Mixture Of Genotypes Research Articles

A Comparison of Mg/Ca Ratios in Globigerinoides ruber (white): sensu stricto versus a Mixture of Genotypes

Abstract: Mg/Ca ratios were measured in core AAS9/21 from the eastern Arabian Sea (EAS), on two sets of planktonic foraminifera, one with a mixture of Globigerinoides ruber genotypes (sensu stricto and sensu lato) and the second which contained only G. ruber sensu stricto, living in the top 30 m of the water column. During the Holocene, sea surface temperatures (SSTs) in set 1 are cooler in comparison to that of set 2. During the deglaciation, SSTs derived using both these sets are similar, probably due to increased upwelling during this period. The drawback in using undifferentiated G. ruber is that the relative proportion of each genotype varies through different time periods as seen in the EAS and may influence the temperature reconstructions.

Genetic composition, genetic diversity and small-scale environmental variation matter for the experimental reintroduction of a rare plant

Aims Reintroduction has become an important tool for the management of endangered plant species. We tested the little-explored effects of small-scale environmental variation, genotypic composition (i.e. identity of genotypes), and genotypic diversity on the population survival of the regionally rare clonal plant Ranunculus reptans. For this species of periodically inundated lakeshores genetic differentiation had been reported between populations and between short-flooded and long-flooded microsites within populations.Methods We established 306 experimental test populations at a previously unoccupied lake shore, comprising either monocultures of 32 genotypes, mixtures of genotypes within populations or mixtures of genotypes between populations. In 2000, three years after planting out at the experimental site, a long-lasting flood caused the death of half of the experimental populations. In 2003, an extreme drought resulted in the lowest summer water levels ever measured.Important findings Despite these climatic extremes, 27 of the established populations survived until the end of the experiment in December 2003. The success of experimental populations largely differed between microsites. Moreover, the success of genotype monocultures depended on genotype and source population. Genetic differentiation between microsites played a minor role for the success of reintroduction. After the flood, populations planted with genotypes from different source populations increased in abundance, whereas populations with genotypes from single source populations and genotype monocultures decreased. We conclude that sources for reintroductions need to be selected carefully. Moreover, mixtures of plants from different populations appear to be the best choice for successful reintroduction, at least in unpredictably varying environments.

Genotypic diversity effects on biomass production in native perennial bioenergy cropping systems.

The perennial grass species that are being developed as biomass feedstock crops harbor extensive genotypic diversity, but the effects of this diversity on biomass production are not well understood. We investigated the effects of genotypic diversity in switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii) on perennial biomass cropping systems in two experiments conducted over 2008–2014 at a 5.4‐ha fertile field site in northeastern Illinois, USA. We varied levels of switchgrass and big bluestem genotypic diversity using various local and nonlocal cultivars – under low or high species diversity, with or without nitrogen inputs – and quantified establishment, biomass yield, and biomass composition. In one experiment (‘agronomic trial’), we compared three switchgrass cultivars in monoculture to a switchgrass cultivar mixture and three different species mixtures, with or without N fertilization. In another experiment (‘diversity gradient’), we varied diversity levels in switchgrass and big bluestem (1, 2, 4, or 6 cultivars per plot), with one or two species per plot. In both experiments, cultivar mixtures produced yields equivalent to or greater than the best cultivars. In the agronomic trial, the three switchgrass mixture showed the highest production overall, though not significantly different than best cultivar monoculture. In the diversity gradient, genotypic mixtures had one‐third higher biomass production than the average monoculture, and none of the monocultures were significantly higher yielding than the average mixture. Year‐to‐year variation in yields was lowest in the three‐cultivar switchgrass mixtures and Cave‐In‐Rock (the southern Illinois cultivar) and also reduced in the mixture of switchgrass and big bluestem relative to the species monocultures. The effects of genotypic diversity on biomass composition were modest relative to the differences among species and genotypes. Our findings suggest that local genotypes can be included in biomass cropping systems without compromising yields and that genotypic mixtures could help provide high, stable yields of high‐quality biomass feedstocks.

Effects of Particle Size, Stem Component and Genotype on Absorbency of Kenaf (<i>Hibiscus Cannabinus</i> L.) Grown in Nigeria for Oil-Spill Clean-Up

The efficacies of different products made from different parts of the stem of different varieties of kenaf grown in Nigeria as oil absorbents were tested. Particle sizes, genotypes and whether mixed or sole stem core or bast differed among the treatments. The fibre component sank in water, and so was not buoyant; the core was partially buoyant while the mixed fibre and core was completely buoyant. Sorption capacity was the highest in the ground whole stem (8.16 g oil/g sorbent), which also retained (6.31 g oil/g sorbent) the highest quantity of oil. Sorption and retention of oil were the lowest in the fibre (1.66 and 0.69 g oil/g sorbent, respectively). After the first use, only the ground stem can be used to re-absorb up to 1.97 g oil/g sorbent of used sorbent. A mixture of genotypes was more absorbent than a single genotype. The implications of these findings in absorbency research are discussed.

Genetic variation in fitness within a clonal population of a plant RNA virus.

A long-standing observation in evolutionary virology is that RNA virus populations are highly polymorphic, composed by a mixture of genotypes whose abundances in the population depend on complex interaction between fitness differences, mutational coupling and genetic drift. It was shown long ago, though in cell cultures, that most of these genotypes had lower fitness than the population they belong, an observation that explained why single-virion passages turned on Muller’s ratchet while very large population passages resulted in fitness increases in novel environments. Here we report the results of an experiment specifically designed to evaluate in vivo the fitness differences among the subclonal components of a clonal population of the plant RNA virus tobacco etch potyvirus (TEV). Over 100 individual biological subclones from a TEV clonal population well adapted to the natural tobacco host were obtained by infectivity assays on a local lesion host. The replicative fitness of these subclones was then evaluated during infection of tobacco relative to the fitness of large random samples taken from the starting clonal population. Fitness was evaluated at increasing number of days post-inoculation. We found that at early days, the average fitness of subclones was significantly lower than the fitness of the clonal population, thus confirming previous observations that most subclones contained deleterious mutations. However, as the number of days of viral replication increases, population size expands exponentially, more beneficial and compensatory mutations are produced, and selection becomes more effective in optimizing fitness, the differences between subclones and the population disappeared.

Complex Genotype Mixtures Analyzed by Deep Sequencing in Two Different Regions of Hepatitis B Virus.

This study assesses the presence and outcome of genotype mixtures in the polymerase/surface and X/preCore regions of the HBV genome in patients with chronic hepatitis B virus (HBV) infection. Thirty samples from ten chronic hepatitis B patients were included. The polymerase/surface and X/preCore regions were analyzed by deep sequencing (UDPS) in the first available sample at diagnosis, a pre-treatment sample, and a sample while under treatment. HBV genotype was determined by phylogenesis. Quasispecies complexity was evaluated by mutation frequency and nucleotide diversity. The polymerase/surface and X/preCore regions were validated for genotyping from 113 GenBank reference sequences. UDPS yielded a median of 10,960 sequences per sample (IQR 16,645) in the polymerase/surface region and 11,595 sequences per sample (IQR 14,682) in X/preCore. Genotype mixtures were more common in X/preCore (90%) than in polymerase/surface (30%) (p<0.001). On X/preCore genotyping, all samples were genotype A, whereas polymerase/surface yielded genotypes A (80%), D (16.7%), and F (3.3%) (p = 0.036). Genotype changes in polymerase/surface were observed in four patients during natural quasispecies dynamics and in two patients during treatment. There were no genotype changes in X/preCore. Quasispecies complexity was higher in X/preCore than in polymerase/surface (p = 0.004).The results provide evidence of genotype mixtures and differential genotype proportions in the polymerase/surface and X/preCore regions. The genotype dynamics in HBV infection and the different patterns of quasispecies complexity in the HBV genome suggest a new paradigm for HBV genotype classification.

Short tandem repeat genotyping for P. jirovecii is more sensitive to mixed genotype than MLST

Objectives The increasing reports of outbreaks in renal transplant and pediatric units highlight the need for reliable genotyping methods to characterize nosocomial acquisition of Pneumocystis jirovecii . PCR-based methods, mainly MLST, have been widely used in this setting. Recently, short tandem repeat (STR) typing was suggested to be more sensitive to mixed infection and detect minority genotypes. Till now, no comparison between MLST and STR have been performed. Methods Thirty-seven samples (one per patient) from two hospitals were analysed (Paris hospital, n = 18, Nantes hospital, n = 19). Each sample was analysed by both a STR typing assay using 6 nuclear markers as recently developed with a MLST typing scheme relying on 4 loci (2 nuclear: SOD and ITS1; and 2 mitochondrial genes: mtLSU , Cytb ) as recently proposed. Analyses were performed blindly for the other typing method. Results Thirty-two and 35 different genotypes were observed by MLST and STR typing, respectively. Mixtures of genotypes were detected in 6/37 (16.2%) samples and 20/37 (54.1%) in MLST and STR typing, respectively. One mixed infection out of 6 detected using MLST was not observed using STR typing. Overall, one STR marker tends to differentiate samples between Paris and Nantes hospitals (Chi 2 ; P = 0.003). We then analyzed the 18/37 (48.6%) samples with a unique P. jirovecii genotype. Out of the 18/37 (48.6%) samples with no mixture, 18 and 17 different genotypes were delineated using MLST and STR respectively (one STR genotype was recovered from 2 patients). Conclusion This study provides evidence that STR typing markers compared well to MLST for molecular typing of P. jirovecii from clinical samples. The present STRs is less time-consuming and better discriminated the geographical origin and detection of minority genotypes, which can impact on the interpretation of epidemiological data.

Ecological connectivity shapes quasispecies structure of RNA viruses in an Antarctic lake.

RNA viruses exist as complex mixtures of genotypes, known as quasispecies, where the evolution potential resides in the whole community of related genotypes. Quasispecies structure and dynamics have been studied in detail for virus infecting animals and plants but remain unexplored for those infecting micro-organisms in environmental samples. We report the first metagenomic study of RNA viruses in an Antarctic lake (Lake Limnopolar, Livingston Island). Similar to low-latitude aquatic environments, this lake harbours an RNA virome dominated by positive single-strand RNA viruses from the order Picornavirales probably infecting micro-organisms. Antarctic picorna-like virus 1 (APLV1), one of the most abundant viruses in the lake, does not incorporate any mutation in the consensus sequence from 2006 to 2010 and shows stable quasispecies with low-complexity indexes. By contrast, APLV2-APLV3 are detected in the lake water exclusively in summer samples and are major constituents of surrounding cyanobacterial mats. Their quasispecies exhibit low complexity in cyanobacterial mat, but their run-off-mediated transfer to the lake results in a remarkable increase of complexity that may reflect the convergence of different viral quasispecies from the catchment area or replication in a more diverse host community. This is the first example of viral quasispecies from natural aquatic ecosystems and points to ecological connectivity as a modulating factor of quasispecies complexity.

Cryptococcosis Serotypes Impact Outcome and Provide Evidence of Cryptococcus neoformans Speciation.

ABSTRACTCryptococcus neoformans is a human opportunistic fungal pathogen causing severe disseminated meningoencephalitis, mostly in patients with cellular immune defects. This species is divided into three serotypes: A, D, and the AD hybrid. Our objectives were to compare population structures of serotype A and D clinical isolates and to assess whether infections with AD hybrids differ from infections with the other serotypes. For this purpose, we analyzed 483 isolates and the corresponding clinical data from 234 patients enrolled during the CryptoA/D study or the nationwide survey on cryptococcosis in France. Isolates were characterized in terms of ploidy, serotype, mating type, and genotype, utilizing flow cytometry, serotype- and mating type-specific PCR amplifications, and multilocus sequence typing (MLST) methods. Our results suggest that C. neoformans serotypes A and D have different routes of multiplication (primarily clonal expansion versus recombination events for serotype A and serotype D, respectively) and important genomic differences. Cryptococcosis includes a high proportion of proven or probable infections (21.5%) due to a mixture of genotypes, serotypes, and/or ploidies. Multivariate analysis showed that parameters independently associated with failure to achieve cerebrospinal fluid (CSF) sterilization by week 2 were a high serum antigen titer, the lack of flucytosine during induction therapy, and the occurrence of mixed infection, while infections caused by AD hybrids were more likely to be associated with CSF sterilization. Our study provides additional evidence for the possible speciation of C. neoformans var. neoformans and grubii and highlights the importance of careful characterization of causative isolates.

Persistence of chloroquine-resistant haplotypes of Plasmodium falciparum in children with uncomplicated Malaria in Lagos, Nigeria, four years after change of chloroquine as first-line antimalarial medicine.

BackgroundIn Nigeria, despite the change in National malaria drug policy to artemisinin combination therapy (ACT) in 2005 due to widespread chloroquine resistance, chloroquine (CQ) is still widely used in the treatment of malaria because it is cheap, affordable and accessible. The use of ACT for the management of uncomplicated malaria is currently being promoted. The employment of genetic markers to track circulating chloroquine-resistant parasites are useful in elucidating likely poor efficacy of chloroquine, especially in settings where it is not recommended for the treatment of uncomplicated falciparum malaria. This study determined the prevalence of pfcrt haplotypes and point mutations in pfmdr1 genes four years after the change in antimalarial treatment policy from CQ to the ACTs in Lagos, a commercial city in South-West, Nigeria.MethodsThis was a cross sectional study on uncomplicated malaria in children less than 12 years that presented with fever and other symptoms suggestive of malaria. Parasite DNA was extracted from 119 patients out of 251 children who were positive for Plasmodium falciparum by microscopy and amplified. The occurrence of haplotypes was investigated in pfcrt gene using probe-based qPCR and single nucleotide polymorphisms in pfmdr1 gene using nested PCR.ResultsOne hundred and nine (109) of the 119 children with P falciparum infection (91.6%) harbourd parasites with the mutant pfcrt haplotype (CVIET). Out of this, 4.2% comprised a mixture of genotypes encoding CVMNK and CVIET, while 4.2% had the wild type (CVMNK). Furthermore, the frequency of point mutations in pfmdr1 was 62.2% and 69.0% for codons Y86 and F184 respectively. There were no mutations at codons 1034, 1042 and 1246 of the Pfmdr1 genes.ConclusionThe high frequency of the CQ-resistant haplotypes (CVIET) and mutations in Pfmdr1 associated with CQ resistance in P. falciparum among these children suggest that CQ-resistant parasites are still in circulation. Continuous use of chloroquine may continue to increase the level of mutations in pfcrt and pfmdr1genes. There is need to strengthen current case management efforts at promoting ACT use as well as urgently restricting access to chloroquine by the National drug regulatory agency, National Agency for Food Drug Administration and Control (NAFDAC).Virtual SlidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2069472010142303

Genotypic diversity of apomictic microspecies of the Taraxacum scanicum group (Taraxacum sect. Erythrosperma)

Populations of polyploid apomictic dandelions consist of a mixture of clonal genotypes that are nearly phenotypically and genetically uniform. Some apomictic taxa are widespread, but many other taxa occur locally or at a single locality. Additionally, certain Central European dandelion populations consist of both diploid sexuals and polyploid apomicts, and the gene flow expected in mixed populations obscures clear genetic and morphological borders among apomictic taxa. In the present study, we investigated genotypic diversity among seven apomictic microspecies coexisting in Central Europe. Using microsatellites, amplified fragment length polymorphism and chloroplast DNA markers, we focused on the clonal structure of apomictic taxa, i.e., whether the studied apomictic taxa are represented by one or a few related genotypes or whether they are represented by many different and unrelated genotypes. The pattern of genotypic diversity within the studied microspecies suggests both intra- and inter-specific genetic diversification. At the intra-specific level, the studied apomictic taxa consist of several related genotypes, and difference among these genotypes is the result of somatic mutations and/or the consequence of repeated origin from different lineages or recent hybridisation. The inter-specific genetic diversity is greater and may reflect a different evolutionary origin of apomictic taxa. Our results show that despite their increased genetic diversity, apomictic dandelions form definable clusters that may be characterised by both molecular markers and morphology.

A Novel Binary Mixture of Helicoverpa armigera Single Nucleopolyhedrovirus Genotypic Variants Has Improved Insecticidal Characteristics for Control of Cotton Bollworms.

The genotypic diversity of two Spanish isolates of Helicoverpa armigera single nucleopolyhedrovirus (HearSNPV) was evaluated with the aim of identifying mixtures of genotypes with improved insecticidal characteristics for control of the cotton bollworm. Two genotypic variants, HearSP1A and HearSP1B, were cloned in vitro from the most pathogenic wild-type isolate of the Iberian Peninsula, HearSNPV-SP1 (HearSP1-wt). Similarly, six genotypic variants (HearLB1 to -6) were obtained by endpoint dilution from larvae collected from cotton crops in southern Spain that died from virus disease during laboratory rearing. Variants differed significantly in their insecticidal properties, pathogenicity, speed of kill, and occlusion body (OB) production (OBs/larva). HearSP1B was ∼3-fold more pathogenic than HearSP1-wt and the other variants. HearLB1, HearLB2, HeaLB5, and HearLB6 were the fastest-killing variants. Moreover, although highly virulent, HearLB1, HearLB4, and HearLB5 produced more OBs/larva than did the other variants. The co-occluded HearSP1B:LB6 mixture at a 1:1 proportion was 1.7- to 2.8-fold more pathogenic than any single variant and other mixtures tested and also killed larvae as fast as the most virulent genotypes. Serial passage resulted in modified proportions of the component variants of the HearSP1B:LB6 co-occluded mixture, suggesting that transmissibility could be further improved by this process. We conclude that the improved insecticidal phenotype of the HearSP1B:LB6 co-occluded mixture underlines the utility of the genotypic variant dissection and reassociation approach for the development of effective virus-based insecticides.

Temporal dynamics and subpopulation analysis of Theileria orientalis genotypes in cattle

In Australia, outbreaks of clinical theileriosis caused by Theileria orientalis have been largely associated with the Ikeda genotype which can occur as a sole infection, or more commonly, as a mixture of genotypes. The most prevalent genotype, Chitose, frequently co-occurs with type Ikeda, however the role of this genotype in clinical disease has not been clearly established. Furthermore, the dynamics of individual genotypes in field infection of cattle have not been examined. In this study we developed quantitative PCR (qPCR) and genotyping methods to examine the role of the Chitose genotype in clinical disease and to investigate the temporal dynamics of T. orientalis Ikeda, Chitose and Buffeli genotypes in naïve animals introduced to a T. orientalis-endemic area. Analysis of the major piroplasm surface protein (MPSP) genes of Chitose isolates revealed the presence of two distinct phylogenetic clusters, Chitose A and Chitose B. A genotyping assay aimed at determining Chitose A/B allele frequency revealed that the Chitose A phylogenetic cluster is strongly associated with clinical disease but nearly always co-occurs with the Ikeda genotype. qPCR revealed that the Chitose genotype (particularly Chitose A), undergoes temporal switching in conjunction with the Ikeda genotype and contributes substantially to the overall parasite burden. The benign Buffeli genotype can also undergo temporal switching but levels of this genotype appear to remain low relative to the Ikeda and Chitose types. Interplay between vector and host immunological factors is presumed to be critical to the population dynamics observed in this study. Genotypic switching likely contributes to the persistence of T. orientalis in the host.

Genetic structure of Daphnia galeata populations in Eastern China.

This study presents the first examination of the genetic structure of Daphnia longispina complex populations in Eastern China. Only one species, D. galeata, was present across the eight investigated lakes; as identified by taxon assignment using allelic variation at 15 microsatellite loci. Three genetically differentiated D. galeata subgroups emerged independent of the type of statistical analysis applied. Thus, Bayesian clustering, discriminant analysis based on results from factorial correspondence analysis, and UPGMA clustering consistently showed that populations from two neighbouring lakes were genetically separated from a mixture of genotypes found in other lakes, which formed another two subgroups. Clonal diversity was high in all D. galeata populations, and most samples showed no deviation from Hardy-Weinberg equilibrium, indicating that clonal selection had little effect on the genetic diversity. Overall, populations did not cluster by geographical origin. Further studies will show if the observed pattern can be explained by natural colonization processes or by recent anthropogenic impact on predominantly artificial lakes.

Influence of genotypic mixtures on field pea yield and competitive ability

Darras, S., McKenzie, R. H., Olson, M. A. and Willenborg, C. J. 2015. Influence of genotypic mixtures on field pea yield and competitive ability. Can. J. Plant Sci. 95: 315–324. Field pea breeding programs have been very successful at improving plant and disease resistance; however, limited success has been achieved in improving the competitive ability of field pea. A study was conducted to determine whether growing field pea in two-way genotypic mixtures could improve the crop's yield and competitive ability. A second objective was to determine if genetic relatedness had any effect on the mixing ability of genotypes. Genotypes were chosen on the basis of pedigree and included two sister lines (CDC1987-3 and CDC1897-14), their common parent (Eclipse), and a distantly related genotype (Midas). The four genotypes were grown as pure stands and as all possible two-way mixtures in field experiments conducted at Lethbridge and St. Albert, Alberta, from 2010 to 2011. The results revealed that CDC1897-3×Eclipse suppressed the model weed (barley); it reduced seed production by 47% (442 kg ha−1) and 61% (391 kg ha−1) compared with the same components within pure stands at Lethbridge 2010 and Lethbridge 2011, respectively. The same mixture also reduced model weed (barley) biomass production by 61% (831 kg ha−1) at St. Albert in 2010, and by 41% (1372 kg ha−1) at Lethbridge in 2010. Although mixtures demonstrated the potential to improve field pea competitive ability, results were not consistent across site-years. However, some mixtures did improve yield and competitive ability over the most poorly competitive genotypes in pure stand.

Influence of genotypic mixtures on field pea yield and competitive ability.

Darras, S., McKenzie, R. H., Olson, M. A. and Willenborg, C. J. 2015. Influence of genotypic mixtures on field pea yield and competitive ability. Can. J. Plant Sci. 95: 315-324. Field pea breeding programs have been very successful at improving plant and disease resistance; however, limited success has been achieved in improving the competitive ability of field pea. A study was conducted to determine whether growing field pea in two-way genotypic mixtures could improve the crop's yield and competitive ability. A second objective was to determine if genetic relatedness had any effect on the mixing ability of genotypes. Genotypes were chosen on the basis of pedigree and included two sister lines (CDC1987-3 and CDC1897-14), their common parent (Eclipse), and a distantly related genotype (Midas). The four genotypes were grown as pure stands and as all possible two-way mixtures in field experiments conducted at Lethbridge and St. Albert, Alberta, from 2010 to 2011. The results revealed that CDC1897-3×Eclipse s...

Establishment and Spread of a Single Parthenogenic Genotype of the Mediterranean Arundo Wasp,Tetramesa romana1, in the Variable Climate of Texas

As part of a biological control program for the invasive weed, Arundo donax L., several genotypically unique populations of the parthenogenetic stemgalling wasp, Tetramesa romana Walker (Hymenoptera: Eurytomidae), from Spain and France were released in an infested riparian zone along the Rio Grande from Brownsville to Del Rio, TX. An adventive population of the wasp of unknown origin with limited distribution in Texas was also discovered, evaluated, and released as part of the program. More than 1.2 million wasps representing the mixture of genotypes were aerially released from 2009 to 2011. Wasps dispersed from their original release locations and now have a continuous distribution along the Rio Grande from Brownsville to Del Rio, and have dispersed throughout most of Central Texas with satellite populations as far west as San Angelo (Tom Green County), north as far as Kaufman (Kaufman County), and east to Navasota (Grimes County). The most successful genotype (#4) represented 390 of the 409 wasps recovered and matched both an imported population from the Mediterranean coast of Spain and an adventive population established in Texas before the start of the biological control program. Several other European genotypes of the wasp released in the program apparently failed to establish. This result demonstrated the benefits of evaluating and releasing the maximum genetic diversity of the biological control agent in the introduced range. Abundance of T. romana on the Rio Grande from Laredo to Del Rio averaged 190% more in 2013–2014 compared to a similar study in 2008–2009 before release of the European wasps. A favorability index was developed that showed that conditions from 1969 to 1977 would have been adverse to the wasp; conditions after 2009 were more favorable. Climate matching predicts the wasp will disperse throughout the southern U.S. and Mexico.

Additive and non‐additive effects of birch genotypic diversity on arthropod herbivory in a long‐term field experiment

Herbivores are important drivers of plant population dynamics and community composition in natural and managed systems. Intraspecific genetic diversity of long‐lived plants like trees might shape patterns of herbivory by different guilds of herbivores that trees experience through time. However, previous studies on plant genetic diversity effects on herbivores have been largely short‐term. We investigated how tree genotypic variation and diversity influence herbivory of silver birchBetula pendulain a long‐term field experiment. Using clones of eight genotypes, we constructed experimental plots consisting of one, two, four or eight genotypes, and measured damage by five guilds of arthropod herbivores twice a year over three different years (four, six and nine years after the experiment was established). Genotypes varied significantly for most types of herbivore damage, but genotype resistance rankings often shifted over time, and none of the clones was more resistant than all others to all types of herbivores. At the plot level, birch genotypic diversity had significant positive additive effect on leaf rollers and negative non‐additive effects on chewing herbivores and gall makers. In contrast, leaf‐mining and leaf‐tying damage was not influenced by birch genotypic diversity. Within diverse plots, the direction of genotypic diversity effects varied depending on birch genotype, some having lower and some having higher herbivory in mixed stands. This research highlights the importance of long‐term studies including different feeding guilds of herbivores to understand the effects of plant genetic diversity on arthropod communities. Different responses of various feeding guilds to genotypic diversity and shifts in resistance of individual genotypes over time indicate that genotypic mixtures are unlikely to result in overall reduction in herbivory over time.

Hepatitis B Virus Core Promoter Mutations in Patients With Chronic Hepatitis B and Hepatocellular Carcinoma in Bucharest, Romania

Background:Accurate and personalized molecular virological diagnosis of hepatitis B virus (HBV) infection is crucial for individualized selection of patients for antiviral therapy in Romania.Objectives:We aimed to investigate HBV mutations in Romanian patients with chronic HBV infection, also to match HBV genotypes with HBV mutations identified and clinical outcomes.Patients and Methods:This was a cross-sectional study. A total of 484 Romanian patients with chronic HBV infection and hepatocellular carcinoma (HCC) were investigated. This was performed in Fundeni Clinical Institute, Bucharest, Romania during January 2005 to August 2010. HBsAg positive patients with chronic HBV infection admitted to Fundeni Clinical Institute were randomly enrolled in the study. Analysis was performed in the Centre for Immunogenetics and Virology, Fundeni Clinical Institute, Bucharest, Romania. Indirect diagnosis was performed with enhanced chemiluminescence method using Architect i2000SR and HBV-DNA was quantified with COBAS TaqMan HBV PCR. Direct sequencing of the PCR-products was performed with the PCR-product sequencing kit. HBV genotyping was performed with INNO-LiPA DR Amplification and INNO-LiPA HBV precore-core.Results:We detected two HBV genotypes; A (8.1%) and D (60.5%), and a mixture of genotypes A and D (31.4%) (P < 0.001). Basal core promoter (BCP) A1762T/G1764A and precore (PC) G1896A mutations were detected in these Romanian patients with chronic HBV infection. HBV chronic carriers had mainly genotype D (54.4%) and HBV WT (64.0%). BCP A1762T, G1764A and PC G1896A were significantly associated with HCC-tissue HBV sequencing (75.3%) (P < 0.001). PC G1896A alone was detected in HCC-serum HBV sequencing group (66.7%).Conclusions:Genotype D was the main genotype detected in Romanian patients with chronic HBV infection. Genotype D presented both BCP and PC mutations more frequently.

Impact of edible chitosan–cassava starch coatings enriched with Lippia gracilis Schauer genotype mixtures on the shelf life of guavas (Psidium guajava L.) during storage at room temperature

The effect of edible chitosan–cassava starch (CH–CS) coatings containing a mixture of Lippia gracilis Schauer genotypes (EOM) on the shelf life of guavas during storage at room temperature for 10days was studied. Sixteen formulations were prepared with a range of chitosan and essential oil mixtures concentrations, and the in vitro antimicrobial activity was tested. Formulations containing 2.0% cassava starch, 2.0% chitosan and 1.0%, 2.0% or 3.0% EOM were most effective in inhibiting the growth of the majority of bacteria. The edible CH–CS coating and CH–CS with 1.0% (CH–CS–EOM1) or 3.0% EOM (CH–CS–EOM3) were added to guavas and the shelf life was evaluated. On the tenth day of storage, total aerobic mesophilic bacteria and mould and yeast counts were statistically lower (p<0.05) in the CH–CS–EOM1- or CH–CS–EOM3-coated fruits than CH–CS-coated fruits. In addition, fruits coated with CH–CS or CH–CS–EOM showed no significant changes of total soluble solids content, while CH–CS–EOM-coated fruits showed lower titratable acidity than CH–CS-coated fruits at the end of storage. CH–CS–EOM3-coated guavas showed lower a∗ and b∗ values and higher L∗ and hue values than those with other coatings.