Variations Among Crop Seasons and Inoculum Availability Have a Higher Impact on Grapevine Rust Epidemics than Training Systems or Plastic Cover.

Canopy management practices can be effective as part of the integrated management of grapevine diseases. This study aimed to determine whether training systems and plastic covers can contribute to prevent Asian grapevine leaf rust (AGLR) development. Additionally, the influence of crop season and inoculum availability on AGLR development was investigated. Six-season experiments were carried out to characterize 16 epidemics that developed from natural inoculum (NI) or artificial inoculum (AI) sources (NI + AI), conducted in different training systems and with or without the plastic cover. The Richards model was fitted to each AGLR disease progress curve to estimate and compare the onsets and intensities of epidemics using eight curve elements. Principal components analysis (PCA) identified the incidence progress rate, the area under the severity progress curve, final disease severity, time to disease onset, and time to reach the inflection point as the main descriptors for AGLR epidemics. The results showed that AGLR epidemic development was related mainly to differences in inoculum availability and climatic conditions throughout the seasons and to a lower extent to the training system and plastic cover. The earliest disease onset was observed in epidemics when the NI was supplemented with an AI source. Differences in AGRL intensity were correlated to accumulated precipitation, being less severe in autumn-winter than in the spring-summer season. The present findings provided a better understanding of the structure and the seasonal variation of AGLR in cultivar 'Niagara Rosada'. The strategies for reducing and/or delaying inoculum buildup among seasons are discussed.

Comparative fitness of Monilinia fructicola isolates with multiple fungicide-resistant phenotypes.

This study characterized 52 isolates of Monilinia fructicola from peach and nectarine orchards for their multi-resistance patterns to thiophanate-methyl (TF), tebuconazole (TEB), and azoxystrobin (AZO) using in vitro sensitivity assays and molecular analysis. The radial growth of M. fructicola isolates was measured on media amended with a single discriminatory dose of 1 µg/ml for TF and AZO and 0.3 µg/ml for TEB. Cyt b, CYP51, and ß-tubulin were tested for point mutations that confer resistance to quinone outside inhibitors (QoIs), demethylation inhibitors (DMIs), and methyl benzimidazole carbamates (MBCs), respectively. Eight phenotypes were identified including isolates with single, double, and triple in vitro resistance to QoI, MBC, and DMI fungicides. All resistant phenotypes to TF and TEB presented the H6Y mutation in ß-tubulin and the G641S mutation in CYP51. None of the point mutations typically linked to QoI resistance were present in the Monilinia isolates examined. Moreover, fitness of the M. fructicola phenotypes was examined in vitro and detached fruit assays. Phenotypes with single-resistance displayed equal fitness in in vitro and fruit assays compared to the wild-type. In contrast, the dual and triple-resistance phenotypes suffered fitness penalties based on osmotic sensitivity and aggressiveness on peach fruit. In this study, multiple resistance to MBC, DMI, and QoI fungicide groups was confirmed in M. fructicola. Results suggest that Monilinia populations with multiple resistance phenotypes are likely to be less competitive in the field than those with single resistance, thereby impeding their establishment over time and facilitating disease management.

Monocyclic Components and Photosynthetic Damage Caused by Myrtle Rust in Guava Leaves.

Austropuccinia psidii, the causal agent of myrtle rust, was, for many years, restricted to the Americas, but since reaching Hawaii in 2005, the pathogen has expanded its global range exponentially. In Brazil, myrtle rust is the main fungal disease in guava plants. Despite this, there are few studies on guava rust epidemiology. The objectives of this study were to quantify the monocyclic components of rust and to evaluate the photosynthetic damage caused by A. psidii in young and old leaves of 'Paluma' guava. The monocyclic components of guava rust and gas exchange in healthy or inoculated (105 ml-1 urediniospores of A. psidii) leaves were quantified over time. Additionally, young leaves were inoculated with varying concentrations of A. psidii inoculum, and leaf gas exchange and chlorophyll fluorescence were measured at 25 days postinoculation. The relationship between the relative CO2 assimilation of a diseased leaf (Px) and a healthy leaf (Po) is related to disease severity (x) by Px/Po = (1 - x)ß. The density of lesions, disease severity, and urediniospore production were high in young leaves, averaging 58 lesions cm-2, 50% leaf area diseased, and 2.5 × 104 urediniospores per lesion, respectively. Rust symptoms were not observed in old leaves, and resistance to infection did not cause any photosynthetic cost to these leaves. On young leaves, ß was 2.13, indicating a reduction on CO2 assimilation at green tissues from symptomatic leaves. Our data revealed that photosynthesis reduction in diseased guava leaves was caused by biochemical and photochemical damage rather than by stomatal limitation.

Prevalent Transmission of 'Candidatus Liberibacter asiaticus' over 'Ca. Liberibacter americanus' in a Long-Term Controlled Environment.

In Brazil, citrus huanglongbing (HLB) is associated with 'Candidatus Liberibacter americanus' (CLam) and 'Ca. Liberibacter asiaticus' (CLas). However, there are few studies about HLB epidemiology when both Liberibacter spp. and its insect vector, the Asian citrus psyllid (ACP, Diaphorina citri), are present. The objective of this work was to compare the transmission of HLB by ACP when both CLam and CLas are present as primary inoculum. Two experiments were performed under screenhouse conditions from April 2008 to January 2012 (experiment 1) and from February 2011 to December 2015 (experiment 2). The experiments were carried out with sweet orange plants infected with CLam or CLas as inoculum source surrounded by sweet orange healthy plants. One hundred Liberibacter-free adult psyllids were monthly confined to the source of inoculum plants for 7 days with subsequent free movement inside the screenhouse. Fortnightly, nymphs and adults of psyllids were monitored. Psyllid and leaf samples were collected periodically for Liberibacter detection by PCR or quantitative PCR. CLas was detected more frequently than CLam in both psyllid and leaf samples. No mixed infections were detected in the psyllids. A clear prevalence of CLas over CLam was observed in both experiments. The final HLB incidences were 16.7 and 14.5% of Liberibacter-positive test plants, and CLas was detected in 92.3 and 93.1% of these infected plants. Mixed infection was observed only in 3.8% of infected test plants in experiment 1. These results endorse the shift in the prevalence of CLam to CLas observed in citrus orchards of São Paulo, Brazil.

First Report of Mulberry Rust Caused by Cerotelium fici on Morus nigra in Brazil.

Leaf rust caused by Cerotelium fici (Cast.) Arth. is the main disease affecting Moraceae family plants, such as Ficus and Morus species (Galleti and Rezende 2016; Srikantaswamy et al. 2006). In August 2020, rust symptoms were observed in 100% of mulberry (Morus nigra L.) trees in an experimental orchard (Piracicaba, SP, Brazil; 22°42'28"S, 47°37'42"W). Mulberry leaves with high rust severity became yellowish and fell-off prematurely. Pustules were light brown with yellowish halo and presented mean size of 0.9 mm2. Uredinial paraphyses (n = 50) measured 42.2 ± 0.67 µm long with wall uniformly ca 0.6-1.1 µm thick. Urediniospores were brownish, echinulate, globoid to broadly ellipsoid, and measured 27.1 ± 0.29 × 21.0 ± 0.27 µm with a wall thickness of 0.6 ± 0.01 µm (n = 100). The morphology of the urediniospores observed in this study was similar to that reported in the literature for C. fici on Morus alba and Ficus spp. (Gupta et al. 1994; McKenzie 1986; Hennen et al. 2005). We used a low-coverage genome-skimming approach to retrieve genetic information of the rRNA cluster and the mtDNA. Genomic DNA was extracted from 3-4 mg of stored urediniospores at -80 °C, macerated in liquid nitrogen, using a modified cetyl trimethylammonium bromide extraction procedure (Lo Piccolo et al. 2012), and sequenced with 150-bp paired-end reads on Illumina NovaSeq 6000 System. Raw data, (45,761,957 X 2 reads) were assembled with SPAdes v3.15.1 (Bankevich et al., 2012) and the output used to create a custom BLAST database. Loci used for the phylogenetic analyses were identified by BLASTn using, as a query, sequences of C. fici from Ficus sp. from Australia publicly available: Accession No. MH047210.1 for the rRNA and MW036502.1 for COX3. The retrieved sequences were deposited in GenBank under accession numbers OM296992 and OP797407 for the partial rRNA cluster and COX3, respectively. The Bayesian inference phylogenetic analysis of the three concatenate loci (18S, 28S, and COX3) revealed that the isolate obtained in this study (MN1) was clustered in a well-supported clade with C. fici type species. Pathogenicity tests were conducted using mulberry potted plants under greenhouse conditions (25 ± 5 °C). The urediniospores suspension (5 × 104 urediniospores ml-1) with 0.05% Tween 20 was sprayed with an airbrush on fully expanded leaves until run-off. As a control, mulberry plants were sprayed with distilled water and kept under the same conditions. Inoculated and mock-inoculated plants were kept in a dark moist chamber at 23 °C (± 2 °C) for 24 h. After this period, plants were moved to the greenhouse. The experimental design was completely randomized with five replicates, each replicate consisted of one potted plant and the experiment was performed twice. At 12 days post-inoculation, all inoculated plants showed rust symptoms identical to those observed in the field, whereas control plants had no symptoms. The first symptoms were small pustules on the abaxial surface of fully expanded leaves. Small chlorotic lesions were observed on the adaxial leaf surface, which evolved into necrotic lesions. The pathogen was re-inoculated into potted plants, where it was maintained through monthly inoculations. To our knowledge, this is the first report of mulberry rust on M. nigra in Brazil. As mulberry leaves are the only natural food for silkworm (Bombyx mori L.), rust poses a significant threat to the sericulture industry because the disease can decrease production and quality of mulberry foliage.

Lack of an Intron in Cytochrome b and Overexpression of Sterol 14α-Demethylase Indicate a Potential Risk for QoI and DMI Resistance Development in Neophysopella spp. on Grapes.

Asian grapevine leaf rust, caused by Neophysopella meliosmae-myrianthae and N. tropicalis, is often controlled by quinone outside inhibitor (QoI) and demethylation inhibitor (DMI) fungicides in Brazil. Here, we evaluated the sensitivity of 55 Neophysopella spp. isolates to pyraclostrobin (QoI) and tebuconazole (DMI). To elucidate the resistance mechanisms, we analyzed the sequences of the cytochrome b (CYTB) and cytochrome P450 sterol 14α-demethylase (CYP51) target proteins of QoI and DMI fungicides, respectively. The CYP51 expression levels were also determined in a selection of isolates. In leaf disc assays, the mean 50% effective concentration (EC50) value for pyraclostrobin was about 0.040 µg/ml for both species. CYTB sequences were identical among all 55 isolates, which did not contain an intron immediately after codon 143. No amino acid substitution was identified at codons 129, 137, and 143. The mean EC50 value for tebuconazole was 0.62 µg/ml for N. tropicalis and 0.46 µg/ml for N. meliosmae-myrianthae, and no CYP51 sequence variation was identified among isolates of the same species. However, five N. meliosmae-myrianthae isolates grew on leaf discs treated at 10 µg/ml tebuconazole, and these were further exposed to tebuconazole selection pressure. Tebuconazole-adapted laboratory isolates of N. meliosmae-myrianthae showed an eight- to 25-fold increase in resistance after four rounds of selection that was not associated with CYP51 target alterations. In comparison with sensitive isolates, CYP51 expression was induced in the presence of tebuconazole in three out of four tebuconazole-adapted isolates tested. These results suggest a potential risk for QoI and DMI resistance development in Neophysopella spp.

Sensitivity of Colletotrichum acutatum Isolates from Citrus to Carbendazim, Difenoconazole, Tebuconazole, and Trifloxystrobin.

Postbloom fruit drop (PFD) of citrus is caused by the Colletotrichum acutatum and C. gloeosporioides species complexes. The disease is important when frequent rainfall occurs during the flowering period of citrus trees. In Brazil, until 2012, PFD was mainly controlled by preventive applications of the methyl-benzimidazole carbamate (MBC) carbendazim and demethylation-inhibitor (DMI) fungicides such as difenoconazole. Since then, mixtures containing the DMI tebuconazole and the quinone-outside inhibitor (QoI) trifloxystrobin have been commonly used. Fungicides are often applied preventively, sometimes even when conditions are not conducive for PFD development. Excessive fungicide applications may favor the selection of resistant populations of Colletotrichum spp. In this study, we assessed the fungicide sensitivity of C. acutatum isolates collected during the two distinct periods of PFD management in Brazil: before and after the trifloxystrobin and tebuconazole mixture became widely employed. The sensitivity of 254 C. acutatum isolates to carbendazim and difenoconazole and of 164 isolates to tebuconazole and trifloxystrobin was assessed. Mycelial growth inhibition of these isolates was evaluated for all the fungicides using either serial dilution of fungicide rates or the spiral gradient dilution method. In addition, inhibition of conidial germination was also assessed for trifloxystrobin. Analysis of partial ß-tub, cytb, and cyp51b gene sequences did not reveal any mutations related to resistance to MBCs, QoIs, and DMIs, respectively. In mycelial growth assays, mean EC50 values were 0.14, 0.11, and 0.21 µg/ml for difenoconazole, tebuconazole, and trifloxystrobin, respectively. The conidial germination inhibition by trifloxystrobin was similar among the tested isolates, and the mean EC50 value was 0.002 µg/ml. All isolates had similar mean mycelial growth inhibition for carbendazim, regardless of the fungicide concentrations. Therefore, based on similar EC50 values and molecular analyses, no shift in the sensitivity of isolates has been observed to the fungicides commonly used in different citrus-producing areas in Brazil.

A Threshold-Based Decision-Support System for Fungicide Applications Provides Cost-Effective Control of Citrus Postbloom Fruit Drop.

Postbloom fruit drop (PFD) of citrus, caused by Colletotrichum acutatum sensu lato and C. gloeosporioides sensu lato, is an important disease in the humid tropics of the American continent. PFD mainly affects flowers, on which typical symptoms are characterized by orange-brown lesions with presence of acervuli. The disease has a sporadic occurrence, but preventative fungicide sprays are applied every season. The objective of this study was to evaluate the effectiveness of a fungicide spray strategy for PFD based on a predictive model of C. acutatum conidium germination linked to weather conditions. Fungicide sprays were performed when the model predicted pre-established thresholds of 10, 15, 20, and 25% of germinated spores (T10, T15, T20, and T25, respectively). Five experiments were conducted in two different seasons in the state of São Paulo, Brazil. PFD control efficacy of the threshold-based treatments was compared with a nontreated control and to a calendar-based spray system. Additionally, an economic analysis was performed to assess the gross income revenues of the fungicide spraying strategies. Disease control in plots treated at T10, T15, and T20 was as effective as the calendar-based strategy. The number of fungicide applications was reduced by 33 to 71% when sprays were applied at T15 and T20, and gross income increased or was comparable to that of the other treatments. Therefore, using a conidium germination model with a threshold of 15 or 20% is recommended as a spraying strategy for PFD management in Brazil.

Sweet Orange Fruit Age and Inoculum Concentration Affect the Expression of Citrus Black Spot Symptoms.

Citrus black spot (CBS), caused by Phyllosticta citricarpa, affects different citrus species worldwide. CBS is mainly expressed as false melanose and hard spot symptoms. There is no consensus in the literature about the period when fruit are susceptible to P. citricarpa infection and the length of the CBS incubation period. Therefore, this study aimed to assess the influence of sweet orange variety, fruit age, and inoculum concentration on the incubation period and the expression of different CBS symptoms. Attached fruit of Hamlin, Pera, and Valencia sweet orange at 1.5, 3.0, 5.0, and 7.0 cm diameter were inoculated with suspensions containing 103 and 105 conidia/ml of P. citricarpa. The percent conidial germination was quantified using scanning electron microscopy. The CBS symptoms on fruit were assessed monthly. The four diameters did not significantly affect conidial germination on the inoculated fruit, although CBS incidences were lower when larger fruit were inoculated. Hard spot symptoms on sweet orange fruit did not develop from the false melanose symptoms and vice versa. The incubation periods for false melanose were shorter than those observed for hard spot. False melanose began to appear 44 days after inoculation, but hard spot only formed at 113 days or later. Incubation periods were shorter and incidences of false melanose were higher following inoculation with higher inoculum concentration and smaller fruit diameter. The incubation period of hard spot varied among varieties and fruit diameters. However, there was no relationship between hard spot incidence and variety. This study provides a better understanding of the factors affecting the variation in the CBS incubation period and disease incidence on fruit.

Reduced Sensitivity to Azoxystrobin of Monilinia fructicola Isolates From Brazilian Stone Fruits is Not Associated With Previously Described Mutations in the Cytochrome b Gene.

Quinone-outside inhibitor (QoI) fungicides are effective tools for preharvest control of brown rot of stone fruit. These fungicides have a very specific site of action so the risk of resistance selection is high. The sensitivity of Monilinia fructicola (G. Winter) Honey isolates to azoxystrobin (QoI) was investigated in 143 isolates collected between 2002 and 2011 from four Brazilian states in orchards with different frequencies of fungicide use (0 to 6 fungicides sprays/season). Sensitivity of the isolates to azoxystrobin was determined in vitro, by inhibition of mycelial growth and spore germination on fungicide-amended media or ex vivo by pathogen inoculation in untreated or treated fruit with azoxystrobin. Potential mutations in codons 143, 137, and 129 of the cytochrome b (Cyt b) gene and the occurrence of an intron immediately after codon 143 were analyzed in a subpopulation of the isolates. The M. fructicola population of São Paulo State was less sensitive to the fungicide than the population from the states of Paraná, Santa Catarina, and Rio Grande do Sul. The low sensitivity of the isolates was confirmed also by comparing to the sensitivity of the baseline isolates. Mutations in G143A, F129L, and G137R in Cyt b gene were not found. In addition, 58 isolates tested showed an intron after codon 143 in Cyt b gene. Our results indicate that other mechanisms of selection for low sensitivity to QoI fungicides should be investigated.

Virus-vector relationship in the Citrus leprosis pathosystem.

Citrus leprosis has been one of the most destructive diseases of citrus in the Americas. In the last decade important progress has been achieved such as the complete genome sequencing of its main causal agent, Citrus leprosis virus C (CiLV-C), belonging to a new genus Cilevirus. It is transmitted by Brevipalpus yothersi Baker (Acari: Tenuipalpidae), and is characterized by the localized symptoms it induces on the leaves, fruits and stems. It occurs in the American continents from Mexico to Argentina. The virus was until recently considered restricted to Citrus spp. However, it was found naturally infecting other plants species as Swinglea glutinosa Merrill and Commelina benghalensis L., and has been experimentally transmitted by B. yothersi to a large number of plant species. Despite these advances little is known about the virus-vector relationship that is a key to understanding the epidemiology of the disease. Some components of the CiLV-C/B. yothersi relationship were determined using the common bean (Phaseolus vulgaris L. cv. 'IAC Una') as a test plant. They included: (a) the virus acquisition access period was 4 h; (b) the virus inoculation access period was 2 h; (c) the latent period between acquisition and inoculation was 7 h; (d) the period of retention of the virus by a single viruliferous mite was at least 12 days; (d) the percentage of viruliferous individuals from mite colonies on infected tissues ranged from 25 to 60%. The experiments confirmed previous data that all developmental stages of B. yothersi (larva, protonymph and deutonymph, adult female and male) were able to transmit CiLV-C and that transovarial transmission of the virus did not occur. CiLV-C can be acquired from lesions on leaves, fruits and stems by B. yothersi. Based on the distribution of lesions produced by single viruliferous B. yothersi on bean leaves, it is concluded that they tend to feed in restricted areas, usually near the veins. The short latent and transmission periods during the larval stage suggest that the CiLV-C/B. yothersi relationship is of the persistent circulative type.

Recent introduction and recombination in Colletotrichum acutatum populations associated with citrus postbloom fruit drop epidemics in São Paulo, Brazil.

Citrus crops in São Paulo State, Brazil, have been severely affected by postbloom fruit drop disease (PFD), which is caused by Colletotrichum acutatum. This disease leads to the drop of up to 100% of young fruits. Previous studies have assumed that this pathogen exhibits a clonal reproductive mode, although no population genetic studies have been conducted so far. Thus, the genetic structure of six C. acutatum populations from sweet orange orchards showing PFD symptoms was determined using nine microsatellite markers, enabling inference on predominant mode of reproduction. C. acutatum populations exhibit a nearly panmictic genetic structure and a high degree of admixture, indicating either ongoing contemporary gene flow at a regional scale or a recent introduction from a common source, since this pathogen was introduced in Brazil only very recently. Sharing haplotypes among orchards separated by 400 km suggests the natural dispersal of fungal propagules, with the possible involvement of pollinators. A significant population expansion was detected, which was consistent with an increase in host density associated with crop expansion toward new areas across the state. Findings of moderate to high levels of haplotypic diversity and gametic equilibrium suggest that recombination might play an important role in these pathogen populations, possibly via parasexual reproduction or a cryptic sexual cycle. This study provides additional tools for epidemiological studies of C. acutatum to improve prevention and management strategies for this disease.

Efficacy of Area-Wide Inoculum Reduction and Vector Control on Temporal Progress of Huanglongbing in Young Sweet Orange Plantings.

Huanglongbing (HLB), caused by 'Candidatus Liberibacter' spp. and transmitted by the Asian citrus psyllid Diaphorina citri (ACP), is an important threat to citrus industries worldwide, causing significant yield loss. The current recommended strategies to manage HLB are to eliminate HLB-symptomatic trees to reduce sources of bacterial inoculum and to apply insecticides to reduce psyllid vector populations. The objective of this study was to assess the effectiveness and the importance of both strategies applied within young citrus plots (local management), in different frequencies and combinations, on HLB temporal progress. Two factorial field experiments, E1 and E2, were initiated in a new plantation of sweet orange in an HLB epidemic region of Sao Paulo, Brazil, in October 2005 and May 2006, respectively. Local inoculum reduction (tree removal) intervals for E1 were every 4, 8, and 16 weeks and, for E2, every 2, 4, 12, and 26 weeks. Local vector control strategies for E1 were no control, program A (PA), and program B (PB); and, for E2, no control and program C (PC), as follows. Psyllids were controlled with two 56-day-interval soil or drench applications of systemic insecticides concurrently with the rainy season each year and, during the rest of the year, with insecticide sprays every 28 days for PA and every 14 days for PB and PC. Regional HLB management was present for E1 and absent for E2. The beginning of the HLB epidemic was delayed for 10 months in E1 compared with appearance of the first diseased tree in E2 but wasn't affected by different local strategies in either experiment. After 60 (E1) and 53 (E2) months, the HLB incidence and progress rates were not affected by different frequencies of local inoculum reduction in either experiment, and were different only in plots with and without local vector control in E2. In E1, the disease incidence was reduced by 90% and the disease progress rate by 50% in plots both with and without vector control. These reductions were explained by smaller psyllid populations and lower frequency of bacterialiferous psyllids in E1 compared with E2. Annual productivity increased over time in E1, as expected for young plantings, but remained stable or decreased in E2. These results confirm that immigration of bacterialiferous ACP vectors plays a critical role in HLB epidemics and suggest that area-wide inoculum reduction and ACP management strongly affect HLB control.

Construction and genetic characterization of an interspecific raspberry hybrids panel aiming resistance to late leaf rust and adaptation to tropical regions.

Raspberries (Rubus spp) are temperate climate fruits with profitable high returns and have the potential for diversification of fruit growing in mid to low-latitude regions. However, there are still no cultivars adapted to climatic conditions and high pressure of diseases that occurs in tropical areas. In this context, our objective was to evaluate the genetic diversity from a 116 raspberry genotypes panel obtained from interspecific crosses in a testcross scheme with four cultivars already introduced in Brazil. The panel was genotyped via genotyping-by-sequencing. 28,373 and 27,281 SNPs were obtained, using the species R. occidentalis and R. idaeus genomes as references, respectively. A third marker dataset was constructed consisting of 41,292 non-coincident markers. Overall, there were no differences in the results when using the different marker sets for the subsequent analyses. The mean heterozygosity was 0.54. The average effective population size was 174, indicating great genetic variability. The other analyses revealed that the half-sibling families were structured in three groups. It is concluded that the studied panel has great potential for breeding and further genetic studies. Moreover, only one of the three marker matrices is sufficient for diversity studies.

Retrospective study of traumatic dental injuries in primary teeth in a Brazilian specialized pediatric practice.

OBJECTIVE: To determine the prevalence of traumatic dental injuries (TDI) in children younger than 7 years and the effect of predisposing clinical factors on their occurrence. MATERIAL AND METHODS: Dental records of 2725 children younger than 7 years seen in a private pediatric dental clinic in Goiânia, Brazil, from February 1993 to December 2008 were analyzed. A group of 150 children without traumatic injuries and with similar characteristics to those of the trauma group were used to study predisposing clinical factors as follows: lip coverage (adequate/inadequate), overbite (normal/negative/deep), and overjet (less or greater than 3 mm). Frequency distribution and a chi-square test were used for data analysis. The level of significance was set at 5%. RESULTS: Clinical and radiographic data of 412 injured deciduous teeth were found in 383 records. TDI prevalence was 11.9%, with no significant difference between sexes. The maxillary central incisors were the teeth most frequently affected (83.3%), and the most frequent type of injury was subluxation (35.1%). TDI was most prevalent in the 13-to-36-month age group (47.7%). Home (43.5%) was the main place of occurrence, followed by school (10.1%); falls (50.3%) were the most common cause, followed by collisions with objects (18.2%). The first examination or treatment occurred in the first 24 h in 46.5% of the cases. The analysis of the association of predisposing clinical factors with TDI revealed no differences in lip coverage; however, overjet >3 mm and deep and negative overbite were significantly associated with TDI. CONCLUSION: TDI in primary dentition should receive careful attention, particularly when it affects children younger than 3 years and in the presence of overjet, overbite, or both.

Effects of traumatic dental injuries to primary teeth on permanent teeth--a clinical follow-up study.

AIM: This study evaluated the prevalence of developmental sequelae to permanent teeth (DSP) after traumatic dental injuries to primary teeth (TDI-1) and their association with age, gender, type of injury, recurrence of injury and post-traumatic damage to primary teeth. MATERIALS AND METHODS: Dental records of 2725 children treated from February 1993 to December 2008 in a private pediatric dental clinic were examined. A total of 308 records had 412 primary teeth that sustained traumatic injuries. Age at the time of injury ranged from 4 months to 7 years. A chi-squared test and logistic regression were used for statistical analyses. RESULTS: One hundred forty-eight children (241 teeth) were followed up until the eruption of the permanent successor. The prevalence of DSP was 22.4%. Discoloration and hypoplasia were the most frequent abnormalities (74.1%), followed by eruption disorders (25.9%). Age at the time of TDI-1 was the only variable significantly associated with DSP. Sequelae were most prevalent among children who suffered an injury between 1 and 3 years of age. CONCLUSIONS: Children who sustain traumatic dental injuries should be followed up regularly for an early diagnosis and treatment of possible DSP.

Photosynthetic Cost Associated With Induced Defense to Plasmopara viticola in Grapevine.

Downy mildew caused by Plasmopara viticola is one of the most destructive diseases of Vitis vinifera worldwide. Grapevine breeding programs have introgressed P. viticola-resistant traits into cultivated V. vinifera genotypes and launched interspecific hybrids with resistance against downy mildew. In general, pathogen infection affects primary metabolism, reduces plant growth and development and modifies the secondary metabolism toward defense responses, which are costly in terms of carbon production and utilization. The objective of this work was to evaluate the photosynthesis impairment by inducible defenses at the leaf level in V. vinifera cultivars resistant to P. viticola. Photosynthetic limitations imposed by P. viticola in susceptible and resistant grapevine cultivars were evaluated. Histochemical localization of hydrogen peroxide and superoxide and the activity of ascorbate peroxidase were assessed. Measurements of leaf gas exchange, chlorophyll fluorescence and the response of leaf CO2 assimilation to increasing air CO2 concentrations were taken, and photosynthetic limitations determined in cultivars Solaris (resistant) and Riesling (susceptible). The net photosynthetic rates were reduced (-25%) in inoculated Solaris plants even before the appearance of cell death-like hypersensitive reactions ("HR"). One day after "HR" visualization, the net photosynthetic rate of Solaris was reduced by 57% compared with healthy plants. A similar pattern was noticed in resistant Cabernet Blanc and Phoenix plants. While the susceptible cultivars did not show any variation in leaf gas exchange before the appearance of visual symptoms, drastic reductions in net photosynthetic rate and stomatal conductance were found in diseased plants 12 days after inoculation. Decreases in the maximum Rubisco carboxylation rate and photochemical impairment were noticed in Riesling after inoculation with P. viticola, which were not found in Solaris. Damage to the photochemical reactions of photosynthesis was likely associated with the oxidative burst found in resistant cultivars within the first 24 h after inoculation. Both chlorophyll degradation and stomatal closure were also noticed in the incompatible interaction. Taken together, our data clearly revealed that the defense response against P. viticola causes a photosynthetic cost to grapevines, which is not reversible even 12 days after the pathogen infection.

The Climate-Driven Genetic Diversity Has a Higher Impact on the Population Structure of Plasmopara viticola Than the Production System or QoI Fungicide Sensitivity in Subtropical Brazil.

Downy mildew, caused by Plasmopara viticola, is the main disease affecting vineyards in subtropical Brazil. Here, we collected 94 P. viticola isolates from four organic and conventional vineyards in the two main grape-growing states of Brazil to evaluate the sensitivity to the quinone outside inhibitor (QoI) azoxystrobin by pheno- and genotyping assays. The impact of location, production system and sensitivity to QoI fungicides on the population genetics and structure of P. viticola was determined using 10 microsatellite markers. Cytochrome b sequencing revealed that 28 and 100% of the isolates from vineyards under organic and conventional management carried the G143A mutation, respectively. The G143A mutation was associated with high levels of azoxystrobin resistance. Three out of the 94 isolates analyzed carried the M125I alteration, not previously described in P. viticola, which was associated with a five-fold reduction in azoxystrobin sensitivity compared to wild-type isolates. Haplotype network analysis based on cytochrome b gene sequences suggested that the Brazilian populations are more closely related to the European than the North American population. A total of six haplotypes were identified, with two of them carrying the G143A mutation. Microsatellite analysis revealed high allelic and genotypic variation among the four populations. Population differentiation analyses indicated that state of origin directly influences the population biology of P. viticola, while production system and QoI sensitivity have little effect. Great genetic diversity, sexual reproduction and high levels of admixture were observed in Rio Grande do Sul State. In contrast, populations in São Paulo State were dominated by a few clonal genotypes, and no admixed genotype was detected between the two genetic pools identified in the state. This study raises the hypothesis that winter weather conditions influence the overwinter survival strategy with profound effects in the population biology of P. viticola.

Persistent Calyxes in Postbloom Fruit Drop: A Microscopy and Microanalysis Perspective.

Citrus postbloom fruit drop, caused by Colletotrichum spp., is an important disease in the Americas. The pathogen infects citrus flowers, produces orange-brown lesions on petals, and may cause the abscission of young fruit. In diseased flowers, the calyxes remain attached to the peduncle after the young fruit drop. No anatomical and microanalysis studies have been conducted to determine whether calyx tissues can be infected by Colletotrichum spp. and why calyxes remain attached to the peduncle. Based on light microscopy, we demonstrate that the ovary abscission zone exhibits a separation region composed of layers of thickened lignified walled cells, indicating that abscission involves the disruption of cell walls. The first layers of the protective zone (PZ) are composed of densely packed cells with suberized walls produced by the wound meristem. Beneath the PZ, there is a compact mass of small cells that accumulate starch grains. X-ray fluorescence microanalysis (µ-XRF) confirmed the increased accumulation of calcium in the receptacle of the persistent calyxes compared to non-inoculated citrus flowers. Moreover, the peduncle pith and the receptacle exhibit hypertrophied cells with thick walls that may be related to calyx retention. Fungal structures are not observed inside the persistent calyx tissues.

HIV/AIDS in small cities in Midwest Santa Catarina, south of Brazil: Clinical and epidemiological aspects, opportunistic infections.

INTRODUCTION: We avaluated the clinical features, epidemiology, opportunistic infections and coinfections of HIV/AIDS patients. METHODS: We analyzed the records of 143 patients receiving antiretroviral therapy at a public center in the Midwest of Santa Catarina, south of Brazil, from December 2014 to September 2015. RESULTS: Most were male, Caucasian, married, with low education level, and aged 31-50 years. Heterosexual transmission was the most common infection route. Regarding coinfection, 3.5% had hepatitis C, 2.1% hepatitis B, 4.2% syphilis, and 4.9% tuberculosis; 38.5% had opportunistic infections. CONCLUSIONS: HIV infection follows the national trend, but hepatitis B and C coinfection rates were higher, while tuberculosis rate was lower.