Effective utilization of available feed resources is the key for economical livestock rearing. Complete feed system is one of the latest developments to exploit the potential of animal feed resources in the best possible way. The complete feed is a quantitative mixture of all dietary ingredients, blended thoroughly to prevent separation and selection, fed as a sole source of nutrients except water and is formulated in a desired proportion to meet the specific nutrient requirements. The concentrate and roughage levels may vary according to the nutrient requirement of ruminants for different production purposes. The complete feed with the use of fibrous crop residue is a noble way to increase the voluntary feed intake and thus animal’s production performance. In this system of feeding, the ruminant animals have continuous free choice availability of uniform feed mixture, resulting in more uniform load on the rumen and less fluctuation in release of ammonia which supports more efficient utilization of ruminal non-protein nitrogen. Feeding complete diet stabilizes ruminal fermentation, thereby improves nutrient utilization. This feeding system allows expanded use of agro-industrial byproducts, crop residues and nonconventional feeds in ruminant ration for maximizing production and minimizing feeding cost, thus being increasingly appreciated. However, to extend the concept extensively to the field and make this technology successful and viable for farmers, more efforts are needed to be taken. Livestock forms an integral part of agriculture sector and encompass a great impact on the national economy; however, nutritional inadequacy for livestock is at currently referred one of the most burning global problems of agricultural countries. Due to higher population growth rate and its consequential pressure, available land for forage production is declining day by day. This wicked scenario of quality feed resource availability has virtually eclipsed the genetic worth of animals. In such circumstances, the extensive use of crop residues in livestock feeding seems to be indispensable to meet the nutritional needs of livestock; however, the major constraint in the utilization of these crop residues is high cellulosic contents and poor nutritive value that even cannot support the maintenance nutrient requirement of the animals. Hence, efforts are being directed toward assisting the animals to utilize these low-grade feedstuffs more efficiently as effective utilization of available feed resources is the key to enhance livestock productivity economically. Efficient utilization of crop residues available locally in appreciable quantum seems to be accomplished by the application of feed technology to maximize advantage from feeds in animal system. In this direction, many technologies have been developed so far, but for several reasons, they have not been adopted by the end-users, especially those having limited finance and poorly skilled . The comBalanced and economical feeding of livestock is extremely important for optimum productivity. Lower livestock production is mainly due to the scarcity of feeds and unbalanced feeding practices. To minimize feed costs and labor, and to maximize production is the need of the time and can be achieved by blending concentrate, mainly comprised locally available by-products and roughage portions of the ration to form complete feed/diet, synonymously termed as total mixed ration (TMR). Complete feed with the use of fibrous crop residue is a noble way to increase the intake and to improve feed utilization and animal production performance. The complete feed system is being increasingly appreciated as it allows expanded the use of agro-industrial byproducts, crop residues and non-conventional feeds in livestock ration for maximizing production and minimizing feeding cost.The merits of complete feed are related to stable rumen environment for optimum fermentation with minimal fermentation losses and fluctuation in the release of ammonia, better utilization of rumen non-protein nitrogen, and stabilization of the acetate to propionate ratio which favors normal fat synthesis and enhancement of utilization of low-grade roughages ,3. FeediComplete feed avoids selective refusal of unpalatable dietary portions, thus has provision for incorporation of nonconventional feed resources to economize animal feeding. Waste materials - such as waste paper , cotton-et al. [et al. [The complete feed has a balanced ratio of concentrate to roughage portions, reduces eating and rumination time, and thus increases resting time. The stall feeding system of livestock with complete feeds during climatic vagaries  has been advocated ,37. Sharet al.  suggeste [et al.  indicate [et al. . Feeding [et al. .Several studies have been carried out for assessing the effects of feeding complete feeds in different ruminant livestock species on various aspects as under:In vitro rumen fermentation technique, a rapid method of analysis has widely been used to study feed degradation, detect small differences in fermentation characteristics between feedstuffs and allow more frequent sampling. This technique enables selection of feed or feed constituents for high efficiency of microbial protein synthesis in rumen along with high DM digestibility and provides a basis for the development of ruminant feeding strategies to maximize substrate fixation into microbial cells [The chemical analysis of feedstuffs provides idea only about nutritive composition but does not give information about their fermentation values , which ial cells .in vitro studies, supplementation of complete feeds with different feed additives has been reported to improve the nutrient degradability by several workers. Mir et al. [in vitro DM degradability (IVDMD) and in vitro organic matter degradability (IVOMD) on supplementation of different herbs at 2% and 3% level in complete feeds for goats. Ganai et al. [in-vitro system was significantly improved when bajra straw-based complete feed was supplemented with Bhringraj herb at 3% DM in comparison to un-supplemented complete feed at 48 h incubation. In another experiment, Ganai et al. [et al. [in vitro fermentation experimentation, Harikrishna et al. [Cellulomonas flavigena in TMR increased in vitro degradation of cellulose from forages that are fed to ruminants [in vitro digestibility and concluded that there was no adverse effect of storage (up to 60 days) and heating (up to 80°C) of TMRs on cellulose and xylanase activities and in vitro fiber digestibility. Grewal et al. [in vitro fermentation of complete feeds and showed that the diet containing ME at 110 or 120% of NRC (2001) with low UDP (24% of dietary CP) supplemented with niacin gave the best response for nutrient digestibility and ME availability. Singh and Srinivas [in vitro fermentation kinetics and reported that in vitro gas production (IVGP) pattern of maize and oat-based concentrate supplement in complete ration mixture were comparable; however, the extent of IVGP on maize-based ration was better than sorghum-based ration and urea-mineral salt fortification yielded better improvement in fermentation pattern on maize-based ration than either sorghum or oat-based rations. Singh [in vitro fermentation studies. Likewise, Grewal et al. [in vitro utilization of complete feed.In r et al.  observedi et al.  also repi et al.  again re [et al.  concludea et al.  reporteduminants . Thakur l et al.  assessedSrinivas  assesseds. Singh  showed tl et al.  reportedin vitro rumen fermentation pattern have been observed. Jakhmola et al. [Calligonum polygonides leaves in sewan (Lasiurus sindicus) grass-based complete diets for ruminants had a potential to positively affect rumen metabolism in vitro. Goswami et al. [Cyamopsis tetragonoloba) meal in concentrate portion of TMRs at all  levels, but depression (p<0.05) was observed in TMR’s containing 50% or more replacement levels. Likewise, Ramachandra et al. [in vitro fermentation.Beneficial effects of incorporating nonconventional feedstuffs in complete feeds on a et al.  reportedi et al.  reporteda et al.  concludeIncreased intake of DM and nutrients in different ruminant species on complete feed system compared to conventional feeding system has been reported by several workers ,25,54-59Lens culinaris) straw-based TMR to growing Barbari kids improved digestibility of various nutrients as well as DCP content (p<0.01) of the diet [et al. [ad libitum. Likewise, Kishore et al. [et al. [Feeding complete diet stabilizes ruminal fermentation that improves efficiency of nutrient utilization in ruminants . Feedingthe diet . In anot [et al.  reportede et al.  reported [et al.  in calveet al. [et al. [et al. [et al. [Tephrosia purpurea based diet possibly due to higher CP content and therefore intake, while cellulose digestibility was higher (p<0.05) in those fed Dactylotennium aegypticum based diet.Utilization of poor quality feedstuffs can be improved by their incorporation in complete diet rather than feeding separately along with concentrate mixture. However, adequate DCP content in crop residue-based complete feeds is necessary to meet the maintenance and growth requirements of the animals. Senani et al.  reported [et al.  conclude [et al. . Likewis [et al.  reported [et al.  reportedet al. [et al. [et al. [et al. [et al. [et al. [Processing of the complete feeds has been reported to improve the nutrient utilization in ruminant animals. Incorporation of crop residues as a roughage source in TMR for ruminants and their further processing (densification/pelleting) is one of the practical ways of improving their utilization. The physical form of the diet can affect potential rate of consumption with densified/pelletized diets ingested more rapidly than those in mash form. In an experiment carried out by Afzal et al.  on Corri [et al.  reported [et al.  reported [et al.  reported [et al.  reported [et al. ,17 and g [et al. , expande [et al. . In cont [et al.  reported [et al.  reportedet al. [et al. [et al. [Orpinomyces joyonii in paddy and wheat straw-based TMRs for lactating buffaloes, improvement (p<0.05) in digestibility of all the nutrients as well as the %TDN content of the diet has been reported.Supplementation of feed additives in complete diets improves their nutritive value and nutrient digestibility in ruminant livestock besides have health promoting and feed economizing effects. Harikrishna et al. , while d [et al.  reported [et al. . In an e [et al.  to asseset al. [et al. [et al. [et al. [et al. [et al. [The roughage:concentrate (R:C) ratio in complete feed may vary in accordance to the production performance of animals. Reddy et al.  reportedet al. . For groet al. ,76. Like [et al.  reported [et al.  while as [et al.  level re [et al.  reported [et al.  to study [et al.  reported [et al.  reportedet al. [2 treatment. Similarly, Samanta et al. [Leucaena leaf meal could replace mustard cake in concentrate portion of complete diets up to 30% safely and inexpensively without affecting nutrient utilization in Barbari goats. In a study carried out by Bhatta et al. [Prosopis cineraria leaves in complete feed mixture on the performance of lambs and kids, reported that feed efficiency ratio was the best in animals fed on complete feed containing 50% leaves. Tiwari et al. [et al. [Feeding complete diet provides scope for incorporation of nutritious non-conventional feeds resources in concentrate or roughage component by avoiding selective eating of palatable components by animals. Mishra et al.  reportedet al.  reportedet al.  and preget al.  goats toa et al.  reporteda et al. , while ai et al.  reportedi et al. . Likewis [et al.  reportedet al. [ad libitum. Nagi et al. [et al. [et al. [et al. [et al. [et al. [adlibitum and concentrate mixture separately in conventional form or the wheat straw-based complete feed in mash and block form, although the calves fed complete mash feed gained higher BW than calves in other dietary treatment groups. The CP level of 15.4% in complete pellet feed has been reported beneficial for commercial goat farming under stall feeding system [Feeding of small ruminants with crop residues-based complete rations appears to be the promising feeding system for improving their productivity in agricultural countries like India. Complete feed system improves nutrient utilization that supports higher growth performance and reduces the cost/kg live weight gain, thus is economical in comparison to conventional feeding system. Several studies have observed the beneficial effect of feeding complete ration on the growth performance of animals. Malisetty et al.  reportedi et al.  reported [et al.  reported [et al.  observed [et al.  observed [et al.  reported [et al. . Sorghum [et al.  recommen [et al. . Similar [et al.  reported [et al. . However [et al.  reportedg system . Likewisg system  observedet al. [et al. [et al. [The level of roughage source in complete feed can be variable depending on production performance of animals; besides, the animal response varies with R:C ratio. Ganai and Beigh  reportedet al.  reportedet al. . Mattoo et al.  reported [et al.  conclude [et al.  were of  [et al. .et al. [et al. [et al. [Acacia nilotica) pods might be incorporated up to 33% in concentrate mixture of TMR for pregnant and lactating goats without affecting growth rate and it might be increased up to 50% by treating with Ca(OH)2 before use. Neem (Azadirachta indica) seed cake could be incorporated into complete diet for lambs after detoxification without any deleterious effect on growth and meat quality [et al. [et al. [D. aegypticum, Cenchrus ciliaris, and T. purpurea) have potential for feeding to kids in the form of complete pellet feed during feed scarcity periods for supporting their optimum growth [et al. [A. indica) leaves could be incorporated up to 40% level replacing groundnut haulms in complete rations without affecting the growth rate in kids.Inclusion of nutritious non-conventional but safe feed resources in concentrate or roughage component of the complete feed has proved to be beneficial for promoting the growth performance of ruminants. Ammoniation of lentil straw for use in TMR might be an effective way for improving nutrient utilization to get better growth response along with reduction in cost of feeding small ruminants ,10. SSB et al.  reported [et al.  reported [et al. ,31 repor [et al.  reported [et al.  reportedm growth . Conventm growth . Rekhate [et al.  reported [et al. . Radhakr [et al.  reportedet al. [et al. [et al. [et al. [et al. [et al. [Various studies have shown enhancement in growth performance of animals fed processed complete feeds. Feeding complete feed in block form to lambs resulted in higher ADG and N retention in comparison to those fed complete mash feed . Furtheret al.  reportedet al.  reported [et al.  reported [et al.  reported [et al.  reported [et al.  reported [et al.  reported [et al. . Expande [et al.  assessed [et al.  or sweet [et al.  might be [et al.  reportedC. flavigena to forage in TMR did not improve digestion, and thus productive performance in lambs at the doses  evaluated [Neocallimastix sp. GR-1) zoospores in wheat straw-based complete feed for Murrah male buffalo calves significantly increased nutrient digestibility and DE value that enhances feed efficiency (by 28.7%) and BW gains (by 16%) [Feed additives supplementation in complete diets improves nutrient utilization promotes higher growth rate in animals. The addition of an enzymatic extract of valuated . However(by 16%) .ad libitum feeding of both the roughage and concentrate portions of the ration separately get reduced. Maximum benefits can be realized by incorporating such feeds as ingredients that are most economically purchased, transported and stored. Nagalakshmi and Reddy [et al. [et al. [et al. [et al. [Blending of roughage and concentrate portions into complete diet increases its energy density  and provnd Reddy  in one o [et al.  reported [et al.  and cros [et al.  as compa [et al.  reported [et al.  reported [et al.  conclude [et al.  reported2 treatment without affecting milk production and its composition [et al. [et al. [et al. [et al. [Feeding of complete diets to dairy animals has variable results on milk composition. Nagalakshmi and Reddy  reportedposition . Similar [et al.  conclude [et al.  reported [et al.  found no [et al.  reportedet al. [et al. [R: C ratio in the complete diet for dairy animals is an important factor regulating the quality of milk. The most common dietary cause of low fat in milk is the diet containing a low level of forage and/or a high level of concentrate. The low milk fat is often confounded by the fact that low forage diets are often associated with high milk production which itself tends to lower fat %. Only 20% roughage in a complete diet could cause a cow to secrete milk with low-fat content, while 40% roughage level in complete ration produced significantly more FCM and milk fat than that of a 30% roughage ration with no significant differences in SNF ,117. Impet al.  reported [et al.  fed dair [et al. .Feeding complete feed to ruminants results in more uniform load on the rumen which is associated with less fluctuation in the release of ammonia so that rumen non-protein nitrogen is more efficiently utilized . CompletL. culinaris) straw-based TMR to growing Barbari kids resulted in higher (p<0.05) total and ammonia-N with comparable pH and TVFA contents in rumen fluid compared to those fed untreated straw-based TMR [et al. [ad libitum. Sharma et al. [et al. [et al. [+ ion and TVFA concentrations were significantly (p>0.01) higher in sheep than in goats while the concentration of total N, trichloracetic acid (TCA) insoluble N, food and protozoal N in SRL was significantly higher in goats than in sheep; however, ammonia and residual N concentrations did not differ significantly between sheep and goats by feeding complete mash diet containing poultry litter (35%). Herbages  could be conserved during monsoon season for feeding to the growing kids as complete pellets diets during fodder scarcity periods without any adverse effect on rumen fermentation characteristics [et al. [in vivo.Several workers have observed no adverse effects of feeding different unconventional feedstuffs based complete rations on rumen fermentation. Feeding ammoniated lentil  culture supplementation to green gram straw-based complete feed blocks promoted rumen fermentation which improved nutrient utilization resulted in stimulated growth of kids. Likewise, yeast supplementation to groundnut straw-based complete feed block could be beneficial to improve the performance of sheep due to a marked improvement in rumen fermentation pattern [et al. [O. joyonii) zoospores incorporated paddy and wheat straw-based TMRs. Likewise, Kumar et al. [et al. [Neocallimastix spp.) zoospores and sulfur (1% sodium sulfate). This technology developed to improve the nutritive value of wheat straw-based diets, can be used at the compound feed industry level for providing a well-balanced wheat straw-based complete feed rations to the animals at farmers door.Supplementation of feed additives in complete feeds promotes rumen fermentation in a favorable direction to enhance animal performance. Nehra et al.  conclude pattern . Sehgal  [et al.  reportedr et al.  reported [et al.  reportedet al. [et al. [et al. [Leucenea leaf meal [et al. [et al. [et al. [et al. [et al. [et al. [Complete ration is the best feeding model as it reduces problems of nutrient deficiencies in livestock fed on poor quality feed resources with no adverse effects on animal health by allowing a synchronous and fractionated supply of all essential nutrients for attaining maximum production potential. Sharma et al.  reported [et al.  reported [et al. . Tripath [et al.  concludeeaf meal . Nagpal eaf meal  observedeaf meal . Kumar e [et al.  also rep [et al. . However [et al.  reported [et al.  reported [et al.  observed [et al.  reported [et al.  conclude [et al.  concludeIn spite of such great importance of complete feed system in ruminant feeding, the technology has remained confined to the organized farms only, as it is not economically feasible for small and marginal livestock farmers to purchase and operate the costly machinery required for blending the ingredients. As such, there is a dire need to extend the concept extensively to the field. In this direction, some limited efforts have been made by providing small-scale forage disintegrator and grinder cum mixer to the villages through local self-help groups on a community basis. Thus, to make this technology successful and viable for farmers, more efforts are needed.Complete feed system for feeding ruminants is comparatively better option than conventional feeding of concentrates and roughages separately or grazing plus supplementation. This has advantages in provision of balanced diet to the ruminants and helping better utilization of the locally available feed resources, resulting in higher productivity along with reduction in feed cost and labor. Thus, the concept of complete feed system is becoming increasingly popular. However, lots of efforts are still needed to be taken for extending the concept extensively to the field.This review is a part of PhD thesis of the first author YAB, who carried out the research under the supervision of AMG. HAA helped in thorough revision of the manuscript. All authors have read and approved the final version of the manuscript.


Eucalyptus is the premier paper pulp, short rotation plantation species grown all over the world. Genetic improvement programs integrating molecular marker tools are in progress in many parts of the globe to increase the productivity. Whole genome sequence and expressed sequence tags (ESTs) of the eucalypts paved way for introduction of molecular genetics and breeding in this genus. Different molecular characterization approaches have been used simultaneously in eucalypts, however, microsatellites or simple sequence repeats (SSRs) with their prolific characteristics could occupy a special niche in Eucalyptus genetic improvement. Further, highly informative SSRs were used for the clonal identity, genetic fidelity and in certification of breeder’s rights. Eucalyptus genetic linkage maps generated with microsatellite loci were used successfully to identify quantitative trait loci (QTLs) for various economically important traits. Progressively more numbers of microsatellites are being linked to genes associated with adaptive and functional variations, therefore making their utility broader in genetic applications. Availability of common SSR markers across the species provides an opportunity to validate the expression of QTLs across variable genetic backgrounds and accurately compare the position of QTLs in other species. Recent evidences suggest that the presence of SSRs in micro RNAs of plant species play a role in the quantitative trait expression. Similar studies in eucalypts may provide new insights into the genetic architecture of transcript-level variations and post transcriptional gene regulation. This review on eucalypts microsatellites, highlights the availability and characteristics of genomic and eSSRs and their potential in genetic analysis of natural and breeding populations and also discusses the future prospects in population genetics and marker assisted selection.The online version of this article (doi:10.1186/s40529-014-0073-3) contains supplementary material, which is available to authorized users. Eucalyptus is the world’s leading industrial plantation species due to its fast growth, wider adaptability and multipurpose utility. Until lately, most of the Eucalyptus plantations all over the world were destined for paper production, however, presently these plantations are looking forth to support solid wood industry including veneer production . Researr (ABRAF ), and unTurnbull ). EfficiTurnbull ). SSRs pTurnbull ; Kalia eTurnbull ). EarlieTurnbull ; Kudapa Turnbull ) which cE. urophylla, E. grandis, E. globulus and E. nitens. However, high synteny of genome existing across eucalypt species has benefited many DNA data deficient eucalypts taxa, thus excellent SSR transferability was witnessed across species . SSRs wa et al. ; Grattapa et al. ). SSRs aa et al. ). Many sa et al. ; Yang eta et al. ). Furthea et al. ; Hudson a et al. ).Eucalyptus. The details on SSR marker code, source species and number of SSRs developed are given in the Table E. grandis and E. urophylla with the prefix as Embra , E. sieberi (8 SSRs), E. globulus (26 SSRs) and E. leucoxylon (13 SSRs) respectively  and 758 validated EST-SSRs (eSSRs), 35 chloroplast SSRs (cpSSRS) and 8 gene based SSRs CG-SS1Rs) have been applied in different species of Rs have bi et al. , 2002],,Eucalypti et al. ; Faria ei et al. , 2011]))Eucalypte et al. ), Corymbs et al. ) and Cord et al. ). The SSe et al. ; Glaubite et al. ; Ottewele et al. ). A set e et al. ). The ISt et al. ). Very fn et al. ; Sexton n et al. ). NGS mel et al. ). A recee et al. ).Table 1E. camaldulensis and E. grandis have been completed by the Kazusa DNA Research Institute, Japan and DOE Joint Genome Institute (JGI), USA in collaboration with members of the Eucalyptus Genome Network (EUCAGEN) respectively . Additii et al. ; http://y et al. ). All thi et al. ; Rabelloi et al. ; Yasodhai et al. ; Rengel i et al. ; He et ai et al. ; Zhou eti et al. ). Recenta et al. ; Bradbura et al. ). Table Eucalyptus for example, 12.9% in NCBI database  13.3% ia et al. ), 25.5% a et al. ; Ceresina et al. ). The tya et al. ; Rabelloa et al. ; Yasodhaa et al. ; Rengel a et al. ; Ranade a et al. ). The dea et al. ). The BAa et al. ). Most oa et al. ; Ottewela et al. ; Brondana et al. ). Uniquea et al. ). A studa et al. ).Table 3Eucalyptus and the transferability rate dropped down in the species of Corymbia and Angophora  to sub genera Symphyomyrtus and Monocalyptus but not to the genus Corymbia. However, modifications in the PCR amplification protocols could improve the transfer rate across genera . Initiae et al. ) tested d et al. ). Many ok et al. ; Glaubitk et al. ; Steane k et al. ; Agrama k et al. ; Ottewelk et al. ; Arumugak et al. ; Subashik et al. ). Similas et al. ; Faria es et al. , 2011];;Eucalypts et al. ; Hudson s et al. ; Petrolis et al. ; Breed es et al. ; He et as et al. ; Bradburs et al. , [b]; Brs et al. ). Recenty et al. ).Table 4Genotyping studies with microsatellites generally preferred dinucleotide repeats because they bring off high polymorphism and display more variation among individuals. In certain occasions the dinucleotide repeats based allelic variation results in shadow bands or stutter bands during electrophoresis thus leading to genotyping errors (Hoffman and Amos ). Hence,E. camaldulensis  and a maximum of 33 alleles (Embra16), with an average of 19.8 × 9.2 alleles per locus . Very hi et al. ). Similat et al. ).E. grandis), 5–12 (E. globulus), 4–10 (E. urophylla), 6–14 , 5–9 (E. dunnii) and 4–14 . Allele size variations favored multiplexing of fluorescent based automated DNA genotyping applications, wherein upto 18 loci were analyzed in a single run with 5 dye format .Eucalyptus microsatellites are usually highly informative as revealed through the statistics of polymorphic information content (PIC) and heterozygosity (He) (Table e) Table . The mose) Table .Table 5The stupendous and multifaceted applications of microsatellites in tree genome analysis are shown in the Figure . The high degree of polymorphism and the clear and simple co-dominant Mendelian inheritance of the SSRs has proven to be an extremely powerful system for the unique identification of Eucalyptus individuals for fingerprinting purposes and parentage testing . Accordt et al. ; Smith et et al. ; Grattapt et al. a; Kirst t et al. ; Ochiengt et al. ; Grossert et al. ; Jones et et al. ; Payn ett et al. ; Rao et t et al. ; Sampsont et al. ; Butchert et al. ; Barbourt et al. ; Faria et et al. ; Shephert et al. ; Ribeirot et al. ; Arumugat et al. ; Subashit et al. ; Wheelert et al. ) Clonal t et al. ). SSR mas et al. ). Similas et al. ), Psidiur et al. ), Olive r et al. ), Tamarii et al. ) and Prub et al. ).E. dunnii  and E. i et al. ) thus rex et al. ). Similatapaglia b). In nas et al. ). On conr et al. ). A smalo et al. ). These o et al. ) showed E. camaldulensis species complex in its natural range was assessed for its genetic structure with 15 microsatellite loci and concluded that the populations belongs to several subspecies with intergrade zones and breeding programs should not treat the species as a single genetic entity . Similas et al. ). The pre et al. ; Yeoh ete et al. ). Wherean et al. ). Such iEucalyptus species with regional distribution were analyzed for their pollen flow and pattern of genetic diversity. These studies involve microsatellite allele diversity largely indicated that in remnant and fragmented populations of E. curtisii , E. leul et al. ), E. wane et al. ), E. benr et al. ), E. gomy et al. , [b]; Bry et al. ), E. incd et al. ) and E. nd Byrne ) the gennd Byrne ; Butchernd Byrne ). Furthend Byrne ). Nucleai et al. ). Effecti et al. ; Barbouri et al. ; Shepheri et al. ).E. grandis, E. urophylla, E. grandis, E. globulus, E. camaldulensis and E. nitens  × (E. urophylla × E. grandis)] explaining 42% of the phenotypic variation . The Adk et al. ). Immedii et al. ). Congru et al. . Vegetas et al. ). Furthea et al. ). With ta et al. ) could ga et al. ). Howevei et al. ). Embra1o et al. ). Recento et al. ).E. globulus putative QTL for Mycosphaerella cryptica resistance was closely associated with microsatellite marker Embra48 . Similay et al. ; Freemany et al. ). Embra1n et al. ). In anon et al. ) in E. ga et al. ).E. nitens and E. globulus  and E. n et al. ). Genic a et al. ). Eucalyn et al. ). Eucalya et al. ).Eucalyptus species to genetic study/breeding in different eucalypt species.Microsatellites have major roles to play in various spheres of eucalypts genetics and improvement. They are one of the three major classes of genetic variations along with SNPs (single nucleotide polymorphisms) and CNVs (copy number variations) and have many important biological functions . RecentGenomic SSRs have a broad range of applications, and in particular being neutral markers, which are not linked to any particular trait, but most probably offer a representation of the underlying genetic diversity in wild populations and to target populations for conservation . FurtheEucalyptus genome. Genic SSR markers could represent the new class of functional markers, finding use in evolutionary studies, comparative mapping, candidate gene association mapping, gene discovery and molecular breeding . In silFurther, enough care should be exercised while handling null alleles, imperfect repeats, and allelic dropout, equally they can lead to an overestimation of observable alleles, a decrease in observed heterozygosity, and an increase in the apparent level of inbreeding. On the basis of microsatellite analysis, spatial genetic structure (SGS) can be estimated to delineate provenances of eucalypts. In addition, non-denaturing FISH (ND-FISH) can be used to compare the distribution of SSRs to determine whether the range of molecular diversity shown by these highly polymorphic sequences is reflected at the chromosome level. They are also ideally used as anchor markers in molecular linkage maps and in generation of consensus maps across species and that can be highly saturated with DArT and SNP/genotyping by sequencing markers (GBS). Development of a high-density consensus genetic map with SSRs in an important chromosomal interval provides eucalypt molecular breeding programs with a better choice regarding the quality of markers and a higher probability of polymorphic markers. They play a major role in aligning linkage map due to their high transferability and have a functional role in trait variation and to see the conservation and diversification of gene order across the species of eucalypts.Microsatellite markers play a major role in eucalypts at different levels of genetic improvement. The inherent potentiality of these marks to distinguish closely related individuals is increasingly encouraging for the mining of more and more SSRs for placing on linkage groups and other genetic studies. Microsatellites containing DNA sequences and their functional role in the eucalypt genome were investigated and detected linkage-to-physical position for a large number of microsatellites. Recently, two hundred and twenty three new microsatellite markers were surveyed for allelic diversity and added to the existing eucalypt SSR map, bringing the total number of genetically mapped SSR loci to >400 and strengthening the comparative genome mapping .Oryza sativa  broughtnd Ganie ). Any prAdditional file 1: Table S1.: Linkage group information for SSR loci in eucalypts. (XLS 546 KB)Authors’ original file for figure 1Authors’ original file for figure 2Below are the links to the authors’ original submitted files for images.


Here, wet al. … gave a relatively thorough overview of the limited existing evidence of the changes that burning can induce in hydrological and aquatic systems. In some places, however, their discussion appears to restate popularly held but unsupported assumptions and to rely heavily on unpublished material. For instance, in the section of their paper concerning fire effects on terrestrial vegetation, they state ‘Burning is considered particularly detrimental to peat-forming Sphagnum species’. Although they do acknowledge that there is contradictory evidence in the scientific literature ‘from a small number of experimental burning plots’, the only citation to support the initial assertion is a report by the Royal Society for the Protection of Birds (RSPB) that has not been formally published or, to our knowledge, peer reviewed.Brown Taking quotations out of context can distort debates , skewingSphagnum species , although some results from a small number of experimental burning plots have contradicted this suggestion . Thus, the processes for changes in Sphagnum cover require study in further detail.Burning is considered particularly detrimental to peat-forming Our review paper , p. 1412et al. [et al. [et al. [Journal of Applied Ecology.The reasons for  criticizet al.  in such  [et al.  is a for [et al.  is publiet al. [et al. [et al. also point to government guidelines that ‘recommend against burning into living moss layers’ but then comment that ‘this level of control is not always achievable’. Notwithstanding the fact that the fuel moisture content of moss layers during the legal burning period are often high enough to make deep combustion physically impossible in all but the most severe droughts, there is good evidence that moss consumption during prescribed burns is very limited and that exposure of bare peat is rare.Brown In a second example, with reference to Brown et al. , Davies  [et al. , p. 9 stet al. [et al. [Yet Davies et al.  have not [et al. , p. 10 set al. [et al. rightly point out that much of our knowledge comes from a single long-term experimental study site , but then they seek to suggest (again on the basis of an unpublished RSPB report) that the results from that location are not generalizable as the fires are ‘extremely controlled’; despite the fact that the use of controlled fire is precisely the aim of prescribed burning. As far as we are aware, no data have actually been published on prescribed burning practices at Hard Hill or the behaviour of the fires burnt there. Furthermore, the inference that at all other sites fire conditions are not ‘extremely controlled’ would perhaps imply that moorland managers are either not very good at, or do not care about, adequate fire control.Brown A third example of misrepresentation by Davies et al. , p. 10 i“However, these plots may not be typical of managed burns elsewhere given their extremely controlled nature” , p. 1413Later, the authors confirmed that they agree with us in this regard:et al. were right to point out that too much of our knowledge comes from a small number of sites and that experimental treatments may not be representative of the variety of management practices on the ground” [“Brown  ground” , p. 12.Thus, the point of their argument is again unclear. We wish to make it clear that we made no insinuation about the ability of moorland managers when using fire as a management tool, directly or implied, contrary to suggestions of Davies et al. [Yet, we did not say that the fires are extremely controlled, nor is it clear why the authors think that we sought to suggest this. Our paper referred to the experimental plot set-up, where, for example, size, shape and treatments are fixed:et al. [et al. [et al. [et al. [et al. [et al. [http://tinyurl.com/zlud2fx, so Davies et al. [et al. [et al. [The criticisms of our review publication , rebutteet al.  presenteet al. . Yet, itet al.  could ha [et al.  because  [et al. , given i [et al.  and Doug [et al.  and thei [et al. , the ass [et al. . For the [et al.  in a cons et al.  could ea [et al.  and Doug [et al. .et al. [et al. [et al. [et al. [Prior to the release of Brown et al. , we prov [et al. , p. 10 a [et al.  have sin [et al.  should het al. [et al. [In summary, the above examples suggest to us that the readers of Davies et al.  have the [et al.  has, the


Surface modification of polymers has become a vibrant field of research on account of a myriad of rationales which stimulated numerous efforts. The current paper serves as a condensed survey of the advances made through different approaches adopted for tuning the surface properties of polyvinyl chloride as a homopolymer extensively used on a large scale. Though it does not address all challenges involved, this paper communicates and highlights, through concise discussion, the findings of the efforts undertaken in recent decades. It is ultimately concluded with a perspective of the huge capacities and promising future directions. Many interactions among different materials in contact take place at surfaces/interfaces. Besides the ambient factors such as pressure, pH and temperature, the interactions are largely controlled by interplay of surface hydrophilicity, topography, and chemical composition. This implies that manipulating the surface characteristics in terms of physicochemical aspects may guide the interactions to a sought-after direction. A broad spectrum of applications are met spanning from simple coatings to complicated biological adsorptions ,2.Polyvinyl chloride (PVC) faces many challenges due to the hydrophobic nature of the polymer surface. The current opinion is that the surface hydrophobicity may bring forth severely adverse consequences, in particular when such materials are exposed to biological systems. This has motivated extensive research during recent decades seeking efficient means to figure out the problem . A clearPVC is the second most widely used polymer in terms of consumption volume accounting for a global production of over 25 million tons per annum. It has mostly found applications in the medical field where it covers around 28% of total plastics use, ranked after polyethylene and before polystyrene . The phtetc. can be regulated to a great extent. This often makes the controlled modification of the surface intended for a certain application a demanding project. In the following paragraphs, a short background is given on different techniques employed for the surface modification purpose. More comprehensive information can be found in notable reviews .,92.91,92et al. [et al. [et al. [in vacuo. Also, the predominant increase in the grain size of the specimen after plasma treatment was attributed to the coalescence of the neighboring grains after plasma treatment.Plasma treatment has been established to strongly affect the PVC surface properties in terms of physical aspects ,59,60,92et al.  learned  [et al. ,47 treat [et al.  found ouIt was almost unanimously accepted that hydrophilicity (wettability) increased after plasma treatment  which coet al. [The extent of wettability enhancement and also thermodynamic recovery have been established to be dependent on plasma power and modification time, as well as the post-plasma storage temperature . This caet al.  modifiedet al. [et al. [et al. [Zhang et al. ,59 used  [et al.  examined [et al.  treated et al. [In connection with the other surface treatment strategies, Jung et al.  used ionet al. [et al. [et al. [et al. [McGinty et al.  showed r [et al.  carried  [et al.  used thi [et al.  found thet al. [et al. [Surface modification of PVC is principally intended for various applications. Thus far, it has been practiced to impart biological activity, ink printability, and ion permeability to PVC. Sowe et al. ,55 treat [et al.  preparedet al. [N,N-dimethylacrylamide) on PVC by well-controlled ATRP and saw that the graft densities of the brushes played an important role in controlling interfacial properties. They found that the blood platelet activation was reduced compared to unmodified PVC especially in higher molecular weights. Liu et al. [2/O2 gas plasma treatment and found that the blood platelets were effectively suppressed on surface sulfonation of PVC film. de Queiroz et al. [et al. [et al. [et al. [et al. [Biocompatibility towards cell and blood as well as biocide activity are all considered most important characteristics which can be granted to PVC via surface modification. Zou et al. ,86 grew u et al.  treated z et al.  evaluate [et al.  coated P [et al.  tried to [et al.  attempte [et al.  examinedet al. [et al. [et al. [et al. [et al. [Asadinezhad et al. ,49,50 im [et al.  used pla [et al.  studied  [et al.  examined [et al.  using phet al. [et al. [+ ions through a pattern mask in order to create patterns of the hydrophilic/hydrophobic regions on the PVC surface. Their results revealed that selective adhesion and proliferation of the cells on the ion-irradiated regions were observed (Khorasani et al.  investig [et al.  studied observed . They beet al. [As for biodegradability, Rios et al.  studied In this brief review, various approaches within the last three decades taken to engineer the PVC surface along with the respective highlights have been sketched. It has been established that the PVC surface can be turned into a hydrophilic surface of high free energy with an enhanced bonding strength and minimized biofouling without affecting the bulk properties. Also, cell and blood compatibility have been observed to increase as a result of appropriate surface modification. The modified PVC exhibited less plasticizer migration, as well. It seems necessary here to describe some general perspectives on this issue. As per the number of publications, it can be claimed that the subject of polymer surface engineering itself is developing quite rapidly. Concerning PVC in particular, a sharp increase is evident over the last decade in the amount of the relevant research. Among various strategies pursued, the solvent-free techniques—especially plasma treatment—are increasingly becoming attractive. Also, wet chemical methods based on the state-of-the-art controlled chemistry are at the center of intense focus. The achievement of enhanced control over the modified surface can also be mentioned as another trend. This concise overview also makes it clear that there are complex phenomena occurring at the surface once the PVC surface is exposed to external bodies. As better understanding of the complex phenomena and aging effects due to the interactions between polymer surface and surroundings is gained, new possibilities of enhanced control over surface properties emerge. Accordingly, a new generation of biomaterials based on PVC is being created to enable the design and production of superior medical devices. In addition to what has already been mentioned, to reach the optimal PVC surface, fine-tuning and the avoidance of overdevelopment, as well as effective, interdisciplinary collaboration among materials scientists, chemists, biologists, and bioengineers will be necessary.


Acta Cryst. (2012), E68, o1640–o1641.Erratum to  et al. , the chemical name in the title is incorrect.In the paper by Tayamon  The scheme and chemical data are correct as given in the original article.In the paper by Tayamon  al. 2012, the che


We investigate the complete convergence of partial sums of randomly weighted extended negatively dependent (END) random variables. Some results of complete moment convergence, complete convergence and the strong law of large numbers for this dependent structure are obtained. As an application, we study the convergence of the state observers of linear-time-invariant systems. Our results extend the corresponding earlier ones. Let us recall the concept of extended negatively dependent (END) random variables which was introduced by Liu .M such that both We call random variables M is 1, then END random variables reduce to NOD random variables which contain NA random variables and NSD random variables . Variou [et al. , Prakasa [et al.  and the  [et al.  studied  [et al.  obtained [et al.  and Hu ementclasspt{minima [et al.  and Yang [et al.  investig [et al.  obtained [et al.  establisg et al.  studied  [et al.  studied  [et al.  obtainedg and Hu  investig [et al.  investig [et al.  obtainedet al. [ρ̃-mixing sequences; Cabrera et al. [et al. [et al. [ρ̃-mixing sequences; Li et al. [In addition, there are many researchers paying attention to the study of the properties of partial sums of randomly weighted random variables. For example, Thanh and Yin  establiset al.  investiga et al.  and Shen [et al.  investig [et al.  and Yao  [et al.  obtained [et al.  obtainedi et al.  studied et al. [et al. [et al. [We aim to investigate the complete convergence of partial sums of randomly weighted END sequences. Some results of complete moment convergence, complete convergence and strong law of large numbers for this dependent structure are obtained. As an application, we study the convergence of state observers of linear-time-invariant systems. We extend some results of Thanh et al. , Wang et [et al.  and Yang [et al.  to the cLiu Let the random variablesbe a sequence of END random variables. Ifis a sequence of all nondecreasing (or nonincreasing) functions, thenis also a sequence of END random variables.M.Let Shen Letandbe an END sequence such thatandfor allThen there exists a positive constantsuch that for allSung Letandbe sequences of random variables. Then, for anyandet al. [Adler and Rosalsky  and Adleet al. Letbe a sequence of random variables which is stochastically dominated by a random variableX, i.e. for some positive constantand allThen, for allandthe following two statements hold: Consequently, one hasfor allHereare some positive constants not depending onn.Letbe a mean zero sequence of END random variables stochastically dominated by a random variablesX.For every, letbe a sequence of independent random variables satisfying thatis independent of.In the following, we list two assumptions: Assume that (A.1) and (A.2) are satisfied. Letandsuch thatThen, for everySo one hasAssume that (A.1) and (A.2) are satisfied. Letandsuch thatfor someThen we also obtain the results of (and (sults of  and (3.3For some Suppose that (A.1) and (A.2) are fulfilled. LetandIfthen, for everyandThus, by the Borel-Cantelli lemma and (the strong law of large numbers is as follows: Let Suppose that (A.1) and (A.2) are fulfilled. LetandIf . Thanh  [et al.  extended [et al.  to randoh et al. ). Yang e [et al.  obtained [et al. ). Genera [et al. , Wang et [et al.  and Yang [et al.  to the cet al. [et al. [et al. [In this paper, we investigate the complete convergence of partial sums of randomly weighted END random variables. Some results of complete moment convergence, complete convergence and strong law of large numbers for this dependent structure are presented , , \documeher with  and  a a\documeFor some Similar to the proof of Theorem proof of , it follproof of , one hasproof of , we obta. So, by  holds. I. So, by  with \doent}p=1,  also holjth component It is easy to check that to prove , it suff2r−1, by , we appli−tn) in  and \docesult of .Moreover, by Assumption tion, by  in Assum′Dn with , we obta′Dn with  immediat


This study was planned to reveal the clinical signs and biochemical alterations in pregnancy toxemic goats.Blood samples were collected from 20 healthy pregnant and 45 pregnancy toxemic goats and analyzed biochemically.The most significant clinical findings were observed in naturally affected goats with pregnancy toxemia included anorexia, recumbency, lethargy, opisthotonos, dropped head, periodic convulsion, sweetish fruity odor from breath, apparent blindness, bloat, grinding of teeth, and frothy salivation. In this study, the level of serum glutamic-oxaloacetic transaminase (SGOT) (84.23±1.44 IU/L), serum glutamic pyruvic transaminase (SGPT) (216.01±4.07 IU/L), blood urea nitrogen (BUN) (22.24±0.31 mg/dl), creatinine (2.13±0.09 mg/dl), β-hydroxybutyric acid (BHBA) (0.46±0.83 mmol/L), and non-esterified fatty acid (NEFA) (1.67±0.71 mmol/L) was significantly higher whereas glucose (30.89±0.38 mg/dl) and calcium (8.10±0.20 mg/dl) levels were significantly decreased in pregnancy toxemic goats as compared to healthy goats.The goats with pregnancy toxemia exhibited clinical signs include anorexia, recumbency, sweetish fruity odor from breath, apparent blindness, bloat, grinding of teeth, and frothy salivation. Biochemically, there were significantly decreased the level of glucose and calcium, and increased level of SGPT, SGOT, BUN, creatinine, BHBA, and NEFA in the pregnancy toxemic goats. Pregnancy toxemia, also known as “twin-lamb” disease, is a metabolic disorder of pregnant small ruminants, caused by an abnormal metabolism of carbohydrates and fats, which occurs at the final stage of pregnancy . Obese eThe early detection of pregnancy toxemia in susceptible animals is essential for successful treatment. In clinical pregnancy toxemia, the diagnosis is based on history, clinical signs of hepatic encephalopathy, and the results of serum biochemical analyses .While clinical pregnancy toxemia in sheep was relatively well studied, there is a paucity of information regarding metabolic changes in the clinical form of the disease, especially in goats. So, this study was planned.Samples were collected from clinical cases coming to Veterinary Hospital at Veterinary College, Anand Agricultural University, Anand. Hence, this particular study did not require ethical approval.2 areas in Gujarat, India, and witnessed a temperature range of 35-38°C with a maximum of 42°C, relative humidity of 57-55% and rainfall about 1-4 inch. The season in this area can be broadly classified into hot and dry summer from March to June, rainy (monsoon) season from July to October, and the winter (mild) season from November to February.Anand district is situated at latitude 22° N and longitude 72° E with 2939.9 kmThe data, recorded from the case records of Department of Teaching Veterinary Clinical Complex, Veterinary College, Anand, were compiled and analyzed for a period of 6-month from June 1, 2015, to December 31, 2015.All the cases of pregnancy toxemia were diagnosed using Rothera’s qualitative test from urine samples . The bloData, obtained from biochemical parameters, were statistically analyzed by Student’s t-test as per the method described by Snedecor and Cochran . T-test:The vital signs like rectal temperature and respiration rate were normal, but heart rate was significantly increased than normal goats. The most significant clinical findings were observed in naturally affected goat with pregnancy toxemia included anorexia 100.00%), recumbency (100.00%), lethargy (86.67%), opisthotonos (73.33%), dropped head 62.22%), periodic convulsion (57.78%), sweetish fruity odor from breath (51.11%), apparent blindness (42.22%), bloat (40.00%), grinding of teeth (37.78%), and frothy salivation (24.44%) (Tables-00.00%, r.22%, perThe study was aimed to evaluate the biochemical indicators in goats positive for naturally occurring pregnancy toxemia in comparison with normal healthy goats . The meaet al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [at al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [In this study, rectal temperature and respiratory rate were found within normal range, which were contrast to Rodolfo et al. , who rep [et al. , Balikci [et al. , and Red [et al. . Anorexi [et al. , El-Seba [et al. , Andrews [et al. , Balikci [et al. , Barakat [et al. , Al-Quda [et al. , Manokar [et al. , Abdelaa [et al. , Souto e [et al. , Reddy e [et al. , Rodolfo [et al. , and Ran [et al. . Depress [et al. , El-Seba [et al. , Andrews [et al. , Balikci [et al. , Barakat [et al. , Hefnawy [et al. , Al-Quda [et al. , Abdelaa [et al. , and Red [et al. . 23 (51. [et al. , Hefnawy [et al. , Abdelaa [et al. , and Ran [et al. . The swe [et al.  and Bara [et al. . Chew mo [et al. , Abdelaa [at al. , Reddy e [et al. , and Ran [et al. . It was  [et al.  and Redd [et al. . It was  [et al. , Andrews [et al. , Barakat [et al. , and Al- [et al. . Nervous [et al. . 18 (40. [et al. , Rodolfo [et al. , and Ran [et al.  and Redd [et al. . Dropped [et al. . 7 (15.5 [et al. , Barakat [et al. , and Ran [et al.  and the et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Increased SGPT and SGOT level of the present study were in agreement with report of Barakat et al. , Gupta e [et al. , Balikci [et al. , Hefnawy [et al. , Abdelaa [et al. , Albay e [et al. , Anoushe [et al. , Reddy e [et al. , Marutso [et al. , and Abb [et al. . In cont [et al.  reportedet al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Increased BUN and creatinine level were in agreement with report of Kolb and Kaskom , Barakatet al. , Hefnawy [et al. , Lima et [et al. , Abdelaa [et al. , Souto e [et al. , Anoushe [et al. , Rodolfo [et al. , and Red [et al. . In thiset al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Decreased glucose level of the present study was in agreement with result of many researcher such as McClymont and Setchell , Robert et al. , Cantley [et al. , El-Seba [et al. , Scott a [et al. , Andrews [et al. , Henze e [et al. , Marteni [et al. , Gupta e [et al. , Barakat [et al. , Ismail  [et al. , Schlumb [et al. , Balikci [et al. , Hefnawy [et al. , Al-Quda [et al. , Hefnawy [et al. , Manokar [et al. , Abdelaa [et al. , Anoushe [et al. , Albay e [et al. , Sharma  [et al. , Reddy e [et al. , Gurdoga [et al.  and Rani [et al. . However [et al.  and Lima [et al.  showed het al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Decreased calcium level of the present findings was in agreement with results of Jopp and Quinlivan , Anousheet al. , Hefnawy [et al. , Manokar [et al. , Souto e [et al. , Albay e [et al. , Rodolfo [et al. , Reddy e [et al. , and Ran [et al. . Decreas [et al.  and Isma [et al. . In cont [et al.  and Rodo [et al.  reportedet al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Increased BHBA level was in agreement with report of Scott and Woodman , Ismail et al. , Balikci [et al. , Al-Quda [et al. , Hefnawy [et al. , Gonzale [et al. , Abdelaa [et al. , Olfati  [et al. , Anoushe [et al. , Gurdoga [et al. , Sharma  [et al. , and Mar [et al. . Increas [et al. , Olfati  [et al. , Anoushe [et al. , and Sha [et al. . The inc [et al.  and Hefn [et al. .The goats with naturally occurring pregnancy toxemia exhibited clinical signs includes anorexia, recumbency, sweetish fruity odor from breath, apparent blindness, bloat, grinding of teeth, and frothy salivation. Characteristic biochemical pattern associated with pregnancy toxemia in goats showed that there were decreased the level of glucose and calcium, and increased levels of SGPT, SGOT, BUN, creatinine, BHBA and NEFA.In the present surveillance, from June to December, the most of animals which shown the clinical sign of recumbency, were suffered from pregnancy toxemia which was confirmed by biochemical tests and urine analysis.This study was a part of PRV’s original research work during M.V.Sc. thesis program. RGJ had designed the plan of work. HVG and FBT helped during sampling, statistical analysis, and manuscript preparation. SDR helped in the laboratory work. All the authors read and approved the final manuscript.


Astrobiology, as the mission draws to a close, is the outcome of our most recent meeting at UC Berkeley in June 2016. Key Words: Enceladus—Enceladus Focus Group—Ocean world—Search for biosignatures. Astrobiology 17, 815–819.The discovery by NASA's Cassini mission at Saturn in 2005 of a large plume of material erupting from the south polar terrain of Enceladus, sourced within a subsurface ocean of salty liquid water laced with organic compounds, has brought together scientists from a diverse range of disciplines over the last decade to evaluate this small moon's potential for extraterrestrial life. The collection of papers published today in  Steel et al.  construcet al. (et al. (et al., we merely scale the bioload at Lost City by the ratio of the geothermal energy flux through the moon's seafloor to that of Earth … a ratio that turns out to be 1. Our results, ∼105 cells/mL, are significantly less optimistic than those of Steel et al. However, we introduce to the study of Enceladus, and describe in some detail, the process of bubble-scrubbing, well known to marine microbiologists, whereby microbes and organic matter in natural bodies of water may be found in significantly enhanced concentrations in the spray produced at the surface by breaking bubbles. This outcome follows from the tendency of bubbles, rising in a column of water, to collect available organic material along the way. It is a process that could very well be at work on Enceladus and could make the job of detecting biosignatures within the plume up to a 1000-fold easier.In the Porco et al.  study, m (et al. , we estiet al. (Mathies et al.  describeet al. (et al.,et al.,Bedrossian et al.  expand oet al. (et al.,The final two papers in our collection make significant advances in the continued analysis of Cassini data and expand our basic knowledge of the geophysical mechanisms underlying the geysers. Teolis et al.  attempt  et al.,. That boe.g., Hurford et al.,et al.,et al.,et al. (et al.,Finally, in all previous attempts to explain the diurnal variation of the plume by modeling the cyclical behavior of tidal stresses across the SPT (,et al.  take a b et al., in satisSo, now, I leave you to it. As Cassini's extraordinary 13 years of exploration conclude, enjoy this up-to-the-minute, far-reaching, wide-ranging look at that little moon at Saturn with the big possibilities.


Among the most contentious management options is ‘tolerance hunting’, or the killing of predators to increase tolerance among groups of people who do not accept the presence of these animals ,2. In 33,4, we uet al.  and Stieet al.  re-visitet al.'s [et al. [et al. [et al. [t > 18 was 69.0% (proportion of posterior that was <0)’ but this concerns all years post-1998, which also include many years without culling. For the relevant period for our paper , we need to interpret the figures ourselves. On figures 3, S2.4, S2.5 and S2.6 in [t > 18 period (1998–2001 approximately) whereas the years with culling seem to show a stable recruitment regardless of the models used [et al. [et al. [et al. [tμ’ implying that more than half the time their estimates failed this relatively undemanding test. Stenglein et al. [et al. [et al. [First, we disagree with Olson et al.'s  and Stieet al.'s  assertioet al.'s , p. 1 ph [et al.  and Stie [et al.  raised t [et al. . In that [et al. ). Stengl [et al. , p. 5 wr S2.6 in , we findels used . Because [et al.  clearly  [et al.  to show  [et al. , p. 5 apn et al.  also didet al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Olson et al.  also arget al.  demonstret al.  was, in et al.  indeed a [et al. . That re [et al.  but did  [et al.  assumed  [et al.  did not  [et al.  found th [et al.  estimate [et al.  also use [et al. . Both Ol [et al.  and Trev [et al. ,13 agree [et al. . Most im [et al.  ignored  [et al.  did not  [et al. ,15.et al. [et al. [et al. [et al. [et al. [et al. [et al.'s [et al. [et al. [et al. [Olson et al.  and Stieet al.  raise otet al.  claims tet al.  suppleme [et al. 's assert [et al. 's hypoth [et al.  because  [et al.  leaves t [et al.  insinuatet al.'s  co-authoet al.'s . The papet al.'s ,17,18 th [et al.  criticiz [et al. . We do n [et al. ,10 is on [et al. . In addi [et al.  seem, inWe appreciate the scrutiny that our analysis and our writing have sparked. Science progresses through invalidation of hypotheses and presentation of new evidence, therefore we welcome scrutiny of our work and collegial discussions. However, we also feel obligated to point out that statements supporting the tolerance hunting hypothesis, either from scientists or governments, seem to be taken for granted and evade scrutiny. A recent illustration is a paper about wolves in Norway bluntly claiming that ‘it is not an unreasonable expectation that allowing legal harvest might prevent some of the illegal killing’ , p. 135.


Astyanax mexicanus, which exhibits both a cave morph and a surface fish morph. As these morphs and their hybrids collectively exhibit zero, one, and two scleral ossicles, they represent a microcosm of teleost scleral ossicle diversity. Our previous research in F2 hybrids of cavefish from Pachón cave and surface fish from Texas suggested that three genes likely influence the formation of scleral ossicles in this group through an epistatic threshold model of inheritance, though our sample size was small. In this study, we expand our sample size using additional hybrids of Pachón cavefish and Mexican surface fish to (1) confirm the threshold model of inheritance, (2) refine the number of genes responsible for scleral ossicle formation, and (3) increase our power to detect quantitative trait loci (QTL) for this trait. To answer these three questions, we scored surface fish and cavefish F2 hybrids for the presence of zero, one, or two scleral ossicles. We then analyzed their distribution among the F2 hybrids using a chi-square (χ2) test, and used a genetic linkage map of over 100 microsatellite markers to identify QTL responsible for scleral ossicle number. We found that inheritance of scleral ossicles follows an epistatic threshold model of inheritance controlled by two genes, which contrasts the three-locus model estimated from our previous study. Finally, the combined analysis of hybrids from both crosses identified two strong QTL for scleral ossicle number on linkage groups 4.2 and 21, and a weaker QTL on linkage group 4.1. Scleral ossification remains a complex trait with limited knowledge of its genetic basis. This study provides new insight into the number and location of genes controlling the formation of scleral ossicles in a teleost fish species.The sclera is the protective outer layer of the eye. In fishes, birds, and reptiles, the sclera may be reinforced with additional bony elements called scleral ossicles. Teleost fish vary in the number and size of scleral ossicles; however, the genetic mechanisms responsible for this variation remain poorly understood. In this study, we examine the inheritance of scleral ossicles in the Mexican tetra,  In mammals, the sclera is a flexible, fibrous tissue; but in fishes, birds, and reptiles, the sclera may be reinforced with an additional ring of cartilage or even small bones. These bony elements, known as scleral ossicles, provide additional support and may even serve as sites of muscle attachment . The forplicated –5.More recent research has focused on the evolution and development of scleral ossicles in teleost fish, which comprise one of the largest and most diverse subsets of vertebrates on earth (reviewed in ). SimilaAstyanax mexicanus, more commonly known as the Mexican tetra, is a teleost fish found throughout Mexico and Texas [Astyanax SF, like other Characiform fishes, possess two elongated scleral ossicles surrounding each eye [Astyanax CF, however, this process is altered. Although CF do form a cartilaginous scleral ring, this ring does not ossify, and thus the scleral ossicles never form [Astyanax CF [Astyanax exhibit microevolutionary patterns of scleral ossicle evolution that mirror the larger macroevolutionary patterns observed in teleosts more generally. Combined with the fact that Astyanax SF and CF are able to successfully breed together and produce viable offspring , this oAstyanax has found that the loss of scleral ossification in members of the Pachón cave population is inherited recessively through the epistatic interaction of three genes [et al. [et al. [n = 196), they could only detect one QTL for overall ossicle size, and none for scleral ossicle number . ,18. 16,1et al. ) or New Astyanax mexicanus CF(Pa) x SF(Mx) F2 hybrids that were originally generated in Protas et al. [et al. [et al. [Astyanax mexicanus CF(Pa) x SF(Tx) F2 hybrids that were originally generated by Yoshizawa et al. [et al. [et al. [The sample size for this study included two different groups of surface fish (SF) and cavefish (CF) hybrids. The first group consisted of 225 individuals of >6 month old adult s et al.  and anal [et al. . The SF  [et al.  were obta et al.  and stai [et al. . The fis [et al.  were als [et al. ), it is et al. [et al. [et al. [et al. [Both hybrid groups used in this study were stained to visualize ossified bone. The 225 CF(Pa) x SF(Mx) F2 hybrids from Gross et al.  were cle [et al.  were sta [et al. . These s [et al.  and O’Qu [et al. .et al. [et al. [To determine the number of scleral ossicles present in each individual, we examined the anterior and posterior regions of the eye for ossification. For the CF(Pa) x SF(Mx) F2 hybrids from Gross et al. , we examet al. , and bec [et al.  since th [et al. . Althouget al. [2 test was performed on the dataset containing the 225 CF(Pa) x SF(Mx) F2 hybrids and then repeated using these 225 individuals in combination with the 196 CF(Pa) x SF(Tx) F2 hybrids from O’Quin et al. [Scleral ossicle loss is controlled by a genetic threshold of approximately three genes . In a geet al.  with  x SF(Mx) F2 hybrids from Gross et al. [et al. [et al. (2014) and O’Quin et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [To identify the genomic location of the genes associated with the number of scleral ossicles, we generated a composite genetic linkage map of the s et al.  and the  [et al. . In orden et al. , we iden [et al.  to match [et al.  and O’Qu [et al. 14 and O’ [et al.  became L [et al.  because  [et al.  were eit [et al. , e.g. LG [et al. . These L [et al. . The finet al. [et al. [et al. [et al. [et al. [We performed three QTL analyses for scleral ossicle number using the combined linkage map in the program R/qtl . For theet al.  using th [et al.  could be [et al. . We do n [et al. , althoug [et al.  and, finscanone and Haley-Knott regression. This analysis calculates the logarithm of the odds (LOD) of association between the average number of scleral ossicles and the genotypes at each genetic marker. We evaluated the statistical significance of the LOD scores by calculating the 95th percentile of genome-wide maximum penalized LOD scores for each phenotype using 1,000 random permutations of the genotypic and phenotypic data. Because scleral ossification is a complex trait controlled by epistatic interactions among multiple loci [scantwo and stepwiseqtl following the same criteria as above. This second analysis is much more powerful, especially for complex traits like scleral ossification, and gives a detailed, LG-level view of individual and interacting QTL. More information on our QTL mapping procedure is available in O’Quin et al. [et al. [et al. [et al. [In each QTL analysis, we performed an initial scan for QTL assuming the presence of a single QTL per linkage group without interactions using the function ple loci , this inn et al. . We did n et al. . In addin et al. ,13, we d [et al.  samples  [et al.  are avaiet al. [We obtained photographs of 225 Pachón cavefish (CF(Pa)) x Mexican surface fish (SF(Mx)) F2 hybrids stained with alizarin red from Gross et al. . We analet al.  found thet al.  Fig 1)..et al. [2 test on the CF(Pa) x SF(Mx) F2 hybrids from Gross et al. [et al. [et al. [2 = 1.85, df = 1, P = 0.168), but not a one, three, or four gene model  x SF(Tx) F2 hybrids with (1+) and without (0) scleral ossicles was consistent with a model of epistatic threshold inheritance caused by three genes . We perfs et al.  and agai [et al.  to confi [et al. , the obs; et al.  and the  [et al.  . This single QTL accounts for 6.86% of the variation in scleral ossicle number among the CF(Pa) x SF(Tx) F2 hybrids from O'Quin et al. [et al. 2015). Although this same region exhibited elevated LOD in the original scan of QTL for scleral ossicle number published in O'Quin et al. [To identify the location of genes and mutations associated with scleral ossicle number, we used genotypes and a composite genetic linkage map for the CF(Pa) x SF(Tx) F2 hybrids from O'Quin et al.  and the  [et al.  to searc [et al. . The ini [et al.  assumed n LG 4.1 . This QTn et al.    x SF(Mx) F2 hybrids from Gross et al.  similarlonal QTL . This QT 0.0002) . Overall ..et al. [et al. [et al. [P < 0.001) and 21  x SF(Tx) F2 hybrids from O'Quin et al.  and 225  [et al.  greatly analysis , indicat; et al.  and the  [et al.  Fig 2),,et al. [ [et al.  Fig 2B)et al. [e; et al.  and Gros [et al.  account et al. [et al. [2 = 50.53, df = 2, P < 0.0001), they are similar in that they are both highly skewed towards F2 hybrids with two scleral ossicles  x SF(Mx) F2 hybrids from Gross et al.  for scle [et al.  found a see also . The slio et al.  and our  [et al.  is likel [et al.  and O’Qu [et al.  and foun [et al. , in whic [et al. , in whic [et al. , but theexicanus . Therefobehavior .et al. [et al. [et al. [et al. [Astyanax mexicanus populations that invaded Central America approximately 2–5 Mya. This ‘old’ lineage of SF gave rise to several CF populations, including Pachón, before going extinct in most regions except Texas [et al. [et al. [By comparing the observed ratio of F2 progeny with (1+) and without (0) scleral ossicles to those expected for threshold inheritance at one to four loci, we were able to estimate the number of genes likely involved in the loss of scleral ossicles. Our analysis of the CF(Pa) x SF(Mx) F2 hybrids sampled in Gross et al.  showed t [et al. , which i [et al.  crossed  [et al.  crossed pt Texas . In contpt Texas . The ‘yopt Texas . Importa [et al.  and Gros [et al.  used CF et al. [Astyanax cave populations, such as those from Curva, Los Sabinos, and Molino caves, remains to be seen since the inheritance of scleral ossicles has not been studied in these groups [It is possible that, since the SF used in each cross are derived from distinct lineages, that different numbers of genetic factors controlling scleral ossification are fixed in each group. For example, the Mexican SF from Río Valles could conceivably be fixed for recessive CF alleles at one of the responsible loci, for example, the weak QTL on LG 4.2 detected only in the CF(Pa) x SF(Tx) F2 hybrids from O'Quin et al. . Since tet al. ,23, and et al. . Whethere groups . Becausee groups ), it wouscaleless mutant in chickens [Scaleless chicks exhibit abnormal keratinization, resulting in an almost complete lack of downfeathers, scales, spurs, and scleral ossicles [scaleless chickens, Abbott and Asmundson [scaleless phenotype was caused by recessive alleles at a single autosomal gene (one-locus threshold model of inheritance). The difference in the number of genes involved in Astyanax versus scaleless chickens may be due to the difference in the types of ossification involved. In chickens, scleral ossification stems directly from a membranous bone [et al. [scaleless to a single nonsense mutation in the gene fgf20. The results of that study indicate that in birds, at least, scleral ossicle formation is either the direct or indirect result of fibroblast growth factor signaling. Other studies of scleral ossicle induction in chicks have identified an important role for hedgehog and bone morphogenic protein signaling as well [This is only the second study to estimate the number of genes responsible for the loss of scleral ossicles among vertebrates . Althougchickens . Scaleleossicles –26. Usinsmundson  determinous bone , while ious bone . Interes [et al.  recently as well .et al. [Astyanax linkage group (LG) 4.1. This same region exhibited elevated LOD in our previous analysis [et al. [et al. [et al. [Quantitative trait loci (QTL) analysis of both datasets, both individually and in combination, allowed us to identify the genomic location of the three genes associated with scleral ossicle number . For theet al. , we founanalysis , but it  [et al. , we foun [et al.  and the  [et al. , but was [et al.   x SF(Mx) F2 hybrid and CF(Pa) x SF(Tx) F2 hybrid linkage maps published in Gross et al.  and O’Qu [et al. .(DOCX)Click here for additional data file.S2 Table(CSV)Click here for additional data file.S3 Table(CSV)Click here for additional data file.S4 TableExpected ratios for 0, 1, and 2 scleral ossicles estimated interactions of two genes caused by epistasis of one (single) or two (duplicate) loci.(DOCX)Click here for additional data file.S1 File(PDF)Click here for additional data file.


Following publication of the original article , the autbLao et al. [15]bBraken et al. [23]bBakhireva et al. [26]bClark et al. [28]These references have already been introduced in Table 1 and should read:Lao et al. [30]Bracken et al. [38]Bakhireva et al. [41]Clark et al. [43]The original article has been updated.


After publication of the original article , it camePate et al.  should h


My first education was in biochemical pathogenesis of disease at the University of Bologna, Italy, University College, London and Max Planck institute, Heidelberg. I entered Virology in 1972 in the University of Bologna, where I still work. I am the head of the molecular virology laboratory in the Department of Experimental Medicine, University of Bologna. The focus of my research is on the molecular mechanisms of herpes simplex virus (HSV) entry into the cell, and on how to modify the HSV tropism in order to generate highly cancer-specific, highly safe, yet fully virulent oncolytic herpesviruses. Academically, I fostered the integration of the Italian Virology in the European Virology.et al.et al.et al. 1998et al.et al.et al.et al.et al.et al.et al.et al.The entry of herpes simplex virus (HSV) into the cell has been my major field of interest for +20 years  Grant.


The review by Klumpp, L. et al. entitled Ion Channels in Brain Metastasis  discusse


A successful endodontic treatment depends on a comprehensive knowledge of the morphology of canal and its variations, an appropriate access cavity, proper cleaning and shaping and adequate root canal filling. The present study was carried out to evaluate the root canal morphology of permanent maxillary first molars in an Iranian population. in vitro study, 80 extracted permanent maxillary first molars from a population in Rafsanjan, Iran were collected. Root canal morphology was evaluated by clearing technique under stereomicroscope under 40× magnification. A combination of Vertucci’s and Sert and Bayirli’s classifications were used to determine the root canal types. Data were analyzed by SPSS 18 software using descriptive statistics. In this . All palatal roots and almost all distobuccal roots had type I configuration. Ten different types of root canal system were found in mesiobuccal roots, among which type I was the most common (38.75%), followed by type II, IV, V, VI, IX, XV, XVI=XIX and VII, respectivelyThe mesiobuccal roots of permanent maxillary first molar had the most complex root configuration. A successful endodontic treatment depends on a comprehensive knowledge of the morphology of canal and its variations, an appropriate access cavity, proper cleaning and shaping and adequate root canal filling. Sometimes a root canal may go undetected, which results in treatment failure . Lack of et al.,  and Sert and Bayirli [7] classification systems were used to determine the root canal morphology . Data were analyzed by SPSS software .In the present study, 80 permanent maxillary first molars were evaluated; 43 teeth (53.75%) from the male patients and 37 samples (46.25%) from the female patients. The patients’ age ranged from 21 to 68 with the mean age of 48 years.Based on “Vertucci” and “Sert and Bayirli” classification in all roots, 10 different root canal types were detected. Type I was the most frequent type (79.16%) followed by type II, IV, V, VI, IX, XV, XVI=XIX and VII, respectively. All of the palatal roots showed type I root canal configuration. In the distobuccal (DB) roots, the most frequent type was type I (98.75%) and only one sample (1.25%) was type V. Moreover, 10 different types of root canals were seen in mesiobuccal (MB) roots. The most frequent type was type I (38.8%) followed by types II, IV, V, VI, IX, XV, XVI=XIX and VII, respectively .et al.  and Khademi et al. . Furthermore, Rouhani et al. , Neelaka [et al.  and Rwen [et al.  studies i et al.  found th [et al. , Naseri  [et al. , Sert et0 % of MB [et al. , the mos [et al.  study, t [et al.  and Khad [et al. , 24, Ind [et al. , Ugandan [et al. , Turkish [et al. , and als [et al. , 21-24 a [et al. , 18. Theet al. [et al. [et al. [et al. [Furthermore, in MB roots, there were ten different root canal types, four of which belonged to Sert and Bayirli’s classification. Unfortunately, most studies , 21-24 a [et al.  and Neel [et al.  were don [et al.  used Ser [et al. , 34, theIn DB roots, type I was the most common root canal type in the present study. All other studies which evaluated DB roots canal types , 21-24 het al. [et al. [et al. [Finally, in palatal roots, all root canal configurations were type I. This finding is in line with Alavi et al. , Rwenyon [et al.  and Fara [et al. . Moreove [et al. , 22-24 s [et al. .Within the limitations of this study, it can be concluded that in the permanent maxillary first molars, mesiobuccal roots have a more complex root canal configuration than palatal and distobuccal roots.


Nanomaterials examines the potential for engineered nanomaterials to negatively impact biological systems and highlights some advances in evaluating key areas of their hazard potential. Nanomaterial science is evolving rapidly with the generation of more complex nanostructures with exciting potential applications. Keeping modern toxicology abreast of this innovation to the point that it guides a safer nanotechnology presents an equally exciting and eminently worthwhile challenge.This Special Issue of et al. [in vitro methods for measurement of nanostructure cytotoxicity. Cohignac et al. [et al. [et al. [et al. [In this issue, Nogueira et al.  provide c et al.  review c [et al.  review t [et al.  that gen [et al.  report oet al. [et al. [et al. [et al. [et al. [et al. [et al. [In medical nano-applications, Qiao et al.  report a [et al.  address  [et al.  review h [et al.  offer ne [et al.  report t [et al.  report t [et al.  report s


Cole .An addendum to the article by  Introduction of Cole et al.Acta Cryst. D73, 234–239][(2017),  is made to recognize the work of Bricogne, Smart and others in the development of methods to make use of Cambridge Structural Database data in protein structure solution.An addendum to the  Introduction to Cole et al. (2017Mogul (Bruno et al., 2004BUSTER and GRADE; Smart et al., 2011Mogul was applied to protein crystallography, and so should also have been recognised.In the  al. 2017 a short


In September 2016, a paper was published that assessed the 27 available reviews and meta-analyses exploring the efficacy of VR in behavioral health , into reality.In their review, Freeman et al.  also sug. et al. agree th.et al. , the lon. et al., but, in


Unfortunately, the original version of this article (Lee et al.


In doing so, they criticize the scientific interpretation and media communication of RSPB [et al. model many of the behaviours of which they accuse others. We illustrate this below, correct scientific misrepresentations of our work by Davies et al. and challenge the view that the current narrative is over-simplified or insufficient to inform policy.Davies et al.  argue throgallus  exemplif of RSPB  and acadet al. [et al. [et al. (2007)’, yet in [et al. 2007)’. A shift in position based on new evidence would be one thing, but use of the same citation to support two different statements on the timing of wildfires is quite another.Davies et al.  question [et al.  state ‘E, yet in , they stet al. [et al. [et al. [et al. [Lagopus lagopus scotica shooting, show a striking seasonal correspondence with the prescribed burning season [et al. [et al. [Regarding scientific criticisms and uses of Douglas et al.  that req [et al.  firstly  [et al. ,5 misint [et al.  state th [et al.  that theet al. of values in [et al. [et al. [annual areas burned. Disregarding of repeat burning renders all the associated calculations in [Secondly, the use by Davies alues in  to estimalues in  are deri [et al.  report t [et al. , also wi [et al. . Davies  [et al.  estimate [et al. , of 0.04tions in  grossly et al. judge that press releases associated with [and the damage it can cause’  does, in the italicized text, exceed the research findings, and we acknowledge that these words should not have been included. However, given Davies et al.'s advice that “…authors ensure that the press releases associated with their findings accurately reflect the content of their research…”, it is unfortunate that their own accepted manuscript was the subject of media coverage (e.g. [Davies ted with  were notge (e.g. ) prior tge , but suew , and the manifest societal and environmental benefits of restoring peat-dominated ecosystems [Throughout their paper, Davies osystems , a more


Acta Cryst. (2006), E62, i150–i151.Erratum to  et al. , the sodium cation was assigned incorrectly.In the paper by Zhang  Choi et al. (1976The authors of Zhang  al. 2006 have ind al. 1976.


Piscine orthoreovirus (PRV) causes acute infection of the red blood cells in salmon  with clal. 2004 ). Palacial. 2013 ). HSMI hal. 2017 ). Hence,et al. [In the correction, Siah et al.  acknowleet al. [Their conclusion appears to be highly dependent on six unique sequences of PRV segment S1, detected by Siah KR478642: collected in May 2001KR478643: collected in Aug. 2001KR478644: collected in Aug. 2001KR347078: collected in Aug. 2001KR347079: collected in Aug. 2001KR347080: collected in Mar. 2005et al. [et al. [et al. [et al. [U) to be between 1 and 0.1 for most RNA viruses, where U is G x u, G is the genome size in nucleotides, and u is the per-nucleotide mutation rate.These six Siah et al.  sequence [et al. ,11 by se [et al. ). Drake  [et al.  estimateO. mykiss) collected in 1977 in support of longer-term PRV presence in BC. This result is cited from Marty et al. [et al. [et al. [et al. 2016 [Siah et al.  cite dety et al. , who pro [et al.  and Gars [et al. . The recFurthermore, this result could not be replicated by a second laboratory (Purcell and Thompson 2014 ) and theTo illustrate our interpretation of the phylogenetic analysis of PRV isolates, we constructed a phylogenetic tree , of the We estimated the divergence time between the genotypes and sub-genotypes  based on the collection time of each isolate described in et al. 2016 [We estimated that the divergence time between sub-genotype Ia and sub-genotype Ib was 2007 or earlier. Because all sequences in et al. [The description of the temporal and spatial distribution of the samples reported on by Siah et al.  is inadeet al. [Furthermore, all but eight of the partial sequences reported by Siah et al.  in theiret al. [et al. 2015 [O. kisutch, from the Copper River in Alaska), and only six Chinook Salmon, O. tshawytscha, were sampled . No other Pacific salmon or trout species were included. Hence, the host species and spatial coverage of PRV sequencing presented by Siah et al. [et al. [There was also inadequate spatial and host-species coverage. Siah et al. , reporteet al. [Furthermore, farm restocking methods could potentially account for at least some of the homogeneity in the Siah et al.  samples.et al. [et al. [et al. [a region where HSMI has never been reported.” (Emphasis added.) However, Kibenge et al. [et al. 2017 [Siah et al.  state, c [et al.  and Mart [et al. , “PRV ise et al.  do cite al. 2017 ).We conclude that the longer-term presence of PRV in BC prior to 2001 has not been adequately described and that the evidence that the virus was introduced from Norway is more robust than the hypothesis that PRV is endemic to the eastern Pacific Ocean.S1 Table(DOC)Click here for additional data file.


Although many orthodontists have no doubts about the effectiveness of functional appliances for mandibular advancement, the impact on the temporomandibular joint (TMJ) is still in dispute. The objective of this systematic review is to examine the main effects on the TMJ of using functional appliances, both in healthy patients and in patients with a pre-existing disorder.A systematic review of the literature was conducted in accordance with the PRISMA guidelines. Only systematic reviews, meta-analyses, randomized clinical trials (RCTs), case-control studies and cohort studies were included. A detailed language-independent electronic search was conducted in the Pubmed, Scopus, Cochrane Library and Embase databases. All studies published between 2000 and 2015 were included.A total of 401 articles were identified. Of these, 159 were duplicates and were excluded. On reading the title and abstract, 213 articles were excluded because they did not answer the research question, leaving a total of 29 articles. These articles were read and assessed. Following critical reading of the full text, eight articles were excluded: seven because they were considered of low quality and one because it published redundant data. As a result, 21 articles were included.After treatment with functional appliances, the condyle was found to be in a more advanced position, with remodelling of the condyle and adaptation of the morphology of the glenoid fossa. No significant adverse effects on the TMJ were observed in healthy patients and the appliances could improve joints that initially presented forward dislocation of the disk.Key words:Temporomandibular joint, TMJ, orthodontic appliances, functional, mandibular advancement, herbst appliance, bionator. The use of a mandibular advancement appliance to correct skeletal malocclusions associated with mandibular retrognathism is indicated during the first stage of orthodontic treatment . The objWhen the functional appliance is inserted, the condyles are moved to a higher position in the articular eminence, which is capable of adaptation, so it could be hypothesized that some morphological changes may take place .Although many orthodontists have no doubts about the effectiveness of functional appliances, their impact on the TMJ is still considered a subject of debate. It is therefore of clinical and scientific interest to investigate the most relevant effects of different functional appliances on the TMJ, particularly with current diagnostic methods. Many authors assert that treatment with these devices does not increase the prevalence of temporomandibular disorders ,7-9. AnoThe main objective of this systematic review is to examine the main effects on the TMJ of different functional appliances for mandibular advancement, both in healthy patients and in patients with a pre-existing disorder.A systematic review of the bibliography was carried out in accordance with the PRISMA  recommendations  and CONS- Eligibility criteriaThe selection criteria for inclusion in the review were: articles, articles in press and reviews. Only the following types of study were accepted: systematic reviews, meta-analyses, randomized clinical trials (RCTs), case-control studies and cohort studies. Both retrospective and prospective studies were included. The articles that met these criteria and studied changes in the temporomandibular joint following the use of functional appliances for mandibular advancement were included in the review.- Search strategy and screeningTo identify the relevant studies, irrespective of language, a thorough electronic search was conducted in the Pubmed, Scopus, Cochrane Library and Embase databases. All studies published between 2000 and 2015 were included. The search was updated on 4 July 2015.The search strategy was implemented through a combination of the following 6 MeSH terms: “Temporomandibular joint”, “TMJ”, “Functional orthodontic appliances”, “Mandibular advancement”, “Herbst appliance”, “Bionator” and “Activator appliances”, and others: “Fränkel Function Regulator”, “Orthodontic Appliances”, “Activator”, “Function Activator”, “Bionator Appliance”, “Functional Orthodontic” and “Twin Block”. The TMJ related terms were combined using the OR Boolean operator, as well as those related to the mandibular advancement devices. Both groups were combined using the AND Boolean operator. Hand-searching in the reference lists of the articles assessed for the review was also carried out.Two independent reviewers assessed the titles and abstracts of all the articles selected. In the event of their disagreeing, a third reviewer was consulted. If the abstract did not provide sufficient information for a decision, the reviewers read the full article before taking the final decision. The reviewers then read the full text of all the resulting articles.- Data extractionet al. , sample size, dropouts, demographic variables , type of appliance used, type of advancement , presence or otherwise of preexisting TMJ disorders, length of treatment, type of radiographic study used to study the changes, follow-up time for each study and quality of the articles accepted . The artet al.  and useds et al. ,14. The s et al. .A thorough search identified 201 articles in Medline, 168 in Scopus, none in the Cochrane Library and 32 in Embase, making a total of 401 articles. 159 duplicates were removed, leaving 242. After reading the title and abstract, 213 were removed because they did not answer the research question, leaving a total of 29 articles. These were read and carefully assessed. Detailed critical reading of the full text resulted in the exclusion of 8 articles, 7 because of their low quality .In relation to the type of diagnostic study, 8 used magnetic resonance, 2 CBCT, 1 SPECT (single-proton emission computed tomography), 6 cephalometric tracing, 1 magnetic resonance together with manual examination, 1 magnetic resonance as well as cephalometric tracing and wrist radiographs, and 1 used magnetic resonance and teleradiography.Regarding the quality of the articles, 19 were of moderate quality and one of high quality. The systematic review was considered of high quality. The main conclusions drawn by the authors of the studies included in the present review are shown in - Changes in disk shape and positionet al. (et al. (et al. (et al. (et al. (et al. (et al. (et al. (et al. (et al. (et al. (According to Wadhawan et al. , the ant (et al. . However (et al.  found no (et al.  observed (et al.  and the  (et al. . Aidar e (et al. , in a pr (et al. , using m (et al. , who als (et al. . Pancher (et al.  performeet al. (et al. (Concerning disk shape, Aidar et al.  reportedet al.  reached  (et al. , using m- Changes in condyle size and positionet al. (et al. (et al. (et al. (et al. (In a case-control study of treatment with the Herbst appliance, Le Cornu et al.  observed (et al.  and Chav (et al. . The lat (et al.  observed (et al.  concludeet al. (et al. (et al. (In contrast to the above observations, Kinzinger et al.  found no (et al. , using m (et al.  also con- Changes in the glenoid fossaet al. (et al. (et al. (et al. (Pancherz et al.  observed (et al.  conclude (et al. . Focusin (et al.  conclude- Changes in the articular eminence and other changeset al. (Reports on adaptations of the articular eminence caused by functional appliances are scarce. Katsavrias et al.  highlighet al. (et al. (With regard to other changes that may take place, according to the study by Le Cornu et al.  the Herb (et al.  observed- Limitations of this reviewThe scientific evidence collected on changes in the TMJ following the use of functional appliances was not abundant despite the thorough systematic search for articles that met the strict inclusion criteria and presented medium to high quality according to the CONSORT criteria.TMJ status is a particularly important subject, yet this review has shown a lack of randomized controlled trials with long-term follow-up using diagnostic tests such as magnetic resonance and computed tomography to establish the effects of the functional appliance in a reliable way.et al. (et al. (et al. (et al. (et al. (et al. (et al. (et al. (The articles included in the review show a lack of methodological homogeneity. For instance, Aidar et al. , Kinzing (et al.  and Wadh (et al.  used mag (et al.  and Aric (et al.  used CBC (et al.  and Güne (et al. , used ce (et al. , preferrThey also employed different appliances: while the great majority used the Herbst, others studied removable appliances such as Twin Block, Bionator ,16, Fränet al. (et al. (et al. (et al. (et al. (et al. (Additionally, the sample size was small in most of these studies: Le Cornu et al.  studied  (et al.  40 patie (et al.  35 patie (et al.  and Kinz (et al.  studied • Following this review, it may be concluded that observations after treatment with functional appliances for mandibular advancement have found the condyle in a more advanced position, condyle remodelling and adaptation of the morphology of the glenoid fossa.• No significant adverse events concerning the temporomandibular joint have been found in healthy patients, and this treatment could improve joints that initially presented forward disk dislocation.


R. Soc. open sci. 4, 170833. (Published 25 October 2017). (doi:10.1098/rsos.170833)et al. [et al. [et al. [Pisanosaurus, which was not made explicit in our manuscript (results shown in fig. 5). Also, owing to an error prior to publication, the full citation to Langer et al. [et al. [In our work, we reanalysed phylogenetic datasets for dinosaurs. This included a dataset assembled by Langer et al.  which wa [et al. . The dat [et al.  to the cr et al.  was not  [et al. , which w