Amarnath Sen   

The apparent absence of drugs in ultra-diluted homeopathic medicines and contested clinical trial results plague homeopathy. Here it is argued that other than drugs, homeopathic medicines contain proteins as components of microbial lysates (products of lysis or disintegration of microbial cells), given that ubiquitous microorganisms from the surrounding environment are unknowingly and unavoidably incorporated into the homeopathic medicines during their preparation and are killed and lysed in ethanol-water drug vehicle forming immunomodulatory microbial lysates during ‘potentization' (dilution and vigorous shaking) of the medicines. The drugs present in the homeopathic medicines bind to the proteins, which are the major ingredients of the microbial lysates. The drug-protein interaction modulates the conformations and in effect, the immunogenicity of the proteins (designated as modulated proteins). In ultra-diluted medicines even in the absence of drugs, unmodulated proteins are modulated through interactions with allosterically coupled modulated proteins (protein-protein interaction). The modulated proteins of characteristic immunogenicity present in the homeopathic medicines mediate antigen-specific mucosal (sublingual) immunotherapy like vaccine therapy via ‘similia principle'. In addition, immunomodulatory microbial lysates present in the homeopathic medicines mediate non-specific immunotherapy and also provide adjuvants for antigen-specific immunotherapy. The proposed hypothesis without invoking any controversial concept can explain the basic ‘laws' of homeopathy. Incidentally, immunomodulatory activities of homeopathic medicines reported by different workers support the hypothesis. As immunotherapy in homeopathy is accidental and hence, in crude form, clinical trial results may occasionally show inconsistencies. However, probing and refining homeopathy from the perspective of immunotherapy may bring forth a simple, reliable and affordable immunotherapy for various diseases.

Pallavi Chattopadhyay   and Ruchi Yadav   

Acetylcholine Receptor (AChR) and Neuronal Acetylcholine Receptor (nAChR) are the proteins which play an important role in neurotransmission. These proteins are widely studied as potential drug targets for neurological study. Structural study of proteins provides functional understanding of proteins. Looking into the importance of AChR and nAChR proteins, the structures of both proteins were predicted by homology modeling method using Swiss model at ExPASy. Structural verification of both modeled proteins was done using Ramachandran Plot analysis by SAVES v5.0 server. To further understand the similarity between AChR and nAChR protein, structural alignment was done between their modeled structures and similarity was thoroughly studied. Pairwise alignment between both structures shows that both the proteins share structural similarity and can have functional similarity too. Interaction study between modeled proteins and ligands was done using molecular docking by Swiss Dock server. Total six antioxidants were selected for docking and docking scores of all the six ligands were compared and the best antioxidant was identified. Docking results show that Glutathione shows the best interaction with both proteins. Glutathione shows the minimum ∆G energy -9.68 kcal/mol with the AChR and -8.93kal/mol with the nAChR. This research signifies the efficiency of AChR proteins as potential drug target and Glutathione as the best ligand. Further studies are required to verify the potentiality of AChR proteins and antioxidants as suitable ligand drug designing and drug development.

Yeri Esther Hien   Fatimata Thiombiano   Denise P. Ilboudo   Donatien Kima   Siaka Kobeane   Gertrude Marilyse Relwende Sawadogo   Y. Caleb Tindano   Guillaume S. Sanou   Jacques Simpore   Lassana Sangare   and Yves Traore   

Malaria and dengue are two important vector‑borne infections that are common in tropical countries. In these areas, simultaneous occurrence of malaria and dengue in an individual cannot be excluded, although published reports of co‑infections are scarce till date. The Aims of this study were to identify dengue and malaria co-infection cases as well as to ascertain the severity of dengue and malaria in co‑infection based on some hematological parameters. A cross-sectional study was conducted from June to November 2019 and 385 subjects with an undifferentiated febrile illness patients were recruited at outpatient clinics and blood sample were collected. All samples were tested for dengue NS1 antigen and for dengue IgM/IgG. Malaria diagnosis was based on the identification of Plasmodium parasites on a thin and thick blood films microscopy. Among the 385 patients included in the study, 23.9% were tested positive for malaria, 32.5% were positive for dengue, and 7.01% showed concurrent infection of both malaria and dengue. Malaria and dengue co-infected patients presented lower leukocytes level (5.024±0.559) than malaria monoinfected patients (7.304± 0.388; p=0.002). A significant reduction in platelet counts was observed in co-infected patients (164.0±10.91 vs 229.1±14.32; p=0.005). Conversely, hemoglobin level was statistically lower in malaria monoinfected patients than dengue monoinfected patients (p=0.014). Thus this study demonstrates that the diagnosis of dengue and malaria should be carried out routinely for better management of fever.

Neha Rauniyar   and Deepa Srivastava   

Human health has been drastically affected due to the emergence of many new and deadly viral infections and also the older that persists, despite much wonderful progress in the area of Biomedical sciences. Some of the best examples which could be cited here is the newly evolving Corona virus (Covid-19) or the virus of 21st century and other several viruses which have been already evolved disturbing the existence of human kind such as Zika virus, Dengue virus [4]. Chikungunya virus, Influenza A virus, SARS, MERS which are known to be transferred in mankind through many arthropods, some animals and several other methods of communication, for this the better understanding of the host pathogen association and relationship with its environment is necessary for the vector control methods which would be used for the prevention of these arboreal infections and this has been proved to be effective in many aspects of controlling the effect of the virus. The main cause of evolution of such pathogenic viruses can be any among some of these such as changing climatic conditions, poor sanitization, increase in population rapidly, rampant construction, uncooked meats etc., and many new evolving anthropophilic vectors which transfer such diseases in humans. Many recent researches have been reported which have shown the immense effort of Medical sciences and scientist to cure various deadly diseases such as HIV, Hepatitis, Chikungunya etc. and to some extent these viruses have been overcome but the nature of viruses is unpredictable and could re-emerge any time in the history which would hamper the control and preventive measures.

Edoga C. O.   Ejo J. E.   Anukwuorji C. A.   Ani C. E.   and Izundu M. I.   

The research was conducted to determine the antioxidant effects of the mixed doses of vitamins B12 and E on male Wistar albino rats infected with T. b. brucei. Fifty-four male Wistar albino rats were randomly divided into six groups of three rats each replicated three times. The rats were marked and kept in stainless wire cages labeled A-F. Groups A, B, and C were normal, negative, and standard controls respectively. Groups D, E and F were infected with 1.0 x 10⁶ trypanosomes and treated with 0.2 + 0.5mg/kg (low-dose), 0.3 + 2.5mg/kg (medium-dose), and 0.4 + 5.0mg/kg (high-dose) of vitamins B₁₂ + E per body weight per day respectively. The experiment lasted for twenty-one days from the day T. b. brucei infection was established. A sample of heart tissue homogenate was collected weekly across the groups and subjected to biochemical determination of malondialdehyde, hydrogen peroxide, nitric oxide, reduced glutathione, and glutathione peroxidase concentrations. There were significant differences in the effect of antioxidant biomarkers which was applied to the duration of the experiment. At post-treatment, the levels of the antioxidant parameters differed significantly (p<0.05) from the negative control. There was a significant increase in the levels of nitric oxide, hydrogen peroxide, and reduced glutathione, and glutathione peroxidase of cardiac tissue homogenate of the experimental rats following the treatments with vitamins B₁₂ + E. The study also showed the pathogenesis of T. b. brucei significantly raised the malondialdehyde concentration and the treatment with vitamins B₁₂ + E decreased the trypanosome-induced elevation of the malondialdehyde level. In conclusion, nitric oxide, hydrogen peroxide, reduced glutathione, and glutathione peroxidase levels significantly increased, while malondialdehyde concentration decreased.

Edoga C. O.   Eyo J. E.   Anukwuorji C. A.   and Nzewi J. I.   

The study was undertaken to check the effects of cyanocobalamin on antioxidant enzymes of male Wistar albino rats infected with T. b. brucei. Fifty-four (54) male Wistar albino rats were divided into six groups of three rats each which were replicated three times. The rats were marked and kept in stainless wire cages labeled A-F. Groups A, B, and C were normal, negative and standard control respectively. Groups D, E and F were infected with 1.0 x 10⁶ trypanosomes and treated with 0.2mg/kg (low-dose), 0.3mg/kg (enriched-dose), and 0.4mg/kg (high-dose) body weight of vitamin B₁₂ respectively. The experiment lasted for twenty-one days from the day T. b. brucei infection was established. A sample of heart tissue homogenate was collected weekly across the groups and subjected to biochemical determination of catalase, superoxide dismutase (SOD), glutathione reductase, and glutathione peroxidase concentrations. There were significant differences in the effect of cyanocobalamin on the concentrations of cardiac tissue antioxidant enzymes which were also dependent on the duration of the experiment. At post-treatment, catalase, superoxide dismutase, and glutathione reductase levels differed significantly (p<0.05) from the negative control. There were significant reductions in the levels of catalase, glutathione reductase, glutathione peroxidase, and rise in the SOD level as infections grow. The result, however, showed that cyanocobalamin caused a significant elevation in the catalase, glutathione reductase, reduction in the concentration of SOD and no change in the level of glutathione peroxidase following treatments with cyanocobalamin. In conclusion, there were significant reductions in the level of SOD, evidenced in an increase in the catalase and glutathione reductase levels, and no change in the glutathione peroxidase concentration following treatments with cyanocobalamin.

Onuorah Samuel   Okafor Mavis   and Odibo Frederick   

Akamu is a popular fermented nutritive porridge made from cereals and is mostly eaten at infancy as a weaning food. Lactic acid bacteria contribute towards the safety, nutritional value, shelf life and acceptability of a wide range of cereal based foods and have been reported to have probiotic potential against gastrointestinal microorganisms, thus in vitro studies of the probiotic properties of lactic acid bacteria isolated from akamu produced with sorghum and maize grains were carried out using standard analytical methods. The pH of the cereal slurries decreased while the titratable acidity and the total lactic acid bacterial counts increased during the period of the studies. The lactic acid bacteria isolated were lactobacillus delbrueckii subspp bulgaricus, L. fermentum, L. brevis, L. plantarum, L. amylovorus, Pediococcus acidilactici acid, P. Pentosaceus. The isolates grew optimally at pH 4.0 and NaCl concentration of 3.0% and survived in fresh bovine bile. The bacteria except Lactobacillus brevis adhered to the intestinal mucosa as shown by the cell surface hydrophobicity assay and were resistant to most of the antibiotics used. This study indicated that the lactic acid bacteria isolated from raw akamu have probiotic characteristics and that raw akamu will be effective in the prevention and treatment of gastrointestinal diseases.

Iheanyi O. Okonko   Anwuli U. Osadebe   Okechukwu Onianwa   and Stella Okereke   

HIV/AIDS is one of the major barriers to community development. It is a global pandemic with high prevalence rates worldwide. This research was carried out to determine the prevalence of HIV among pregnant women attending the antenatal clinic at the University College Hospital, Ibadan using Enzyme Linked Immunosorbent Assay (ELISA) Gen Screen ^TM ULTRA HIV Ag-Ab Kit. HIV prevalence was found to be 26.4%. A total of 91 samples were tested, 24 (26.4%) were found to be positive and 67 (73.6%) negative. Women between the ages 25 – 31 had the highest prevalence rate of 16.5% while those between the ages of 39 – 45 years had the least prevalence rate of 1.1%. The results revealed a relatively high prevalence rate for HIV among pregnant women which suggests the need for active surveillance and implementation of preventive measures to avoid further spread of the virus. Pregnant women need to be continually tested for HIV and those infected monitored closely to prevent mother to child transmission.

Onuorah Samuel   Igwemadu Nkiruka   and Odibo Frederick   

Ogbaru communities in Anambra State, Nigeria experience heavy flooding during the wet season thereby exposing the boreholes sited there to the risk of contamination by microorganisms. Water samples from fifteen drinking water boreholes in the area were analysed for the presence of fungi during the dry and wet seasons using standard analytical methods. The fungal counts were 3-25/cfu/ml and 4-30cfu/ml during the dry and wet seasons respectively. The fungi isolated during both seasons were Penicillium chrysogenum, Aspergillus versicolor, Cladophialophora carrioni, Aspergillus terreus, Candida albicans and Rhodotorula glutinis. C. albicans and A. terreus, each were isolated from 20.0%, R. glutinis and C. carrioni each in 13.3%, P. chrysogenum in 33.3% and A. versicolor in 26.7% of the boreholes studied during the dry season while C. albicans, R. glutinis and A. terreus each were detected in 26.7%, P. chrysogenum in 40.0%, A. versicolor in 33.3% and C. carrioni in 20.0% of the boreholes investigated during the wet season. Aspergillus terreus occurred most frequently (31.4%) in the borehole water samples during the dry season while Penicillium chrysogenum had the highest frequency of occurrence of 28.3% during the wet season. All the borehole water samples analysed were contaminated by fungi and must undergo adequate treatment such as sand filtration chlorination, aeration and boiling before use.

Mohammad Showkat Mahmud   Eusha Islam   Md. Giasuddin   Mohammed Abdus Samad   Md. Rezaul Karim   and Md. Zulfekar Ali   

Foot and Mouth Disease (FMD) is an important viral as well as transboundary disease affecting almost all cloven-hoofed animals. The aim of the present study is the focused on determination of biological titer by tissue culture infective dose₅₀ (TCID₅₀) assay of currently available FMD virus serotype in Bangladesh. For adaptation of FMD virus (FMDV), BHK-21 cell line was used. BHK-21 cell subculture was done from preserved cultured bottle of Foot and Mouth Disease (FMD) Research Laboratory of Bangladesh Livestock Research Institute (BLRI), Savar, Dhaka. RT-PCR confirmed selected three different positive serotypes (O, A and Asia 1) of FMD virus were inoculated into BHK-21 cell culture and cytopathic effects (CPE) were observed after adaptation into BHK-21 cell from 3^rd to 5^th passages. Clear infectious BHK-21 cell culture fluid was collected and preserved at -80℃ temperature. The TCID₅₀ assay was performed to determine the biological titer of the three positive serotype of FMDV. The biological titer of this study was found 10^6.5/ml viral titer for O type, 10^6.75/ml viral titer for A type, 10^6.66/ml viral titer for Asia-1 type. These three specific serotypes can be used as vaccine seed against FMD virus. The effective vaccination of susceptible animals is considered to be the corner stone to the disease in our country Bangladesh. The findings of this study can be helpful for the trivalent vaccine development in Bangladesh and it may effective in limiting the spread of FMD.

María Cecilia Yubini   Caterina Contreras   María Alejandra Cerda   Gonzalo Diaz   María Antonieta Guzmán   Dannette Guiñez   and Rodrigo Cornejo   

Multifocal metastatic infection by Staphylococcus aureus is severe and uncommon. We describe the case of a previously asymptomatic 63-year-old man diagnosed with complicated staphylococcal endocarditis of the mitral valve along with cervical and lumbar spondylodiscitis, and multiple pre/para-vertebral abscesses; establishing a persistent flaccid tetraparesis, neurogenic bladder and anal sphincter dysfunction. The patient overcame infection after an 8-week Cloxacillin/ Rifampicin treatment associated to multiple surgical procedures. Thereafter, immunological assessment was performed, driving to the diagnosis of selective IgM deficiency. We propose a physiopathological relationship between this rare dysgammaglobulinemia and such fulminant staphylococcal invasive infection; that would, perhaps, justify humoral immunity assessment in patients undergoing staphylococcemia.

Prashant Kumar Dhakad   Pramod Kumar Sharma   and Sokindra Kumar   

Capparis decidua has been traditionally used in the Ayurveda to treat rheumatoid arthritis and it is reported to have anti-inflammatory and analgesic activity. Considering its anti-inflammatory activity the present research work has been designed to assess the anti-arthritic activity in Wistar rats. The anti-arthritic activity of hydroalcoholic extract of C. decidua root, stem and leaves was evaluated using Freund's complete adjuvants (FCA) induced arthritic models in Wistar rats. Oral administration of C. decidua extract at the dose of 100mg/kg, 200mg/kg, per se group (only 200mg/kg of C. decidua) was subjected to Wistar rats for 28 days. Standard drug Diclofenac sodium at the dose of 5mg/kg and FCA at 1mg/ml was used in the study. The normal control group was administered only distilled water at 1ml/kg without induction of arthritis. The arthritic investigation was carried out on basis of parameters including changes in body weight, paw volume, hematological studies like ESR count, RBC count, WBC count, Hemoglobin count, platelet count. At the end of study period, animals were sacrificed and histological parameters were evaluated. Phytochemical analysis of C. decidua extract was done to assess the various constituents present in C. decidua. The results of C. decidua extract administration significantly (P<0.001) attenuated the body weight, paw volume, hematological alteration induced by the FCA in dose-dependent manner. The tarsal joint was extracted for histopathological studies. The overall results indicate that C. decidua extract (100mg/kg and 200mg/kg) showed a potent protective effect against FCA induced arthritic rats which could be attributed to phytoconstituents present in C. decidua and its effect is comparable to the standard drug diclofenac sodium.

Reazul Karim   and M. A. Hakim   

Urinary tract infections (UTIs) are serious health affecting problems worldwide. Urinary tract infection (UTI) can be caused by a lot of bacteria. Among them, E. coli is one of the main causes of both nosocomial and community acquired Urinary tract infections in humans. Drug resistance of E. coli is becoming alarming with the passage of time worldwide in general and in third world countries in particular. The study was intended to collect and analyze the samples from the patients suffering from Urinary tract infection of two medical of southern region of Bangladesh to reveal the antibiotic resistance pattern of E. coli responsible for Urinary tract infection. 50 samples from each hospital were taken. 36 and 41 samples respectively were identified as E. coli. Antibiotic sensitivity testing was done on Mueller-Hinton agar by Kirby–Bauer disc diffusion method. All samples exhibited a different percentage of resistance where ciprofloxacin was showed evidence of higher resistance rate, which was 63.88% and 70.73% respectively. Higher resistance (63% & 75%) against Ampicillin was also found in patients of both medical. On the other hand, 16% resistance against ceftriaxone and 23.53% resistance against cefotaxime suggesting, they were comparatively more effective.

Okafor Arthur C.   Aquaowo Uwakmfon A.   Ojiagu Kingsley D.   and Agu Kingsley C.   

Microbial contamination is considered the most significant hazard associated with food. Data on pathogens present in foods are needed to provide valuable information on the exposure of consumers to certain food borne pathogens. Also, consumption habits among consumers may increase the risk of health hazards. This study aimed to assess some practices, signs and symptoms associated with consumption of soaked garri among university students, to also determine the presence and virulence characteristics of some bacteria in garri on sale in the market. A questionnaire was designed and distributed to a hundred university students to ascertain their perception on safety of garri and handling practices. Fifty samples of garri were randomly collected from the market and assessed for presence of bacteria. Isolates of selected genera were then screened for some virulence-associated properties. The results showed that majority of the students (91%) affirmed they do consume soaked garri at least once in a while, even though almost 70% think garri is nutritious. The students (54%) reported to have experienced at least a symptom of ill health after consuming garri soaked in water. The most commonly reported signs and symptoms were: stomach upset > malaise > fever > nausea > vomitting. There was a direct relationship between the number of students that suspect garri could be a source of illness (57.1%) and those that have actually experienced at least a symptom after consuming garri (54%). Results of bacteriological analysis of garri samples from the market indicated mean total aerobic plate count to be 5.42 log cfu/g. Staphylococcus aureus and Klebsiella were found in 60% of the samples collected. Most isolates of Staphylococcus aureus, Klebsiella and Pseudomonas were found to be positive for haemolysin and biofilm formation tests which indicate their pathogenic potential. The conclusion is that bacteria with pathogenic potential are common in processed garri being sold in the market and that university students in the habit of consuming soaked garri are probably at risk of health hazards associated with pathogens in garri. Therefore, new food safety regulations should be formulated to incorporate food safety management systems during processing of garri in order to reduce the prevalence of food borne pathogens.

Mohammad Shahidul Islam   Mohammed Ahasan Habib   Mohammad Rafiqul Islam   Mohammad Showkat Mahmud   Provat Chandra Saha   Tahmina Ruba   Priya Mohan Das   and Mohammad Abu Hadi Khan   

Foot and Mouth Disease (FMD) is a highly contagious viral disease of farm animals. This study was aimed to identify symptoms, characteristics pathology and viral serotypes involved in infected cattle of Bangladesh. Dominant signs observed were salivation, vesicular eruption, lameness and maggot infestations in 66%, 80%, 36% and 14% cattle respectively. Out of 202 infected cattle investigated, 35 were below six months of age. A total of 18 young calves and 8 adult cattle were died due to FMD. Seven young calves and two adult cattle were examined at necropsy, 'Tiger heart disease' was commonly seen both in the young and adult cattle. The 'currant jelly clot' was common in the atrium, ventricles, aorta and veins of the heart of adult cattle. Congested and consolidated lungs, atrophied spleen, and fibrosed udder were grossly seen in the infected and dead cattle. Microscopically, hyalinization of cardiac muscle in young calf and degeneration, necrosis and multifocal lymphocytic infiltration in heart muscle were suggestive for infectivity due to FMD viruses. There were bronchiolitis, pulmonary emphysema, interstitial pneumonia and thickening of interlobular septa of lungs. Cattle survived from the acute infection developed respiratory distress in 15% cattle. Out of 202 samples tested in reverse transcriptase polymerase chain reaction (RT-PCR), 187 samples found to generate positive amplicons. Single infectivity due to FMD viral Serotypes O, A and Asia 1 were seen in 115 (62%), 26 (14%) and 34 (18%) cases, respectively. Seven cattle were co-infected with FMD viral Serotypes O and Asia 1 and four with FMD viral Serotypes O and A. FMD viral serotype O was dominating in cattle all over the country and was related to death of infected young and adult cattle. Cardiac myonecrosis and respiratory failure were the dominant lesions may be related to death of infected animals.

Mohammad Showkat Mahmud   Eusha Islam   Mohammed Abdus Samad   Md. Rezaul Karim   Amal Kumar Saha   and Md. Giasuddin   

Baby Hamster Kidney-21(BHK-21) cells are the most commonly used cell for viral seed preparation and vaccine production. The aim of the present study was to adapt BLRI three different field isolates of foot and mouth disease virus (FMDV) serotypes (O, A, Asia 1) into baby hamster kidney-21 cell line for FMD trivalent master seed preparation. For this, samples were collected from the tongue epithelium, saliva, milk and inter digital tissues of the suspected animals from different districts of Bangladesh during the year 2011-2014. All samples were subjected to RNA extraction and RT-PCR for detection and serotyping of FMDV. BHK-21 cell subculture was done from a previously cultured flask containing BHK-21 cells. Prepared inocula (positive for serotype O, A, Asia 1) were inoculated into BHK-21 cell culture and cytopathic effects (CPE) were observed in BHK-21 cell line characterized by rounding and flattening of the cells, multinucleated giant cells formation, breaking down of the intracellular bridges and finally cell death indicated the presence of FMDV. Adaptation of the virus into baby hamster kidney-21 cell line was confirmed by detection of viral infection in infected cells by RT-PCR with serotype specific primers. The cell adapted foot and mouth disease virus in the present study provides a scope to undertake further research on foot and mouth disease virus in Bangladesh towards development of trivalent FMD vaccine for use in the country.

Mahbubur Rahman   AKM Fahmidul Haque   Iztiba Mallik Deeba   Dilruba Ahmed   Tanha Zahidi   Afrina Hossain Rimu   Mahmuda Akter   Fatema Akter   and K A Talukder   

The objective of the study was to investigate current species distribution and growing antimicrobial resistance (AMR) of Shigella isolates for proper treatment. Shigellae, isolated from faecal samples in International Centre for Diarrhoeal Disease research, Bangladesh, Dhaka hospital in 2015, were tested for antimicrobial susceptibility by disc diffusion method to ampicillin, co-trimoxazole, ciprofloxacin, azithromycin, mecillinam, ceftriaxone/cefixime and meropenem. Extensively drug-resistant (XDR, resistant to 5 or 6 of 7 useful anti-Shigella drugs tested) Shigella isolates resistant to 6 drugs were analyzed for ESBL and AmpC phenotypes, plasmid profiles, R-plasmids transfer, bla_SHV, bla_TEM, bla_CTX-M, bla_OXA; and mphA, mphB, ermA, ermB, ermC, ereA, ereB, mefA and msrA genes by PCR; and clonality of S. sonnei by PFGE. Of 134 isolates cultured from 3722 (3.6%) diarrhoeal faecal samples, 46% were S. sonnei, 37% S. flexneri, 4% S. boydii, 5% S. dysenteriae and 7% non-typeable. Multidrug-resistant (MDR, resistant simultaneously to ≥3 drugs) S. sonnei were 95% compared to 66% (P<0.01) MDR S. flexneri including 18% & 14% XDR types, respectively. All isolates were susceptible to meropenem. Four (6%) S. sonnei, 2 (4%) S. flexneri and 1 (17%) S. boydii (total of 7 isolates) were six-drugs XDR; 5 of them had ESBL phenotypes. Three S. sonnei and 1 S. flexneri had bla_TEM and bla_CTX-M; 1 S. boydii had bla_SHV, bla_TEM and bla_CTX-M; 1 S. sonnei had bla_TEM β-lactamase. All but one S. flexneri had only mphA gene on 62-MDa conjugative-R-plasmid coding azithromycin resistance. PFGE identified MDR-S. sonnei Global III clade. Thus, MDR-S. sonnei replaced S. flexneri as predominant isolate in Dhaka, Bangladesh; many emerged as XDR strains requiring treatment by meropenem. The findings demand judicial use of antibiotics to contain emergence and spread of resistance locally and globally. Physicians should be informed about MDR and XDR Shigella for judicious prescribing of antimicrobial therapy.

Sylvia Grune Loffler   Carlos Leiva   Exequiel Scialfa   Leandro Redondo   Mónica Florin-Christensen   Mara Martínez   Graciela Romero   and Bibiana Brihuega   

Leptospirosis is a worldwide distributed zoonosis, caused by pathogenic strains of spirochetes belonging to the genus Leptospira spp. So far, the only gold standard diagnostic method for this disease has been the Micro-Agglutination Test (MAT). The purpose of this study was to test whether a correlation exists between serological diagnosis by MAT and the presence of Leptospira spp. DNA traces in serum. Anti-Leptospira spp. antibody titers were determined in dog serum samples obtained at a canine shelter from Buenos Aires Province, Argentina (n= 80) by MAT using ten Leptospira spp. serovars. DNA extracted from these samples using Chelex-100 resin was used as template for Leptospira spp. specific PCR and loop-mediated isothermal amplification (LAMP) tests. In the latter case, product detection by incubation with malachite green dye was set up, obtaining optimal results at a dye concentration of 0.02% (w(v)) and a pH of 6.1. PCR and LAMP showed the same sensitivity, detecting leptospiral DNA in 12 out of the 48 MAT- positive sera. No detection was attained by either method in MAT-negative samples. These results show that it is possible to use LAMP for the detection of leptospirosis in non-human mammals. It also shows that, in the case of serum samples, MAT has higher diagnostic sensitivity than the currently available PCR and LAMP tests.

Jude Ogechukwu Okoye   Precious Enyinnaya Okorie   Nnodimele Onuigbo Atulomah   Folahan Sholeye   Ukpai Agwu Eze   Esther Chinenye Okoro   and Gideon Taiwo Faloye   

This questionnaire-laboratory based cohort study evaluated some socio-demographic characteristics and urothelial integrity among pulmonary tuberculosis (TB) patients. A total of 49 confirmed TB positive patients (males= 17 and females =32) within the age range of 10-79 years were recruited from Abeokuta and Ilishan metropolises, and divided into 2 groups: patients on anti-TB drugs and those not on anti-TB drugs. Smears made using cell deposits from fresh clean catch urine samples were stained by the Papanicolaou and Giemsa techniques. In this study, high frequency of TB infection was associated with gender, ethnicity, residency, marital status, occupation, and income status. The prevalence of urothelial lesion and breast tuberculosis was 24.5% and 8.2%, respectively with high frequency among patients within the age range of 30 to 39 years. A significant difference between patients on anti-TB drugs and those not on anti-TB drugs was observed in relation to low grade urothelial lesions (p<0.05). Subgroup comparison showed that approximately 47%, 45%, 34% and 17% of the urothelial lesions were associated with lack of TB therapy, HIV and TB co-infection, females and breast tuberculosis, respectively when compared with other subgroups. Interestingly, polyomavirus (45%), HIV (43%) fungal infections (39%), parasites (24%) and crystals (18%) were associated with the urothelial lesions (p<0.05). This study suggests that lack of TB therapy, age, female-sex and some co-infections may increase the risk of urothelial lesions and breast tuberculosis. Thus, female patients living in TB endemic areas and within the age range of 30 to 59 years should be screened pre- and post treatment.

Liuber Y Machado   Orlando Martínez   Héctor M Díaz   Enrique Noa   Madeline Blanco   Dayamí Martín   Dervel F Díaz   María L Sánchez   and Marta Dubed   

Background: Infection with human HIV-2 is endemic in West Africa. The virus originated from West African sooty mangabeys during the first half of the 20th century and an epidemic initiation in Guinea Bissau that coincides with the independence war (1963-1974). The HIV-2 group A is categorized as epidemic group. The presence of HIV-2 group A in Cuba has been previously documented. However, the evolutionary history of HIV-2 group A in the Cuban epidemic is unknown. The aim of this work is to estimate the origin and evolutionary history of the HIV-2 group A in Cuba. Methods: We used a Bayesian coalescent method to analyze the env gene of Cuban HIV-2 group A. The rate of nucleotide substitution was determined and was used to date the phylogenies and reveal the evolutionary history of HIV-2 group A in Cuba. Results: Multiple introductions of HIV-2 group A, mainly from Guinea Bissau and Portugal were detected. The most recent common ancestor of Cuban HIV-2 groups A was dated back to about 1972 (95 % HPD: 1966-1978). The rate of nucleotide substitutions was 5.02 x 10^-3 substitutions per site per years (95 % HDP: 4.51-5.52 x 10^-3). Conclusions: The results of this study allowed for the first time to estimate the evolutionary history of HIV-2 in Cuba and establish the basis for phylogeographic and phylodynamics studies.

Aileen Antonio-Santos M.D.   Eric R. Eggenberger   Fiona Costello M.D.   and Laura Balcer M.D.   

Objective to investigate whether optical coherence tomography (OCT) could demonstrate axonal loss through thinning of the retinal nerve fiber layer (RNFL) in multiple sclerosis (MS) patients. Furthermore, the degree of RNFL loss was compared in the different MS subgroups (with or without optic neuritis, affected or fellow eye, single episode or recurrent optic neuritis, relapsing remitting or progressive MS). RNFL thinning was also determined in MS patients who had serial OCT without any intervening clinical optic neuritis. Design: Retrospective chart review. Setting: Academic tertiary care MS centers. Participants: 177 patients (334 eyes) with multiple sclerosis, with or without optic neuritis, and 159 healthy controls that underwent OCT RNFL measurements. Main Outcome: Retinal nerve fiber layer measurements by OCT. Results: Average RNFL measurements were thinner in MS patients (90 μm) compared to controls (105 μm), p <0.0001. RNFL was significantly reduced in MS patients with optic neuritis (87 μm) versus those without optic neuritis (94 μm), p <0.0001. Among the different quadrants, the degree of RNFL loss was greatest in the temporal quadrant of MS patients (22%). Progressive (primary and secondary progressive) MS patients had thinner RNFL (82 μm) compared to all relapsing remitting MS patients (90 μm), p <0.0001. Greater RNFL loss was seen in SPMS patients (77 μm) versus PPMS (88 μm), p = 0.004. In the 45 MS patients without any intervening clinical optic neuritis, serial OCT (mean of 2 OCT scans per patients, averaging 11 months apart) showed that RNFL decreased by 3.7 μm per year. Conclusion: Retinal nerve fiber layer is significantly reduced in patients with multiple sclerosis. Progressive MS subtypes showed more marked RNFL thinning than relapsing remitting MS. This study reflects the role of OCT in MS patient monitoring and its potential as a surrogate marker in MS therapeutic trials.

Chetan Datta Poduri   

As per UNAIDS database, India has the World's third largest population of people living with HIV infection. Nigeria is second in position in this list of people living with HIV. This observation is significant for the simple reason that Nigeria is also presently affected with the recent Ebola epidemic (Ebola-2014). The West African region, to which Nigeria belongs, suffers the Ebola-2014 from a different strain of Ebola virus as compared to a concurrent epidemic in Equatorial African nation of Democratic Republic of Congo (DRC). Presented in this article is an analysis of the documented literature on this multi-dimensional Ebola-2014 epidemic. The mystifying phenomenon of persistence of the epidemic in West Africa is explained through various aspects attributable to the geopolitics of the region, biology of the virus, funeral customs in the region, fiction, economics and conspiratorial theories.

Chetan Datta Poduri   

In the past there were at least 23 epidemics with Ebola virus (EbV). Presently there are pandemics with Hepatitis C virus (HCV) and Human Immunodeficiency virus (HIV). The recent epidemic of EbV has taken an year to get contained. Infections with HCV and HIV are continuing to increase. At present, except for EbV, neither HCV nor HIV have a vaccine to prevent future infections. Here, a snapshot of the three viruses with regard to their characteristics and prevalence (both global and Indian) are generated with an emphasis to study the infection dynamics of these three RNA viruses in relation to each other. An emphasis is made towards the common or shared features of the three viruses.

Nfor E.   K. Kakanda   G. Buki   A. Kane   and J. Dongang   

Background: The Government of Cameroon and its partners have made major investments in the last decade in prevention, treatment, and care of HIV-infected patients. However, unmet need for antiretroviral therapy (ART) among HIV-positive pregnant women remains high at 66%. Critical to satisfying this need is ensuring adequate availability of prevention of mother-to-child transmission (PMTCT) commodities for rollout of new Option B+ guidelines. The Cameroon supply system consists of a cost recovery system for essential medicines and other health commodities and a free-of-charge system for priority commodities including those for PMTCT and ART. This study examines options for improving the supply and availability of these commodities. Method: Supply chain (SC) operational data was collected in July 2014 from central (CENAME) and 4 regional warehouses (CAPRs); 10 district stores; and 30 service delivery points (SDPs), including ART and PMTCT sites. The study also included seven central private-sector logistics firms. In addition, SC cost data was obtained from CENAME and CAPRs financial statements audited in 2013. Data collected served for analysis of three options to improve effectiveness of delivering PMTCT commodities, based on the four variables detailed in Figure 2.4.1. Results: Asset utilization within the cost recovery system ranged between 73% and 89% while inventory turnover was at 1.5. Therefore, a reliable supply of medicines to SDPs is ensured. However, for PMTCT and ART commodities, distribution to the SDPs was unreliable (in 2013, 40% of prescriptions remained unfilled). Meanwhile, results of the options analysis indicated that the model of CAPRs delivering PMTCT commodities to SDPs was the most desirable. Although the distance traveled was higher, the need for network storage space was minimal. Moreover, its total cost and human resource requirements were more favorable. Conclusion: As a result of disseminating the findings, the Ministry of Health adopted Option 2. PMTCT free-of-charge commodities are also amenable to being managed within the existing effective cost recovery system.

Md. Rezaul Karim   Mohammad Showkat Mahmud   and Md. Giasuddin   

Fasciolosis is an economically important parasitic disease of cattle in Bangladesh. The aim of the study was to determine the prevalence of fasciolosis in cattle and identifying the potential risk factors associated with the disease. A cross-sectional survey was carried out from November 2012 to October 2013 on bovine fasciolosis in and around Shahjadpur upazila of Bangladesh. For this, a total of 762 fecal samples were collected from crossed breed cattle and all samples were subjected to standard sedimentation test at Animal Health Laboratory of BLRI regional station, Shahjadpur, Bangladesh. The overall prevalence of bovine fasciolosis was found to be 66.14% from 504 positive samples. From the various risk factor analysed, age categories of cattle, sex and seasonal influence were found to be significantly associated (p<0.01) with the prevalence of bovine fasciolosis. When considered the relationship with sex, it was observed that female (70.3%) cattle had significantly (p<0.01) higher than male (55.23%). As regards to age, bovine fasciolosis was significantly (p<0.01) higher in old cattle (76.43%) compared to adult (68.69%) and young (48.62%). The seasonal influence of outbreak of this disease was significantly (p<0.01) higher in summer season (72.44%) followed by winter (67.75%) and rainy season (58.55%). It can be concluded that these risk factors need to be taken in consideration when designing effective fasciolosis control management system for cattle in Bangladesh.

Taslima Yesmin   Mohammed Abdun Nur Sayam   Md. Manjurul Quader   Abul Hossain Khan   and Md. Abdullah Yusuf   

Background: The detection of extended-spectrum β-lactamase-producing (ESBL) bacteria is of importance for infection control and epidemiological surveillance. Objective: The purpose of the present study was to compare two phenotypic methods for the detection of ESBL-positive Enterobacteriaceae and Pseudomonas species. Methodology: This cross sectional study was carried out in the Department of Microbiology at Mymensingh Medical College, Bangladesh from January 2011 to June 2011. Patients of all ages and genders presented with UTI or wound infection were taken as study population. Gram negative bacilli (GNB) were analyzed by two methods used for routine susceptibility testing which were Disk diffusion methods and MIC reduction methods. Two methods designed for the detection of ESBL production Ceftazidime and Ceftazidime plus Clavulinic acid, (CAZ/CAZC) and Cefotaxime and Cefotaxime plus Clavulinic Acid (CTX/CTXC) were used and the PCR was considered as gold standard for evaluation of the other test methods. Result: A total number of 300 GNB were isolated and identified of which 214(71%) were ESBL positive. For the disk diffusion method, resistant to third generation Cephalosporins were the highest 87.0%, when tested by ceftazidime and by MIC reduction methods were 67%. For the phenotypic confirmatory methods, specificities were 64% by (CAZ/CAZC), 59% by (CTX/CTXC) and by both method 52%. Among the phenotypic confirmatory ESBL positive strains by Genotypic method ESBL positive were 50.46% TEM, 18.69 % SHV and 46.72% CTX-M gene. Conclusion: Two-step strategies using both DDDT phenotypic methods are useful diagnostic tools for the detection of ESBL from the Gram negative bacilli.

Ayman Rihawi   Glen Huang   and Royce Miller   

This article discusses Streptococcus salivarius meningitis in a 39-year old woman. She was admitted to the hospital for general fatigue, headache, and fever. Cerebrospinal fluid analysis revealed leukocytosis and cultures were positive for Streptococcus salivarius. Streptococcus salivarius meningitis is rare and has been documented to be associated with procedures involving the spinal column such as epidural anesthesia. In this patient the meningitis caused by Streptococcus salivarius was thought to be spontaneous.

Sharif Neowajh   Md. Mer Mosharraf Hossain   Ibrahim Kholil   Sumana Nasrin Mona   Sherajul Islam   and Mitheela Kabi   

This study was intended to address, the effect of oxytetracycline, erythromycin, and streptomycin on the antimicrobial resistance of Aeromonas sp. isolated during the production process, from infected common carp (Cyprinous carpio). Antimicrobial resistance patterns were defined by determination of minimal inhibitory concentrations (MIC) of three antimicrobial drugs, and bacteria. Results indicated that In vitro bactericidal activity, oxytetracycline showed lowest number of bacterial colonies (1×10^-4 c.f.u./ml) than erythromycin (3×10^-4) and streptomycin (4.5×10^-4) compared to the control (9×10^-4). Oxytetracycline showed the highest maximum zone of inhibition (20 mm) than erythromycin (18 mm) and streptomycin (15 mm) in antibiotic susceptibility test. In infection inhibition assay, serum from oxytetracycline fed diets inhibited Aeromonas infection to common carp. The susceptibility of the fish fed antibiotics (50 µg/mg feed) and challenge with Aeromonas sp. (1×10^-4) was examined for 30 days. There was high relative percent of survivility (RPS) in oxytetracycline was 73%. Very low RPS 63% and 47.37% was found in erythromycin and streptomycin against Aeromonas sp. infection. These results demonstrated that the oxytetracycline might be a potential for Aeromonas sp. infection to common carp (Cyprinous carpio).

Mohammed Hafizur Rahman   Mohammed Giasuddin   Mohammed Rafiqul Islam   Mahmudul Hasan   Mohammad Showkat Mahmud   Mohammed Ahsanul Hoque    Paritosh Kumar Biswas   Emdadul Haque Chowdhury   Bidhan Chandra Das   Mohammed Ziaur Rahman   Nitish Chandra Debnath   Garba Ahmed   and Mat Yamage   

Highly pathogenic avian influenza (HPAI) causes enormous losses to the poultry industry of Bangladesh every year since the detection of first outbreak in 2007. In this study biomolecular diagnosis of avian influenza virus in Bangladesh from different species of birds was carried. A total of 1135 samples including 280 from migratory birds, 680 from live bird market (LBM) chickens, 118 from ducks and 57 from pigeons were collected from different locations of Bangladesh during the period from July 2013 to June 2014. Out of 1135 samples, 69 were found positive for influenza A by real time RT-PCR. In addition, 741 Influenza A positive samples collected by different institutes from different species of birds under the FAO-led surveillance program were also received at Bangladesh Livestock Research Institute (BLRI) for further analysis. The real time RT-PCR positive samples were inoculated in 10-day-old embryonated chicken eggs. Out of 810 (741+ 69) influenza A positive samples, 110 were selected on the basis of species and region to cover the whole country and were sent to the OIE reference laboratory for detailed study. The bio-molecular results revealed that in case of migratory bird samples one was H5 (HPAI), two were H9 and eight were of undetermined subtype. Among the LBM chicken samples, seven were H5 (HPAI), 51 were H9, six were mixed infection with H5 & H9 and three were of undermined subtype. In case of duck samples, one was H2, six were H4, six were H5 (HPAI), one with mixed infection with H5 & H2, one with both H5 & H4, two were H9 and one was H7N5, though H7 was genetically distinct from that of the novel H7N9 virus of China. Among quail samples, one was H5 (HPAI) and one was H9. In case of pigeon samples, all three contained H9. From this study it was revealed that HPAI (H5N1) is still circulating in our poultry population. The novel avian influenza virus H7N9 was not found during this study.

Girum Tefera   

Background: Intestinal parasitic infections are among the most common infections worldwide. High prevalence is found in people with low socio-economic status, poor living condition, overcrowded areas, poor environmental sanitation, improper garbage disposal, unsafe water supply and unhygienic personal habits. Thus, the objective of this retrospective study was to determine the prevalence of intestinal parasitosis among patients with diarrhea who visited Wonago health center Southern Ethiopia. Methodology: Institution based retrospective study design was employed. Stool examination records of all patients with diarrhea who visited Wonago health center from September 2010 to September 2012 were included. A total of 10694 stool examination results of those diarrheic patients were analyzed for this study. Samples were examined using direct saline wet mount technique. Statistical analysis was done by SPSS version 16 and results were shown in percent and prevalence rates. Results: The overall prevalence of intestinal parasite for at least one parasite in this retrospective study was 1776/10694(16.6%). Furthermore, 5.7% of those positive patients were infected by two species of parasites and 4.6%of patients had triple infections. The most prevalent parasites were E. histolytica/dispar 956/1776(53.8%) followed by G. lamblia 420/1776(23.7%) and Taenia species 140/1776(7.9%) respectively. Conclusion: Intestinal parasitosis is highly prevalent in varying magnitude among diarrheic patients who visited Wonago health Center, Southern Ethiopia. E. histolytica/dispar was the most prevalent parasite infecting patients followed by, G. lamblia, Taenia species and H. nana. Because of absence of molecular diagnosis for E. histolytica, over diagnosis might be possible for the high prevalence of amoebiasis.

Girum Tefera   

Back ground: Anemia is a common and serious problem in pregnancy. Parasitic infections contribute to iron deficiency anemia in pregnant women. The objective of the study was to assess determinants of anemia among pregnant women with emphasis on intestinal helminthic infection at Sher- Ethiopia hospital, Ziway, southern Ethiopia. Method: A cross-sectional study was carried out among pregnant women who came for their antenatal care follow up at Sher-Ethiopia hospital, Ziway, Southern Ethiopia from April, 1/2014 – June 30/ 2014. A total of 748 pregnant women were screened for intestinal parasitic infection using direct microscopy and formol ether concentration techniques and level of hemoglobin was determined using hematocrit screening technique. Other determinants of anemia during pregnancy were also assessed by using structured questionnaire. Data were analyzed using SPSS for windows version 16.0. Statistical tests were performed at the level of significance of 5%. Result: Prevalence of intestinal parasite infection in pregnant women was 58.2%. Out of 436 intestinal parasite infected women 176(40.4%) had more than one intestinal parasitic infection. Prevalence of anemia in this study subjects was 51.9% and the mean hematocrit level was 34%. Anemic women were 10 times likely to have hookworm plus other intestinal helminthic infection (p value = .000), 2 times likely to have birth interval less than two years (p value = .018), 2 times likely not to have shoe wearing habit (p value = .045) and 2 times likely not having taken iron during pregnancy (p value = .025). Conclusion: From the result it can de concluded that all antenatal care attendants should be screened for anemia and intestinal helminthic infection at their first visit and those anemic and helminthic infected women should be treated properly.

Suresh J. Antony   

Rhinoscleroma is an unusual disease that is seen in many parts of the tropics. It is caused by a bacteria called Klebsiella rhinoscleromatis and can leave the patient with a chronic inflammatory lesion of the upper respiratory airway and much disfigurement. In the age of global migration and travel physicians must learn to recognize this disease especially in immigrants and patents that come from countries endemic of the disease. It is endemic in parts of Mexico, Central and South America, Central and Eastern Europe, Southeast Asia and Africa. Humans are the only identifiable host and the transmission is thought to be airborne. This disease is mostly seen in the poor rural areas of the world where overcrowding and poor hygiene are common.

Girum Tefera   

Back ground: Malaria is usually an important hazard in order to open health and economic progress inside the African continent. Throughout sub-Saharan the African continents, 90% of all demise largely below 5 youngsters tend to be due to malaria. Yearly you can find in relation to 270-480 million circumstances associated with malaria leading to in relation to one million demise, largely inside The African continent as well as especially within 5 youngsters. Almost all of the full division of Ethiopia is actually malarious, doing malaria the best open health problem inside Ethiopia. The purpose of this particular review seemed to be to look into your frequency as well as associated factors among patients attending at Hallaba health center based on thick and thin blood film examination. Thus, this research was launched to determine the prevalence of malaria and associated factors among patients who attended at Hallaba health center. Methods: Health center based cross sectional study was conducted from April 19 to June 21, 2009 at Hallaba health center on a total of 204 individuals who were selected conveniently. Structured questionnaire was used to obtain socio-demographic and other data. Then the data were entered and analyzed by using SPSS software. Statistical tables and graphs were used to present the result. Chi-square tests and OR (95% CI) were used to test the association. Results: The analyses showed that the highest prevalence of malaria was found between age group of 0-19 year. There was association between age group and prevalence of malaria (X2= 77.07, P<0.005) and also there was association between prevalence of malaria and sign and symptom like fever (OR=11.84%, 95%CI, 4.62-30.27, P<0.005). Vomiting (OR=8.29, 95% CI 3.39-20.49, P<0.005).Chills (OR=8.76, 95% CI 3.71-20.70, P<0.005) Joint and back pain (OR=3.85, 95%CI 1.26-11.94, P<0.005), headache (OR=75.17, 95% CI 21.12-270.43, P<0.005). Conclusion: Those with inadequate socio-economic conditions are really linked to malaria contamination. On the result it can be concluded that most of the respondents were infected with Plasmodium vivax. The respondents' knowledge or having information about malaria has no association with the prevalence of malaria. Age, altitude, rainfall and socio-economic factors contribute to high prevalence of malaria at Hallaba health Center. This kind of facts is vital to style increased strategic involvement with the lessening connected with malaria pandemic in Halaba town. Generally, from the result of this study it can concluded that the prevalence of Plasmodium vivax and Plasmodium falciparum is different from the national figure.

Razi Syed   Daniel H. Havlichek   Gary E. Stein   and Curtis L. Smith   

OBJECTIVES: To determine the usefulness of Procalcitonin (PCT) in detecting bacterial infection in hospitalized elderly patients with COPD exacerbation. DESIGN: Prospective observational study to compare PCT levels in patients with and without pneumonia. SETTING: Acute care community hospital in United States. PARTICIPANTS: 18 elderly patients age 65 years and older admitted to an internal medicine ward. MEASUREMENTS: Demographic characteristics, general signs and symptoms, laboratory and chest radiographic results, PCT levels, antibiotic use and duration, length of hospital stay and 90-day readmission rates. RESULTS: Eight (44%) patients had a diagnosis of pneumonia. Procalcitonin levels suggestive of a bacterial infection were found in only four patients and each of these patients had pneumonia. Three patients with pneumonia had positive sputum cultures without an elevated PCT level. No patient without pneumonia had an elevated PCT level. The sensitivity and specificity for predicting a bacterial pneumonia in our patients was 50% (95% CI, 16-84%) and 100% (95% CI, 69-100%), respectively. This biomarker had a positive predictive value of 100% (95% CI, 40-100%) and a negative predictive value of 71% (95% CI, 42-91%) CONCLUSION: Our study results suggest that PCT is not sufficiently sensitive to be used as a diagnostic test of bacterial infection in elderly patients with COPD exacerbation. It may assist clinicians in identifying patients without pneumonia who do not require antibiotics due to its high specificity and negative predictive value.

Paras Singh   Mradula Singh   Devika Tayal   Vithal Prasad Myneedu   Manpreet Bhalla  Puncham Adlakha   and Rohit Sarin   

Background and Objectives: In spite of higher incidence and prevalence of tuberculosis, the diagnosis of extra-pulmonary tuberculosis (EPTB) in various clinical specimens (such as pleural fluid, ascitic fluids, CSF, lymph node aspirate etc), remains true challenge. Current tools for the diagnosis of tuberculosis in various body fluids are suboptimal. Clinicians underestimate these diseases, and use of insensitive conventional analytical method has contributed to the difficulties in managing patient with extra pulmonary tuberculosis. It is important to develop rapid, sensitive and specific test for early diagnosis of extra pulmonary tuberculosis because of the lack of sensitivity & specificity of the conventional methods as AFB smear by ZN technique and culture on LJ media. Pleural tuberculosis (TB) diagnosis often requires invasive procedures such as pleural biopsy. The study was undertaken to evaluate the combined utility of polymerase chain reaction (PCR) for different gene targets (IS6110, MPB64 and protein antigen b; Pab ) especially in pleural fluid specimens with adenosine deaminase (ADA) levels in the diagnosis of pleurisy. Methods: Total 430 clinical specimens (412 extra-pulmonary and 18 pulmonary tuberculosis) were recruited from the outdoor and indoor Department of National Institute of Tuberculosis and Respiratory Diseases, New Delhi during the 2011-2013 periods. All specimens were further processed for AFB smear, culture on LJ media, ADA level and conventional PCR (IS6110 & MPB 64 and Pab gene targets). Results: The PCR positivity IS6110, MPB64 &Pab gene targets was found to be 90.3% (N=28/31) in smear negative & culture positive (as gold standard) extra pulmonary tuberculosis cases. However the total PCR positivity of Pab (43.9%; 181/412) gene target was found to be significantly low as compared to the IS6110 (65.3%; 269/412; X2=37.058; pc=0.000; Odds ratio 2.401; 95% CI=1.795-3.213) & MPB 64 (63.6%; 262/412; X2=31.245; pc=0.000; Odds ratio 2.229; 95% CI=1.669-2.978) gene targets in extra pulmonary tuberculosis cases. Further we have analyzed the combined utility of PCR with ADA levels among the body fluids (165 pleural fluid, 15 ascitic fluid, 1 lymph node and 1 cerebrospinal fluid; CSF). Our results indicated that the PCR alone can detect total 72.5% (132/182) TB cases, whereas ADA alone can detect 61.5% (112/182; considering cutoff value >40IU/L or confirmed cases of TB on clinic-radiological findings), M.tuberculosis in body fluids. Further data was compared in between single, two and three gene targets considering cut off value ADA >40IU/L levels in body fluids. Our observation showed that the positivity of tuberculosis cases were significantly higher through three gene targets (N=48/83; 57.8%; Mean of ADA >40IU/L =127.3) as compared to single gene target (N=10/83; 12.1%; Mean of ADA >40IU/L =68.2; X2=36.27; pc=0.000; Odds ratio 10.011; 95% CI=4.272-24.008) utilizing conventional PCR technology. No significant difference has been observed in other body fluids. The combined evaluation of both techniques (PCR and ADA) raised 14-15 % additional diagnosis of tuberculosis in body fluids (158/ 182; 86.8%; in pleural fluid= 147/165; 89.1%).Interpretation and Conclusion: Our results suggested that Protein antigen b (Pab) gene target showed less sensitivity as compared to IS6110 & MPB64. This study demonstrated the combined utility of both techniques (multigene target PCR with ADA level), enhanced the sensitivity of diagnosis of tuberculosis in body fluids. The study also confirmed the high diagnostic utility of PCR and ADA methods in diagnosis of tuberculosis in various paucibacillary body fluids in Indian scenario.

Danila De Vito   Emilio Jirillo   Andrea Ballini   and Giuseppe Mastrorillo   

Since we face the problem of rapidly growing rates of antimicrobial resistance, autovaccination may provide a treatment alternative at least in those patients which suffer from treatment refractory infections. Interest is turning towards the therapy of infectious diseases by stimulation of the immune defence mechanisms. In fact there are reports of drug resistance in a wide range of bacterial diseases. In our experience, autovaccine immunization has the potential to treat chronic infections such as osteomyelitis unresponsive to antimicrobial therapy.

Meshgi B   and Khodaveisi M.   

Dicrocoeliasis caused by the small liver fluke Dicrocoelium dendriticum (syn. D.lanceolatum), which live in the bile ducts and gall bladder of wild and domesticated mammals, particularly livestock farming. Parasitological methods are not reliable enough for diagnosis of this trematode infection. In the present study, the sera from rabbits immunized with somatic and excretory-secretory antigens of D.dendriticum were assayed with immunoblot technique. Adult worms were collected from infected livers of sheep at a local abattoir. The excretory-secretory product and somatic antigens were prepared through homogenization and incubation of trematodes, respectively. Polyclonal antibodies were raised in rabbits after inoculation of the two antigens with emulsified Freunds adjuvant followed by booster injection. Comparative study between D.dendriticum antigens and those of Fasciola, cysticercus tenuicollis and hydatid cyst fluids antigens was carried out. SDS-PAGE of antigens was performed and for immunoblotting analysis, hyperimmune sera were obtained from immunized rabbits of case and control groups. Immunoblot analysis of rabbits antisera against somatic and excretory-secretory antigens of D.dendriticum were showed six and four protein bands ranging from 25 to more than 170 and 25 to 70 kDa molecular weights, respectively. However cross-reaction with other specific proteins was observed in Fasciola excretory-secretory, hydatid cyst and cysticercus tenuicollis fluid antigens. The results demonstrated that the 25-27 kDa proteins were strongly recognized by both hyperimmune sera and was common in both somatic and excretory-secretory antigens. According to the results of this study performed with two antigens of D.dendriticum shows that the 25-27 kDa polypeptide as immunodominant protein, could be considered for the immunodiagnosis of dicrocoeliasis and it probably induces protective immunity against Dicrocoelium infection.

ShailenMohan Tripathi   and Neeti Mishra   

Here, we are reporting a case of late onset mania in a 61 year old male secondary to dengue fever. The past and family history of the mood disorder was absent in the patient. Episode of mania had a temporal correlation with the dengue fever. Psychiatric co-morbidities are mentioned in the literature following dengue fever but the literature on the mania following dengue is sparse. The late onset mania in dengue fever is unreported in the literature.

Waheedha Basheer   Dale Kunde   and Rajaraman Eri   

Chemokines are important players in the immune system with individual chemokine axes demonstrating significant associations with numerous inflammatory disorders. The chemokine receptor CCR6 and its ligand CCL20 are reported to be involved in the pathogenesis of inflammatory bowel disease, however the exact mechanism remains elusive. The Ccr6gene has been identified as a susceptibility gene in Crohn’s disease, while the expression of its ligand, CCL20 is up regulated following inflammatory stimulus in the intestine. The identification of the role of CCR6-CCL20 axis during the inflammation will shed valuable light into the pathogenesis of IBD as well as providing a potential therapeutic target for treatment for IBD.

Amit Shankar Singh   

Human Immunodeficiency Virus (HIV) disease pose a significant health hazard in developing countries like India which is carrying pressure of world second most population burden. Both visible and covert factors dominate in the failure of control of this disease. But despite that intensified efforts are needed to surmount this hurdle of misery and debility.

Paras Singh   Puncham Adlakha   Pusphendra Verma   Vithal Prasad Myneedu    and RohitSarin   

Background & Objectives: Undiagnosed and mismanaged tuberculosis (TB) continues to fuel the global epidemic. Rapid, accurate and early diagnosis of tuberculosis is a major health concern especially in developing country like India. It is important to develop rapid, sensitive and specific test (i.e. Polymerase chain reaction; PCR) for early diagnosis of tuberculosis because the conventional methods like Ziehl-Neelsen (ZN) smear, lack the sensitivity & specificity, cultures on Lowenstein-Jensen (LJ) media is time consuming and cumbersome techniques. Current study evaluates the significance of In house PCR with commercially available multiplex PCR kit (IS6110 and MPB64 gene targets) in clinical M. tuberculosis samples. Methods: The performance of In-House PCR for the detection of M. tuberculosis (IS6110 & MPB 64 gene target) was compared with multiplex M.tuberculosis PCR kit (Seeplex^RMTB ACE Detection Kit; www. Seegene.com). Fifty samples were processed for ZN smear, culture on LJ media and for PCR.Results: Overall combined PCR positivity for both PCR (In House PCR and multiplex M. tuberculosis PCR kit) was observed 76% (38/50) and 70.7% (29/50) in smear negative (S^-ve), culture negative (C^-ve) extra-pulmonary tuberculosis samples. No significant difference was observed between the positivity rate of both PCR (p = 0.832; χ² =0.045). However the smear ZN (6%) and culture positivity (14%) in LJ media was observed very low in extra-pulmonary paucibacillary samples.Interpretation & Conclusion: Current study evaluated the significance of In-House PCR (IS6110 & MPB 64 gene target) with multiplex M. tuberculosis PCR kit in rapid diagnosis of tuberculosis in clinical tuberculosis samples particularly in extra-pulmonary smear negatives.

Dhelia M. Williamson. Ruth Ann Marrie Allison Ashley-Koch and Glen A. Satten 

This study was conducted to determine whether single nucleotide polymorphisms (SNPs) in nine genes (human leukocyte antigen (HLA), T cell receptor beta (TCA receptor β), tumor necrosis factor α (TNF α), tumor necrosis factor β (TNF β), apolipoprotein E (APOE), interleukin 7 receptor alpha chain (IL7RA) interleukin 2 receptor alpha chain (IL2RA) myelin basic protein (MBP) and vitamin D receptor (VDR)) associated with multiple sclerosis (MS) could be replicated in a population-based sample, and to determine if these associations are modified by presence of HLA DRB1*1501. DNA was available from 722 individuals (223 with MS and 499 controls) who participated in a population-based case-control study. Cases and controls were matched on ancestry, age, gender and geographic area. HLA DRB1*1501 risk allele (T) was confirmed in this population using a genotypic test, controlling for multiple comparisons. Examining the effect of each SNP in the presence or absence of the HLA DRB1*1501 risk allele identified significant associations with TNF α-1031 (rs1799964) among those without the HLA risk allele. No additional interactions were significant in a cases-only analysis. Our results indicate that an interaction between SNPs in TNF α and HLA DRB1*1501 may influence the risk of developing MS.

Mradula Singh Paras Singh Vithal Prasad Myneedu Manpreet Bhalla Puncham Adlakha Pusphendra Verma and Rohit Sarin 

Background & Objectives: Globally, tuberculosis (TB) still remains a major public health problem. India is a high TB burden country contributing to 26 per cent of global TB burden. Pulmonary tuberculosis (PTB) cases are more common (~ 90% of cases) while extra pulmonary tuberculosis (EPTB) constitutes around 10 to 20% of all tuberculosis cases in India. The diagnosis of the EPTB cases is difficult because of few bacilli and consequently is associated with low sensitivity of Zhiel-Neelson (ZN) smear and culture on LJ media. The present study evaluates the utility of PCR for the detection of M. tuberculosis in paucibacillary extra pulmonary and pulmonary tuberculosis samples. Methods: A total of 561 samples (553 EPTB & 8 PTB cases) were collected from the extra pulmonary and pulmonary tuberculosis patients which were processed for ZN smear, culture on LJ media and conventional PCR using two gene targets (IS6110 and MPB64). Results: The PCR positivity of IS6110 and MPB64 gene targets was found to be 91.3% (N=63/69) and 89.9% (N= 62/69) in majority of smear negative & culture positive (as a gold standard) extra pulmonary cases, respectively. However the PCR positivity was observed 100% in smear positive, culture positive Line probe assay tested MDR PTB cases (true positive controls; N=34). Further the PCR specificity was determined >95% (true negative healthy controls; N=26). The positivity of M. tuberculosis by IS6110 & MPB 64 gene targets was found to be range of 88% to 100% in various clinical paucibacillary extra pulmonary samples i.e. pleural fluid, ascitic fluid, lymph node, pus, CSF and others. Our data on 64 samples (non respiratory, n=63 & respiratory samples, n=1) revealed 40.6% positivity by Cobas TaqMan Real Time PCR (utilizing 16S rRNA probe; Roche, USA). Interpretation & Conclusion: Our data revealed that utility of both PCR and Real Time PCR in rapid diagnosis of M. tuberculosis in paucibacillary extra pulmonary tuberculosis samples in Indian scenario.