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Fahr's syndrome is a rare neurodegenerative disorder characterized by symmetric bilateral calcifications in the basal ganglia. While this is largely a hereditary disease with autosomal dominant inheritance, a small percentage is sporadic in nature with no metabolic or other underlying causes identified. Fahr's syndrome has both neurological and psychiatric manifestations that include movement abnormalities, seizures, psychosis, and depression. Approximately 40% of patients with basal ganglia calcification present with psychiatric symptoms including mania, apathy, or psychosis. We present a case of a 50-year-old woman with no previous medical or psychiatric history who presented with an altered mental status that progressed to psychosis over three years. On one admission, the patient was found to have elevated liver enzymes and a positive antinuclear antibody panel but was without electrolyte abnormalities or movement disturbances. The patient was subsequently diagnosed with unspecified psychosis in the emergency department, which was later revised to Fahr's syndrome confirmed by neuroimaging. This report discusses her presentation, clinical symptoms, and management of Fahr's syndrome. Above all, it underscores the importance of complete workup and adequate follow-up of middle-aged and elderly patients with cognitive and behavioral disturbances, as Fahr's syndrome can be elusive in the early stages.
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INTRODUCTION: Microvascular free flaps (MVFF) are the current standard of care for reconstruction of oral ablative defects; however, pedicled myocutaneous flaps (PMCF) are still used widely in India. The rationale behind the preference for selecting PMCF in the present era is not well understood. The associated complications and swallowing outcomes are variable. METHODS: We retrospectively analysed the records of patients who underwent reconstructive surgery for oral cancer ablative defects over a 3-year period. RESULTS: Ninety-seven pedicled myocutaneous flaps [89 pectoralis major myocutaneous (PMMC) flaps, eight lower trapezius island myocutaneous (TMC) flaps] and 113 MVFFs were performed. The reasons for selecting PMCF were financial constraints 38.7%, MVFF salvage 22.5%, medically compromised 10.7%, vessel-depleted neck 6.4%, old age with PS2 + 5.3%, early recurrence 5.3%, borderline resectable 4.3%, palliative resection 2.1%. Overall complication rate was 20.4%. Of patients, 50.7% and 34.7% were on regular and semisolid diet, respectively; 66.6% had acceptable swallowing-related social well-being. CONCLUSION: PMCFs have an important role in developing countries with patients having financial constraints. The other potential reasons driven by patient factors were discussed. The swallowing outcomes are good, with majority of the people having socially acceptable swallowing function.
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BACKGROUND: The health of women is of particular concern because, in many societies, they are disadvantaged by discrimination rooted in sociocultural factors. Women need to breach many social barriers to empower and to get access for quality health care services. Health seeking behavior is one of the important determinants of women health. OBJECTIVES: To assess healthcare seeking behavior among rural women in Telangana. METHODS: Cross-sectional study with sample size of 200 was conducted in three villages attached to a medical college. Women of aged 20 years and above were included in the study. Data was collected by predesigned pretested semi-structured questionnaire. Data was presented in proportions with confidence interval and Chi-square test was applied to find the association between variables by using SPSS ver. 23. RESULTS: Only 34.5% [95% CI: 27.9, 41.5] of the subjects seek medical care as soon as symptoms appear and 69% [95% CI: 62.1, 75.3] of the participants were aware of nearby functioning health centres. Majority (60.5%) of the subjects Visits qualified medical practitioner during illness. CONCLUSIONS: The present study found that there is still a need to create awareness about the importance of healthcare and available health centers as significant proportion of women population approached unqualified medical practitioners and seeking home remedies as first consultancy source for their health remedies.
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OBJECTIVES: Adequate vitamin B12 is a requisite during pregnancy and its deficiency is linked with increased risk for adverse outcomes, likely mediated by impaired placental angiogenesis. Thus, we aimed to test associations of maternal vitamin B12 status with the placental expression of angiogenesis-associated genes ENG, VEGF, and FLT. SUBJECTS/METHODS: In this retrospective case-control study, placental and maternal trimester 1 blood samples (n = 104) were collected from small for gestational age (SGA) and appropriate for gestational age (AGA) full-term singleton pregnancies. Maternal trimester 1 vitamin B12 status was measured. Placentae and neonates were weighed at birth. Realtime quantitative PCR was performed to assess placental transcript abundance of ENG, VEGF, and FLT normalized to a panel of reference genes. Associations of placental transcript abundance of the genes with maternal trimester 1 vitamin B12 status were evaluated. RESULTS: Placental ENG transcript abundance associated negatively with maternal trimester 1 vitamin B12 status (ß = -0.461, P = 0.017, n = 104). This association was specific to the female births (ß = -0.590, P = 0.014, n = 60). Placental VEGF transcript levels were negatively associated with maternal trimester 1 vitamin B12 status only in the female births (ß = -1.995, P = 0.029). Placental FLT transcript levels were not associated with maternal trimester 1 vitamin B12 status. CONCLUSION: Maternal trimester 1 vitamin B12 status was associated negatively with placental ENG and VEGF expression predominantly in the female births. Therefore, we hypothesize that the placenta adapts to low maternal vitamin B12 status by up-regulating angiogenic pathways in a gender-specific manner.
Subject(s)
Vascular Endothelial Growth Factor A , Vitamin B 12 , Case-Control Studies , Endoglin , Female , Humans , Infant, Newborn , Placenta , Pregnancy , Retrospective Studies , Vascular Endothelial Growth Factor A/genetics , VitaminsABSTRACT
A major challenge to control the vector mosquitoes and their diseases. The discovery of bacteria like Bacillus sphaericus Neide (Bs) and B. thuringiensis serovar israelensis de Barjac (Bti), which are highly toxic to dipterans larvae, have opened up the possibility of their use as potential bio-larvicides in mosquito eradication programmes the world over. These bacteria have some important advantages over conventional insecticides in mosquito control operations, besides being safe for non-target organisms including human beings. But, the recent researchers have been reported mosquito resistant against these biological agents. Bacillus cereus VCRC B540 is one of the most potential bio-pesticides which were isolated from the gut contents of the marine fish (Lutjanuas sanguineus) collected in east coastal zone of the Bay of Bengal (Union Territory of Pondicherry, India) to control Culex, Anopheles and followed by Aedes species. The isolated strain was confirmed as Bacillus cereus based on the biochemical characteristics and 16S rDNA gene sequence. The larvicidal activity of B. cereus VCRC B540 was further characterized.
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Ovarian cancer (OC) is the most deadly gynecological cancer and unlike most other neoplasms, survival rates for OC have not significantly improved in recent decades. We show that RAD6, an ubiquitin-conjugating enzyme, is significantly overexpressed in ovarian tumors and its expression increases in response to carboplatin chemotherapy. RAD6 expression correlated strongly with acquired chemoresistance and malignant behavior of OC cells, expression of stem cell genes and poor prognosis of OC patients, suggesting an important role for RAD6 in ovarian tumor progression. Upregulated RAD6 enhances DNA damage tolerance and repair efficiency of OC cells and promotes their survival. Increased RAD6 levels cause histone 2B ubiquitination-mediated epigenetic changes that stimulate transcription of stem cell genes, including ALDH1A1 and SOX2, leading to a cancer stem cell phenotype, which is implicated in disease recurrence and metastasis. Downregulation of RAD6 or its inhibition using a small molecule inhibitor attenuated DNA repair signaling and expression of cancer stem cells markers and sensitized chemoresistant OC cells to carboplatin. Together, these results suggest that RAD6 could be a therapeutic target to prevent and treat acquired chemoresistance and disease recurrence in OC and enhance the efficacy of standard chemotherapy.
Subject(s)
Neoplasms, Glandular and Epithelial/enzymology , Ovarian Neoplasms/enzymology , Ubiquitin-Conjugating Enzymes/physiology , Antineoplastic Agents/pharmacology , Carboplatin/pharmacology , Carcinoma, Ovarian Epithelial , Cell Line, Tumor , DNA Repair , DNA Replication , Disease-Free Survival , Drug Resistance, Neoplasm , Female , Humans , Neoplasms, Glandular and Epithelial/drug therapy , Neoplasms, Glandular and Epithelial/mortality , Neoplasms, Glandular and Epithelial/pathology , Neoplastic Stem Cells/enzymology , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/mortality , Ovarian Neoplasms/pathology , Prognosis , Signal TransductionABSTRACT
BACKGROUND: Subacute subdural hematoma (SASDH) is an entity which is yet to capture the popular imagination among the neurosurgeons. Its management is often equated clinically to that of the chronic subdural hematoma (CSDH). However, their neurological deterioration is usually rapid, which seems to align them with acute subdural hematoma (ASDH). We proceed for their epidemiological evaluation. The advantages of a novel "double barrel technique (DbT)" over the conventional burrhole drainage are also presented. METHODS: This retrospective study was conducted on all the patients having clinical and radiological evidence of SASDH, admitted to a tertiary care referral institute, during the period August 2013 to December 2015. Postoperatively, patients were followed-up for 3-24 months. RESULTS: 46.87% of the patients belonged to the 35-54 year age group with a male predominance (3.6:1); 68.7% had a history of alcohol abuse, whereas aspirin users were 25%. 87.5% cases were unilateral, 18.75% were hemispheric, and 46.87% were present on the left side. Altered consciousness (100%) followed by headache (37.5%) were the most common presenting clinical features. CONCLUSION: SASDH is an uncommon neurosurgical entity (0.89% of traumatic brain injury cases in our study) and mimics both CSDH as well as ASDH. The true incidence of SASDH may have been underestimated due to its clinical imitation with CSDH. This study in a South Asian nation also provides the epidemiological data of this rare neurosurgical entity. Outcome of surgery is good; our retrospective study confirms that "DbT" is an adequate and safe treatment. However, a better designed, randomized control trial will be needed to reinforce our findings.
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BACKGROUND: Intramedullary spinal cord metastases (IMSCM) are typically drop lesions from intracranial metastases and are a rare manifestation of systemic malignancy (8.5% of central nervous system metastases). They arise from primaries such as the lungs, breast, kidney, melanoma, or lymphoma. On the other hand, they arise very rarely from papillary thyroid carcinoma (PTC), even though it is the most common type of primary thyroid malignancy. CASE DESCRIPTION: A 54-year-old male presented with pain in the lower back along with tingling, numbness, and weakness in the bilateral lower limbs. This was associated with urine incontinence for 1½ months. In the previous month, he developed a left-sided solitary thyroid nodule. Fine needle aspiration cytology and ultrasonography were suggestive of metastasis. Furthermore, the thoracolumbar magnetic resonance imaging showed T1-hypo and T2-hyper-intense D11-D12 level intramedullary lesion, with intense enhancement, which was consistent with an intramedullary lesion involving the conus. At surgery, a firm, brownish yellow, friable, vascular tumour was removed en toto. Upon discharge, the patient was neurologically intact except for residual bladder incontinence. CONCLUSION: In an extensive literature review (pubmed), IMSCM metastasis from PTC primary is confirmed as a rarity and this may be the fourth documented case. Moreover, this may be the first report of a case of PTC metastatic neurological deterioration "even before the treatment of the primary was undertaken." Early diagnosis and microsurgical resection can result in improvement of neurological deficits and in the quality of life of patients with IMSCM.
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BACKGROUND: Isolated orbital neurofibroma unassociated with systemic neurofibromatosis is relatively rare and may be difficult to clinically differentiate from other orbital tumors. Sensory branches of the trigeminal nerve-namely lacrimal, nasociliary, and frontal-are the most common nerves of origin for intraorbital neurofibroma, but we discovered a neurofibroma arising out of the right trochlear nerve, in absence of clinical stigmata of neurofibromatosis type 1, which is rare. CASE DESCRIPTION: A 41-year-old adult presented with painless progressive proptosis of the right eye for 10 years without history of visual problems or diplopia. The right eye had axial proptosis with periorbital swelling. On magnetic resonance imaging (MRI), a right orbital extraconal, expansile, lobulated, cystic space-occupying lesion was seen with an enhancing component, pushing the lateral rectus with T1 isointensity and T2 hyperintensity, suggesting a preoperative working diagnosis of pseudotumor or lymphoproliferative tumor. Intraoperatively, a rudimentary slender, white, elongated structure was passing through the length of the tumor. The elongated tumor engulfing the trochlear nerve was traced up to the lateral part of the superior orbital fissure. The tumor was excised completely and was found to be a neurofibroma. CONCLUSION: Isolated trochlear nerve neurofibromas, in the absence of clinical stigmata of NF1, are rare. Multiplicity, multilobulation, ring-configured contrast enhancement, and heterogenous MRI signal intensities help in the accurate preoperative imaging diagnosis. A possible cure is thus achievable with complete excision without damaging important adjacent neurovasculo-musculotendinous structures in the orbit. To the best of our knowledge, this is the fourth reported case of isolated trochlear nerve neurofibroma.
Subject(s)
Cranial Nerve Neoplasms/diagnostic imaging , Neurofibroma/diagnostic imaging , Trochlear Nerve Diseases/diagnostic imaging , Adult , Cranial Nerve Neoplasms/pathology , Humans , Magnetic Resonance Imaging , Male , Neurofibroma/pathology , Neurofibromatosis 1 , Trochlear Nerve Diseases/pathologyABSTRACT
Control of mosquitoes is the most important aspect of public health, as mosquitoes transmit many human diseases, including the fatal infection, Japanese encephalitis. This paper addresses the isolation of new mosquitocidal bacteria from soil samples in the Union Territory of Pondicherry, India, where, no clinical cases of vector borne infections have been reported. Bacterial isolates from soil samples were screened for potential mosquitocidal strains and bioassays against mosquito vectors (Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti) were carried out. Genomic DNA of potential mosquitocidal isolates was amplified and species identification was carried out using BLASTn program (NCBI). Phylogenetic analysis of 16S rRNA sequences of mosquitocidal bacteria revealed seven potential isolates. SDS-PAGE results have shown that there was considerable difference in the protein profiles. Numerical analysis revealed 4 distinct groups at similarity level 25%. The relationship between VBDs and prevalence of soil mosquitocidal bacteria in the study sites has elicited considerable interest in the diversity of mosquitocidal bacteria and their application for mosquito borne diseases control.
Subject(s)
Bacteria/classification , Bacteria/isolation & purification , Culicidae/microbiology , Culicidae/physiology , Encephalitis, Arbovirus/epidemiology , Soil Microbiology , Animals , Bacteria/chemistry , Bacteria/genetics , Bacterial Proteins/analysis , Biological Assay , Cluster Analysis , Cross-Sectional Studies , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Incidence , India/epidemiology , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Survival AnalysisABSTRACT
Mosquitoes transmit major communicable diseases such as dengue, malaria, filariasis, Japanese encephalitis, chikungunya, and so on. Vector control is important in epidemic disease situations as there is an urgent need to develop new and improved mosquito control methods that are economical and effective yet safe for non-targeted organisms. In the present study, silver nanoparticles (AgNPs) were synthesized from the aqueous leaf extract of neem plant (Azadirachta indica), and their effects on mosquito vectors (Aedes aegypti and Culex quinquefasciatus) were assessed. The synthesised AgNPs were characterized by UV-vis spectroscopy, scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction analysis (XRD). The nanoparticles have maximum absorption at 442 ± 1.5 nm with an average size of 41-60 nm. The XRD data showed six well-defined diffraction peaks, corresponding to a relative intensity of the crystal structure of metallic silver 36.42, 100.00, 53.70, 14.20, 16.05, and 6.79, respectively. The FT-IR data showed strong prominent peaks in different ranges, reflecting its complex nature. The mosquito larvae were exposed to varying concentrations of AgNPs synthesized from the neem leaves under investigation (0.07-25 mg/l) for 24 h; this revealed larvicidal activity of AgNPs with LC50 and LC90 values of 0.006 and 0.04 mg/l for A. aegypti, respectively. Further, the LC50 and LC90 values were also identified as 0.047 and 0.23 mg/l for Cx. quinquefasciatus, respectively. The result obtained from this study presents biosynthesized silver nanoparticle from A. indica as the biolarvicidal agent with the most potential for mosquito control.
Subject(s)
Azadirachta , Insecticides/chemistry , Metal Nanoparticles/chemistry , Mosquito Control/methods , Silver/chemistry , Aedes/drug effects , Animals , Culex/drug effects , Larva/drug effects , Metal Nanoparticles/ultrastructure , Microscopy, Electron, Scanning , Plant Extracts/chemistry , Plant Leaves/chemistry , Silver/pharmacology , Spectroscopy, Fourier Transform Infrared , X-Ray DiffractionABSTRACT
Control of mosquitoes is the most important aspect of public health, as mosquitoes transmit many human diseases, including the fatal infection, Japanese encephalitis. This paper addresses the isolation of new mosquitocidal bacteria from soil samples in the Union Territory of Pondicherry, India, where, no clinical cases of vector borne infections have been reported. Bacterial isolates from soil samples were screened for potential mosquitocidal strains and bioassays against mosquito vectors (Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti) were carried out. Genomic DNA of potential mosquitocidal isolates was amplified and species identification was carried out using BLASTn program (NCBI). Phylogenetic analysis of 16S rRNA sequences of mosquitocidal bacteria revealed seven potential isolates. SDS-PAGE results have shown that there was considerable difference in the protein profiles. Numerical analysis revealed 4 distinct groups at similarity level 25%. The relationship between VBDs and prevalence of soil mosquitocidal bacteria in the study sites has elicited considerable interest in the diversity of mosquitocidal bacteria and their application for mosquito borne diseases control.
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Mosquitocidal bacteria are environmentally friendly alternatives to chemical insecticides for controlling mosquitoes and therefore, there have been tremendous world-wide efforts to identify novel mosquitocidal bacteria from natural environment. In the present study, excreta from arid-birds were analyzed for identifying mosquitocidal bacteria. The selection of sample for bacterial screening is significant, because, arid-birds are the unique living species and gathering the foods from variety of sources from environment. Out of 1000 samples examined, twelve bacterial strains were identified as mosquitocidal and the 16S rRNA gene sequence alignment depicted that these isolates belonged to Bacillus species (Bacillus thuringiensis, B.sphaericus and B. cereus). Toxicity assay against mosquito vectors have shown that these isolates are potential. The B. sphaericus VCRC-B547 (NCBI: JN377789) has shown a higher toxicity against Cx. quinquefasciatus, An. stephensi, and Aed. aegypti. Result from SDS-PAGE has shown that there was considerable difference in the protein profiles among the new bacterial isolates. Phylogenetic tree with branch length 0.05 revealed three distinct groups with homology among the closely related Bacillus strains. This study therefore throws considerable interest on the diversity of microbial organisms from arid birds and its application in mosquito control.
Subject(s)
Bacillus/isolation & purification , Birds/parasitology , Culicidae/drug effects , Culicidae/parasitology , Feces/parasitology , Mosquito Control/methods , Animals , Bacillus/classification , Bacillus/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Bacterial Toxins/toxicity , DNA, Bacterial/genetics , Electrophoresis, Polyacrylamide Gel , Larva/parasitology , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/geneticsABSTRACT
Every day, food processing industries release wastes, which are environmental menance. Chicken feathers have been discarded in bulk as waste from poultry industries, globally. Degrading these wastes, as unused disposals, without acquiring any additional benefits has led to an idea to develop a new technology. We have reported earlier that Bacillus thuringiensis serovar israelensis (Bti) can be used for biodegradation of feather waste for biopesticide production. In the present study, purification and characterization of keratinase from feather degrading bacterium (Bti) is reported. Protein precipitate obtained at Ammonium sulphate saturation at 60% level and Sephacryl S-200 column chromatography resulted in 2.3 and 11.68 fold purification of the enzyme respectively. The purity was revealed in SDS-PAGE by a single band of molecular weight of 40 kDa and it was characterized. The optimum pH of the enzyme shifted to a more neutral range (6.0-8.0) with the highest activity (7.0). The optimum temperature of the reaction was determined to be 30ºC. The keratinase enzyme retained 51% residual activity (303 U/mg protein) at 70ºC (60 min) and the half-lives of the enzyme were 130 minutes at 40ºC, 90 min at 50ºC and of 60 min at 70ºC, respectively. Keratinase activity was enhanced by calcium and magnesium ions while EDTA, PMSF, ß- mercaptoethanol and manganese inhibited the activity. This is the first report investigating the keratinase from Bti degraded chicken feathers for the bio-synthesis of mosquitocidal toxins.
Subject(s)
Bacillus thuringiensis/enzymology , Chickens/microbiology , Mosquito Control/methods , Peptide Hydrolases/isolation & purification , Animals , Enzyme Stability , Feathers/microbiology , Hydrogen-Ion Concentration , Hydrolysis , Molecular Weight , Peptide Hydrolases/metabolism , Protease Inhibitors , Substrate Specificity , TemperatureABSTRACT
Bacterial insecticides like, Bacillus sphaericus and Bacillus thuringiensis serovar israelensis, have been used for the control of nuisance and vector mosquitoes for more than two decades. For many years, it was assumed that the use of microbial larvicides based on B. sphaericus would not lead to resistance in mosquitoes. However, recent reports have shown that B. sphaericus toxins are not free from this problem. Therefore, the resistance of mosquito populations to be will seriously threaten the sustainability of current mosquito control programme using these microbial insecticides. In the present study, we have characterised a novel protein responsible for resistance development in the filariasis vector of Culex quinquefasciatus. Laboratory selection experiments with B. sphaericus against the larvae were carried out up to 17 generations, and the occurrence of resistance was reported (resistance ratio (RR) at lethal concentration (LC)50 and LC90 = 1,987 and 2,051 folds, respectively). The protein profiles of B. sphaericus-resistant and susceptible population have confirmed with the expression of a new polypeptide (80 kDa) in the resistant strain only. Sequence result revealed that the newly expressed protein was 'hexamerin', and this factor might conceivably be responsible for the inheritance of resistance. This study is therefore valuable for comprehending the underlining factor and management of B. sphaericus resistance problem in mosquito population.
Subject(s)
Bacillus/physiology , Bacterial Toxins/pharmacology , Culex/immunology , Drug Resistance/genetics , Insect Proteins/genetics , Amino Acid Sequence , Animals , Bacillus/pathogenicity , Bacterial Toxins/biosynthesis , Bacterial Toxins/isolation & purification , Biological Control Agents , Breeding , Culex/genetics , Culex/microbiology , Female , Gene Expression Regulation/drug effects , Insect Proteins/biosynthesis , Larva/genetics , Larva/immunology , Larva/microbiology , Male , Molecular Sequence Data , Mosquito ControlABSTRACT
Entomopathogenic bacteria to control mosquitoes are a promising environmentally friendly alternative to synthetic pesticides. In the present study, a novel mosquitocidal bacterium was isolated from marine soil collected from east coastal areas at Pondicherry (India). 16S rRNA gene sequence alignment depicted that this isolate belonged to Bacillus cereus VCRC-B520 (NCBI: KC-119192). Biochemical studies on bacterial growth, biomass, and toxin production have revealed that this strain could possibly be helpful in the production of a biopesticide in mosquito control. Toxicity assay with B. cereus against mosquito larvae has shown that the filariasis vector, Culex quinquefasciatus, is more susceptible than the other two species (Anopheles stephensi and Aedes aegypti). The LC50 and LC90 values for C. quinquefasciatus were 0.30 and 2.21 mg/L, respectively. No effect of B. cereus was found on nontargeted organisms. SDS-PAGE analysis and protein purification result from the cell mass of B. cereus have shown that a well-perceptible polypeptide was the dependable factor (85 kDa) for mosquitocidal action. Protein characterization (M/S MALDI-TOF) has shown that it is an endotoxin-specific insecticidal protein, namely "Cry4Aa". Phylogenetic analyses of 16S rDNA gene sequence from this marine isolate have revealed the presence of homology among closely related Bacillus strains. Therefore, considerable interest has been shown on the identification of a potential mosquitocidal bacterium from marine environment (B. cereus), which was not reported earlier in view of the current scenario of the rapid development of resistance to Bacillus sphaericus in mosquito vector control program.
Subject(s)
Bacillus cereus/isolation & purification , Biological Control Agents , Culicidae/microbiology , Mosquito Control , Aedes/microbiology , Animals , Anopheles/microbiology , Bacillus cereus/classification , Bacillus cereus/genetics , Bacterial Proteins/toxicity , Culex/microbiology , DNA, Bacterial/genetics , Endotoxins/toxicity , India , Larva/microbiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Soil MicrobiologyABSTRACT
Sugarcane bagasse is a renewable resource that can be used to produce biopesticide for the control of mosquito vectors. In the present study, we demonstrated that cane processed bagasse could be used to produce Bacillus thuringiensis serovar israelensis (Bti) for control of mosquito vectors viz: Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti. Biochemical studies indicated that the Bti spore/crystal toxins produced from the test culture medium (Bagasse, BG + Soybean, SB) are higher than that from the conventional medium (Nutrient Yeast Extract Salt Medium, NYSM). The bacteria produced in these media (NYSM, BG, SB, BG+SB) were bioassayed against the mosquito species and the toxic effect was found to be effective. Cost-effective analysis indicates that the use of BG and SB, as bacterial culture medium, is successful and economical, for production of this mosquito pathogenic bacillus.
Subject(s)
Bacillus thuringiensis/growth & development , Bacillus thuringiensis/metabolism , Bacterial Proteins/metabolism , Cellulose/metabolism , Culture Media/chemistry , Endotoxins/metabolism , Hemolysin Proteins/metabolism , Industrial Microbiology/methods , Industrial Waste , Aedes/microbiology , Aedes/physiology , Animals , Anopheles/microbiology , Anopheles/physiology , Bacillus thuringiensis Toxins , Biological Assay/methods , Culex/microbiology , Culex/physiology , Insecticides/metabolism , Insecticides/pharmacology , Saccharum , Survival AnalysisABSTRACT
In the present study, to comprehend the genetic diversity of mosquitocidal bacteria, the genotypic analysis of 30 strains of Bacillus species isolated newly from diverse environmental sources has been conducted. Randomly amplified polymorphic DNA polymerase chain reaction was conducted to characterize the genotype diversity between the bacterial strains. The matrix of scores from each bacterial DNA was analyzed, and the correlation between the co-efficients and the similarity matrix data was computed. Clusters from dendrogram showing diversity among isolates could be distinguished genetically based on their origin of isolates. The first major cluster consists of 43 strains grouped under various subclusters (91.489 %). A second cluster contains only four strains (8.511 %). An average similarity value of 0.36 revealed the dendrogram split into 28 distinct "groups" or "clusters," allowing almost a complete separation of strains within the Bacillus group isolated from various sources and thus facilitating assessment of genetic diversity of species and subspecies level. The conclusion from the result was that there was broad diversity among the mosquitocidal strains, and cluster analysis revealed the associations among the isolates based on their origin. A high level of polymorphism with distinct genetic lineages consequent to the source of origin of bacterial strains is the significant impact of this study.
Subject(s)
Bacillus cereus/genetics , Bacillus thuringiensis/genetics , Culicidae/microbiology , Genetic Variation , Animals , Culicidae/genetics , DNA, Bacterial/genetics , Genotype , PhylogenyABSTRACT
Fruit rot disease (FRD), an emerging problem of tinda (Praecitrullus fistulosus) in India. FRD epidemics begin during rainy and warm weather and often spoil marketable produce. Symptoms appear as numerous, pale brown-to-dark brown, deeply penetrating circular soft rot lesions on fleshy fruit tissues. Noneffervescent bacterial exudates occasionally form on lesions. Repeated isolations from FRD-affected tinda fruits consistently yielded the same bacterial species. Inoculation of the isolated bacterium into asymptomatic tinda fruits produced identical soft rot symptoms. Fruits were inoculated with the isolate ITCC B0030 (0.1 OD) by removing a 2.0-cm deep tissue plug with a sterile cork borer (5 mm in diameter) and injecting the inoculum with a syringe in the cylindrical cavity. After inoculation, the plug (upper 5 mm) was reinserted, sealed with sterile paraffin, and covered with a small piece of wet absorbent cotton to prevent dehydration. High humidity (>90%) and 30 to 33°C temperature was maintained after inoculation in a glasshouse. After 4 to 10 days, fruits showed FRD symptoms. The reisolated bacterium from artificially inoculated symptomatic fruits was identical with the original inoculated bacterium. Identity of the bacterial pathogen for FRD was confirmed by phenotypic and genotypic methods. The causal bacterium was a gram-negative, non-sporing motile rod with a single polar flagellum. The bacterium produced yellowish green and blue-green diffusible pigments on King's B (KB) medium. On yeast dextrose calcium carbonate agar at 30°C, the colonies produced abundant, blue, diffusible pigment within 48 h. The bacterium grew at temperatures up to 42°C but not at 4°C. Excellent growth occurred on Salmonella-Shigella agar and MaConkey's medium, as reported also for Pseudomonas aeruginosa strain P8. The bacterium produced ammonia, hydrogen sulfide, arginine dihydrolase, urease, lipase, catalase, gelatinase, and casinase but not amylase, indole, or acetyl methyl carbinol. The bacterium was identified as P. aeruginosa using Biolog based Bacterial Identification System version 4.2 (Biolog Inc., Hayward, CA). The bacterium did not utilize cellobiose, dulcitol, maltose, sorbitol, sucrose, arabinose, and starch. Upon infiltration on tobacco leaves (Nicotiana tabacum cv. Xanthi) at 107 or more cells ml-1, the bacterium gave a strong hypersensitive reaction within 24 h. Transmission electron micrographs (TEM, KYKY 1000B, Japan) of the causal bacterium revealed a single, polar flagellum. Identity was further confirmed as P. aeruginosa based on 16S rRNA sequence (1,491 nt) analysis with universal primers F1 (5'-GAGTTTGATCCTGGCTCAG-3') and R13 (5'-AGAAAGGAGGTGATCCAGCC-3'). A blastN search of GenBank revealed a >99% nt identity with P. aeruginosa strain TAUC 7 (HQ914782). The 16S rRNA gene sequence (1,491 nt) was deposited in Bankit GenBank (JF797204). To our knowledge, this is the first report of fruit rot of tinda caused by P. aeruginosa in India (ITCC B0030) and a new record of bacterial rot of Praecitrullus fistulosus induced by a fluorescent and blue-green pigment producing P. aeruginosa. To date, P. syringae pv. lachrymans and a nonfluorescent P. pseudoalcaligenes subsp. citrulli were reported to infect Citrullus lanata (1) and Praecitrullus fistulosus (2), respectively. References: (1) D. L. Hopkins and N. C. Schenck. Phytopathology 62:542, 1972. (2) N. W. Schaad et al. Int. J. Syst. Bacteriol. 28:117, 1978.