Targeted (PCR-based) screening of antibiotic resistance genes' prevalence in the gut microbiota of tribal people of Nabarangpur, Odisha, India.

Antibiotic resistance is a major public health concern worldwide. The gut microbiota harbours multiple antibiotic resistant genes (ARGs) that contribute to the existing and future microbial population in a community or ecosystem. This study aimed to investigate the prevalence of 35 antibiotic resistance genes (ARGs) in the gut microbiota of the tribal people of Nabarangpur, Odisha, India. A total of 83 faecal samples were collected from three different tribes (Bhatra, Gond, and Paraja). Total faecal DNA was extracted, and the simplex polymerase chain reaction was performed to detect selected ARGs. Further analysis was done to estimate the incidence of these ARGs across these tribes based on alcohol consumption habits. We identified a higher prevalence of tetracycline resistance genes (tetW, tetQ and tetM) in the gut microbiota among three populations. Furthermore, a significant (P = 0·024) difference in ARG prevalence against vancomycin in individuals with and without alcohol consumption habits was noticed. The overall distribution of ARGs among the three major tribes of this location was found to be very similar. Together, irrespective of the tribes, the people of this location have gut microbiota harbouring different kinds of ARGs and tetracycline-resistant genes are the most commonly found ARGs.

Impact of mineral deposition on shrimp, Penaeus monodon in a high alkaline water.

This study compares water quality parameters, shrimp growth and mortality rates, and biomass at harvest in two ponds of equal size, seeded with the same density (7 m2) of White Spot Syndrome Virus (WSSV) and Monodon Baculo Virus (MBV) negative post-larvae (PL)-20 of shrimp, Penaeus monodon in the Vellar estuary of South India. The primary difference between the ponds was the water source; one was filled from the estuary and the second with water from bore wells with high alkalinity. Temperature in both ponds was similar and reached 320C after 185 days of culture. Dissolved oxygen (DO) levels were within the acceptable range although levels in the alkaline pond were near the lower limit for the last 90 days before harvest. Salinity levels were similar in both ponds, above optimal levels, and increased over the 185 days. Alkalinity in the estuarine water was typically <50 ppm and again 200-320 ppm in the alkaline pond. In the alkaline pond, beginning on the 75th day mineral deposits was observed covering all parts of the shrimp including the eye and the inner gill chambers, and by harvest, 42% of the shrimp showed this coating. Elemental analysis identified the major constituents as calcium, phosphorus and manganese. Survival rates in the estuarine-water-fed pond was 92% with a total pond biomass at harvest of 1.65 tons ha-1 compared to survival of 79% in the alkaline pond and a biomass at harvest of 1.020 tons ha-1. When well water must be used, its alkalinity should be monitored and diluted with water from other sources.

Comparative histochemical localization of secondary metabolites in seed-raised and in vitro propagated plants of Excoecaria agallocha Linn. (Euphorbiaceae), the milky mangrove tree of historical significance.

Mangroves synthesize novel secondary chemicals that are poorly understood. Among the euphorbiaceous mangrove species, Excoecaria agallocha Linn. produces novel terpenoids and alkaloids of medicinal importance. We conducted a comparative tissue level histochemical study of E. agallocha L. to determine whether in vitro propagation alters the content of phytochemicals within the plant parts. Transverse sections of the root, stem and leaves of seed-raised saplings and in vitro propagated plants stained with 10% vanillin-perchloric acid revealed accumulation of terpenoids in the cork cambium. Alkaloids were localized using Dragendorf's reagent in the cortex of the root sections as brown layers. Methylene blue staining revealed that seed-raised plants possessed more lignified cells, distinct latex ducts and ellipsoidal guard cells compared to the plants propagated in vitro, which revealed abnormal, circular guard cells. The phytochemical content of E. agallocha propagated by the in vitro method was comparable to the seed-raised plants. Phytochemical studies of the species of E. agallocha propagated in vitro would confirm whether the species could be used for its medicinal compounds.

Delayed occurrence of restenosis in drug eluting stents: an evidence of delayed healing.

Drug eluting stents have made a significant impact on restenosis. However, there are concerns regarding delayed "catch-up" of restenosis. In this case report we present two such patients with delayed occurrence of restenosis after drug eluting stent implantation.

Genetic diversity in cultivars and landraces of Oryza sativa subsp. indica as revealed by AFLP markers.

Genetic diversity among 49 Indian accessions of rice (Oryza sativa subsp. indica), including 29 landraces from Jeypore, 12 modern cultivars, and 8 traditional cultivars from Tamil Nadu, was investigated using AFLP markers. In total, nine primer combinations revealed 664 AFLPs, 408 of which were found to be polymorphic. The percentage of polymorphic AFLPs was approximately the same within the cultivars and landraces. Similar results were obtained when genetic diversity values were estimated using the Shannon-Weiner index of diversity. Genetic diversity was slightly higher in the modern cultivars than in the traditional cultivars from Tamil Nadu. Among the landraces from Jeypore, the lowland landraces showed the highest diversity. The present study showed that the process of breeding modern cultivars did not appear to cause significant genetic erosion in rice. Cluster analysis and the first component of principle component analysis (PCA) both showed a clear demarcation between the cultivars and landraces as separate groups, although the genetic distance between them was narrow. The modern cultivars were positioned between the landraces from Jeypore and the traditional cultivars from Tamil Nadu. The second component of PCA further separated medium and upland landraces from lowland landraces, with the lowland landraces found closest to the traditional and modern cultivars.

Species identification in seven small millet species using polymerase chain reaction-restriction fragment length polymorphism of trnS-psbC gene region.

The chloroplast trnS-psbC gene regions from total genomic DNA of 119 accessions from seven small millet species were amplified by polymerase chain reaction (PCR) and digested with eight restriction enzymes individually as well as in combinations of two enzymes to generate restriction fragment length polymorphism (PCR-RFLP). PCR-RFLP with individual enzymes revealed polymorphism between only some species. However, all the species could be distinguished by using a combination of two enzymes, specifically HaeIII and MspI. PCR-RFLP of 11 to 20 accessions with the same enzyme combination showed no intraspecific variation, which established that the differential banding patterns were species specific. In contrast, the same enzyme combination was not useful for differentiating different species of the genera Cajanus, Rhyncosia, Abies, Rhizophora, Ceriops, and Bruguiera, and it also revealed intraspecies variation in three species of Abies. The present study indicated that digestion of trnS-psbC with two four-base recognizing enzymes reveals more variation than with either enzyme alone and that it may be a method of choice for species identification in some genera.

Molecular phylogeny of mangroves. VI. Intraspecific genetic variation in mangrove species Excoecaria agallocha L. (Euphorbiaceae).

Genomic DNA from 84 individuals of Excoecaria agallocha from seven mangrove populations were analysed for random amplified polymorphic DNAs (RAPDs) using 16 random 10-mer primers. Polymorphism within populations varied from 20% to 31%. At the interpopulation level, 111/149 (74%) of RAPDs were polymorphic. Restriction fragment length polymorphism (RFLP) analysis of 21 individuals (3 individuals randomly selected from the 7 populations) using 30 probe-enzyme combinations revealed a high level of interpopulation polymorphism (62.2%) indicating interpopulation genetic divergence. The polymorphic RAPDs and RFLPs were pooled, and clustering was carried out based on mean similarity for individual populations. The dendrogram showed groupings of populations from the West and East Coasts of India into separate clusters, at 60% similarity level. Further, RAPD and RFLP analysis of male and female plants showed approximately the same level of variation in both sexes, and no sex-linked markers were found. These results demonstrate that considerable intrapopulation and interpopulation genetic variations exist in E. agallocha, and that lack of genetic variation is not the reason for the morphological uniformity observed across the range of the species.

Molecular phylogeny of mangroves II. Intra -and inter-specific variation in Avicennia revealed by RAPD and RFLP markers.

Random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers were used to estimate intra- and inter-specific variation in three species of an exclusive mangrove genus, Avicennia. Intrapopulation polymorphism among the 10 populations of Avicennia marina, as measured by percentage of polymorphic RAPDs, varied between 17.8 and 38.9%, with a standard deviation of 7.28, and the coefficient of variation was 26.5%. Polymorphism in Avicennia officinalis (Pichavaram population, 32.3%) and Avicennia alba (Coringa population, 37.8%) was greater than the intrapopulation variation observed in the populations of A. marina from each of the respective locations. It was greater than the average percentage of polymorphism at the intrapopulation level (27.47%) but far less than the variation measured at the interpopulation level in A. marina. Interpopulation variation in A. marina (76.7% for RAPDs and 66% for RFLPs) was greater than the variation in any individual population of this species, indicating a high degree of divergence between the populations. Interpopulation variation as revealed by RAPD and RFLP markers did not indicate the existence of more than one distinct entity in this species in India. The implications of these observations in genetic sampling and conservation are discussed. Statistical analysis of 109 RAPDs and 84 RFLPs observed in one representative genotype from each species showed that the widely distributed A. marina was more closely related to A. alba (genetic distance (1 − F) = 0.22) than to A. officinalis (genetic distance (1 − F) = 0.37). RAPD analysis of six randomly selected genotypes in each species and principal component analysis of the data also favoured this observation.

Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh.

RAPD markers were used to assess genetic fidelity of 23 micropropagated plants of a single clone (L34) of Populus deltoides. Eleven arbitrary 10-base primers were successfully used to amplify DNA from in vivo and in vitro material. Of these, 5 distinguished a total of 13 polymorphisms common across 6 micropropagated plants. Apart from these 6 plants, the amplification products were monomorphic across all the micropropagated plants, the mother plant and 4 additional field-grown control plants. Our results show that RAPD markers can be used to gain rapid and precise information about genetic similarities or dissimilarities in micropropagation systems that might not be so easily evident from other commonly used techniques.

Associated chromosomal DNA changes in polyploids.

The 2C and 4C nuclear DNA amounts were estimated in eight diploid species, belonging to three diverse genera (Vicia, Tephrosia, and Phlox) and their corresponding colchitetraploids. In P. drummondii, T. purpurea, and T. oxygona tetraploids the deviation from the expectation was highly significant. The DNA in P. drummondii was further discarded in subsequent (C1, C2) generations, thus attaining an overall reduction of about 25%. The DNA content in the subsequent generations was the same as that of C2. It is concluded that rapid DNA loss in the first and subsequent generations was not only associated with the substantial increase (30-66%) in the seed set, but it also helped in the establishment and stabilization of the tetraploid. The possible relationship between such a nucleotypic change and success of polyploids is discussed. The DNA change from the expected value in the P. drummondii tetraploid was achieved by equal decrement to each chromosome independent of size, i.e., small chromosomes loose the same amount of DNA as the large chromosomes.

Ploidy-dependent genomic stability in the tissue cultures of ornamental Phlox drummondii Hook.

Comparisons of the chromosome numbers, 2C nuclear DNA amounts and karyomorphology were made in explant cultures of diploid (2n = 2x = 14) and autotetraploid (2n = 4x = 28) Phlox drummondii. In 6-36 week old calli derived from diploid internodal segment explants, and in cells of root tips regenerated from such callus, marked differences were observed in chromosome number. The chromosome numbers ranged from 2n = 14 to 2n = 100 and DNA amounts from 8.20 to 63.20 pg in the diploid derived callus, while the extent of variation was much reduced in the regenerated roots. In contrast, the autotetraploid cultures were characterised by the maintenance of the same chromosome number and DNA amounts as the mother plant. Modified chromosome structures were not apparent in any of the cultures. The possible reasons for the chromosomal instability at the diploid level and stability at the tetraploid level are discussed.