Diagnosis of Childhood Leprosy – Changing Trends

Leprosy, a chronic infectious disease caused by mycobacterium leprae, mainly involves the skin, respiratory mucosa and the peripheral nervous system. Leprosy continues to remain a public health problem. In 2011, the global new case detection was 219075 and in India it was 127295. Thus, India accounts for > 58% of total cases of leprosy worldwide. Pediatric leprosy accounts for around 10% of the total disease burden.The main source of transmission of leprosy is from the untreated lepromatous patients and the most common route is through the nasal secretions. From the nasal mucosa, the bacteria spreads by hematogenous route to skin and the peripheral nerves. The disease has a long incubation period of 3-5 yrs (can be upto 20 yrs).After infection, the child first develops indeterminate leprosy which can either get cured spontaneously or on treatment or it can progress to one of the several clinical forms (tuberculoid, borderline or lepromatous). The clinical spectrum varies from tuberculoid, where there are a few, large, anesthetic skin patches with thickened peripheral nerves and no detectable bacilli to lepromatous type where there are multiple, small skin lesions with intact sensation and high bacillary load. In our study spanning over 20 years, we have observed no significant change in the clinical profile.Early diagnosis of leprosy requires a high index of suspicion on the part of the clinician. It is based on detection of 2 of the following features, namely, characteristic skin lesion, loss of sensation and thickened peripheral nerves or the detection of AFB in skin or nasal smear.We conducted a number of studies, evaluating various newer techniques for early detection of the disease. In one study, we found the FLA-ABS and Lepromin tests, to be of immense value for identification of "at risk" population in the community and for detecting subclinical infection. We also studied antibody response against 35k Da antigen by SACT and found that nearly 50% smear negative, 42% lepromin +ve and 70% lepromin -ve cases showed positive antibody response with no false positive response. Gene probes developed at our institute were tried on 100 patients. AII smear +ve cases, lepromin +ve cases and majority of smear- ve cases were detected by this method. 9 cases (4 indeterminate & 5 nonspecific) with inconclusive histopathology were also detected. In another study on 22 children, in-situ hybridization technique helped in diagnosing the children with negative skin smear and non specific histopathology. It also permitted the concomitant study of tissue pathology.Again, in our pioneer study, evaluation of the in-situ PCR technique revealed that histopathology detected 45% of total cases, in-situ PCR detected as much as 60% of the total cases. Thus, In-situ PCR offered excellent structural correlation permitting concomitant study of tissue pathology. As contamination by foreign DNA/RNA does not exist, it is a valuable tool for diagnosis of childhood leprosy.RLEP based PCR is yet another useful tool to detect cases where skin smears are -ve and skin biopsy is not feasible. In our study involving 73 patients, Z-N staining for AFB was positive in 17/73 (23.28%) cases and RLEP PCR in 56/73 (76.71%) cases. All 30 controls showed negative results. RLEP PCR technique had a significantly greater positivity (especially in early stages of leprosy) than ZN staining (p< 0.001).Suggested algorithm for diagnosis, whenever there is clinical suspicion, we can either go for smear for AFB or histology to confirm the diagnosis. Apositive smear for AFB is confirmatory. If it is negative then, we can subject the specimen for gene probes or PCR/In-Situ PCR/RLEP PCR. If the result is positive, it is diagnostic of leprosy. On the other hand, if histology shows characteristic features then it is confirmatory; if it is not characteristic, we can go for in-situ hybridization. Apositive in-situ hybridization is diagnostic of leprosy; if it is negative then we can opt for in-situ PCR.To conclude, leprosy often poses a diagnostic dilemma. It is important that after a good clinical assessment, new diagnostic tests be used to diagnose the condition at an early stage & prevent complications/ deformities.Â

Présence et devenir des hormones stéroïdiennes dans les stations de traitement des eaux usées.

Différents produits chimiques naturels et d’origine industrielle présentent une activité endocrinienne. Ces composés sont susceptibles d’agir sur le système endocrinien des animaux et des êtres humains (altération ou déficiences des fonctions reproductrices, occurrence de tumeurs malignes, etc.). De nos jours, il reste de nombreuses questions en suspens (ampleur de la contamination, devenir dans l’environnement, etc.) et le caractère persistant de ces substances en fait un problème majeur pour les générations futures. De plus, plusieurs études ont montré que les estrogènes naturels (tels que l’estrone, le 17β‑estradiol et l’estriol) et l’estrogène synthétique (17α‑éthinylestradiol) sont les composés les plus oestrogéniques présents dans les effluents de stations de traitement des eaux usées. En conséquence, ces estrogènes peuvent rejoindre les eaux de surfaces et les contaminer. En effet, des concentrations aussi faibles que 0,1 ng/L peuvent provoquer des effets oestrogéniques chez les organismes aquatiques exposés. Cette revue s’est donc concentrée sur la présence de ces types de composé dans les eaux de surface et usées, et plusieurs procédés de traitement sont discutés par rapport à leur capacité d’enlèvement des estrogènes. Ainsi, le traitement primaire seul ne permet pas l’enlèvement des estrogènes des eaux usées, alors qu’un traitement secondaire à boue activée réduit généralement les quantités d’estrogènes. Les procédés dits avancés montrent un potentiel élevé pour l’enlèvement des estrogènes, bien qu’ils peuvent former des sous-produits oestrogéniques et/ou cancérigènes. Finalement, les systèmes enzymatiques pourraient être des procédés innovateurs pour le traitement des estrogènes, puisqu’ils montrent un potentiel élevé pour l’enlèvement des composés aromatiques dans les eaux usées.Various natural chemicals and certain contaminants from industries present an endocrine activity. These substances are likely to act on animal and human endocrine system (deterioration or reproductive functions deficiencies, malignant tumor occurrences, etc). Nowadays, many questions related to these compounds are not resolved and the persistent character of these compounds makes it a major problem for future generations. Furthermore, diverse studies determined that natural estrogens (e.g., estrone, 17β-estradiol and estriol) and synthetic estrogen (17α-ethinylestradiol) are the most estrogenic compounds in wastewater treatment plant effluents. Thus, these estrogens can reach surface water and contaminate it. Indeed, concentrations as low as 0.1 ng/L can induce estrogenic effects in exposed aquatic organisms. This review concentrated on this type of compounds in surface water and wastewater, and a number of treatment processes are discussed with regard to their potential on endocrine disrupting chemicals removal. In the primary treatment effluent, limited removal of estrogens has been observed, while secondary treatment involving activated sludge generally reduces all estrogens effluents concentrations. The advanced processes display a high potential for removing estrogens, although they may produce estrogenic and/or carcinogenic by-products. Enzymatic processes could be an innovative strategy for estrogens removal since they show a high potential to remove aromatic compounds from wastewater

Pyocolpos in a 4 year old: a rare scenario

Pyocolpos is cystic dilatation of vagina due to accumulation of pus resulting from genital tract obstruction. Hydrocolpos or pyocolpos are usually encountered either in the neonatal period due to stimulation from maternal hormones or at menarche. Their occurrence beyond infancy and before puberty is rare. We report this rare case of pyocolpos in a 4 year old child

Evaluation of cartridge-based nucleic acid amplification test for diagnosis of tuberculosis in children in various body fluids

Introduction: The paucibacillary nature presents a major challenge in the diagnosis of tuberculosis (TB) in children. The utilization of cartridge-based nucleic acid amplification test (CBNAAT) for the diagnosis of TB presents itself with added advantages such as detection to resistance to rifampicin and short turnaround time. Objectives: The aim of the study is to evaluate the diagnostic yield of CBNAAT in various body fluids and to compare with BACTEC-MGIT 960 and acid-fast bacilli (AFB) microscopy in children with suspected TB and to see the prevalence of rifampicin resistance in the study population using CBNAAT. Materials and Methods: This cross-sectional study included participants <14 years with suspected TB. Gastric aspirate samples obtained from pulmonary TB cases and body fluid specimens obtained from extrapulmonary TB cases were processed for the detection of Mycobacterium tuberculosis (MTB) using CBNAAT, BACTEC-MGIT 960, and AFB microscopy. The results obtained using CBNAAT were compared to other laboratory tests using an appropriate statistical method. Results: Fifty patients diagnosed with TB (34 pulmonary, 10 pleural effusion, and 6 abdominal) were included in the study, and clinical fluid specimens obtained from study participants were processed for the detection of MTB. Out of 34 gastric aspirate samples, 28 (82%) were positive by CBNAAT which was statistically higher than BACTEC-MGIT 960 (P < 0.05). Among extrapulmonary TB cases, only 2 pleural fluid specimens were positive by CBNAAT, whereas BACTEC-MGIT 960 and AFB microscopy could not detect MTB. Out of 34, 4 (11.76%) patients with newly diagnosed pulmonary TB were found to be rifampicin resistant using CBNAAT. Conclusions: CBNAAT showed promising results as a diagnostic tool in detecting MTB and rifampicin resistance in pulmonary TB using gastric aspirate. It, however, did not show good results in children with extrapulmonary TB in the clinical fluid specimen. The present study also showed the presence of high rifampicin resistance in treatment naïve pulmonary TB patients

Diagnostic value of in situ polymerase chain reaction in childhood leprosy

Objective: Our aim was to assess the diagnostic value of in situ polymerase chain reaction (PCR) in leprosy, particularly for enhancing histopathological diagnosis. Methods: We prospectively studied 20 children (aged <16 years) with leprosy. Clinical examination of each case was performed, and skin smear for acid-fast bacillus was prepared. A biopsy of the lesion site was performed for histopathological examination and in situ PCR testing. Results: Histopathological examination confirmed the clinical diagnosis in only 45% of the cases; nonspecific histopathology was reported for the remaining 55% of the cases. In situ PCR showed a positivity of 57.1% in the early/localized form of leprosy (indeterminate/borderline tuberculoid) and 61.5% in the borderline borderline/borderline lepromatous group. When compared with the histopathological examination, a significant enhancement of 15% in diagnosis was seen. With in situ PCR, the diagnosis could be confirmed in 4 (36.3%) of 11 cases with nonspecific histopathological features (which is common in early disease) in addition to confirmation of 8 (88.8%) of 9 histopathologically confirmed tissue sections. Histopathology and in situ PCR combined together confirmed the diagnosis in 13 (65%) of the 20 cases. Conclusions: In situ PCR is an important diagnostic tool, especially in early and doubtful cases of leprosy

Pre-treatment Technologies to Enhance Anaerobic Digestion

Sustainable energy production is the major priority in the world due to global warming, climate change, and fossil fuels depletion. Anaerobic digestion (AD) of sludge is the sustainable process producing the energy and minimizing the fossil fuel usage. However, conventional AD of sludge is not sustainable since it takes longer time for digestion which increases the energy input and greenhouse emissions. Therefore, pretreatment technologies have emerged to enhance methane production and thus the energy output from the AD process. In this chapter, pre-treatment technologies adopted mainly physical, chemical, thermal, and other advanced processes to enhance methane production in the last decade are elaborated. In addition, energy balance of the process and the feasibility of the pre-treatment technologies and their current status are discussed

IN VITRO ASSESSMENT OF BIOPESTICIDE BACILLUS THURINGIENSIS VAR. KURSTAKI HD- 1 EFFECTIVENESS ON PHYTOPHTHORA PALMIVORA, AGENT OF COCOA BLACK POD ROT IN CÔTE D’IVOIRE

Phytophthora Palmivora is responsible for the cocoa black pod rot in Côte d'Ivoire, with production losses varying from 20 to 40% depending on the regions. Effectiveness of the environmentally friendly biopesticide Bacillus thuringiensis var. kurstaki HD-1 was evaluated in vitro on the pathogen. The medium pea supercooled has been incorporated with the bioproduct at different concentrations: C1 = 100%, C2 = 50%, C3 =25%, C4 = 12.5% and C5 = 6.25%. This medium into Petri dishes was inoculated either by spreading, with 0.1 mL of P. palmivora inoculum at103 zoospores /mL, or with a mycelial disc of P. palmivora calibrated at 6 mm diameter in the center of the box. The results showed that the germination of zoospores was inhibited at the rate of 100% in the presence of the concentrations C1 and C2. This rate was 74.82 ± 21.53%, 73.17 ± 20.88% and 59.12 ± 39.05%, respectively, for C3, C4 and C5. The efficacy of the bioproduct is revealed by a significant reduction of mycelia growth ranging between 57.77± 2.4 to 100% in the presence of the concentrations C5 to C1. The IC50 of germination was 3.36% of concentration and the IC90 was 43.22%; while those which inhibit 50% and 90% of mycelial growth were 5.67% and 77.94% respectively. The biopesticide Btk HD-1 has a proven fungistatic effect against P. palmivora. Its effectiveness increases with its concentration. So it can be a valuable component of an integrated cocoa black pod management

IN VITRO ASSESSMENT OF BIOPESTICIDE BACILLUS THURINGIENSIS VAR. KURSTAKI HD- 1 EFFECTIVENESS ON PHYTOPHTHORA PALMIVORA, AGENT OF COCOA BLACK POD ROT IN CÔTE D’IVOIRE

Phytophthora Palmivora is responsible for the cocoa black pod rot in Côte d'Ivoire, with production losses varying from 20 to 40% depending on the regions. Effectiveness of the environmentally friendly biopesticide Bacillus thuringiensis var. kurstaki HD-1 was evaluated in vitro on the pathogen. The medium pea supercooled has been incorporated with the bioproduct at different concentrations: C1 = 100%, C2 = 50%, C3 =25%, C4 = 12.5% and C5 = 6.25%. This medium into Petri dishes was inoculated either by spreading, with 0.1 mL of P. palmivora inoculum at103 zoospores /mL, or with a mycelial disc of P. palmivora calibrated at 6 mm diameter in the center of the box. The results showed that the germination of zoospores was inhibited at the rate of 100% in the presence of the concentrations C1 and C2. This rate was 74.82 ± 21.53%, 73.17 ± 20.88% and 59.12 ± 39.05%, respectively, for C3, C4 and C5. The efficacy of the bioproduct is revealed by a significant reduction of mycelia growth ranging between 57.77± 2.4 to 100% in the presence of the concentrations C5 to C1. The IC50 of germination was 3.36% of concentration and the IC90 was 43.22%; while those which inhibit 50% and 90% of mycelial growth were 5.67% and 77.94% respectively. The biopesticide Btk HD-1 has a proven fungistatic effect against P. palmivora. Its effectiveness increases with its concentration. So it can be a valuable component of an integrated cocoa black pod management

Utilisation du broyat de péricarpes des cabosses de cacao comme milieu de culture alternatif pour la production de Bacillus thuringiensis var. kurstaki HD-1

La recherche de matières premières alternatives pour la production de biopesticides microbiens suscite beaucoup d’intérêts. La présente étude s’est proposée d’utiliser le broyat de péricarpes des cabosses de cacao comme substrat de fermentation pour produire Bacillus thuringiensis var. kurstaki HD-1 (Btk HD-1). Les échantillons de péricarpes des cabosses ont été collectés dans une plantation cacaoyère de Toumokro à Yamoussoukro en Côte d’Ivoire. Les paramètres physico-chimiques ont été déterminés selon les méthodes standards. Le substrat de fermentation a été autoclavé, 2 fois successivement, à 121 °C pendant 30 min et ensemencé avec 4% de l’inoculum de Btk HD-1. Il ressort de cette étude que les teneurs en carbone et en azote total ont été respectivement de 79.7% et 1.4%. Les concentrations en minéraux du substrat ont été plus élevées en calcium (8406.5 mg/kg) et en potassium (8248.8 mg/kg) qu’en magnésium (632.6 mg/kg), sodium (471.5 mg/kg), fer (57 mg/kg), cuivre (52.8 mg/kg), zinc (51.8 mg/kg) et manganèse (44.3 mg/kg). Le dénombrement des cellules et des spores viables a donné respectivement 3.2.1011 et 3.02.1011 UFC/ml, pour le surnageant, et 6.5.1013 et 5.02.1013 UFC/ml, pour le culot. Ce substrat végétal offre une bonne perspective pour la production d’un biopesticide à base de Btk HD-1.© 2016 International Formulae Group. All rights reserved.Mots-clés: Péricarpes des cabosses de cacao, biopesticide, Bacillus thuringiensis var. kurstaki HD-1, fermentation en milieu solide, Côte d’IvoireEnglish Title:  Use of crushed cocoa hulls as an alternative culture medium for Bacillus thuringiensis var. kurstaki HD-1 productionEnglish AbstractResearch for alternative raw material for microbial biopesticides production raises outstanding interests.This study is proposed to use the crushed pericarps cocoa pods as fermentation substrate to produce Bacillus thuringiensis var. kurstaki HD-1 (Btk HD-1). The samples pericarps pods were collected in a cocoa plantation of Toumokro in Yamoussoukro area, Côte d'Ivoire. The physico-chemical parameters were determined using standard methods. The fermentation substrate was autoclaved 2 times successively at 121 °C for 30 minutes and inoculated with 4% of Btk HD-1 inoculum. It appears from this study that the carbon and total nitrogen were 79.7% and 1.4% respectively. Mineral concentrations in substrate were higher in calcium (8406.5 mg/kg) and potassium (8248.8 mg/kg) than magnesium (632.6 mg/kg), sodium (471.5 mg/kg), iron (57 mg/kg), copper (52.8 mg/kg), zinc (51.8 mg/kg) and manganese (44.3 mg/kg). Cell counts and viable spores gave respectively 3.2.1011 and 3.02.1011 CFU/ml, to the supernatant, and 6.5.1013 and 5.02.1013 CFU/ml, for the pellet. This vegetal substrate offers a good prospect for Btk HD-1 based biopesticide production.© 2016 International Formulae Group. All rights reserved.Keywords: Cocoa hulls pod, biopesticide, Bacillus thuringiensis var. kurstaki HD-1, solid medium fermentation, Côte d’Ivoir