Kluyveromyces marxianus: a potential biocatalyst of renewable chemicals and lignocellulosic ethanol production.

Kluyveromyces marxianus is an ascomycetous yeast which has shown promising results in cellulosic ethanol and renewable chemicals production. It can survive on a variety of carbon sources under industrially favorable conditions due to its fast growth rate, thermotolerance, and acid tolerance. K. marxianus, is generally regarded as a safe (GRAS) microorganism, is widely recognized as a powerhouse for the production of heterologous proteins and is accepted by the US Food and Drug Administration (USFDA) for its pharmaceutical and food applications. Since lignocellulosic hydrolysates are comprised of diverse monomeric sugars, oligosaccharides and potential metabolism inhibiting compounds, this microorganism can play a pivotal role as it can grow on lignocellulosic hydrolysates coping with vegetal cell wall derived inhibitors. Furthermore, advancements in synthetic biology, for example CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, will enable development of an engineered yeast for the production of biochemicals and biopharmaceuticals having a myriad of industrial applications. Genetic engineering companies such as Cargill, Ginkgo Bioworks, DuPont, Global Yeast, Genomatica, and several others are actively working to develop designer yeasts. Given the important traits and properties of K. marxianus, these companies may find it to be a suitable biocatalyst for renewable chemicals and fuel production on the large scale. This paper reviews the recent progress made with K. marxianus biotechnology for sustainable production of ethanol, and other products utilizing lignocellulosic sugars.

Detection of atypical porcine pestivirus in Brazil in the central nervous system of suckling piglets with congenital tremor.

Atypical porcine pestivirus (APPV) has been detected in piglets with congenital tremor (CT) from three different continents including North America, Europe and Asia. Thirteen piglets from four farms in two different states in Brazil with CT were sampled. Viral RNA was detected by quantitative real-time PCR in the cerebellum or cerebellum and spinal cord in the 100% of the piglets with CT, and APPV was not detected in any tissue sample from clinically non-affected piglets with the exception of the cerebellum of one piglet from Farm A. Piglets with CT had an odds ratio of 99.0 (95% CI 3.4, 2823.8; p = .0072) compared to piglets without CT to test positive for APPV by qRT-PCR. A subset of positive samples was selected for sequencing of the NS3 gene. Phylogenetic analysis revealed that Brazilian sequences of the NS3 formed an independent cluster and had the highest sequence identity with a sequence from the United States. This is the first identification of APPV infection in piglets with CT in South America.

Detection of atypical porcine pestivirus in semen from commercial boar studs in the United States.

Atypical porcine pestivirus (APPV) has recently been identified as a cause of congenital tremor (CT) in pigs and has been detected in semen and preputial swabs from boars that were known to be clinically affected with CT. Accordingly, the objectives of this study were to 1) detect the presence of APPV in semen, preputial fluids and preputial swabs from adult boars by quantitative reverse transcription PCR (qRT-PCR) and 2) genetically characterize a subset of positive samples to better understand the ecology of APPV in commercial boar studs and the potential risk of transmission of APPV via semen. A total of 597 samples of semen, preputial fluid and preputial swabs each representing a different boar were obtained from four commercial boar studs located in three different states in the United States. Viral RNA was detected by qRT-PCR in 90 samples (15.08%; 90/597), with the greatest per cent positive from preputial swabs (23.81%; 5/21) followed by preputial fluid (22.81%; 26/114) and semen (12.91%; 59/457). The mean cycle quantification (Cq) between sample types was similar while eleven semen samples had Cq values lower than 27.0 corresponding to approximately 2 × 106  copies/ml. Based on phylogenetic analysis of the Npro gene, different viral strains can be on the same farm at the same and different times. This is the first report of detection of APPV in semen from commercial boar studs. Studies investigating the role of semen in the transmission of APPV and production of CT are needed.

Detection of a novel sapelovirus in central nervous tissue of pigs with polioencephalomyelitis in the USA.

An approximately 3,000 finishing swine operation in the United States experienced an outbreak of an atypical neurologic disease in 11-weeks-old pigs with an overall morbidity of 20% and case fatality rate of 30%. The clinical onset and progression of signs in affected pigs varied but included inappetence, compromised ambulation, ataxia, incoordination, mental dullness, paresis, paralysis and decreased response to environmental stimuli. Tissues from affected pigs were submitted for diagnostic investigation. Histopathologic examination of the cerebrum, cerebellum and spinal cord revealed severe lymphoplasmacytic and necrotizing polioencephalomyelitis with multifocal areas of gliosis and neuron satellitosis, suggestive of a neurotropic viral infection. Bacterial pathogens were not isolated by culture of neurologic tissue from affected pigs. Samples tested by polymerase chain reaction (PCR) were negative for pseudorabies virus and atypical porcine pestivirus. Immunohistochemistry for porcine reproductive and respiratory syndrome virus, porcine circovirus and Listeria was negative. Porcine sapelovirus (PSV) was identified in spinal cord by a nested PCR used to detect porcine enterovirus, porcine teschovirus and PSV. Next-generation sequencing of brainstem and spinal cord samples identified PSV and the absence of other or novel pathogens. In addition, Sapelovirus A mRNA was detected in neurons and nerve roots of the spinal cord by in situ hybridization. The PSV is genetically novel with an overall 94% amino acid identity and 86% nucleotide identity to a recently reported sapelovirus from Korea. This is the first case report in the United States associating sapelovirus with severe polioencephalomyelitis in pigs.

Avaliação da depressão miocárdica por meio do feature tracking bidimensional em cadelas com sepse / Assessment of myocardial depression by two-dimensional feature tracking in dogs with sepsis

O objetivo deste estudo prospectivo foi analisar a função do ventrículo esquerdo de cadelas que apresentavam sepse ou não e diagnosticar a depressão miocárdica de forma precoce, por meio do feature tracking imaging bidimensional (FTI-2D).Vinte e nove cadelas diagnosticadas com piometra, patologia usada como modelo experimental para sepse, foram avaliadas. Os animais foram submetidos ao exame ecocardiográfico convencional e ao FTI-2D e divididos em três grupos: grupo I (controle), grupo II (piometra sepse) e grupo III (piometra não sepse). Em relação aos parâmetros da ecocardiografia convencional, não houve diferença estatística entre os grupos avaliados. Quanto ao strain radial, os grupos II e III apresentaram função ventricular inferior no momento do diagnóstico (22,805,80 grupo I; 18,455,96 grupo II; 18,719,45 grupo III; P=0,032) e o grupo III manteve essa redução no retorno. Já no strain rate radial, apenas o grupo III obteve função ventricular inferior no momento do diagnóstico (1,900,57 grupo I; 1,910,70 grupo II, 1,570,53 grupo III; P=0,021). Para o strain longitudinal, observou-se função ventricular mais baixa nos grupos II e III no momento do diagnóstico (11,043,38 grupo I; 8,593,97 grupo II; 8,211,99; P=0,048) e após 10 dias de pós-cirúrgico (11,043,38 grupo I; 9,002,16 grupo II; 8,122,27 grupo III; p=0,048). O FTI-2D foi capaz de detectar alterações precoces da função ventricular esquerda, que não foram observadas no exame ecocardiográfico convencional, sugerindo quadro de depressão miocárdica, até mesmo em cadelas com piometra sem quadro clínico evidente de sepse. Assim, essa modalidade pode ser útil no diagnóstico, pois pode detectar alterações cardíacas precoces, possibilitando uma intervenção terapêutica rápida.(AU)

The aim of this prospective study was to analyze the left ventricular function of dogs who had sepsis or not and diagnose myocardial depression early on through the two-dimensional feature tracking imaging (2D-FTI). Twenty-nine dogs diagnosed with pyometra, a pathology used as an experimental model for sepsis were evaluated. The animals were subjected to conventional echocardiography and 2D-FTI, and divided into three groups: Group I (control), group II (sepsis pyometra) and group III (no sepsis pyometra). Regarding the parameters of conventional echocardiography, there was no statistical difference between the groups. As for the radial Strain, group II and III had lower ventricular function at diagnosis (22.805.80 Group I; 18.455.96 Group II; 18.719.45 Group III; P = 0.032) and group III maintained this reduction in return. Regarding the radial Strain rate, only the III group had lower ventricular function at diagnosis (1.900.57 Group I; 1.910.70 Group II, 1.570.53 Group III; P = 0.021). For the Longitudinal Strain we observed lower ventricular function in groups II and III at diagnosis (11.043.38 Group I, Group II 8.593.97, 8.211.99; P = 0.048) and ten days after surgery (11.043.38 Group I; 9.002.16 Group II; 8.122.27 Group III; P = 0.048). The 2D-FTI was able to detect early changes in left ventricular function, suggesting myocardial depression frame, even in female dogs with pyometra without clear clinical picture of sepsis, which were not seen in conventional echocardiography. Thus, this model can be useful in diagnosis, because it can detect early cardiac changes, enabling a rapid therapeutic intervention.(AU)

Effects of different lipid sources on intake, digestibility and purine derivatives in hair lambs.

An experiment was conducted to evaluate the effects of different lipid sources on the nutrient intake, digestibility and purine derivative excretion of lambs. Thirty-five 60-day-old, male, non-castrated Santa Ines lambs with an initial average body weight (BW) of 13.00 ± 1.80 kg were used in a randomized complete block design with seven blocks and five treatments. The experimental treatments consisted of a control diet without supplemental lipids and four test diets with different lipid supplements, selected according to the degree of ruminal protection from hydrogenation: supplementation, being supplementation with whole cottonseed (WC), supplementation with cashew nut meal (CNM), supplementation with both cottonseed and cashew nut meal (WC-CNM) and supplementation with calcium salts of long-chain fatty acids (Ca-LCFA). The lambs were slaughtered after reaching 28 kg average BW for each treatment. The ether extract intake (EEI) was higher (p < 0.01) for the lipid supplemented compared to control diet lambs. Supplementation with WC decreased the digestibility of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF) and total carbohydrate (TC) (p < 0.01), whereas supplementation with CNM, WC-CNM and Ca-LCFA reduced non-fibrous carbohydrate (NFC) digestibility (p < 0.01). The ether extract (EE) digestibility coefficient was higher with CNM, followed by Ca-LCFA and WC, when compared to WC-CNM and control diets. Nitrogen balance (NB) was not influenced (p > 0.05) by the different lipid sources. A lower purine derivative (PD) excretion and thus lower microbial protein supply (MPS) was observed for animals supplemented with Ca-LCFA (p < 0.01) compared to the WC-CNM and control diets. In conclusion, WC, CNM and WC-CNM supplementation did not have negative effects on MPS, although negative effects have been observed on nutrient digestibility.

Characterization of Porcine Epidemic Diarrhea Virus Isolate US/Iowa/18984/2013 Infection in 1-Day-Old Cesarean-Derived Colostrum-Deprived Piglets.

Porcine epidemic diarrhea virus (PEDV) was first recognized in North America in April 2013 and has since caused devastating disease. The objective of this study was to characterize disease and viral detection associated with an original North American PEDV isolate inoculated in neonatal piglets. Thirty-six 1-day-old cesarean-derived and colostrum-deprived piglets were randomly assigned to the control (n = 16) or challenged group (n = 20); the latter were orogastrically inoculated with 1 ml of US/Iowa/18984/2013 PEDV isolate titered at 1 × 10(3) plaque-forming units per milliliter. Rectal swabs were collected from all piglets prior to inoculation and every 12 hours postinoculation (hpi) thereafter, with 4 control and 5 challenged piglets euthanized at 12, 24, 48, and 72 hpi. One piglet had a positive real-time quantitative polymerase chain reaction test on rectal swab at 12 hpi, and all remaining piglets were positive thereafter, with highest viral quantities detected at 24 and 36 hpi. Diarrhea was evident in 30% and 100% of challenged piglets at 18 and 24 hpi, respectively. Viral antigen was detected in enterocytes by immunohistochemistry in the duodenum and ileum of piglets euthanized at 12 hpi and was apparent throughout the small intestine of all piglets thereafter, with villus height:crypt depth ratios consistently below 4:1. Viremia was confirmed in 18 of 20 pigs at euthanasia. Clinical disease was severe and developed rapidly following infection with an original North American PEDV isolate, with lesions, viremia, and antigen detection possible by 12 hpi.

Análise produtiva e econômica de suínos criados nos sistemas wean-to-finish e convencional de produção / Productive and economic analysis of pigs raised in wean-to-finish and conventional production systems

O modelo de produção de suínos denominado Wean-to-Finish (WF) é considerado uma alternativa que busca alcançar melhor desempenho produtivo, redução de estresse aos animais e melhorias na logística. Nesse sistema, os leitões são desmamados e transferidos diretamente a um galpão WF (creche-crescimento-terminação), onde permanecem até o abate. Com o objetivo de avaliar os comportamentos produtivo e econômico de suínos alojados no sistema WF em comparação ao sistema convencional de produção (CC), foram avaliados dois grupos de suínos provenientes das mesmas unidades produtoras de leitões, nascidos e desmamados na mesma semana. Um grupo foi alojado em uma creche do sistema convencional de produção (grupo CC), e o outro em um galpão WF adaptado (grupo WF), ambos em baias coletivas. Para análise de produção, foram calculados o consumo médio diário de ração (CMDR), o ganho de peso médio diário (GPMD) e a conversão alimentar (CA). Para análise econômica, foi determinado o custo por quilograma de ganho de peso vivo. Os dados foram submetidos à análise de variância utilizando-se o pacote estatístico SAS, previamente testados para normalidade dos resíduos pelo teste de Shapiro-Wilk, e as médias dos grupos comparadas pelo teste de Tukey (5%). Na fase de creche, o GPMD foi de 0,468 e 0,449, e a CA de 1,48 e 1,42 para os Grupos WF e CC, respectivamente. Nas fases de crescimento e terminação, o GPMD foi de 0,924kg no grupo WF e de 0,909kg no grupo CC. A conversão alimentar foi de 2,41 para ambos os grupos, não sendo diferentes estatisticamente entre si (P>0,05) nas diferentes fases de produção nos parâmetros analisados. No aspecto econômico, o grupo WF apresentou os menores custos, com diferença de R$ 0,08 por quilograma de ganho de peso vivo em comparação ao grupo CC. Os resultados do desempenho produtivo associado ao aspecto econômico indicam que o sistema WF pode ser uma alternativa viável dentro da cadeia produtiva de suínos.

The pig production model called "Wean-to-Finish" (WF) is considered an alternative to better productive performance, reducing stress to the animals and improvements in logistics. In this system, the piglets are weaned and transferred directly to a WF shed (nursery-grower-finisher), where they remain until slaughter. Aiming to evaluate the productive and economic behavior of pigs housed in the WF system compared to conventional production system (CC), two groups of pigs from the same sow farm, born and weaned in the same week were evaluated. One group was housed in a conventional nursery production system (CC group) and the other in a WF shed adapted (WF group), both in collective pens. For analysis of the average production, average daily feed intake (ADFI), average daily weight gain (ADWG) and feed conversion ratio (FCR) were calculated. For economic analysis the cost per kilogram of live weight gain was determined. Data were subjected to analysis of variance using the SAS statistical package, previously tested for normality of the residuals using the Shapiro-Wilk test, and the means of groups were compared by the Tukey test (5 %). In the nursery phase ADWG was 0.468 and 0.449, and the FCR was 1.48 and 1.42 for the WF and CC groups respectively. During the growing and finishing ADWG was 0.924kg in the WF group and 0.909 in the CC group. The FCR was 2.41 for both groups, not being statistically different from each other (P>0.05) at different stages of production in the analyzed parameters. In the economic aspect, the WF group had the lowest costs, with a difference of R$ 0.08 per kilogram of live weight gain compared to the CC group. The results of productive performance associated with the economic aspect indicate that the WF system can be a viable alternative in the pig production chain.

Pathogenesis of porcine epidemic diarrhea virus isolate (US/Iowa/18984/2013) in 3-week-old weaned pigs.

Porcine epidemic diarrhea virus (PEDV) is associated with clinical diarrhea in naïve swine of all ages. This report describes timing of antibody generation and disease progression following infection with a US PEDV isolate by assessing fecal viral shedding, morphometric analysis of intestinal lesions, and magnitude of immunohistochemical staining. Sixty-three, 3-week-old pigs were randomly allocated into control (n=27) and challenged (n=36) groups. Challenged pigs were administered 1 mL of 1 × 10(3) PFU/mL of US/Iowa/18984/2013 PEDV isolate by oro-gastric gavage. Three control and four challenged pigs were necropsied on days post-inoculation (dpi) 1, 2, 3, 4, 7, and weekly thereafter, until study termination on dpi 35. Clinical disease, fecal shedding, body weight, and temperature were monitored during the study period. Diarrhea was observed in challenged pigs beginning for some on dpi 2, affecting a majority of pigs by dpi 6 and subsiding by dpi 10. Average daily gain was significantly lower (P<0.001) for one week post-infection in challenged pigs. PEDV was detected in feces by PCR on dpi 1 and continued in a subset of pigs until dpi 24. PEDV-specific antigen was detected in villous enterocytes of challenged pigs by immunohistochemistry (IHC) on dpi 1, 2, 3, 4, 7, and 14. Microscopic lesions included severe diffuse atrophic enteritis with significantly reduced (P<0.001) villous length observed on dpi 3, 4, and 7. Under the conditions of this study, fecal shedding of PEDV and IHC staining can precede and continue beyond the observation of clinical signs, thus increasing the risk of viral transmission.

Cessation of clinical disease and spirochete shedding after tiamulin treatment in pigs experimentally infected with "Brachyspira hampsonii".

With the emergence of "Brachyspira hampsonii" associated with swine dysentery in North America, identification of effective treatments and interventions is a pressing need. Denagard® (tiamulin hydrogen fumarate) Liquid Concentrate 12.5% is approved in the United States for treatment of dysentery caused by Brachyspira hyodysenteriae at 0.006% in the water. In this study, the effectiveness of tiamulin in resolving clinical disease, eliminating viable spirochete shedding, and reducing neutrophilic colitis following infection with either "B. hampsonii" or B. hyodysenteriae was evaluated. Seventy-eight 7-week-old crossbred pigs were divided into three groups [sham-inoculated (n = 18), "B. hampsonii"-inoculated (n = 30), and B. hyodysenteriae-inoculated (n = 30)]. Each inoculum group was divided into three subgroups which received either 0.006% tiamulin, 0.018% tiamulin, or no medication. Both levels of tiamulin resolved clinical disease within 24 h of treatment initiation, eliminated spirochete shedding within 72 h of treatment initiation, and resolved and/or prevented histologic lesions in pigs infected with either Brachyspira spp.

Comparison of sesion severity, distribution, and colonic mucin expression in pigs with acute swine dysentery following oral inoculation with "Brachyspira hampsonii" or Brachyspira hyodysenteriae.

Swine dysentery is classically associated with infection by Brachyspira hyodysenteriae, the only current officially recognized Brachyspira sp. that consistently imparts strong beta-hemolysis on blood agar. Recently, several strongly beta-hemolytic Brachyspira have been isolated from swine with clinical dysentery that are not identified as B. hyodysenteriae by PCR including the recently proposed species "Brachyspira hampsonii." In this study, 6-week-old pigs were inoculated with either a clinical isolate of "B. hampsonii" (EB107; n = 10) clade II or a classic strain of B. hyodysenteriae (B204; n = 10) to compare gross and microscopic lesions and alterations in colonic mucin expression in pigs with clinical disease versus controls (n = 6). Gross lesions were similar between infected groups. No histologic difference was observed between infected groups with regard to neutrophilic inflammation, colonic crypt depth, mucosal ulceration, or hemorrhage. Histochemical and immunohistochemical evaluation of the apex of the spiral colon revealed decreased expression of sulphated mucins, decreased expression of MUC4, and increased expression of MUC5AC in diseased pigs compared to controls. No difference was observed between diseased pigs in inoculated groups. This study reveals significant alterations in colonic mucin expression in pigs with acute swine dysentery and further reveals that these and other microscopic changes are similar following infection with "B. hampsonii" clade II or B. hyodysenteriae.

Preparo e caracterização de tinturas das folhas de chá verde [Camellia sinensis (L.) O. Kuntze] Theaceae / Preparation and characterization of Theaceae green tea [Camellia sinensis (L.) O. Kuntze] leaf tinctures

Tinturas preparadas com as folhas de Camellia sinensis (chá verde), foram caracterizadas visando a obtenção de extratos enriquecidos em polifenóis. As tinturas foram obtidas por meio de maceração estática da droga pulverizada, com diferentes misturas de 60, 70, 80 e 94,5% de etanol em água. Em seguida foram filtradas e, após 8 dias de extração, realizaram-se as analises: organoléptica, pH, densidade, determinação do resíduo seco, perfil cromatográfico por camada delgada, e teor de polifenóis. Os resultados demonstraram que o solvente (mistura hidroalcoólica a 60 ou 70%) extraiu a maior quantidade de constituintes químicos do chá verde; também foi o mais seletivo ao extrair, especificamente os polifenóis. Estes dados sugerem que a utilização de álcool etílico a 60 ou 70% conduz à obtenção de tinturas mais ricas em polifenóis, a partir das folhas do chá verde (C. sinensis).

In this work, we prepared and characterized Camellia sinensis leaf tinctures aiming to obtain a polyphenol-enriched extract. The tinctures were prepared through the static maceration process of the powdered drug, at different mixtures 60, 70, 80 and 94.5% of ethanol in water. The tinctures were filtered, and after 8 days of extraction, submitted to organoleptic analysis, determination of pH, density analysis, dry residue analysis, thin layer chromatographic profile and polyphenol percentage analysis. The results showed that the 60 or 70% alcohol-water mixture has the best extraction of the constituents of green tea and it was more selective to extract, specifically, the polyphenols of the plant. These data suggest the use of 60 or 70% ethanol to carry out polyphenol-enriched tinctures from green tea leaves (C. sinensis).

Application of a simulated annealing optimization to a physically based erosion model.

A major risk concerning the calibration of physically based erosion models has been partly attributable to the lack of robust optimization tools. This paper presents the essential concepts and application to optimize the erosion parameters of an erosion model using data collected in an experimental basin, with a global optimization method known as simulated annealing (SA) which is suitable for solving optimization problems of large scales. The physically based erosion model that was chosen to be optimized here is the Watershed Erosion Simulation Program (WESP), which was developed for small basins to generate the hydrograph and the respective sedigraph. The field data were collected in an experimental basin located in a semiarid region of Brazil. On the basis of these results, the following erosion parameters were optimized: the soil moisture-tension parameter (N(s)) that depends also on the initial moisture content, the channel erosion parameter (a), the soil detachability factor (K(R)), and the sediment entrainment parameter by rainfall impact (K(I)), whose values could serve as initial estimates for semiarid regions within northeastern Brazil.

Evaluation of sorghum straw hemicellulosic hydrolysate for biotechnological production of xylitol by Candida guilliermondii

A preliminary study on xylitol production by Candida guilliermondii in sorghum straw hemicellulosic hydrolysate was performed. Hydrolysate had high xylose content and inhibitors concentrations did not exceed the commonly found values in other hemicellulosic hydrolysates. The highest xylitol yield (0.44 g/g) and productivity (0.19 g/Lh) were verified after 72 hours.

Evaluation of sorghum straw hemicellulosic hydrolysate for biotechnological production of xylitol by Candida guilliermondii.

A preliminary study on xylitol production by Candida guilliermondii in sorghum straw hemicellulosic hydrolysate was performed. Hydrolysate had high xylose content and inhibitors concentrations did not exceed the commonly found values in other hemicellulosic hydrolysates. The highest xylitol yield (0.44 g/g) and productivity (0.19 g/Lh) were verified after 72 hours.

High-lysine maize: the key discoveries that have made it possible.

Forty-five years ago, a paper published by Mertz et al. (Science 145:279-280, 1964) initiated a revolution in the history of plant protein quality and affected dramatically the study of cereal crop storage proteins. The observation of the high lysine content of the endosperm of the opaque-2 (o2) maize mutant was a key factor in bringing about a new concept in the production of cereal seeds with a high nutritional value. It has been a long and very interesting road with astonishing results over these 45 years. We are now probably about to see the release of commercially engineered high-lysine maize lines. We have decided to pinpoint some key contributions to the science behind high-lysine plants and concentrated on the research done on maize, which is possibly the most complete and simple example to illustrate the advances achieved. However, studies on other plant species such as barley and model species such as tobacco are totally relevant and will be briefly addressed.

ESTs as a source for sequence polymorphism discovery in sugarcane: example of the Adh genes.

Expressed sequence tags (ESTs) have proven to be a valuable tool to discover single nucleotide polymorphism (SNP) in human genes but their use for this purpose is still limited in higher plants. Using a database of approximately 250,000 sugarcane ESTs we have recovered 219 sequences encoding alcohol dehydrogenases ( Adh), which tagged 178 distinct cDNAs from 27 libraries, constructed from at least four different cultivars. The partitioning of these ESTs into paralogous genes revealed three Adh genes expressed in sugarcane, one Adh2 and two Adh1. The soundness of the partition was carefully checked by comparison to external data, especially from the closely related sorghum. Analysis of polymorphism in the alignments of EST sequences revealed a total of 37 highly reliable SNPs in the coding and untranslated regions of the three Adh genes. In the coding regions, the mean occurrence of SNPs was one for every 122 base pair. A total of eight insertion-deletions was observed, their occurrence being limited to untranslated regions. These results show that EST data constitute an invaluable source of sequence polymorphism for sugarcane that is worth carefully collecting for the future development of new marker tools.

Molecular characterisation and evolutionary relationships of a potyvirus infecting Crotalaria in Brazil. Brief report.

The 3' terminal genomic region of a potyvirus causing mosaic disease in several Crotalaria species has been cloned and sequenced. Comparisons of the nucleotide and deduced amino acid (aa) sequences of the cloned cDNA with those from other potyviruses show that the Crotalaria-infecting virus (designated Crotalaria mosaic virus; CrMV) is closely related to Cowpea aphid-borne mosaic virus (CABMV). Maximum identity (95.4%) at the coat protein (CP) aa level was observed between CrMV and a Brazilian strain of CABMV. Phylogenetic analyses derived from the sequence alignments of the CP and 3' untranslated region confirmed the identification of CrMV as a strain of CABMV and the name CABMV-Cr is suggested.