De novo transcriptome sequencing of drought tolerance-associated genes in little millet (Panicum sumatrense L.).

The genome size of the little millet Panicum sumatrense is unknown, although its genome is fairly diploid (2n = 4x = 36). Despite tremendous nutritional value and adaptability to adverse climatic conditions, P. sumatrense use was limited by their low palatability, coarse grain, and lack of variety of culinary preparations. Hence, understanding how to vary their usage to offer food and nutritional security in the continuously changing modern world, the proposed study was aimed to determine potential genes and metabolites implicated in drought resistance. The drought-resistant genotype of tiny millet OLM-203/Tarini was offered in pots under both relaxed and demanding circumstances. The experimental seedlings were 32 days old and had been under water stress for 23 days. A total of 7606 genes were compared between 23 and 32 days for roots and 7264 total genes were compared between 23 and 32 days for leaves, according to a research on differential expression genes (DEGs). Twenty essential genes for drought tolerance were up-or down-regulated in the control and treated roots of the OLM-203 genotype. For instance, the genes RS193 and XB34 were up-regulated in leaves while, WLIM1 was found to be down-regulated. Gene SKI35 was up-regulated in roots, whereas MPK6 and TCMOp1 were down-regulated in root samples. The roots and leaves of the tiny millet OLM-203 genotype expressed 36 up-regulated and 21 down-regulated serine transcripts, respectively. Gene annotations for leaf samples were classified as having "molecular function" (46%), "cellular component" (19%), and "biological process" (35%), while root sample gene annotations were categorized as having "biological process" (573 contigs), "molecular function" (401 contigs), and "cellular components" (166 contigs). Noteworthy, polyamines play a crucial role in drought stress tolerance in the genotype, and it was found that top ten DEGs encoding for polyamines were common in two tissues (leaf and root). Collectively, transcriptomics profiling (RNA-seq) unveiled transcriptional stability drought stress provide a new insight in underlying modus of operandi in little millet genotype "OLM-203/Tarini" in response to heat stress.

LCCNN: a Lightweight Customized CNN-Based Distance Education App for COVID-19 Recognition.

In the global epidemic, distance learning occupies an increasingly important place in teaching and learning because of its great potential. This paper proposes a web-based app that includes a proposed 8-layered lightweight, customized convolutional neural network (LCCNN) for COVID-19 recognition. Five-channel data augmentation is proposed and used to help the model avoid overfitting. The LCCNN achieves an accuracy of 91.78%, which is higher than the other eight state-of-the-art methods. The results show that this web-based app provides a valuable diagnostic perspective on the patients and is an excellent way to facilitate medical education. Our LCCNN model is explainable for both radiologists and distance education users. Heat maps are generated where the lesions are clearly spotted. The LCCNN can detect from CT images the presence of lesions caused by COVID-19. This web-based app has a clear and simple interface, which is easy to use. With the help of this app, teachers can provide distance education and guide students clearly to understand the damage caused by COVID-19, which can increase interaction with students and stimulate their interest in learning.

Randomized Trial of Metformin With Anti-Tuberculosis Drugs for Early Sputum Conversion in Adults With Pulmonary Tuberculosis.

BACKGROUND: Metformin, by reducing intracellular Mycobacterium tuberculosis growth, can be considered an adjunctive therapy to anti-tuberculosis treatment (ATT). We determined whether metformin with standard ATT reduces time to sputum culture conversion and tissue inflammation in adults with pulmonary tuberculosis (PTB). METHODS: In a randomized, 8-week, clinical trial, newly diagnosed, culture-positive PTB patients were randomized to standard ATT (HREZ = control arm) or standard ATT plus daily 1000 mg metformin (MET-HREZ = Metformin with Rifampicin [METRIF] arm) for 8 weeks during 2018-2020 at 5 sites in India. The primary end point was time to sputum culture conversion by liquid culture during 8 weeks of ATT. Plasma inflammatory markers were estimated in a subset. A Cox proportional hazard model was used to estimate time and predictors of culture conversion. RESULTS: Of the 322 patients randomized, 239 (74%) were male, and 212 (66%) had bilateral disease on chest radiograph with 54 (18%) showing cavitation. The median time to sputum culture conversion by liquid culture was 42 days in the METRIF arm and 41 days in the control arm (hazard ratio, 0.8; 95% confidence interval [CI], .624-1.019). After 8 weeks of ATT, cavitary lesions on X-ray (7, 5.3% vs 18, 12.9%; relative risk, 0.42; 95% CI, .18-.96; P = .041) and inflammatory markers were significantly lower in the METRIF arm. Higher body mass index and lower sputum smear grading were associated with faster sputum culture conversion. CONCLUSIONS: The addition of metformin to standard ATT did not hasten sputum culture conversion but diminished excess inflammation, thus reducing lung tissue damage as seen by faster clearance on X-ray and reduced inflammatory markers. CLINICAL TRIALS REGISTRATION: Clinical Trial Registry of India (CTRI/2018/01/011176).

A Seven-Layer Convolutional Neural Network for Chest CT-Based COVID-19 Diagnosis Using Stochastic Pooling.

(Aim) COVID-19 pandemic causes numerous death tolls till now. Chest CT is an effective imaging sensor system to make accurate diagnosis. (Method) This article proposed a novel seven layer convolutional neural network based smart diagnosis model for COVID-19 diagnosis (7L-CNN-CD). We proposed a 14-way data augmentation to enhance the training set, and introduced stochastic pooling to replace traditional pooling methods. (Results) The 10 runs of 10-fold cross validation experiment show that our 7L-CNN-CD approach achieves a sensitivity of 94.44±0.73, a specificity of 93.63±1.60, and an accuracy of 94.03±0.80. (Conclusion) Our proposed 7L-CNN-CD is effective in diagnosing COVID-19 in chest CT images. It gives better performance than several state-of-the-art algorithms. The data augmentation and stochastic pooling methods are proven to be effective.

Covid-19 Opens up domestic market for Indian shrimp.

COVID-19 pandemic presents both a challenge and an opportunity to the Indian shrimp sector. With revitalizing the institutional arrangements and redirecting the focus, the Indian shrimp industry can flourish just by adapting to the needs of the local demand, even when the export prospects are uncertain. This paper takes a historical perspective of Indian shrimp farming and exports and suggests a domestic alternative/supplementary market for Indian farmed shrimp, resulting from COVID-19.

SARS-Net: COVID-19 detection from chest x-rays by combining graph convolutional network and convolutional neural network.

COVID-19 has emerged as one of the deadliest pandemics that has ever crept on humanity. Screening tests are currently the most reliable and accurate steps in detecting severe acute respiratory syndrome coronavirus in a patient, and the most used is RT-PCR testing. Various researchers and early studies implied that visual indicators (abnormalities) in a patient's Chest X-Ray (CXR) or computed tomography (CT) imaging were a valuable characteristic of a COVID-19 patient that can be leveraged to find out virus in a vast population. Motivated by various contributions to open-source community to tackle COVID-19 pandemic, we introduce SARS-Net, a CADx system combining Graph Convolutional Networks and Convolutional Neural Networks for detecting abnormalities in a patient's CXR images for presence of COVID-19 infection in a patient. In this paper, we introduce and evaluate the performance of a custom-made deep learning architecture SARS-Net, to classify and detect the Chest X-ray images for COVID-19 diagnosis. Quantitative analysis shows that the proposed model achieves more accuracy than previously mentioned state-of-the-art methods. It was found that our proposed model achieved an accuracy of 97.60% and a sensitivity of 92.90% on the validation set.

Impact of culture condition modulation on the high-yield, high-specificity and cost-effective production of terpenoids from microbial sources: A review.

Recent years have seen a remarkable increase in the non-natural production of terpenoids from microbial route. This is due to the advancements in synthetic biology tools and techniques, which have overcome the challenges associated with the non-native production of terpenoids from microbial hosts. Although, microbes in their native form have ability to grow in wide range of physicochemical parameters such as, pH, temperature, agitation, aeration etc; however, after genetic modifications, culture conditions need to be optimized in order to achieve improved titers of desired terpenoids from engineered microbes. The physicochemical parameters together with medium supplements, such as, inducer, carbon and nitrogen source, and cofactor supply not only play an important role in high-yield production of target terpenoids from engineered host, but also reduce the accumulation of undesired metabolites in fermentation medium, thus facilitate product recovery. Further, for the economic production of terpenoids, the biomass derived sugars can be utilized together with the optimized culture conditions. In the present mini-review, we have highlighted the impact of culture conditions modulation on the high-yield and high-specificity production of terpenoids from engineered microbes. Lastly, utilization of economic feedstock has also been discussed for the cost-effective and sustainable production of terpenoids.

Detection of COVID-19 from speech signal using bio-inspired based cepstral features.

The early detection of COVID-19 is a challenging task due to its deadly spreading nature and existing fear in minds of people. Speech-based detection can be one of the safest tools for this purpose as the voice of the suspected can be easily recorded. The Mel Frequency Cepstral Coefficient (MFCC) analysis of speech signal is one of the oldest but potential analysis tools. The performance of this analysis mainly depends on the use of conversion between normal frequency scale to perceptual frequency scale and the frequency range of the filters used. Traditionally, in speech recognition, these values are fixed. But the characteristics of speech signals vary from disease to disease. In the case of detection of COVID-19, mainly the coughing sounds are used whose bandwidth and properties are quite different from the complete speech signal. By exploiting these properties the efficiency of the COVID-19 detection can be improved. To achieve this objective the frequency range and the conversion scale of frequencies have been suitably optimized. Further to enhance the accuracy of detection performance, speech enhancement has been carried out before extraction of features. By implementing these two concepts a new feature called COVID-19 Coefficient (C-19CC) is developed in this paper. Finally, the performance of these features has been compared.

Molecular characterization and antibiotic resistance of Vibrio parahaemolyticus from Indian oyster and their probable implication in food chain.

Vibrio parahaemolyticus is one of the leading causes of diarrhoea and gastroenteritis in human on consumption of raw or insufficiently cooked seafood. This study was aimed at isolating and characterizing the pathogenic and pandemic V. parahaemolyticus from oysters (n = 90) in coastal parts of West Bengal, India; their antibiotic resistance and potential for involvement in the food chain. During bacteriological culture, typical V. parahaemolyticus colony was recovered in 88.9% samples followed by presumptive identification in 71 (78.9%) samples by characteristic biochemical (K/A) test. All the presumptive isolates (n = 71) were confirmed by species specific Vp-toxR PCR assay. Of these, 10 (14.08%) were tdh+ and none for the trh. Further, 5 (50%) of these tdh+ isolates were found to carry the pandemic potential gene in PGS-PCR assay; however, none in GS-PCR. Majority (80%) of these pathogenic (tdh+) isolates belonged to pandemic serovars (OUT: KUT; OUT: K24; O1: KUT; O1:K25; O10: KUT) and only 20% to non-pandemic serovars (OUT: K15; O9:K17). All the isolates (100%) exhibited resistance to cefpodoxime followed by ampicillin and cefotaxime (90%), ceftizoxime (60%), tetracycline (50%), ceftriaxone (40%), ciprofloxacin and nalidixic acid (10% each). Overall, the study findings suggested that 11.1% (10/90) of commonly marketed oysters in this area were harbouring pathogenic V. parahaemolyticus. Moreover, 5.5% (5/90) of the oyster population were harbouring pandemic strains of this pathogen. Besides, the pathogenic isolates from oysters were exhibiting a considerable genetic relatedness (53 to 70%) to human clinical isolates in PFGE analysis that relates to a substantial public health risk. Further, their multidrug resistance added gravity to the antimicrobial resistance (AMR), a globally growing public health threat and this is a critical area of concern especially during the treatment of foodborne gastroenteritis.

Association of CYP19A1 gene variations with adjuvant letrozole-induced adverse events in South Indian postmenopausal breast cancer cohort expressing hormone-receptor positivity.

PURPOSE: Musculoskeletal adverse events (MS-AEs) and vasomotor symptoms (VMSs) are the major side-effects of newer generation non-steroidal aromatase inhibitor (AI), letrozole. Single-nucleotide polymorphisms (SNPs) in CYP19A1 gene coding for the enzyme aromatase are related to AI treatment-associated adverse drug reactions. Therefore, we aimed to determine whether SNPs in the CYP19A1 gene are associated with adjuvant letrozole-induced 'specific' AEs in postmenopausal hormone receptor-positive (HR+) breast cancer patients. METHODS: Genomic DNA was isolated from 198 HR+ breast cancer patients by the phenol-chloroform method, and eleven SNPs in the CYP19A1 gene were genotyped by TaqMan genotyping assays on the qRT-PCR system. Toxicity was assessed according to the National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0, and the data were analyzed using SPSS v19.0 and Haploview v4.2 statistical software. RESULTS: Subjects carrying the genetic variants of CYP19A1 gene SNP rs700519 had significantly higher odds (OR 2.33; 95% CI [1.29-4.20], P = 0.0057) of MS-AEs under dominant statistical effect. The frequency of the two distinct haplotypes that include the variant allele 'T' at rs700519 locus, H5-GCTATCTGGCG (P = 0.042) and H11-GCTATTGCACG (P = 0.013) were significantly higher in patients with musculoskeletal toxicity than in those without MS-AEs and thus predisposing to MS-AEs. Similarly, H6-GCCAGCTGGCG (P = 0.037) haplotype exhibited higher frequencies in patients presented with VMSs. However, no such association was observed between CYP19A1 genotypes and VMSs. CONCLUSIONS: To the best of our knowledge, this is the first study assessing the impact of CYP19A1 genetic variations with adjuvant letrozole treatment-associated AEs in Indian women. Genetic variations in the CYP19A1 gene is associated with letrozole-induced AEs and warrants further investigation in larger cohorts to validate this finding.

Pouch of Douglas internal hernia successfully treated laparoscopically.

A rare case of internal herniation of a peritoneal defect in the pouch of Douglas is being reported. It presented as a case of intestinal obstruction, which after investigation, on laparoscopic exploration was found to be a case of internal hernia getting obstructed in the pouch of Douglas. It was successfully treated by marsupialisation of the defect laparoscopically. A 33-year-old female presented with pain abdomen and vomiting. On investigations, she was found to be having a small intestinal obstruction. Conservative trial failed and then diagnostic laparoscopy was done, which revealed a peritoneal defect in the pouch of Douglas with the incarcerated distal ileal loop. Contents were reduced, and laparoscopic marsupialisation of the peritoneal defect was done. A rare case of defect in peritoneum with no defect in muscular layer in the pouch of douglas. Internal hernia is being reported and successfully treated laparoscopically by marsupialisation for the first time.

3-Methyl-3-buten-1-ol (isoprenol) confers longevity and stress tolerance in Caenorhabditis elegans.

The present investigation demonstrates the longevity-promoting effects of 3-methyl-3-buten-1-ol (isoprenol) in the animal model Caenorhabditis elegans that might be served as a lead nutraceutical in geriatric research. Our results showed that 0.5 mM isoprenol extended the mean lifespan of worms by 25% in comparison to control worms. Isoprenol also significantly enhanced survival of the worms under various stress conditions. It was found that the longevity-promoting effects of isoprenol were associated with improved age-associated physiological behaviour and reduced intracellular reactive oxygen species (ROS) accumulation. Finally, studies with gene-specific mutants revealed the involvement of pro-longevity transcription factors (TFs) DAF-16 and SKN-1 with simultaneous over-expression of GST-4 and SOD-3 in isoprenol treated worms. In silico analysis revealed the binding affinity of isoprenol with DAF-16 and SKN-1 TFs. Together, the findings suggest that isoprenol is able to enhance the lifespan of C. elegans and embarks its potential in the developments of formulations for age-related ailments.

Pretreatment loss to follow-up of tuberculosis patients in Chennai, India: a cohort study with implications for health systems strengthening.

BACKGROUND: Pretreatment loss to follow-up (PTLFU) is a barrier to tuberculosis (TB) control in India's Revised National TB Control Programme (RNTCP). PTLFU studies have not been conducted in India's mega-cities, where patient mobility may complicate linkage to care. METHODS: We collected data from patient registries for May 2015 from 22 RNTCP designated microscopy centers (DMCs) in Chennai and audited addresses and phone numbers for patients evaluated for suspected TB to understand how missing contact information may contribute to PTLFU. From November 2015 to June 2016, we audited one month of records from each of these 22 DMCs and tracked newly diagnosed smear-positive patients using RNTCP records, phone calls, and home visits. We defined PTLFU cases as including: (1) patients who did not start TB therapy within 14 days and (2) patients who started TB therapy but were lost to follow-up or died before official RNTCP registration. We used multivariate logistic regression to identify factors associated with PTLFU. RESULTS: In the audit of May 2015 DMC registries, out of 3696 patients evaluated for TB, 1273 (34.4%) had addresses and phone numbers that were illegible or missing. Out of 344 smear-positive patients tracked from November 2015 to June 2016, 40 (11.6%) did not start TB therapy within 14 days and 36 (10.5%) started therapy but were lost to follow-up or died before official RNTCP registration, for an overall PTLFU rate of 22.1% (95%CI: 17.8%-26.4%). Of all PTLFU patients, 55 (72.4%) were lost to follow-up and 21 (27.6%) died before starting treatment or before RNTCP registration. In the regression analysis, age > 50 years (OR 2.9, 95%CI 1.4-6.5), history of prior TB (OR 3.9, 95%CI 2.2-7.1), evaluation at a high patient volume DMC (OR 3.2, 95% CI 1.7-6.3), and absence of legible patient contact information (OR 4.5, 95%CI 1.3-15.1) were significantly associated with PTLFU. CONCLUSIONS: In an Indian mega-city, we found a high PTLFU rate, especially in patients with a prior TB history, who are at greater risk for having drug-resistance. Enhancing quality of care and health system transparency is critical for improving linkage of newly diagnosed patients to TB care in urban India.

Gonadotropin receptors of Labeo rohita: Cloning and characterization of full-length cDNAs and their expression analysis during annual reproductive cycle.

Follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh), secreted from pituitary, stimulate gonadal function by binding to their cognate receptors FSH receptor (FSHR), and LH/choriogonadotropin receptor (LHCGR). Rohu (Labeo rohita) is a commercially important seasonal breeder freshwater fish species, but till date, the regulation of expression of gonadotropins and their receptors gene during different phases of annual reproductive cycle has not been investigated. We envisaged the critical role of these molecules during seasonal gonadal development in this carp species. We cloned full- length cDNAs of fshra and lhcgrba from rohu testis using RACE (Rapid amplification of cDNA ends) and analyzed their expression along with fsh and lh by quantitative real time PCR (qRT-PCR) assay at various gonadal developmental stages of the annual reproductive cycle. Full-length rohu fshra and lhcgrba cDNA encodes 670 and 716 amino acids respectively, and in adult fish, they were widely expressed in brain, pituitary, gonad, liver, kidney, head kidney, heart, muscle, gill, fin, eye and intestine. In male, both fsh and fshra transcripts showed high level of expression during spermatogenesis, however, in female, expression level was found to be higher in the fully grown oocyte stages. The expression of rohu lh and lhcgrba mRNA increased with increment of gonadosomatic index and showed highest level during spermiation stage in male and fully matured oocyte stage in female. These results together may suggest the involvement of fshra and lhcgrba in regulating function of seasonal gonadal development in rohu.

Diabetes, Hypertension and Kidney Disease Combination "DHKD Syndrome" is common in India.

Objectives: The study was designed to find out frequency of (i) Diabetes mellitus (DM) as a cause Chronic Kidney Disease (CKD), (ii) Association between diabetic-CKD (diabetic patients who subsequently developed CKD as complication), hypertension (HT) and obesity. Further assessment was made to (iii) Identify percentage of diabetics attending medical and nephrology OPD had prior testing for proteinuria and or creatinine. Methods: After ethical consideration this prospective observational study was conducted on consecutive 6175 patients who gave consent to participate in two major referral hospitals one in Delhi and other in Bhubaneswar (BBSR). Primary hypertension was defined as blood pressure of ≥140/90 mmHg detected before onset of DM or detected together in the absence of CKD (elevated serum creatinine S Cr ≥1.7 mg/dL and or proteinuria > 0.3g/24H). Upper limit of serum creatinine was kept at 1.7 mg for this study. Mean value of three estimations on different days was recorded. Detail clinical history of DM and HT was taken. Body Mass Index (BMI), ocular fundi examination, urine analysis, serum creatinine, lipid profile, blood glucose, HbA1C tests were conducted in all patients. They were regularly followed up in renal clinic at about 2 month interval for repeat investigations. Blood pressure in nondiabetic-CKD patients was recorded for comparison. Further, consecutive diabetic patients attending general medicine OPD for first time were examined, their previous investigations were carefully scrutinized and recorded. Urine for albuminuria and serum creatinine were tested every month over a period of one year. Results: In Delhi diabetic-CKD was observed in 68.4% and the same was 56.2% in BBSR giving a combined figure of 62.3 percent. On close analysis of past record primary hypertension was observed in 75.4% who subsequently developed diabetes and CKD. Frequency of association between diabetic-CKD and HT were 88.2% and 69.3% in two cities respectively, combined frequency being 78.7 percent. Association of diabetic-CKD and obesity was 55.1 % and 55.9% in two cities respectively with combined frequency of 55.5 percent. In contrast obesity in non-diabetic-CKD patients in Delhi and BBSR was found in 43.1% and 18.5% respectively, combined frequency being 30.8%. Fifty four percent of diabetic patients who attended medical OPD for the first time were found to have proteinuria and elevated serum creatinine. However, they were not earlier tested for those parameters. Hence, they were unaware of CKD. Conclusion: Diabetes was found to be a bigger cause (62.3%) of CKD than what has been reported thus far in India. At presentation association of diabetic-CKD with HT was recorded higher (78.7%) in India. Hence use of the syndrome "DHKD", (complex of diabetes, hypertension and kidney disease) is justifiable. Our study shows 54.4% of diabetic patients attending medicine OPD were uninvestigated by either physician or GP for CKD because urine albumin and serum creatinine tests were lacking. Thus, progression to CKD in many patients went unnoticed. Syndromic diagnosis of "DHKD" therefore in our view is important to create general awareness for early detection and effective treatment of diabetic nephropathy.

Anaplastic Carcinoma Thyroid in a Young Child - an Extremely Rare Occurrence.

Anaplastic thyroid carcinoma (ATC), one of the most aggressive malignancies, is extremely rare in childhood. We present a case of 5-yearold child who presented with rapidly progressing thyroid swelling and stridor, for which she underwent emergency tracheostomy and biopsy. Histopathological features were suggestive of ATC and the patient died within two months after diagnosis. ATC, though very rare in childhood, should be kept in the differential diagnoses of rapidly enlarging neck masses in children. To the best of our knowledge, this is the youngest case of ATC reported in literature.

Molecular characterization of locus of enterocyte effacement pathogenicity island in shigatoxic Escherichia coli isolated from human & cattle in West Bengal, India.

BACKGROUND & OBJECTIVES: Shigatoxic Escherichia coli (STEC) recovered from dairy animals of Kolkata, India, harboured the putative virulence genes; however, the animals did not exhibit clinical symptoms. Similarly, human isolates in this locality also showed variations in degree of symptoms. Hence, this study was designed to know the presence of recognized gene(s) in the locus of enterocyte effacement (LEE) pathogenicity island in these STEC isolates and functional status of the cardinal gene (eae) related to pathogenicity. METHODS: Genes were characterized using polymerase chain reaction (PCR) assays, and functional status of cardinal gene (eae) was evaluated by fluorescent actin staining (FAS) assay. Variation in eae gene was determined by intimin PCR. RESULTS: Cattle STEC isolates carried 22 genes in LEE pathogenicity island in different frequencies ranging from 5.63 to 47.88 per cent of the isolates. In human isolates, the genes namely ler, escRSTU, orf 2, esc C, esc V, orf 3 and tir that are associated with secretory function, were found to be absent and rest of the genes were present in lower frequency. Further, the cardinal gene (eae) responsible for initiation of pathogenesis was in a very low frequency in human (n=2; 10.5%) and cattle (n=11; 15.5%) isolates. None of theseeae+ STEC isolates from human and cattle revealed positivity in FAS assay. INTERPRETATION & CONCLUSIONS: Majority of human STEC isolates lacked the cardinal virulence gene (eae), and genes for secretory function that are essential for facilitating pathogenesis. This may partially be attributed to low occurrence of STEC in human clinical diarrhoea in this area. Although a few isolates (11 of 71) from cattle had eae gene, they did not express phenotypically. This could be one of the reasons for not appearing of clinical symptoms in the hosts.

Molecular characterization and antimicrobial resistance profile of Clostridium perfringens type A isolates from humans, animals, fish and their environment.

The study was aimed to characterize, and determine antibiogram of C. perfringens type A isolated from the feces of human and animal diarrhoeal cases, as well as healthy animals, meat of pigs and goats, gills and intestine of fish and samples from fish pond. A total of 460 samples, including human diarrhoeal cases (n = 130); diarrhoeal cases of pig (n = 52) and goat (n = 50); fecal samples from healthy pig (n = 50) and goat (n = 50); meat samples viz. pork meat (n = 52); goat meat (n = 50) and fish including their environmental sources (n = 26) were used for isolation and identification of C. perfringens type A. All the biochemically confirmed isolates were positive for species-specific 16S rRNA and cpa genes by PCR assays. Toxinotyping of C. perfringens type A isolates showed that overall prevalence of C. perfringens type A with only cpa+ gene was 43.2%; with cpa+ and cpb2+ genes was 45.4%; with cpa+ and cpe+ genes was 4.9%; however, with cpa+, cpb2+and cpe+ genes was 6.6%. Antimicrobial susceptibility testing revealed that 83.7% of isolates were resistant to three or more antibiotics.

Isoprenoid-Based Biofuels: Homologous Expression and Heterologous Expression in Prokaryotes.

Enthusiasm for mining advanced biofuels from microbial hosts has increased remarkably in recent years. Isoprenoids are one of the highly diverse groups of secondary metabolites and are foreseen as an alternative to petroleum-based fuels. Most of the prokaryotes synthesize their isoprenoid backbone via the deoxyxylulose-5-phosphate pathway from glyceraldehyde-3-phosphate and pyruvate, whereas eukaryotes synthesize isoprenoids via the mevalonate pathway from acetyl coenzyme A (acetyl-CoA). Microorganisms do not accumulate isoprenoids in large quantities naturally, which restricts their application for fuel purposes. Various metabolic engineering efforts have been utilized to overcome the limitations associated with their natural and nonnatural production. The introduction of heterologous pathways/genes and overexpression of endogenous/homologous genes have shown a remarkable increase in isoprenoid yield and substrate utilization in microbial hosts. Such modifications in the hosts' genomes have enabled researchers to develop commercially competent microbial strains for isoprenoid-based biofuel production utilizing a vast array of substrates. The present minireview briefly discusses the recent advancement in metabolic engineering efforts in prokaryotic hosts for the production of isoprenoid-based biofuels, with an emphasis on endogenous, homologous, and heterologous expression strategies.