of 1558 donors were screened for the study. On the basis of inclusion/ exclusion criteria, we enrolled 1531subjects for the study.Twenty seven donors were excluded from the study, out of which 6 were detected HIV +ve, 11 were positive for HBsAg and 10 were found positive for HCV.Mean age was 30.51 ± 7.75 years.Of 1531subjects, 18 (1.18%) had a past history of typhoid fever, 28 (1.83%) had chikungunya fever, 9 (0.59%) had malaria and 43 subjects (2.81%) had a past history of symptomatic dengue infection.About 2.22% (34) of subjects were found to have sero-positive for NS1 Ag with a peak point prevalence of 7.14% in the month of October and sero-positive of IgM Ab was observed about 5.49% (84)with a peak point prevalence of 14.29% in the month of October. Sero-prevalnce of IgGwas detected in about 64.21% (983) of subjects. Conclusion: Acute asymptomatic dengue (NS1 Ag+ve) was observed in 7.14%, as the subjects were having no symptoms at the time of sampling. This group of subjects poses a potential public health threat for transmitting dengue infection through blood transfusion (TTI) in the community as evident by presence of active viral infection due to NS1Ag +VE. Therefore a policy may be implemented in the blood bank for testing NS1 Ag to look for active dengue infection for preventing dengue transmission through blood transfusion (TTI). Acute or Subacute dengue infection ( IgM Ab+ve) was observed from 5.49% to 14.29% which is a peak point prevalence in the month of October. About 64.21% of the population were immunized by natural dengue infection ( IgG Ab+ve) in theNorthern province of India. This might be helpful for implementing the dengue vaccine in a region. Blood samples in blood banks should be tested for dengue before transfusion to any other person to prevent transfusion transmitted dengue infection as we estimated upto 7.14% positivity of NS1 Ag in our study which indicates presence of dengue virus in blood donors’ samples.

Keywords: Dengue Burden, Seroprevalance, Asymptomatic dengue, Dengue transmission through blood transfusion

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21 Data Quality and Associated Factors on Regular Immunization Programme at Ararso District: Somali Region- Ethiopia

Authors: Eyob Seife, Molla Alemayaehu, Tesfalem Teshome, Bereket Seyoum, Behailu Getachew

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Globally, immunization averts between 2 and 3 million deaths yearly, but Vaccine-Preventable Diseases still account for more in Sub-Saharan African countries and takes the majority of under-five deaths yearly, which indicates the need for consistent and on-time information to have evidence-based decision so as to save lives of these vulnerable groups. However, ensuring data of sufficient quality and promoting an information-use culture at the point of collection remains critical and challenging, especially in remote areas where the Ararso district is selected based on a hypothesis of there is a difference in reported and recounted immunization data consistency. Data quality is dependent on different factors where organizational, behavioral, technical and contextual factors are the mentioned ones. A cross-sectional quantitative study was conducted on September 2022 in the Ararso district. The study used the world health organization (WHO) recommended data quality self-assessment (DQS) tools. Immunization tally sheets, registers and reporting documents were reviewed at 4 health facilities (1 health center and 3 health posts) of primary health care units for one fiscal year (12 months) to determine the accuracy ratio, availability and timeliness of reports. The data was collected by trained DQS assessors to explore the quality of monitoring systems at health posts, health centers, and at the district health office. A quality index (QI), availability and timeliness of reports were assessed. Accuracy ratios formulated were: the first and third doses of pentavalent vaccines, fully immunized (FI), TT2+ and the first dose of measles-containing vaccines (MCV). In this study, facility-level results showed poor timeliness at all levels and both over-reporting and under-reporting were observed at all levels when computing the accuracy ratio of registration to health post reports found at health centers for almost all antigens verified. A quality index (QI) of all facilities also showed poor results. Most of the verified immunization data accuracy ratios were found to be relatively better than that of quality index and timeliness of reports. So attention should be given to improving the capacity of staff, timeliness of reports and quality of monitoring system components, namely recording, reporting, archiving, data analysis and using information for decisions at all levels, especially in remote and areas.

Keywords: accuracy ratio, ararso district, quality of monitoring system, regular immunization program, timeliness of reports, Somali region-Ethiopia

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20 Evolutionary Analysis of Influenza A (H1N1) Pdm 09 in Post Pandemic Period in Pakistan

Authors: Nazish Badar

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In early 2009, Pandemic type A (H1N1) Influenza virus emerged globally. Since then, it has continued circulation causing considerable morbidity and mortality. The purpose of this study was to evaluate the evolutionary changes in Influenza A (H1N1) pdm09 viruses from 2009-15 and their relevance with the current vaccine viruses. Methods: Respiratory specimens were collected with influenza-like illness and Severe Acute Respiratory Illness. Samples were processed according to CDC protocol. Sequencing and phylogenetic analysis of Haemagglutinin (HA) and neuraminidase (NA) genes was carried out comparing representative isolates from Pakistan viruses. Results: Between Jan2009 - Feb 2016, 1870 (13.2%) samples were positive for influenza A out of 14086. During the pandemic period (2009–10), Influenza A/ H1N1pdm 09 was the dominant strain with 366 (45%) of total influenza positives. In the post-pandemic period (2011–2016), a total of 1066 (59.6%) cases were positive Influenza A/ H1N1pdm 09 with co-circulation of different Influenza A subtypes. Overall, the Pakistan A(H1N1) pdm09 viruses grouped in two genetic clades. Influenza A(H1N1)pdm09 viruses only ascribed to Clade 7 during the pandemic period whereas viruses belong to clade 7 (2011) and clade 6B (2015) during the post-pandemic years. Amino acid analysis of the HA gene revealed mutations at positions S220T, I338V and P100S specially associated with outbreaks in all the analyzed strains. Sequence analyses of post-pandemic A(H1N1)pdm09 viruses showed additional substitutions at antigenic sites; S179N,K180Q (SA), D185N, D239G (CA), S202A (SB) and at receptor binding sites; A13T, S200P when compared with pandemic period. Substitution at Genetic markers; A273T (69%), S200P/T (15%) and D239G (7.6%) associated with severity and E391K (69%) associated with virulence was identified in viruses isolated during 2015. Analysis of NA gene revealed outbreak markers; V106I (23%) among pandemic and N248D (100%) during post-pandemic Pakistan viruses. Additional N-Glycosylation site; HA S179N (23%), NA I23T(7.6%) and N44S (77%) in place of N386K(77%) were only found in post-pandemic viruses. All isolates showed histidine (H) at position 275 in NA indicating sensitivity to neuraminidase inhibitors. Conclusion: This study shows that the Influenza A(H1N1)pdm09 viruses from Pakistan clustered into two genetic clades, with co-circulation of some variants. Certain key substitutions in the receptor binding site and few changes indicative of virulence were also detected in post-pandemic strains. Therefore, it is imperative to continue monitoring of the viruses for early identification of potential variants of high virulence or emergence of drug-resistant variants.

Keywords: Influenza A (H1N1) pdm09, evolutionary analysis, post pandemic period, Pakistan

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19 Monitoring of Serological Test of Blood Serum in Indicator Groups of the Population of Central Kazakhstan

Authors: Praskovya Britskaya, Fatima Shaizadina, Alua Omarova, Nessipkul Alysheva

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Planned preventive vaccination, which is carried out in the Republic of Kazakhstan, promoted permanent decrease in the incidence of measles and viral hepatitis B. In the structure of VHB patients prevail people of young, working age. Monitoring of infectious incidence, monitoring of coverage of immunization of the population, random serological control over the immunity enable well-timed identification of distribution of the activator, effectiveness of the taken measures and forecasting. The serological blood analysis was conducted in indicator groups of the population of Central Kazakhstan for the purpose of identification of antibody titre for vaccine preventable infections (measles, viral hepatitis B). Measles antibodies were defined by method of enzyme-linked assay (ELA) with test-systems "VektoKor" – Ig G ('Vektor-Best' JSC). Antibodies for HBs-antigen of hepatitis B virus in blood serum was identified by method of enzyme-linked assay (ELA) with VektoHBsAg test systems – antibodies ('Vektor-Best' JSC). The result of the analysis is positive, the concentration of IgG to measles virus in the studied sample is equal to 0.18 IU/ml or more. Protective level of concentration of anti-HBsAg makes 10 mIU/ml. The results of the study of postvaccinal measles immunity showed that the share of seropositive people made 87.7% of total number of surveyed. The level of postvaccinal immunity to measles in age groups differs. So, among people older than 56 the percentage of seropositive made 95.2%. Among people aged 15-25 were registered 87.0% seropositive, at the age of 36-45 – 86.6%. In age groups of 25-35 and 36-45 the share of seropositive people was approximately at the same level – 88.5% and 88.8% respectively. The share of people seronegative to a measles virus made 12.3%. The biggest share of seronegative people was found among people aged 36-45 – 13.4% and 15-25 – 13.0%. The analysis of results of the examined people for the existence of postvaccinal immunity to viral hepatitis B showed that from all surveyed only 33.5% have the protective level of concentration of anti-HBsAg of 10 mIU/ml and more. The biggest share of people protected from VHB virus is observed in the age group of 36-45 and makes 60%. In the indicator group – above 56 – seropositive people made 4.8%. The high percentage of seronegative people has been observed in all studied age groups from 40.0% to 95.2%. The group of people which is least protected from getting VHB is people above 56 (95.2%). The probability to get VHB is also high among young people aged 25-35, the percentage of seronegative people made 80%. Thus, the results of the conducted research testify to the need for carrying out serological monitoring of postvaccinal immunity for the purpose of operational assessment of the epidemiological situation, early identification of its changes and prediction of the approaching danger.

Keywords: antibodies, blood serum, immunity, immunoglobulin

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18 Concepts of the Covid-19 Pandemic and the Implications of Vaccines for Health Security in Nigeria and Diasporas

Authors: Wisdom Robert Duruji

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The outbreak of SARS-CoV-2 serotype infection was recorded in January 2020 in Wuhan City, Hubei Province, China. This study examines the concepts of the COVID-19 pandemic and the implications of vaccines for health security in Nigeria and Diasporas. It challenges the widely accepted assumption that the first case of coronavirus infection in Nigeria was recorded on February 27th, 2020, in Lagos. The study utilizes a range of research methods to achieve its objectives. These include the double-layered culture technique, literature review, website knowledge, Google search, news media information, academic journals, fieldwork, and on-site observations. These diverse methods allow for a comprehensive analysis of the concepts and the implications being studied. The study finds that coronavirus infection can be asymptomatic; it may be the antigenicity of the leukocytes (white blood cells), which produce immunogenic hapten or interferons (α, β and γ) that fight infectious parasites, was an immune response that prevented severe virulence in healthy individuals; the reason healthy patients of coronavirus infection in Nigeria naturally recovered after two to three weeks of on-set of infection and test negative. However, the fatality data from the Nigerian Centre for Disease Control (NCDC) is incorrect in this study’s finding; it perused that the fatalities were primarily due to underlying ailments, hunger, and malnutrition in debilitated, comorbid, or compromised patients. This study concluded that the kits and Polymerase Chain Reaction (PCR) machine currently used by the Nigerian Centre for Disease Control (NCDC) in testing and confirming COVID-19 in Nigeria is not ideal; it is programmed and negates separating the strain to its specific serotypes amongst its genera coronavirus, and family Coronaviridae; and might have confirmed patients with the symptoms of febrile caused by cough, catarrh, typhoid and malaria parasites as Covid-19 positive. Therefore, it is recommended that the coronavirus species infected in Nigeria are opportunistic parasites that thrive in human immuno-suppressed conditions like the herpesvirus; it cannot be eradicated by vaccines; the only virucides are interferons, immunoglobulins, and probably synthetic antiviral guanosine drugs like copegus or ribavirin. The findings emphasized that COVID-19 is not the primary pandemic disease in Nigeria; the lockdown was a mirage and not necessary; but rather, pandemic diseases in Nigeria are corruption, nepotism, hunger, and malnutrition caused by ineptitude in governance, religious dichotomy, and ethnic conflicts.

Keywords: coronavirus, corruption, Covid-19 pandemic, lock-down, Nigeria, vaccine

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17 Approaching a Tat-Rev Independent HIV-1 Clone towards a Model for Research

Authors: Walter Vera-Ortega, Idoia Busnadiego, Sam J. Wilson

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Introduction: Human Immunodeficiency Virus type 1 (HIV-1) is responsible for the acquired immunodeficiency syndrome (AIDS), a leading cause of death worldwide infecting millions of people each year. Despite intensive research in vaccine development, therapies against HIV-1 infection are not curative, and the huge genetic variability of HIV-1 challenges to drug development. Current animal models for HIV-1 research present important limitations, impairing the progress of in vivo approaches. Macaques require a CD8+ depletion to progress to AIDS, and the maintenance cost is high. Mice are a cheaper alternative but need to be 'humanized,' and breeding is not possible. The development of an HIV-1 clone able to replicate in mice is a challenging proposal. The lack of human co-factors in mice impedes the function of the HIV-1 accessory proteins, Tat and Rev, hampering HIV-1 replication. However, Tat and Rev function can be replaced by constitutive/chimeric promoters, codon-optimized proteins and the constitutive transport element (CTE), generating a novel HIV-1 clone able to replicate in mice without disrupting the amino acid sequence of the virus. By minimally manipulating the genomic 'identity' of the virus, we propose the generation of an HIV-1 clone able to replicate in mice to assist in antiviral drug development. Methods: i) Plasmid construction: The chimeric promoters and CTE copies were cloned by PCR using lentiviral vectors as templates (pCGSW and pSIV-MPCG). Tat mutants were generated from replication competent HIV-1 plasmids (NHG and NL4-3). ii) Infectivity assays: Retroviral vectors were generated by transfection of human 293T cells and murine NIH 3T3 cells. Virus titre was determined by flow cytometry measuring GFP expression. Human B-cells (AA-2) and Hela cells (TZMbl) were used for infectivity assays. iii) Protein analysis: Tat protein expression was determined by TZMbl assay and HIV-1 capsid by western blot. Results: We have determined that NIH 3T3 cells are able to generate HIV-1 particles. However, they are not infectious, and further analysis needs to be performed. Codon-optimized HIV-1 constructs are efficiently made in 293T cells in a Tat and Rev independent manner and capable of packaging a competent genome in trans. CSGW is capable of generating infectious particles in the absence of Tat and Rev in human cells when 4 copies of the CTE are placed preceding the 3’LTR. HIV-1 Tat mutant clones encoding different promoters are functional during the first cycle of replication when Tat is added in trans. Conclusion: Our findings suggest that the development of an HIV-1 Tat-Rev independent clone is challenging but achievable aim. However, further investigations need to be developed prior presenting our HIV-1 clone as a candidate model for research.

Keywords: codon-optimized, constitutive transport element, HIV-1, long terminal repeats, research model

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16 Missed Opportunities for Immunization of under Five Children in Calabar South County Cros River State, Nigeria, the Way Forward

Authors: Celestine Odigwe, Epoke Lincoln, Rhoda-Dara Ephraim

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Background; Immunization against the childhood killer diseases is the cardinal strategy for the prevention of these diseases all over the world in under five children, these diseases include; Tuberculosis, Measles, Polio, Tetanus, Diphthria, Pertusis, Yellow Fever, Hepatitis B, Haemophilus Influenza type B. 6.9 million children die before their fifth birthday , 80% of the worlds death in children under 5 years occur in 25 countries most in Africa and Asia and 2 million children can be saved each year with routine immunization Therefore failure to achieve total immunization coverage puts several children at risk. Aim; The aim of the study was to ascertain the prevalence, Investigate the various reasons and causes why several under five children in a suburb of calabar municipal county fail to get the required immunizations as at and when due and possibly the consequences, so that efforts can be re-directed towards the solution of the problems so identified. Methods; the study was a community based cross sectional study. The respondents were the mothers/guardians of the sampled children who were all aged 0-59 months. To be eligible for recruitment into the study, the parent or guardian was required to give an informed consent, reside within the Calabar South County with his/her children aged 0-59 months. We calculated our sample size using the Leslie-Kish formula and we used a two-staged sampling method, first to ballot for the wards to be involved and then to select four of the most populated ones in the wards chosen. Data collection was by interviewer administered structured questionnaire (Appendix I), Data collected was entered and analyzed using Statistical Package for the Social Sciences (SPSS) Version 20. Percentages were calculated and represented using charts and tables Results; The number of children sampled was 159. We found that 150 were fully immunized and 9 were not, the prevalence of missed opportunity was 32% from the study. The reasons for missed opportunities were varied, ranging from false contraindications, logistical problems resulting in very poor access roads to health facilities and poor organization of health centers together with negative health worker attitudes. Some of the consequences of these missed opportunities were increased susceptibility to vaccine preventable diseases, resurgence of the above diseases and increased morbidity and mortality of children aged less than 5 years. Conclusion; We found that ignorance on the part of both parents/guardians and health care staff together with infrastructural inadequacies in the county such as- roads, poor electric power supply for storage of vaccines were hugely responsible for most missed opportunities for immunization. The details of these and suggestions for improvement and the way forward are discussed.

Keywords: missed opportunity, immunization, under five, Calabar south

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15 Tuberculosis (TB) and Lung Cancer

Authors: Asghar Arif

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Lung cancer has been recognized as one of the greatest common cancers, causing the annual mortality rate of about 1.2 million people in the world. Lung cancer is the most prevalent cancer in men and the third-most common cancer among women (after breast and digestive cancers).Recent evidences have shown the inflammatory process as one of the potential factors of cancer. Tuberculosis (TB), pneumonia, and chronic bronchitis are among the most important inflammation-inducing factors in the lungs, among which TB has a more profound role in the emergence of cancer.TB is one of the important mortality factors throughout the world, and 205,000 death cases are reported annually due to this disease. Chronic inflammation and fibrosis due to TB can induce genetic mutation and alternations. Parenchyma tissue of lung is involved in both diseases of TB and lung cancer, and continuous cough in lung cancer, morphological vascular variations, lymphocytosis processes, and generation of immune system mediators such as interleukins, are all among the factors leading to the hypothesis regarding the role of TB in lung cancer Some reports have shown that the induction of necrosis and apoptosis or TB reactivation, especially in patients with immune-deficiency, may result in increasing IL-17 and TNF_α, which will either decrease P53 activity or increase the expression of Bcl-2, decrease Bax-T, and cause the inhibition of caspase-3 expression due to decreasing the expression of mitochondria cytochrome oxidase. It has been also indicated that following the injection of BCG vaccine, the host immune system will be reinforced, and in particular, the rates of gamma interferon, nitric oxide, and interleukin-2 are increased. Therefore, CD4 + lymphocyte function will be improved, and the person will be immune against cancer.Numerous prospective studies have so far been conducted on the role of TB in lung cancer, and it seems that this disease is effective in that particular cancer.One of the main challenges of lung cancer is its correct and timely diagnosis. Unfortunately, clinical symptoms (such as continuous cough, hemoptysis, weight loss, fever, chest pain, dyspnea, and loss of appetite) and radiological images are similar in TB and lung cancer. Therefore, anti-TB drugs are routinely prescribed for the patients in the countries with high prevalence of TB, like Pakistan. Regarding the similarity in clinical symptoms and radiological findings of lung cancer, proper diagnosis is necessary for TB and respiratory infections due to nontuberculousmycobacteria (NTM). Some of the drug resistive TB cases are, in fact, lung cancer or NTM lung infections. Acid-fast staining and histological study of phlegm and bronchial washing, culturing and polymerase chain reaction TB are among the most important solutions for differential diagnosis of these diseases. Briefly, it is assumed that TB is one of the risk factors for cancer. Numerous studies have been conducted in this regard throughout the world, and it has been observed that there is a significant relationship between previous TB infection and lung cancer. However, to prove this hypothesis, further and more extensive studies are required. In addition, as the clinical symptoms and radiological findings of TB, lung cancer, and non-TB mycobacteria lung infections are similar, they can be misdiagnosed as TB.

Keywords: TB and lung cancer, TB people, TB servivers, TB and HIV aids

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14 Microbial Contamination of Cell Phones of Health Care Workers: Case Study in Mampong Municipal Government Hospital, Ghana

Authors: Francis Gyapong, Denis Yar

Abstract:

The use of cell phones has become an indispensable tool in the hospital's settings. Cell phones are used in hospitals without restrictions regardless of their unknown microbial load. However, the indiscriminate use of mobile devices, especially at health facilities, can act as a vehicle for transmitting pathogenic bacteria and other microorganisms. These potential pathogens become exogenous sources of infection for the patients and are also a potential health hazard for self and as well as family members. These are a growing problem in many health care institutions. Innovations in mobile communication have led to better patient care in diabetes, asthma, and increased in vaccine uptake via SMS. Notwithstanding, the use of cell phones can be a great potential source for nosocomial infections. Many studies reported heavy microbial contamination of cell phones among healthcare workers and communities. However, limited studies have been reported in our region on bacterial contamination on cell phones among healthcare workers. This study assessed microbial contamination of cell phones of health care workers (HCWs) at the Mampong Municipal Government Hospital (MMGH), Ghana. A cross-sectional design was used to characterize bacterial microflora on cell phones of HCWs at the MMGH. A total of thirty-five (35) swab samples of cell phones of HCWs at the Laboratory, Dental Unit, Children’s Ward, Theater and Male ward were randomly collected for laboratory examinations. A suspension of the swab samples was each streak on blood and MacConkey agar and incubated at 37℃ for 48 hours. Bacterial isolates were identified using appropriate laboratory and biochemical tests. Kirby-Bauer disc diffusion method was used to determine the antimicrobial sensitivity tests of the isolates. Data analysis was performed using SPSS version 16. All mobile phones sampled were contaminated with one or more bacterial isolates. Cell phones from the Male ward, Dental Unit, Laboratory, Theatre and Children’s ward had at least three different bacterial isolates; 85.7%, 71.4%, 57.1% and 28.6% for both Theater and Children’s ward respectively. Bacterial contaminants identified were Staphylococcus epidermidis (37%), Staphylococcus aureus (26%), E. coli (20%), Bacillus spp. (11%) and Klebsiella spp. (6 %). Except for the Children ward, E. coli was isolated at all study sites and predominant (42.9%) at the Dental Unit while Klebsiella spp. (28.6%) was only isolated at the Children’s ward. Antibiotic sensitivity testing of Staphylococcus aureus indicated that they were highly sensitive to cephalexin (89%) tetracycline (80%), gentamycin (75%), lincomycin (70%), ciprofloxacin (67%) and highly resistant to ampicillin (75%). Some of these bacteria isolated are potential pathogens and their presence on cell phones of HCWs could be transmitted to patients and their families. Hence strict hand washing before and after every contact with patient and phone be enforced to reduce the risk of nosocomial infections.

Keywords: mobile phones, bacterial contamination, patients, MMGH

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13 In vivo Estimation of Mutation Rate of the Aleutian Mink Disease Virus

Authors: P.P. Rupasinghe, A.H. Farid

Abstract:

The Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1) causes persistent infection, plasmacytosis, and formation and deposition of immune complexes in various organs in adult mink, leading to glomerulonephritis, arteritis and sometimes death. The disease has no cure nor an effective vaccine, and identification and culling of mink positive for anti-AMDV antibodies have not been successful in controlling the infection in many countries. The failure to eradicate the virus from infected farms may be caused by keeping false-negative individuals on the farm, virus transmission from wild animals, or neighboring farms. The identification of sources of infection, which can be performed by comparing viral sequences, is important in the success of viral eradication programs. High mutation rates could cause inaccuracies when viral sequences are used to trace back an infection to its origin. There is no published information on the mutation rate of AMDV either in vivo or in vitro. The in vivo estimation is the most accurate method, but it is difficult to perform because of the inherent technical complexities, namely infecting live animals, the unknown numbers of viral generations (i.e., infection cycles), the removal of deleterious mutations over time and genetic drift. The objective of this study was to determine the mutation rate of AMDV on which no information was available. A homogenate was prepared from the spleen of one naturally infected American mink (Neovison vison) from Nova Scotia, Canada (parental template). The near full-length genome of this isolate (91.6%, 4,143 bp) was bidirectionally sequenced. A group of black mink was inoculated with this homogenate (descendant mink). Spleen sampled were collected from 10 descendant mink after 16 weeks post-inoculation (wpi) and from anther 10 mink after 176 wpi, and their near-full length genomes were bi-directionally sequenced. Sequences of these mink were compared with each other and with the sequence of the parental template. The number of nucleotide substitutions at 176 wpi was 3.1 times greater than that at 16 wpi (113 vs 36) whereas the estimates of mutation rate at 176 wpi was 3.1 times lower than that at 176 wpi (2.85×10-3 vs 9.13×10-4 substitutions/ site/ year), showing a decreasing trend in the mutation rate per unit of time. Although there is no report on in vivo estimate of the mutation rate of DNA viruses in animals using the same method which was used in the current study, these estimates are at the higher range of reported values for DNA viruses determined by various techniques. These high estimates are logical based on the wide range of diversity and pathogenicity of AMDV isolates. The results suggest that increases in the number of nucleotide substitutions over time and subsequent divergence make it difficult to accurately trace back AMDV isolates to their origin when several years elapsed between the two samplings.

Keywords: Aleutian mink disease virus, American mink, mutation rate, nucleotide substitution

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12 Challenges in the Last Mile of the Global Guinea Worm Eradication Program: A Systematic Review

Authors: Getahun Lemma

Abstract:

Introduction Guinea Worm Disease (GWD), also known as dracunculiasisis, is one of the oldest diseases in the history of mankind. Dracunculiasis is caused by a parasitic nematode, Dracunculus medinensis. Infection is acquired by drinking contaminated water with copepods containing infective Guinea Worm (GW) larvae). Almost one year after the infection, the worm usually emerges out through the skin on a lower, causing severe pain and disabilities. Although there is no effective drug or vaccine against the disease, the chain of transmission can be effectively prevented with simple and cost effective public health measures. Death due to dracunculiasis is very rare. However, it results in a wide range of physical, social and economic sequels. The disease is usually common in the rural, remote places of Sub-Saharan African countries among the marginalized societies. Currently, GWD is one of the neglected tropical diseases, which is on the verge of eradication. The global Guinea Worm Eradication Program (GWEP) was started in 1980. Since then, the program has achieved a tremendous success in reducing the global burden and number of GW case from 3.5 million to only 28 human cases at the end of 2018. However, it has recently been shown that not only humans can become infected, with a total of 1,105 animal infections have been reported at the end of 2018. Therefore, the objective of this study was to identify the existing challenges in the last mile of the GWEP in order To inform Policy makers and stakeholders on potential measures to finally achieve eradication. Method Systematic literature review on articles published from January 1, 2000 until May 30, 2019. Papers listed in Cochrane Library, Google Scholar, ProQuest PubMed and Web of Science databases were searched and reviewed. Results Twenty-five articles met inclusion criteria of the study and were selected for analysis. Hence, relevant data were extracted, grouped and descriptively analyzed. Results showed the main challenges complicating the last mile of global GWEP: 1. Unusual mode of transmission; 2. Rising animal Guinea Worm infection; 3. Suboptimal surveillance; 4. Insecurity; 5. Inaccessibility; 6. Inadequate safe water points; 7. Migration; 8. Poor case containment measures, 9. Ecological changes; and 10. New geographic foci of the disease. Conclusion This systematic review identified that most of the current challenges in the GWEP have been present since the start of the campaign. However, the recent change in epidemiological patterns and nature of GWD in the last remaining endemic countries illustrates a new twist in the global GWEP. Considering the complex nature of the current challenges, there seems to be a need for a more coordinated and multidisciplinary approach of GWD prevention and control measures in the last mile of the campaign. These new strategies would help to make history by eradicating dracunculiasis as the first ever parasitic disease.

Keywords: dracunculiasis, eradication program, guinea worm, last mile

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11 Effect of Graded Level of Nano Selenium Supplementation on the Performance of Broiler Chicken

Authors: Raj Kishore Swain, Kamdev Sethy, Sumanta Kumar Mishra

Abstract:

Selenium is an essential trace element for the chicken with a variety of biological functions like growth, fertility, immune system, hormone metabolism, and antioxidant defense systems. Selenium deficiency in chicken causes exudative diathesis, pancreatic dystrophy and nutritional muscle dystrophy of the gizzard, heart and skeletal muscle. Additionally, insufficient immunity, lowering of production ability, decreased feathering of chickens and increased embryo mortality may occur due to selenium deficiency. Nano elemental selenium, which is bright red, highly stable, soluble and of nano meter size in the redox state of zero, has high bioavailability and low toxicity due to the greater surface area, high surface activity, high catalytic efficiency and strong adsorbing ability. To assess the effect of dietary nano-Se on performance and expression of gene in Vencobb broiler birds in comparison to its inorganic form (sodium selenite), four hundred fifty day-old Vencobb broiler chicks were randomly distributed into 9 dietary treatment groups with two replicates with 25 chicks per replicate. The dietary treatments were: T1 (Control group): Basal diet; T2: Basal diet with 0.3 ppm of inorganic Se; T3: Basal diet with 0.01875 ppm of nano-Se; T4: Basal diet with 0.0375 ppm of nano-Se; T5: Basal diet with 0.075 ppm of nano-Se, T6: Basal diet with 0.15 ppm of nano-Se, T7: Basal diet with 0.3 ppm of nano-Se, T8: Basal diet with 0.60 ppm of nano-Se, T9: Basal diet with 1.20 ppm of nano-Se. Nano selenium was synthesized by mixing sodium selenite with reduced glutathione and bovine serum albumin. The experiment was carried out in two phases: starter phase (0-3 wks), finisher phase (4-5 wk) in deep litter system. The body weight at the 5th week was best observed in T4. The best feed conversion ratio at the end of 5th week was observed in T4. Erythrocytic catalase, glutathione peroxidase and superoxide dismutase activity were significantly (P < 0.05) higher in all the nano selenium treated groups at 5th week. The antibody titers (log2) against Ranikhet diseases vaccine immunization of 5th-week broiler birds were significantly higher (P < 0.05) in the treatments T4 to T7. The selenium levels in liver, breast, kidney, brain, and gizzard were significantly (P < 0.05) increased with increasing dietary nano-Se indicating higher bioavailability of nano-Se compared to inorganic Se. The real time polymer chain reaction analysis showed an increase in the expression of antioxidative gene in T4 and T7 group. Therefore, it is concluded that supplementation of nano-selenium at 0.0375 ppm over and above the basal level can improve the body weight, antioxidant enzyme activity, Se bioavailability and expression of the antioxidative gene in broiler birds.

Keywords: chicken, growth, immunity, nano selenium

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