Increased in vitro production of bovine embryos resulting from oocyte maturation in the presence of triciribine, a specific inhibitor of AKT.

The aim of this study was to evaluate the effect of different concentrations of triciribine, a selective Akt inhibitor, on various aspects of oocyte maturation and on the IVF of bovine embryos. Cumulus-oocyte complexes (COCs) were matured in vitro in medium supplemented with: 0 (control), 1, 5, 10, and 20 µM of triciribine. The nuclear maturation was assessed by staining with acetic orcein, while the cytoplasmic maturation was evaluated by mitochondrial (MitoTracker® Red CMXRos) and lipid droplets distribution (LipidTOX). COCs were fertilized in vitro and cultured for nine days. Cleavage rates, blastocyst production, and hatching rates were determined on days three, seven, and nine of in vitro culture, respectively. Oocytes from COCs treated with 1 µM of triciribine were stained at 3, 6, and 9 h of IVM to determine the inhibitor's involvement in germinal vesicle breakdown. Analysis of variance (ANOVA) of the data was performed and the means were compared using the SNK test at a 5 % significance level. Exposure of COCs to 1, 5, and 10 µM of triciribine did not alter the number of matured oocytes (P < 0.05), a concentration of 20 µM reduced the number of oocytes in MII with a consequent increase in oocytes in MI (P < 0.05). This concentration markedly reduced the number of oocytes with peripheral cortical granules and the rates of cleavage and blastocysts (P < 0.05). On the other hand, when COCs were matured in the presence of 1 µM, there was an increase in the blastocyst rate (P < 0.05), but without altering the timing of meiosis resumption (P < 0.05). It is concluded that the Akt pathway participates in the nuclear and cytoplasmic events of in vitro maturation of bovine oocytes, but through mechanisms that do not interfere with germinal vesicle breakdown. Modulation of Akt activity in bovine COCs during IVM with 1 µM of triciribine increases the in vitro production of bovine embryos.

Pathogens and microorganisms in the mangrove oyster Crassostrea gasar cultivated in an estuarine environment in Northeast Brazil.

Estuaries are important ecosystems due to the ecological services they provide, acting as nurseries for many species of fish and invertebrates, and are also used as environments for the extraction and cultivation of mollusks. Oysters are animals that filter water to obtain oxygen and nutrients. In this process, they can bioaccumulate microorganisms and chemical substances in their tissues. The growth of mollusk culture in Northeastern Brazil requires the health identification of cultivated oysters through the quantification of the potentially harmful microbiota accumulated in the animals. Therefore, the present work aims to quantify and identify bacteria and possible pathogens found in the tissues of cultivated oysters and their culture waters. The Most Probable Number of Coliforms (MPN) in oysters and water were considered suitable according to the Brazilian current legislation, Vibrio sp. obtained low colonization and Salmonella sp. was not observed. The prevalence of microorganisms potentially pathogenic to oysters was 33.7%, highlighting metazoans and Nematopsis sp., however, the intensity of the infestation of these organisms was moderate. The low contamination of oysters demonstrates that this culture environment is promising for this activity. However, continuous environmental and sanitary monitoring is fundamental to guarantee the safety of the culture waters and the sustainability of aquaculture activities.

Impact of parental attributes on the oral health-related quality of life of Brazilian preschool children.

PURPOSE: Considering the importance of parental attitudes and their influence on children's oral health, it is important to know which characteristics may impact it. The aim of this population study was to evaluate the impact of the parental sociodemographic characteristics on the oral health-related quality of life (OHRQoL) of preschool children from southeastern Brazil. METHODS: A population-based study was conducted with a sample of 321 children aged 2 to 5 years old who underwent a calibrated clinical oral examination to assess oral health status and their parents were asked to answer the two following questionnaires: (1) sociodemographic conditions and (2) Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS). Descriptive analysis, Mann-Whitney and Kruskal-Wallis tests and adjusted Poisson regression models were used. RESULTS: Younger mothers (RP: 1.47; 95% CI: 1.11-1.95), lower household income (RP: 1.47; 95% CI: 1.11-1.96) and the presence of dental caries (RP: 2.28; 95% CI: 1.74-3.0) were more likely to have a negative impact on the OHRQoL B-ECOHIS total scores (P < 0.01). CONCLUSION: The characteristics of mothers, such as age, schooling and salary, influenced the mean B-ECOHIS total score; however, paternal attributes were not associated with a negative impact on the children's quality of life. Also, dental caries and household income influenced the children's oral health-related quality of life.

Antifungal activity of poly(ε-caprolactone) nanoparticles incorporated with Eucalyptus essential oils against Hemileia vastatrix.

Coffee (Coffea L.) is one of the main crops produced globally. Its contamination by the fungus Hemileia vastatrix Berkeley and Broome has been economically detrimental for producers. The objective of this work was to extract and characterize the essential oils from Eucalyptus citriodora Hook, Eucalyptus camaldulensis Dehn and Eucalyptus grandis Hill ex Maiden, produce and characterize nanoparticles containing these essential oils and evaluate the in vivo and in vitro antifungal activity of free and nanoencapsulated essential oils. The principal constituent of the essential oil from E. citriodora was citronellal; that from E. grandis was α-pinene; and that from E. camaldulensis was 1,8-cineol. The in vitro antifungal activity against the fungus H. vastatrix was 100% at a concentration of 1000 µl l-1 for all the oils and nanoparticles containing these natural products. The sizes of the nanoparticles produced with the essential oils from E. citriodora, E. camaldulensis and E. grandis were 402·13 nm, 275·33 nm and 328·5 nm, respectively, with surface charges of -11·8 mV, -9·24 mV and - 6·76 mV, respectively. Fourier transform infrared analyses proved that the encapsulation of essential oils occurred in the polymeric matrix of poly(ε-caprolactone). The incorporation of essential oils into biodegradable poly(ε-caprolactone) nanoparticles increased their efficiency as biofungicides in the fight against coffee rust, decreasing the severity of the disease by up to 90·75% after treatment with the nanoparticles containing the essential oil from E. grandis.

Mid-pregnancy poly(I:C) viral mimic disrupts placental ABC transporter expression and leads to long-term offspring motor and cognitive dysfunction.

Limited information is available about the effect of mid-pregnancy viral infections on the placental expression of efflux transporters and offspring behavior. We hypothesized that maternal exposure to polyinosinic-polycytidylic acid [poly(I:C)], a synthetic double-stranded RNA viral mimic, would impair placental cell turnover, the expression of selected ABC transporters and adult offspring behavior. C57BL/6 mice were administered poly(I:C) (10 mg/Kg;ip) or vehicle at gestational day (GD) 13.5 (mid-pregnancy). Dams were euthanized for blood collection 4 h after injection, fetal and placental collection at GD18.5 or allowed to deliver spontaneously at term. At GD 13.5, poly(I:C) induced an acute pro-inflammatory response characterized by an increase in maternal plasma levels of IL-6, CXCL-1 and CCL-2/MCP-1. At GD 18.5, poly(I:C) decreased cell proliferation/death in the labyrinthine and increased cell death in the junctional zones, characterizing a disruption of placental cell turnover. Abca1 and Abcg1 immunolabelling was decreased in the labyrinthine zone, whereas Abca1, Abcg1 and breast cancer resistance transporter (Bcrp) expression increased in the junctional zone. Moreover, adult offspring showed motor and cognitive impairments in the Rotarod and T-water maze tests. These results indicate that viral infection during mid-pregnancy may disrupt relevant placental efflux transporters, as well as placental cell turnover and offspring behavior in adult life.

Association of coronary calcification with prognosis of Covid-19 patients without known heart disease.

Risk factors that determine the severity of Covid-19 have not been fully elucidated. The aim of this study was to evaluate the role of coronary artery calcification (CAC) as a risk factor for death or mechanical ventilation (MV) of patients without known heart disease infected with Covid-19. We analyzed 283 consecutive in-patients with acute respiratory symptoms with chest computed tomography (chest-CT), without previous heart disease, and criteria for Covid-19 (RT-PCR positive and/or typical clinical and chest-CT findings). CAC was classified by the number of coronary segments affected as absent (0), mild (1-3), and severe calcification (more than 3). The association between CAC, CAC severity, and death or MV due to severe respiratory failure was assessed by logistic regression. The mean age was 58.7±15.7 years and 54.1% were men. Patients with CAC were older, more likely to have hypertension, and less likely to be obese. CAC was present in 75 patients (26.5%), of which 42 had a mild calcification and 33 had severe calcification, and was associated with death (OR=2.35, 95%CI: 1.01-5.48) or MV (OR=2.72, 95%CI: 1.20-6.20) adjusted for multiple confounders, with significant and increased odds ratio for the severe form of CAC (death: OR=3.70, 95%CI: 1.20-11.42; MV: OR=3.30, 95%CI: 1.09-9.95). We concluded that CAC was an independent risk factor for death or MV in Covid-19 patients without previous heart disease, particularly for those with severe calcification. CAC can be easily visualized on common chest-CT, widely used in evaluation of moderate to severe Covid-19.

New insights on the removal of diclofenac and ibuprofen by CWPO using a magnetite-based catalyst in an up-flow fixed-bed reactor.

This research has been focused on the removal of two anti-inflammatory drugs, diclofenac (DCF) and ibuprofen (IBU), by a continuous catalytic wet peroxide oxidation (CWPO) process using a lab-synthesized nanomagnetic catalyst (Fe3O4/MWCNTs). The central composite rotatable design (CCRD) method was used to study the effect of DCF and IBU concentration (expressed as theoretical oxygen demand (ThOD) between 0 and 52.5 mg L-1) and of the feed stream pH (from 3 to 7) on the removal of total organic carbon (TOC) and the concentration of aromatic compounds (Arm) and total phenolic compounds (TP) by CWPO. It could be observed that DCF was preferably removed from the DCF-IBU aqueous mixture at pH values ranging from 3 to 5. In addition, feed stream pH had a significant effect on the pollutants removal, as well as on TOC, TP and aromatic compounds removal, observing an increasing in the pollutants degradation when feed stream pH decreased from 7 to 3. Quadratic models predicted for response variable, such as TOC, TP and aromatic compounds removal, and their maximum model-predicted removal values were of 90.0, 80.2 and 90.0%, respectively. Finally, as a proof of concept, three environmentally-relevant aqueous matrices, spiked with DCF-IBU mixture, were treated. In this case, relatively high TOC degradation values were found after 20 h reaction time (ca. 57.7, 73.9 and 54.5% in surface water, WWTP effluent and hospital wastewater, respectively). This work deals the first study about DCF-IBU removal in aqueous solution by CWPO, as well as a continuous study using real wastewater that allow to extend the experimental results to a real scenario.

Association of coronary calcification with prognosis of Covid-19 patients without known heart disease

Risk factors that determine the severity of Covid-19 have not been fully elucidated. The aim of this study was to evaluate the role of coronary artery calcification (CAC) as a risk factor for death or mechanical ventilation (MV) of patients without known heart disease infected with Covid-19. We analyzed 283 consecutive in-patients with acute respiratory symptoms with chest computed tomography (chest-CT), without previous heart disease, and criteria for Covid-19 (RT-PCR positive and/or typical clinical and chest-CT findings). CAC was classified by the number of coronary segments affected as absent (0), mild (1-3), and severe calcification (more than 3). The association between CAC, CAC severity, and death or MV due to severe respiratory failure was assessed by logistic regression. The mean age was 58.7±15.7 years and 54.1% were men. Patients with CAC were older, more likely to have hypertension, and less likely to be obese. CAC was present in 75 patients (26.5%), of which 42 had a mild calcification and 33 had severe calcification, and was associated with death (OR=2.35, 95%CI: 1.01-5.48) or MV (OR=2.72, 95%CI: 1.20-6.20) adjusted for multiple confounders, with significant and increased odds ratio for the severe form of CAC (death: OR=3.70, 95%CI: 1.20-11.42; MV: OR=3.30, 95%CI: 1.09-9.95). We concluded that CAC was an independent risk factor for death or MV in Covid-19 patients without previous heart disease, particularly for those with severe calcification. CAC can be easily visualized on common chest-CT, widely used in evaluation of moderate to severe Covid-19.

Breast cancer resistance protein (Bcrp/Abcg2) is selectively modulated by lipopolysaccharide (LPS) in the mouse yolk sac.

Bacterial infection alters placental ABC transporters expression. These transporters provide fetal protection against circulating xenobiotics and environmental toxins present in maternal blood. We hypothesized that lipopolysaccharide (LPS-bacterial mimic) alters the yolk sac morphology and expression of key ABC transporters in a gestational-age dependent manner. Yolk sac samples from C57BL/6 mice were obtained at gestational ages (GD) 15.5 and GD18.5, 4 or 24 h after LPS exposure (150ug/kg; n = 8/group). Samples underwent morphometrical, qPCR and immunohistochemistry analysis. The volumetric proportions of the histological components of the yolk sac did not change in response to LPS. LPS increased Abcg2 expression at GD15.5, after 4 h of treatment (p < 0.05). No changes in Abca1, Abcb1a/b, Abcg1, Glut1, Snat1, Il-1ß, Ccl2 and Mif were observed. Il-6 and Cxcl1 were undetectable in the yolk sac throughout pregnancy. Abca1, breast cancer resistance protein (Bcrp, encoded by Abcg2) and P-glycoprotein (P-gp/ Abcb1a/b) were localized in the endodermal (uterine-facing) epithelium and to a lesser extent in the mesothelium (amnion-facing), whereas Abca1 was also localized to the endothelium of the yolk sac blood vessels. LPS increased the labeling area and intensity of Bcrp in the yolk sac's mesothelial cells at GD15.5 (4 h), whereas at GD18.5, the area of Bcrp labeling in the mesothelium (4 and 24 h) was decreased (p < 0.05). Bacterial infection has the potential to change yolk sac barrier function by affecting Bcrp and Abcg2 expression in a gestational-age dependent-manner. These changes may alter fetal exposure to xenobiotics and toxic substances present in the maternal circulation and in the uterine cavity.

Resistive switching of silicon-silver thin film devices in flexible substrates.

Novel applications for memory devices demand nanoscale flexible structures. In particular, resistive switching (RS) devices are promising candidates for wearable and implantable technologies. Here, the Pt/Si/Ag/TiW metal-insulator-metal structure was fabricated and characterized on top of flexible substrates using a straightforward microfabrication process. We also showed that these substrates are compatible with sputtering deposition. RS was successfully achieved using both commercial cellulose cleanroom paper and bacterial cellulose, and polymer (PET) substrates. The bipolar switching behavior was observed for both flat and bent (under a radius of 3.5 mm) configurations. The observed phenomenon was explained by the formation/rupture of metallic Ag filaments in the otherwise insulating Si host layer.

Benquerencia (La Serena - Spain) rock art: An integrated spectroscopy analysis with FTIR and Raman.

La Serena region is a large plateau with open landscapes bounded in the south by a mountain chain formed by the Benquerencia, Tiros and La Rinconada Sierras. There are more than 300 painted and engraved sites in the region. Cueva Grande, Cueva de En medio and Cueva Pequeña are three Schematic rock art shelters located in the municipality of Benquerencia de la Serena, Badajoz, Spain. Over their panels have been documented more than a hundred of painted schematic figures (anthropomorphic figures, eye-shape figures and symbols) (Neolithic - Copper Age). Paintings are monochromatic with red or black coloration. A total of 13 samples (10 red and 3 black samples) from different panels were collected and analyzed using micro-Raman spectroscopy and ATR-FTIR. Micro-Raman spectroscopy was able to characterize the main mineral component, respectively hematite for the red figures and charcoal for the black paintings. ATR-FTIR was useful to possible ochre and possible organic identification. These latest results are particularly important for understanding manufacturing processes and addressing conservation problems.

Genomic characterization of MDR/XDR-TB in Kazakhstan by a combination of high-throughput methods predominantly shows the ongoing transmission of L2/Beijing 94-32 central Asian/Russian clusters.

BACKGROUND: Kazakhstan remains a high-burden TB prevalence country with a concomitent high-burden of multi-drug resistant tuberculosis. For this reason, we performed an in depth genetic diversity and population structure characterization of Mycobacterium tuberculosis complex (MTC) genetic diversity in Kazakhstan with both patient and community benefit. METHODS: A convenience sample of 700 MTC DNA cultures extracts from 630 tuberculosis patients recruited from 12 out of 14 regions in Kazakhstan, between 2010 and 2015, was independently studied by high-throughput hybridization-based methods, TB-SPRINT (59-Plex, n = 700), TB-SNPID (50-Plex, n = 543). DNA from 391 clinical isolates was successfully typed by two methods. To resolve the population structure of drug-resistant clades in more detail two complementary assays were run on the L2 isolates: an IS6110-NTF insertion site typing assay and a SigE SNP polymorphism assay. RESULTS: Strains belonged to L2/Beijing and L4/Euro-American sublineages; L2/Beijing prevalence totaled almost 80%. 50% of all samples were resistant to RIF and to INH., Subtyping showed that: (1) all L2/Beijing were "modern" Beijing and (2) most of these belonged to the previously described 94-32 sublineage (Central Asian/Russian), (3) at least two populations of the Central Asian/Russian sublineages are circulating in Kazakhstan, with different evolutionary dynamics. CONCLUSIONS: For the first time, the global genetic diversity and population structure of M. tuberculosis genotypes circulating in Kazakhstan was obtained and compared to previous local studies. Results suggest a region-specific spread of a very limited number of L2/Beijing clonal complexes in Kazakhstan many strongly associated with an MDR phenotype.

Microcosm evaluation of the impact of oil contamination and chemical dispersant addition on bacterial communities and sediment remediation of an estuarine port environment.

AIM: To evaluate the interactive effects of oil contamination and chemical dispersant application on bacterial composition and sediment remediation of an estuarine port environment. METHODS AND RESULTS: A multifactorial controlled microcosm experiment was set up using sediment cores retrieved from an estuarine port area located at Ria de Aveiro lagoon (Aveiro, Portugal). An oil spill with and without chemical dispersant addition was simulated. Sediment oil hydrocarbon concentrations and benthic bacterial community structure were evaluated by GC-MS and 16S rRNA high-throughput sequencing respectively. Although initially (first 10 days) chemical dispersion of oil enhanced the concentrations of the heavier polycyclic aromatic hydrocarbons and of the C22 -C30 alkane group, with time (21 days), no significant differences in hydrocarbon concentrations were detected among treatments. Moreover, no significant changes were detected in the structure of sediment bacterial communities, which mainly consisted of operational taxonomic units related to hydrocarbon-contaminated marine environments. We hypothesize that the environmental background of the sampling site preconditioned the communities' response to additional contamination. CONCLUSION: This experimental microcosm study showed that the chemical dispersion of oil did not influence sediment remediation or bacterial community composition. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study showed that chemical dispersion of oil may not improve the remediation of port sediments. Further studies are needed to investigate the impact of chemical dispersants in combination with bioremediation strategies on the process of sediment remediation in port areas.

Energy requirements for growth of pubertal female Saanen goats.

Previous research on energy requirements of female Saanen goats, using the factorial approach, has not considered the specific requirements for maintenance and growth during the pubertal phase. Thus, the purpose of this study was to estimate energy requirements for maintenance (Trial 1) and growth (Trial 2) of non-pregnant and non-lactating female Saanen goats at the pubertal phase from 30 to 45 kg. In Trial 1, the net energy requirements for maintenance (NEm ) were estimated using 18 female Saanen goats randomly assigned to three levels of intake: ad libitum, and 70% and 40% of ad libitum intake. These animals were pair-fed in six slaughter groups, each consisting of one animal for each level of intake. In Trial 2, the net energy requirements for growth (NEg ) were estimated using 18 female Saanen goats, which were fed ad libitum and slaughtered at targeted BW of 30, 38 and 45 kg. The NEm was 52 kcal/kg(0.75) of BW. The NEg increased from 3.5 to 4.7 Mcal/kg of BW gain as BW increased from 30 to 45 kg. Our results suggest that the guidelines of the major feeding systems for the entire growth phase may not be adequate for females at pubertal phase.

Low frequency electric current noise in glioma cell populations.

Measuring the electrical activity of large and defined populations of cells is currently a major technical challenge to electrophysiology, especially in the picoampere-range. For this purpose, we developed and applied a bidirectional transducer based on a chip with interdigitated gold electrodes to record the electrical response of cultured glioma cells. Recent research determined that also non-neural brain glia cells are electrically active and excitable. Their transformed counterparts, e.g. glioma cells, were suggested to partially retain these electric features. Such electrophysiological studies however are usually performed on individual cells and are limited in their predictive power for the overall electrical activity of the multicellular tumour bulk. Our extremely low-noise measuring system allowed us to detect not only prominent electrical bursts of neuronal cells but also minute, yet constantly occurring and functional, membrane capacitive current oscillations across large populations of C6 glioma cells, which we termed electric current noise. At the same time, tumour cells of non-brain origin (HeLa) proved to be electrically quiescent in comparison. Finally, we determined that the glioma cell activity is primarily caused by the opening of voltage-gated Na+ and K+ ion channels and can be efficiently abolished using specific pharmacological inhibitors. Thus, we offer here a unique approach for studying electrophysiological properties of large cancer cell populations as an in vitro reference for tumour bulks in vivo.

Rendimentos de cortes, proporção tecidual da carcaça e composição centesimal da carne de caprinos jovens em função do grupo racial e do peso corporal de abate / Cut yields, carcass and tissue composition proportion in meat from young goats depending on the breed and slaughter body weight

O objetivo do presente trabalho foi avaliar as proporções de cortes comerciais, tecidos do lombo e composição da carne de cabritos de cinco grupos raciais: Alpino, ½ Boer + ½ Alpino (½ BA), ½ Anglo Nubiano + ½ Alpino (½ ANA), ¾ Boer + » Alpino (¾ BA), » Boer + » Alpino + ½ Anglo Nubiano (Tricross), submetidos a três pesos de abate (25, 30 e 35kg) em sistema de confinamento, com utilização de dieta completa. O cruzamento das raças Boer e Anglo Nubiana com fêmeas Alpinas apenas melhorou, nos cabritos, os rendimentos de cortes considerados de terceira categoria, como pescoço e costela. Pesos corporais de abate entre 30 e 35kg devem ser preferidos por aumentarem a proporção de lombo, reduzirem a proporção de pescoço nos Alpinos e aumentarem a de perna em Alpinos e ¾ Boer, além de proporcionarem quantidades adequadas de músculo e gordura intermuscular.

The purpose of this study was to evaluate the commercial cuts, loin tissues and composition proportions in meant from five racial groups: Alpine, ½ Boer + ½ Alpine (½ BA), ½ Anglo Nubian + ½ Alpine (½ ANA), ¾ Boer + » Alpine (¾ BA), » Boer + » Alpine + ½ Anglo Nubian (Tricross), submitted to three slaughter weights (25, 30 and 35kg) in feedlot, using a complete diet. The crossing of Boer and Anglo Nubian with Alpine females only improved, in kids, the yield of cuts considered third category such as neck and rib. Slaughter body weights between 30 and 35kg should be preferred due to increase in the proportion of the loin, reduction in the proportion of the neck in Alpine and increase in the leg in Alpine and ¾ Boer, beyond the appropriate amounts of intermuscular fat and muscle.

Assessment of oxidative status markers and NO bioavailability in hypertensive disorders of pregnancy.

The systemic oxidative status in hypertensives disorders of pregnancy (HDP) and its association with endothelial dysfunction is controversial. In the present study, we evaluated systemic plasma levels of oxidative stress markers (TBARS (thiobarbituric acid-reactive substances) and carbonyl) and total antioxidant status (FRAP (ferric reducing ability of plasma (ferric reducing/antioxidant power) and reduction of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide))), as well as assessed the impact these markers have on nitric oxide (NO) status in healthy pregnant (HP, n=38), gestational hypertensive (GH, n=33) and preeclamptic pregnant women (PE, n=28). We found similar values of TBARS among all groups, and reduced carbonyl levels in HDP between the PE and GH. Conversely, significant increases in plasma activity of antioxidant status were observed in the GH and PE groups compared to the HP group (using both MTT or FRAP method). Importantly, HDP present significantly lower nitrite levels compared to HP women. In Conclusion, our findings show a compensatory antioxidant mechanism against reactive oxygen species (ROS) generation in HDP, which is not associated with nitrite levels restoration.

Understanding the relationship between Mycobacterium bovis spoligotypes from cattle in Latin American countries.

Spoligotyping is the most frequently used method for genotyping isolates of Mycobacterium bovis worldwide. In the current work, we compared spoligotypes from 1684 M. bovis isolates from Argentina (816), Brazil (412), Chile (66), Mexico (274) and Venezuela (116), obtained from cattle, humans, pigs, wild boars, farmed deer, goats, buffaloes, cats, and wild animals. A total of 269 different spoligotypes were found: 142 (8.4%) isolates presented orphan spoligotypes, whereas 1542 (91.6%) formed 113 different clusters. In cattle, SB0140 was the most representative spoligotype with 355 (24.6%) isolates, followed by SB0121 with 149 (10.3%) isolates. Clustering of spoligotypes ranged from 95.2% in Argentina to 85.3% in Mexico. Orphan spoligotypes were also variable, ranging from 23.7% in Mexico to 4.1% in Brazil. A large proportion of spoligotypes were common to the neighboring countries Argentina, Brazil and Chile. In conclusion, despite the diversity of spoligotypes found in the five countries studied, there are major patterns that predominate in these neighboring countries. These clusters may reflect a long-lasting active transmission of bovine tuberculosis or common historical origins of infection.

Tuberculosis caused by RDRio Mycobacterium tuberculosis is not associated with differential clinical features.

BACKGROUND: We recently described the Mycobacterium tuberculosis RD(Rio) genotype, a clonally derived sublineage within the Latin American-Mediterranean (LAM) family. Genetic diversity of M. tuberculosis likely affects the clinical aspects of tuberculosis (TB). Prospective studies that address this issue are scarce and remain controversial. OBJECTIVE: To determine the association of differential clinical features of pulmonary TB with the RD(Rio) M. tuberculosis etiology. METHODS: Culture-proven pulmonary TB patients (n = 272) were clinically evaluated, including history, physical examination, chest X-ray and anti-human immunodeficiency virus serology. Isolates were classified as RD(Rio) or non-RD(Rio) M. tuberculosis by multiplex polymerase chain reaction and further spoligotyped. Clinical and M. tuberculosis genotype data were analyzed. RESULTS: RD(Rio) M. tuberculosis caused disease in 26.5% (72/270) of all TB cases. The LAM genotype, of which RD(Rio) strains are members, was responsible for 46.0% of the TB cases. Demographic data, major signs and symptoms, radiographic presentation, microbiological features and clinical outcomes were not significantly different among patients with TB caused by RD(Rio) and non-RD(Rio) strains. CONCLUSIONS: Disease caused by M. tuberculosis RD(Rio) strains was not clinically distinctive or more severe than disease caused by non-RD(Rio) strains in this series of TB patients. Larger prospective studies specifically designed to disclose differential clinical characteristics of TB caused by specific M. tuberculosis lineages are needed.