Sensitivity of hyperparasitic fungi to alternative products for use in the control of papaya black spot / Sensibilidade de fungos hiperparasitas a produtos alternativos para uso no controle da pinta preta do mamoeiro

Abstract The use of more than one control technique can maximize the reduction of the damages caused by the fungus Asperisporium caricae causal agent of the black spot in the papaya crop. The objective of this work was to evaluate the sensitivity of the fungi Hansfordia pulvinata and Acremonium spp. to alternative products with potential for use in the control of the black-spotted ptarmigan. Three isolates of Acremonium spp. (A-598, A-602 and A-617) and an isolate of H. pulvinata (H-611) were grown in BDA medium containing Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separately. The Amistar 500WG ® fungicide was used as a positive control and pure BDA as a negative control. The toxicity of the tested products was determined based on the values of the biological index, derived from the means of mycelial growth, sporulation and germination of conidia, in each experimental unit. In this way it was possible to select the products classified as compatible for all isolates, and to test them in vivo. In the greenhouse, only the isolates and isolates with selected products, were applied in papaya plants with foliar symptoms of black-spotted. The incidence of leaves with hyperparasites and the percentage of black-painted lesions colonized by the tested isolates were evaluated. Thus, the H-611 isolate proved to be compatible with most of the alternative products tested, except with Hortifos®. Bion® and Matrix® products were compatible with all tested isolates and could be used in conjunction with Acremonium spp. and H. pulvinata to control the papaya black spot, since these products did not present toxicity on the hyperparasitic fungi.

Resumo O uso de mais de uma técnica de controle pode maximizar a redução dos danos causados pelo fungo Asperisporium caricae agente causal da pinta preta na cultura do mamoeiro. O objetivo deste trabalho foi avaliar a sensibilidade dos fungos Hansfordia pulvinata e Acremonium spp. a produtos alternativos com potencial para uso no controle da pinta-preta. Três isolados de Acremonium spp. (A-598, A-602 e A-617) e um isolado de H. pulvinata (H-611) foram cultivados em meio BDA contendo Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separadamente. Fungicida Amistar 500WG® foi utilizado como controle positivo e BDA puro, como controle negativo. A toxicidade dos produtos testados foi determinada com base nos valores do índice biológico, provenientes das médias do crescimento micelial, esporulação e germinação de conídios, em cada unidade experimental. Dessa forma foi possível selecionar os produtos classificados como compatíveis para todos os isolados, e testá-los in vivo. Em casa de vegetação, somente os isolados e os isolados com produtos selecionados, foram aplicados em plantas de mamoeiro com sintomas foliares de pinta-preta. Avaliaram-se a incidência de folhas com hiperparasitas e a porcentagem de lesões de pinta-preta colonizadas pelos isolados testados. Assim o isolado H-611 mostrou-se compatível com a maioria dos produtos alternativos testados, exceto com Hortifos®. Os produtos Bion® e Matriz® apresentaram compatibilidade com todos os isolados testados, podendo ser utilizados em conjunto com o Acremonium spp. e H. pulvinata para controlar a pinta-preta do mamoeiro, uma vez que estes produtos não apresentaram toxidade sobre os fungos hiperparasitas.

Sensitivity of hyperparasitic fungi to alternative products for use in the control of papaya black spot.

The use of more than one control technique can maximize the reduction of the damages caused by the fungus Asperisporium caricae causal agent of the black spot in the papaya crop. The objective of this work was to evaluate the sensitivity of the fungi Hansfordia pulvinata and Acremonium spp. to alternative products with potential for use in the control of the black-spotted ptarmigan. Three isolates of Acremonium spp. (A-598, A-602 and A-617) and an isolate of H. pulvinata (H-611) were grown in BDA medium containing Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separately. The Amistar 500WG ® fungicide was used as a positive control and pure BDA as a negative control. The toxicity of the tested products was determined based on the values of the biological index, derived from the means of mycelial growth, sporulation and germination of conidia, in each experimental unit. In this way it was possible to select the products classified as compatible for all isolates, and to test them in vivo. In the greenhouse, only the isolates and isolates with selected products, were applied in papaya plants with foliar symptoms of black-spotted. The incidence of leaves with hyperparasites and the percentage of black-painted lesions colonized by the tested isolates were evaluated. Thus, the H-611 isolate proved to be compatible with most of the alternative products tested, except with Hortifos®. Bion® and Matrix® products were compatible with all tested isolates and could be used in conjunction with Acremonium spp. and H. pulvinata to control the papaya black spot, since these products did not present toxicity on the hyperparasitic fungi.

Age-related oxidative modifications to uterine albumin impair extravillous trophoblast cells function.

Advanced maternal age is associated not only with a significant reduction in fertility but also with an additional risk of developing pregnancy-related disorders. Most of these disorders are now believed to be the clinical manifestation of an incorrect placentation, namely deficient transformation of maternal spiral arteries and ineffective trophoblast invasion through uterine stroma. In the present study it was hypothesized that an age-related loss in uterine redox homeostasis interferes with the function of extravillous trophoblasts (EVTs) and placentation. To test this hypothesis, relative levels of oxidatively modified proteins were evaluated in human samples from placenta and placental bed, and the role of specific oxidative modifications to proteins in placentation was studied using a cell culture model of EVTs. In the placental bed, the carbonylation level of a 66 kDa protein (identified as albumin) presented a strong, positive and significant correlation with maternal age. Albumin was immunodetected preferentially in endothelial cells and connective tissue between muscle fascicles. In vitro results showed that carbonylated albumin overload did not alter cell viability, but reduced EVTs motility and triggered cell stress response pathways. Moreover, EVTs presented decreased ability to adhere to and invade a collagen extracellular matrix pre-treated with carbonylated albumin. In conclusion, reproductive ageing is accompanied by an increase in maternal uterine carbonylated albumin, that may have a deleterious role in the modulation of EVTs function.

In vitro method to evaluate virus competition between BVDV-1 and BVDV-2 strains using the PrimeFlow RNA assay.

Bovine viral diarrhea viruses (BVDV), segregated in BVDV-1 and BVDV-2 species, lead to substantial economic losses to the cattle industry worldwide. It has been hypothesized that there could be differences in level of replication, pathogenesis and tissue tropism between BVDV-1 and BVDV-2 strains. Thus, this study developed an in vitro method to evaluate virus competition between BVDV-1 and BVDV-2 strains. To this end the competitive dynamics of BVDV-1a, BVDV-1b, and BVDV-2a strains in cell cultures was evaluated by a PrimeFlow RNA assay. Similar results were observed in this study, as was observed in an earlier in vivo transmission study. Competitive exclusion was observed as the BVDV-2a strains dominated and excluded the BVDV-1a and BVDV-1b strains. The in vitro model developed can be used to identify viral variations that result in differences in frequency of subgenotypes detected in the field, vaccine failure, pathogenesis, and strain dependent variation in immune responses.

New polyomavirus species identified in nutria, Myocastor coypus polyomavirus 1.

A novel polyomavirus (PyVs) comprising 5,422 bp was identified by high-throughput sequencing (HTS) in pooled organs of nutria (Myocastor coypus). The new genome displays the archetypal organization of PyVs, which includes open reading frames for the regulatory proteins small T antigen (sTAg) and large T antigen (LTAg), as well as for the capsid proteins VP1, VP2 and VP3. Based on the International Committee on Taxonomy of Viruses (ICTV) Polyomaviridae Study Group criteria, this genome comprises a new PyVs species for the Alphapolyomavirus genus and is putatively named "Myocastor coypus Polyomavirus 1" . The complete genome sequence of this Myocastor coypus Polyomavirus 1 (McPyV1) isolate is publically available under the GenBank accession no. MH182627.

Serological survey for antibodies against pestiviruses in Wyoming domestic sheep.

Pestiviruses including Bovine viral diarrhea virus type 1 (BVDV-1), BVDV-2 and Border disease virus (BDV) have been reported in both sheep and cattle populations, together with the HoBi-like, an emerging group of pestiviruses. Pestivirus control programs in the United States have focused on the control of BVDV-1 and 2. The incidence of pestivirus infection in sheep in the United States and the risk of transmission between cattle and sheep populations are unknown. The aim of this study was to perform serological surveillance for pestivirus exposure in sheep from an important sheep producing state in the Unites States, Wyoming. For this, sera from 500 sheep, collected across the state of Wyoming (US) in 2015-2016, were examined by comparative virus neutralization assay against four species/proposed species of pestiviruses: BVDV-1, BVDV-2, BDV and HoBi-like virus. Rates of exposure varied between geographic regions within the state. The overall pestivirus prevalence of antibodies was 5.6%. Antibodies were most frequently detected against BVDV-1 (4%), and the highest antibody titers were also against BVDV-1. Data from this study highlights understanding of the dynamics of sheep pestivirus exposure, consideration of reference strains used for VN assays, transmission patterns, and potential vaccination history should be taken into account in implementation of control measures against pestiviruses in sheep and for successful BVDV control programs in cattle.

Comprehensive evolutionary and phylogenetic analysis of Hepacivirus N (HNV).

Hepaciviruses (HVs) have been detected in several domestic and wild animals and present high genetic diversity. The actual classification divides the genus Hepacivirus into 14 species (A-N), according to their phylogenetic relationships, including the bovine hepacivirus [Hepacivirus N (HNV)]. In this study, we confirmed HNV circulation in Brazil and sequenced the whole genome of two strains. Based on the current classification of HCV, which is divided into genotypes and subtypes, we analysed all available bovine hepacivirus sequences in the GenBank database and proposed an HNV classification. All of the sequences were grouped into a single genotype, putatively named 'genotype 1'. This genotype can be clearly divided into four subtypes: A and D containing sequences from Germany and Brazil, respectively, and B and C containing Ghanaian sequences. In addition, the NS3-coding region was used to estimate the time to the most recent common ancestor (TMRCA) of each subtype, using a Bayesian approach and a relaxed molecular clock model. The analyses indicated a common origin of the virus circulating in Germany and Brazil. Ghanaian sequences seemed to have an older TMRCA, indicating a long time of circulation of these viruses in the African continent.

Backyard pigs are a reservoir of zoonotic hepatitis E virus in southern Brazil.

Background: Hepatitis E virus (HEV) is the causative agent of acute hepatitis worldwide. There is no seroprevalence study in backyard farms, which are characterized by suboptimal hygienic conditions in Brazil. We aimed to determine the seroprevalence and genetic diversity of HEV in backyard pigs in Brazil. Methods: Swine serum samples collected in 2012 (n=731) and 2014 (n=713) were analysed. The presence of anti-HEV immunoglobulin G in pig serum was evaluated by indirect enzyme-linked immunosorbent assay. Reverse transcription polymerase chain reaction was performed and phylogenetic analyses were carried out based on the partial ORF1 and ORF2 coding regions. Results: Anti-HEV antibodies were detected in 77.6% (567/731; 95% confidence interval [CI] 74.5 to 90.6%) of serum samples in 2012 and 65.5% (467/713; 95% CI 62.0 to 69.0%) in 2014. The herd seroprevalence was 91.7% (187/204; 95% CI 91% to 99%) in 2012 and 83.7% (164/196; 95% CI 78% to 89%) in 2014. Further, HEV RNA was detected in 0.8% (6/713) of samples from 2014. Phylogenetic analysis showed three different genotype 3 subtypes with high similarity to human HEV strains. Conclusions: This study showed that backyard pigs are a reservoir of HEV and alerts us to the need to control infection and spillover from backyard farms. GenBank accession numbers: MF438128-MF438135.

HoBi-like is the most prevalent ruminant pestivirus in Northeastern Brazil.

The ruminant pestiviral species BVDV-1, BVDV-2 and BDV, along with the putative species HoBi-like, may cause substantial economic losses in cattle, sheep and goats. Brazil's large size, variable biomes and wide range of ruminant animal production within different geographic regions suggest that the presence and prevalence of ruminant pestivirus may differ by regions within Brazil. This study investigated the genetic diversity of ruminant pestiviruses and determined the frequency of active infections within two states of the Northeast Region of Brazil, Maranhão and Rio Grande do Norte. Serum samples from 16,621 cattle and 2,672 small ruminants from 569 different herds residing in this region were tested by RT-PCR followed by DNA sequencing. Seventeen positive cattle were detected (0.1%) from fifteen different herds (2.64%). All isolates were classified as HoBi-like pestiviruses based on phylogenetic analysis. All small ruminant samples tested negative. The findings presented herein suggest that the Northeast Region of Brazil has a uniquely high prevalence of HoBi-like viruses. The increasing reports of HoBi-like viruses detected in cattle in the field suggest that natural infection with these viruses may be more widespread than previously thought. The identification of HoBi-like viruses as the most prevalent type of ruminant pestivirus circulating in the Northeast Region of Brazil indicates the need for both continued monitoring and determination of the extent of economic losses associated with HoBi-like virus infections. In addition, it must be taken into account in the choice of diagnostic tests and in vaccine formulations.

Rendimento industrial, eficiência de fabricação e características físico-químicas de queijo colonial produzido de leite com dois níveis de células somáticas / Industrial yield, manufacturing efficiency and physical and chemical characteristics of colonial cheese produced from milk with two levels of somatic cells

Com base em dois níveis de contagem de células somáticas (CCS) do leite utilizado como matéria-prima, determinou-se influência no rendimento e na eficiência de fabricação do queijo colonial, em escala industrial. Os dois níveis de CCS foram formados do leite captado pela indústria, baixa CCS (<500.000 células/mL) e alta CCS (≥500.000 células/mL), sendo produzidos sete lotes de queijo para cada nível. O rendimento foi avaliado com rendimento simples (litros de leite/kg de queijo produzido) e rendimento seco (litros de leite/kg de sólidos totais de queijo produzido). A eficiência de fabricação do queijo foi avaliada por meio das perdas de sólidos no soro de queijo. Mediante análises multivariadas, demonstrou-se a relação entre altas CCS e a diminuição no rendimento simples e seco de queijo colonial, que apresentou também menor eficiência de fabricação, sem alterar a composição do queijo. Além disso, determinaram-se dados relativos às características físico-químicas de queijo colonial, o qual ainda não possui um regulamento técnico de identidade e qualidade. Conclui-se que a qualidade do leite tem influência direta na produção dos queijos e que, ao utilizar matéria-prima com alta CCS, a indústria assume os prejuízos do menor rendimento e da menor eficiência de fabricação dos queijos coloniais.(AU)

From two somatic cells count levels (SCC) of the milk used as raw material, the influence on the yield and cheese-making efficiency of colonial cheese was determined on an industrial scale. Both SCC levels were formed from milk received by the industry, being low SCC (<500.000 cells/mL) and high SCC (≥500.000 cells/mL), and seven batches of cheese were produced for each level. The yield was evaluated as simple yield (liters of milk/kg of cheese produced) and dry yield (liters of milk/kg of total solids of cheese produced). The cheese-making efficiency was evaluated by the loss of solids in cheese whey. Multivariate analysis showed a relationship between high SCC and decrease in simple and dry colonial cheese yield, which also had lower manufacturing efficiency, without altering the composition of the cheese. Moreover, data on the physical and chemical characteristics of colonial cheese were determined, that does not have Technical Regulation of Identity and Quality. We conclude that the quality of milk has a direct influence on the production of cheeses and, when using raw materials with high SCC, the industry takes the costs of worsening in performance and manufacturing efficiency in colonial cheese.(AU)

Incidence and risk factors for Dengue virus (DENV) infection in the first 2 years of life in a Brazilian prospective birth cohort.

This study assessed the incidence and risk factors for dengue virus (DENV) infection among children in a prospective birth cohort conducted in the city of Recife, a hyperendemic dengue area in Northeast Brazil. Healthy pregnant women (n = 415) residing in Recife who agreed to have their children followed were enrolled. Children were followed during their first 24 months of age (May/2011-June/2014), before the 2015 Zika virus outbreak. DENV infection was detected by reverse-transcriptase polymerase chain reaction and/or serology (anti-DENV IgM/IgG). The incidence rates per 1000 person-years (py) and its association with risk factors by age bands (0-12, >12-30 months) were estimated through Poisson regression models. Forty-nine dengue infections were detected; none progressed to severe forms. The incidence rates were 107·6/1000py (95% CI 76·8-150·6) and 93·3/1000py (95% CI 56·1-154·4) in the first and second years of age, respectively. Male children (risk ratios (RR) = 2·33; 95% CI 1·09-4·98) and those born to DENV-naïve mothers (RR = 2·42; 95% CI 1·01-5·80) were at greater risk of infection in the first year of age. In the second year, children born to Caucasian/Asian descent skin colour mothers had a threefold higher risk of infection (RR = 3·34; 95% CI: 1·08-10·33). These data show the high exposure of children to DENV infection in our setting and highlight the role of biological factors in this population's susceptibility to infection.

Methods for classifying obesity in spinal cord injury: a review.

STUDY DESIGN: Narrative review. OBJECTIVES: Review methods used to measure and classify obesity in individuals with spinal cord injuries (SCI). Outline the strengths and weaknesses of each method used to measure obesity in individuals with SCI. SETTING: International. METHODS: PubMed was used to identify articles before 2016. Search terms ('obesity' or 'weight status' and 'spinal cord injury'). Filters: adults, English and human. Studies were retained that (1) included participants, 18 years or older, with SCI; (2) took place in inpatient, outpatient or community-based settings and (3) measured obesity status. Unique methods for classifying individuals with SCI as obese were identified and examples are presented. RESULTS: Methods identified for classifying obesity were as follows: World Health Organization body mass index (BMI) cutoff⩾30 kg m-2, BMI cutoff ⩾25-29 kg m-2, and SCI-specific BMI cutoff ⩾22 kg m-2, waist circumference cutoff (women >102 cm, men >88 cm), percent body fat cutoffs ⩾25% using bioelectrical impedance analysis and dual-energy X-ray absorptiometry, computerized tomography scan visceral fat area ⩾100 cm2 and percentage of ideal body weight. CONCLUSIONS: BMI is the most widely used measure of obesity in the SCI literature. Although some studies identified alternative cutoffs or other metrics, there is no standardized obesity classification in SCI. However, research is needed to determine and validate obesity classification specific to SCI due to physiological changes that occur following injury. We recommend that researchers and clinicians proceed with caution and use methodology based on the purpose of measurement.

Association among practice frequency on depression and stress among competitive US male wheelchair rugby athletes with tetraplegia.

STUDY DESIGN: Cross-sectional. OBJECTIVES: To determine whether frequency of training is related to self-reported lower psychological distress, defined as depressive symptomology and perceived stress, among the US male wheelchair rugby athletes with tetraplegia. SETTING: United States. METHODS: Survey data were collected on a convenience sample at wheelchair rugby tournaments from January-April 2016. Participants self-reported depressive symptomology (CES-D-10), perceived stress scale (PSS), and frequency of rugby practice. Covariate-adjusted regression models were conducted among the full sample and a subsample of individuals who reported spinal cord injury (SCI) as the nature of their disability. RESULTS: Participants included 150 males with tetraplegia, and 87% identified the nature of their disability as SCI. Participants were primarily Caucasian with an average age of ~35 years. Participants scored low on measures of depressive symptomology (mean=5.63; s.d.=4.35) and perceived stress (mean=4.63; s.d.=2.73). Sixty-seven percent of the participants practiced two or more times per week. Results of the main analyses indicated that practicing wheelchair rugby two times or more (compared to once a week or less) was significantly associated with lower depressive symptomology and perceived stress among the full sample and subsample of individuals with SCI. CONCLUSIONS: Greater frequency of wheelchair rugby participation was associated with lower levels of psychological distress. Future research should examine the directional and mechanistic relationship between frequency of sports participation and psychological distress to inform the benefits of adaptive sport.

Phylogeography and population structure analysis reveals diversity by mutations in Lasiodiplodia theobromae with distinct sources of selection.

Lasiodiplodia theobromae is a plant pathogen with a worldwide distribution, with low host specificity, causing stem cankers, dieback diseases, and fruit rot in several species of plants. In coconut, this pathogen is reported as the etiological agent of "coconut leaf blight" (CLB) disease, causing several losses in fruit production. The CLB is an important disease for this crop in Brazil. In our study, we used a phylogeographic approach through the molecular characterization of the translation elongation factor 1-α (TEF1-α) to elucidate the pathogen distribution in Brazil and other countries, besides, search information about diversity sources of this pathogen in coconut palm tree at Brazilian northern, northeast, and southeast. We found that L. theobromae diversity is within populations (locations), and populations that are located closest to the center of the tropical zone have more variability as Central Africa, Brazilian Southeast, and Northeast. The widespread distribution could be in part related with long-distance dispersal via global trade of plants and plant products. The entrance route of L. theobromae in Brazil probably occurred from Africa route and not occurred once. In Brazil, the diversity of this pathogen in coconut tree could be linked to two agents of selection: high host diversity (in Northeast) and distinct management measures adopted in Southeast. These different sources of selection, mainly the mutations, could be one of the reasons that we found distinct reactions to "coconut leaf blight" chemical control in these regions.

Selecting black-spot resistant papaya genotypes derived from backcrossing and hybrids.

Papaya crop is important to Brazilian agribusiness. However, the expansion of papaya cultivation in the country is affected by the absence of commercial cultivars presenting good disease resistance. The black-spot caused by the fungus Asperisporium caricae is the most damaging foliar disease affecting Brazilian papaya crops. The use of genetically resistant cultivars is a promising strategy to reduce the dependence of papaya crops on fungicides. A field split-plot experiment was carried out in the municipality of Linhares, Espírito Santo State, and included 20 hybrids derived from the cross between 14 superior lines and four elite genotypes ('SS72/12', 'SEKATI', 'JS/12' and '41/7'), two commercial cultivars ('Golden' and 'Tainung 01'), and the superior line '36/7', which were evaluated for resistance to black-spot in the fruits and leaves. The treatments were arranged in a randomized block design with six repetitions of three plants per plot. The incidence and severity of black spot in the fruits and leaves were evaluated at three different times in the 2015-2016 crop season. Lines 4, 9, 21, and the parent SEKATI were notable for their capacity to reduce disease severity in the leaves and fruits. Lines 1, 2, 9, 16, and 19, and the parents 'SEKATI' and 'SS-72/12' had reduced disease incidence in their fruits. The most resistant hybrids 'SS-72/12 X 4', 'SS-72/12 X 6', 'SEKATI X 1', 'SEKATI X 2', 'SEKATI X 6', 'SEKATI X 9', and 'SEKATI X 20' presented negative heterosis values for improved black-spot resistance. The current study allowed the selection of black-spot resistant genotypes and hybrids, which presented a significantly reduced disease index in the field.

Genetic Diversity of Brazilian Bovine Pestiviruses Detected Between 1995 and 2014.

Pestivirus infections in ruminants result in significant economic losses worldwide. The aetiological agents are three species from the genus Pestivirus, family Flaviviridae, including bovine viral diarrhoea virus type 1 (BVDV-1), BVDV-2, border disease virus (BDV), and an atypical pestivirus named HoBi-like pestivirus. In this study, eighty-nine pestivirus isolates that were collected in Brazil between 1995 and 2014 and that originated from either cattle, fetal bovine serum (FBS) or as cell culture contaminants were genotyped based on a comparison of gene sequences from their 5' untranslated regions (5'UTR), N-terminal autoprotease (Npro ) and envelope glycoprotein 2 (E2). Of these isolates, 53.9% of the sequences were genotyped as BVDV-1, 33.7% as BVDV-2 and 12.4% as HoBi-like pestivirus. The prevalence of subgenotypes within the species was as follows: BVDV-1a (35.9%), BVDV-2b (31.4%), BVDV-1b (10.1%), BVDV-1d (6.7%), BVDV-2c (2.2%) and BVDV-1e (1.1%). BVDV-2c and BVDV-1e were detected for the first time in Brazil. This study revealed extensive genetic diversity among Brazilian pestivirus isolates, and the combination of pestiviruses that was detected is unique to Brazil. This information may serve as a foundation for designing and evaluating diagnostic tools and in the development of more effective vaccines; therefore, it may potentially contribute to pestivirus control and eradication.

Natural Outbreak of BVDV-1d-Induced Mucosal Disease Lacking Intestinal Lesions.

Bovine viral diarrhea virus (BVDV) belongs to the Pestivirus genus, which is further divided into subgenotypes (1a-1u and 2a-c). When persistent infection occurs, the calf will be immunotolerant to BVDV and possibly develop mucosal disease. This study describes an outbreak of BVDV-1d-induced mucosal disease lacking intestinal lesions. Eleven calves presented with anorexia, sialorrhea, lameness, recumbency, and death. Three calves were necropsied, showing ulceration of the interdigital skin and the oral and nasal mucosa; linear ulcers in the tongue, esophagus, and rumen; and rounded ulcers in the abomasum. Microscopically, mucosa and skin had superficial necrosis, with single-cell necrosis and vacuolation in epithelial cells, and severe parakeratosis. Immunohistochemistry (IHC) showed BVDV antigen in the cytoplasm of epithelial cells in skin and mucosa. All 11 dead calves were positive upon reverse transcription-polymerase chain reaction (RT-PCR) for the detection of Pestivirus along with another 11 live calves from the herd, which were positive again by RT-PCR and IHC after a 4-week interval. Sequencing of the 5' untranslated region and N-terminal protease showed that viruses from these 22 calves were homologous and of subgenotype BVDV-1d. Cytopathic BVDV was isolated from 8 of 11 dead calves, but only noncytopathic BVDV was isolated from the 11 live animals. The findings indicate that this was an outbreak of mucosal disease caused by BVDV-1d, with high morbidity, and lesions restricted to the upper alimentary system and skin and absent from intestine. Thus, the epidemiological and pathological features in this form of mucosal disease may be similar to vesicular diseases, including foot and mouth disease.

Sensitivity of hyperparasitic fungi to alternative products for use in the control of papaya black spot

Abstract The use of more than one control technique can maximize the reduction of the damages caused by the fungus Asperisporium caricae causal agent of the black spot in the papaya crop. The objective of this work was to evaluate the sensitivity of the fungi Hansfordia pulvinata and Acremonium spp. to alternative products with potential for use in the control of the black-spotted ptarmigan. Three isolates of Acremonium spp. (A-598, A-602 and A-617) and an isolate of H. pulvinata (H-611) were grown in BDA medium containing Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separately. The Amistar 500WG ® fungicide was used as a positive control and pure BDA as a negative control. The toxicity of the tested products was determined based on the values of the biological index, derived from the means of mycelial growth, sporulation and germination of conidia, in each experimental unit. In this way it was possible to select the products classified as compatible for all isolates, and to test them in vivo. In the greenhouse, only the isolates and isolates with selected products, were applied in papaya plants with foliar symptoms of black-spotted. The incidence of leaves with hyperparasites and the percentage of black-painted lesions colonized by the tested isolates were evaluated. Thus, the H-611 isolate proved to be compatible with most of the alternative products tested, except with Hortifos®. Bion® and Matrix® products were compatible with all tested isolates and could be used in conjunction with Acremonium spp. and H. pulvinata to control the papaya black spot, since these products did not present toxicity on the hyperparasitic fungi.

Resumo O uso de mais de uma técnica de controle pode maximizar a redução dos danos causados pelo fungo Asperisporium caricae agente causal da pinta preta na cultura do mamoeiro. O objetivo deste trabalho foi avaliar a sensibilidade dos fungos Hansfordia pulvinata e Acremonium spp. a produtos alternativos com potencial para uso no controle da pinta-preta. Três isolados de Acremonium spp. (A-598, A-602 e A-617) e um isolado de H. pulvinata (H-611) foram cultivados em meio BDA contendo Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separadamente. Fungicida Amistar 500WG® foi utilizado como controle positivo e BDA puro, como controle negativo. A toxicidade dos produtos testados foi determinada com base nos valores do índice biológico, provenientes das médias do crescimento micelial, esporulação e germinação de conídios, em cada unidade experimental. Dessa forma foi possível selecionar os produtos classificados como compatíveis para todos os isolados, e testá-los in vivo. Em casa de vegetação, somente os isolados e os isolados com produtos selecionados, foram aplicados em plantas de mamoeiro com sintomas foliares de pinta-preta. Avaliaram-se a incidência de folhas com hiperparasitas e a porcentagem de lesões de pinta-preta colonizadas pelos isolados testados. Assim o isolado H-611 mostrou-se compatível com a maioria dos produtos alternativos testados, exceto com Hortifos®. Os produtos Bion® e Matriz® apresentaram compatibilidade com todos os isolados testados, podendo ser utilizados em conjunto com o Acremonium spp. e H. pulvinata para controlar a pinta-preta do mamoeiro, uma vez que estes produtos não apresentaram toxidade sobre os fungos hiperparasitas.

Micropropagation of Calophyllum brasiliense (Cambess) from nodal segments / Micropropagação de Calophyllum brasiliense (Cambess) a partir de segmentos nodais

Abstract Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture™ were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture™ incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and α-naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM α-naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax® substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.

Resumo A micropropagação de Calophyllum brasiliense Cambess. (Clusiaceae) é uma maneira de superar dificuldades para sua produção em larga escala, devido à natureza recalcitrante das sementes, frutificação irregular e ausência de propagação vegetativa natural da espécie. Culturas foram estabelecidas utilizando segmentos nodais com 2 cm de comprimento, obtidos de plantas com 1 a 2 anos de idade, mantidas em casa de vegetação. Cloreto de mercúrio e Plant Preservative Mixture™ foram utilizados durante a etapa de desinfestação, com melhores resultados alcançados com a incorporação de Plant Preservative Mixture™ ao meio de cultura. Polivinilpirrolidona, carvão ativado, cisteína, ácido ascórbico ou ácido cítrico foram adicionados ao meio de cultura para evitar a oxidação dos explantes. Após 30 dias de cultivo, o uso de polivinilpirrolidona ou ácido ascórbico proporcionou melhores resultados, eliminando a oxidação na maioria dos explantes. Para multiplicação das brotações, benzilaminopurina foi usada em concentrações de 4.4 e 8.8 µM em meio WPM, resultando em uma média de 4.43 e 4.68 brotações por explante, respectivamente, após 90 dias. Ácido indol-3-butírico e ácido α-naftaleno acético foram usados para a indução de raízes, alcançando um enraizamento máximo de 24% com o uso de 20µM de ácido α-naftaleno acético. As plantas enraizadas foram transferidas para substrato Plantmax® e cultivadas em casa de vegetação, alcançando 79% de sobrevivência após 30 dias e 60% após um ano.