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Coxiella burnetii is the etiologic agent of Q fever, a worldwide zoonosis. Cattle, sheep and goats are considered the main reservoirs of the disease. Transmission to humans occurs mainly through the inhalation of infectious aerosols from milk, faeces, urine, and birth products from infected ruminants. In this study, a 2-year longitudinal approach was performed to ascertain the excretion of C. burnetii in bulk tank milk samples of sheep from a mountain plateau in central Portugal, with sampling conducted during the years 2015 and 2016. From a total of 156 bulk tank milk samples tested by qPCR, only one showed to be positive for C. burnetii (1.28% [95%CI: 0.03-6.94]), from 2015, the first year of collection. Bidirectional sequencing and phylogenetic analysis of IS1111 transposase partial region confirmed the presence of C. burnetii DNA. The presence of C. burnetii in raw milk samples highlights the necessity for additional research to determine if raw milk is a potential source for human infection. Animal health surveillance and prevention measures against this zoonotic disease should be considered., (© 2024. The Author(s).)

Crimean-Congo hemorrhagic fever virus (CCHFV) is a widespread zoonotic pathogen that can cause mild to severe hemorrhagic disease in humans. CCHFV may be transmitted through direct contact with tissue or blood of viremic animals; however, the primary transmission route is through infected tick bites. CCHFV RNA has been detected in ticks feeding on domestic and wild animals in western Spain, suggesting an established circulation of CCHFV in Western Europe. Ruminants have been recognized as important CCHFV reservoirs and have been linked to human cases in endemic regions. Given the emergence of CCHF in neighboring Spain, and a report of two CCHFV seropositive humans in southern Portugal in 1985, we investigated the potential circulation of this virus in the country by performing a nationwide anti-CCHFV IgG serosurvey in sentinel sheep of Portugal. Sera (n = 459) randomly selected from widely distributed farms (n = 20) of Portugal were tested using a commercial double-antigen enzyme-linked immunosorbent assay, yielding an overall seroprevalence of 0.4% (95% confidence interval [CI] 0.04-1.56%). Positive sheep were from the southern region of Portugal (Alentejo region), which raise the seroprevalence of this region to 0.74% (95% CI 0.09-2.66%). This is the first study reporting the presence of CCHFV antibodies in sheep of Portugal, thus suggesting a geographical expansion of CCHFV to this country. It seems likely that CCHFV may exist focally in southern Portugal., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Rickettsiosis is considered an emerging/re-emerging vector-borne disease that causes significant public health threats. Ticks are reservoirs and vectors of Rickettsia having a significant role in the transmission of rickettsiae. In Portugal, little is known about tick-borne Rickettsia species in sheep. Therefore, this study aimed to investigate rickettsiae infection in ticks and their sheep host from 27 farms in four districts of central Portugal, to clarify the role of the sheep host in the circulation of this zoonotic agent. Between March and May 2021, EDTA blood samples (n = 100) of healthy grazing sheep and their ticks (n = 100, one tick per animal) were collected during a herd health program in central Portugal. Obtained ticks were identified as Rhipicephalus sanguineus sensu lato by PCR targeting a partial sequence of 16S rRNA gene followed by sequence analysis. Rhipicephalus sanguineus s.l. and host sheep blood were tested for the presence of Rickettsia spp. by PCR targeting a partial sequence of ompB and ompA genes. From a total of 100 paired R. sanguineus s.l. and host sheep, Rickettsia massiliae was detected in 62 ticks and 35 grazing sheep blood samples, collected in central Portugal, 2021. All 35 positive sheep had attached positive R. sanguineus s.l., with matching nucleotidic sequences. These findings suggest that sheep may develop rickettsiemia and are likely capable of transmitting and amplifying the infection to uninfected ticks maintaining rickettsiae in circulation in the domestic cycle., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aimed to compare the efficiency of two different sizes of the Surber sampler to assess benthic macroinvertebrates in headwater streams in two Amazonian regions. Two Surber samplers of different sizes were used, one measuring 20 × 20 cm and the other 30 × 30 cm, both with a 0.25-mm net. The number of replicates taken was 6 for the smaller sampler and 3 for the bigger one, maintaining approximately the same total sampled area. The study was carried out in 12 headwater streams with different environmental conditions. Biological metrics were calculated for each size at each site and compared within each stream health category. A two-way analysis of similarities test was performed to compare the community structure assessed by each method at each stream. A normalized sampling effort was used to quantify the number of samples required to correctly sample each site. The data did not show a significant difference between the two sizes regarding the taxonomic recruitment and the community structure sampled at each stream, but differences were found between the two sizes in dominance values and in Shannon index scores for the natural sites. Furthermore, the smaller Surber was able to assess 70% of the estimated richness in all sites, which suggests that it is better to assess benthic macroinvertebrates than the larger Surber. Moreover, the smaller Surber is easier to transport in the field, reducing the effort of the technician, and takes less time to sort the material collected with it, which can reduce the sample processing effort, therefore reducing the cost of the project.

Objectives: To describe and validate an artificial intelligence (AI)-driven structured reporting system by direct comparison of automatically generated reports to results from actual clinical reports generated by nuclear cardiology experts., Background: Quantitative parameters extracted from myocardial perfusion imaging (MPI) studies are used by our AI reporting system to generate automatically a guideline-compliant structured report (sR)., Method: A new nonparametric approach generates distribution functions of rest and stress, perfusion, and thickening, for each of 17 left ventricle segments that are then transformed to certainty factors (CFs) that a segment is hypoperfused, ischemic. These CFs are then input to our set of heuristic rules used to reach diagnostic findings and impressions propagated into a sR referred as an AI-driven structured report (AIsR). The diagnostic accuracy of the AIsR for detecting coronary artery disease (CAD) and ischemia was tested in 1,000 patients who had undergone rest/stress SPECT MPI., Results: At the high-specificity (SP) level, in a subset of 100 patients, there were no statistical differences in the agreements between the AIsr, and nine experts' impressions of CAD (P = .33) or ischemia (P = .37). This high-SP level also yielded the highest accuracy across global and regional results in the 1,000 patients. These accuracies were statistically significantly better than the other two levels [sensitivity (SN)/SP tradeoff, high SN] across all comparisons., Conclusions: This AI reporting system automatically generates a structured natural language report with a diagnostic performance comparable to those of experts.

Q fever is a zoonosis caused by Coxiella burnetii, and transmission to humans is often associated with contact with ovine and caprine livestock. Those exposed to sheep are particularly at high risk of infection. Recent studies show that Q fever is increasing in sheep farms in Portugal raising alerts on spillover to humans. We detected anti-C. burnetii IgG in shepherds and sheep milk cheesemakers (27 [28.1%] in a total of 96; 95% confidence interval [CI] 19.4-38.2%) and in controls (21 [8.1%] in a total of 260; 95% CI 5.1-12.1%), pointing to an increased risk of C. burnetii infection (P = 0.0001), with an odds ratio for anti-C. burnetii of 4.45 (95% CI 2.4-8.4%; P = 0.0001), in individuals with occupational contact with sheep in Portugal.

Q fever is a zoonotic disease caused by Coxiella burnetii that is highly prevalent across the world. In this study, a prospective serosurvey was performed to study C. burnetii circulation in a population of sheep in the central region of Portugal. Blood from a representative sample of 168 animals was drawn in both 2015 and 2016, and sera were tested for IgG anti-C. burnetii by EIA. In 2015, 7.7% (13/168) animals tested positive for IgG anti-C. burnetii, while in 2016, 17.3% (29/168) tested positive, showing a statistically significant (P = 0.008) increase in anti-C. burnetii seroprevalence. Results support the notion that Q fever is emerging in central Portugal.

Worries are common in surgical patients, especially in children. The present study analyzed the factor structure and the psychometric properties of a Portuguese version of the Child Surgery Worries Questionnaire (CSWQ-P) in a Portuguese sample of 490 children. Exploratory factor analysis, conducted via principal axis factoring with oblimin rotation, provided evidence for a four-factor structure of the 21 item questionnaire. A confirmatory factor analysis was also conducted, showing the good fit of this solution. The CSWQ-P proved to have one more subscale than the original Spanish version CSWQ. Correlations with the children's trait anxiety provided evidence of convergent validity for the CSWQ-P. Females also scored higher on worries than males on all subscales. Psychometric properties of this revised version of the CSWQ provided support for use with young children, and indicate the CSWQ-P has value for use in healthcare practice and in clinical research.

Pneumocystis pneumonia (PcP) is a major HIV-related illness caused by Pneumocystis jirovecii. Definitive diagnosis of PcP requires microscopic detection of P. jirovecii in pulmonary specimens. The objective of this study was to evaluate the usefulness of two serum markers in the diagnosis of PcP. Serum levels of (1-3)-beta-d-glucan (BG) and lactate dehydrogenase (LDH) were investigated in 100 HIV-positive adult patients and 50 healthy blood donors. PcP cases were confirmed using indirect immunofluorescence with monoclonal anti-Pneumocystis antibodies and nested-PCR to amplify the large subunit mitochondrial rRNA gene of P. jirovecii in pulmonary specimens. BG and LDH levels in serum were measured using quantitative microplate-based assays. BG and LDH positive sera were statistically associated with PcP cases (P ≤ 0.001). Sensitivity, specificity, positive/negative predictive values (PPV/NPV), and positive/negative likelihood ratios (PLR/NLR) were 91.3 %, 61.3 %, 85.1 %, 79.2 %, 2.359, and 0.142, respectively, for the BG kit assay, and 91.3 %, 35.5 %, 75.9 %, 64.7 %, 1.415 and 0.245, respectively, for the LDH test. Serologic markers levels combined with the clinical diagnostic criteria for PcP were evaluated for their usefulness in diagnosis of PcP. The most promising cutoff levels for diagnosis of PcP were determined to be 400 pg/ml of BG and 350 U/l of LDH, which combined with clinical data presented 92.8 % sensitivity, 83.9 % specificity, 92.8 % PPV, 83.9 % NPV, 5.764 PLR and 0.086 NLR (P < 0.001). This study confirmed that BG is a reliable indicator for detecting P. jirovecii infection. The combination between BG/LDH levels and clinical data is a promising alternative approach for PcP diagnosis.

Background: Left ventricular dyssynchrony is an adverse consequence of ST-elevation myocardial infarction (STEMI) and bears an unfavorable prognosis. Mechanical dyssynchrony as measured by phase analysis from gated single photon emission computed tomography (GSPECT) correlates well with other imaging methods of assessing dyssynchrony but has not been studied in STEMI. We hypothesized that systolic dyssynchrony as measured by GSPECT would correlate with adverse remodeling after STEMI., Methods: In 28 subjects suffering STEMI, GSPECT with technetium-99m sestamibi was performed immediately after presentation (day 5) and remotely (6 months). Parameters of left ventricular dyssynchrony (QRS width, histogram bandwidth (HBW) and phase standard deviation (PSD)) were measured from GSPECT using the Emory Cardiac Toolbox. Left ventricular volumes, ejection fraction (LVEF) and infarct size were also assessed., Results: After successful primary percutaneous coronary intervention to the infarct-related artery, subjects had an LVEF of 46.4% ± 11% and a resting perfusion defect of 27.4% ± 16% at baseline. Baseline QRS width was normal (91.5 ± 17.5 ms). Subjects with STEMI had dyssynchrony compared with a cohort of 22 normal subjects (age 57.2 ± 10.6 years, <5% perfusion defect) by both HBW (100.3° ± 70.7° vs 26.5° ± 5.3°, P < .0001) and PSD (35.3° ± 16.9° vs 7.9° ± 2.1°, P < .0001). Baseline HBW correlated with resting perfusion defect size (r = 0.67, P < .001), end-systolic volume (r = 0.72, P < .001), end-diastolic volume (r = 0.63, P = .001), and inversely with LVEF (r = -0.74, P < .001). HBW and PSD improved over the follow-up period (-24.1 ± 35.9 degrees, P = .003 and -8.7° ± 14.6°, P = .006, respectively), and improvement in HBW correlated with reduction in LV end-systolic volumes (r = 0.43, P = .034). Baseline HBW and PSD, however, did not independently predict LVEF at 6 months follow-up., Conclusions: After STEMI, subjects exhibit mechanical dyssynchrony as measured by GSPECT phase analysis without evidence of electrical dyssynchrony. Improvement in mechanical dyssynchrony correlates with beneficial ventricular remodeling. The full predictive value of this measure in post-infarct patients warrants further study.

Nucleotide variation in the Pneumocystis jirovecii upstream conserved sequence (UCS) and kexin-like serine protease (Kex1) loci was studied in pulmonary specimens from Portuguese HIV-positive patients. DNA was extracted and used for specific molecular sequence analysis. The number of UCS tandem repeats detected in 13 successfully sequenced isolates ranged from three (9 isolates, 69%) to four (4 isolates, 31%). A novel tandem repeat pattern and two novel polymorphisms were detected in the UCS region. For the Kex1 gene, the wild-type (24 isolates, 86%) was the most frequent sequence detected among the 28 sequenced isolates. Nevertheless, a nonsynonymous (1 isolate, 3%) and three synonymous (3 isolates, 11%) polymorphisms were detected and are described here for the first time.

Background: Cardiac resynchronization therapy (CRT) is approved for the treatment of patients with advanced systolic heart failure and evidence of dyssynchrony on electrocardiograms. However, a significant percentage of patients do not demonstrate improvement with CRT. Echocardiographic techniques have been used for more accurate determination of dyssynchrony. Single photon emission computed tomography (SPECT) myocardial perfusion imaging has not previously been used to evaluate cardiac dyssynchrony. The objective of this study is to evaluate mechanical dyssynchrony as described by phase analysis of gated SPECT images in patients with left ventricular dysfunction, conduction delays, and ventricular paced rhythms., Methods and Results: A novel count-based method is used to extract regional systolic wall thickening amplitude and phase from gated SPECT images. Five indices describing the phase dispersion of the onset of mechanical contraction are determined: peak phase, phase SD, bandwidth, skewness, and kurtosis. These indices were determined in consecutive patients with left ventricular dysfunction (n = 120), left bundle branch block (n = 33), right bundle branch block (n = 19), and ventricular paced rhythms (n = 23) and were compared with normal control subjects (n = 157). Phase SD, bandwidth, skewness, and kurtosis were significantly different between patients with left ventricular dysfunction, left bundle branch block, right bundle branch block, and ventricular paced rhythms and normal control subjects (all P < .001) Peak phase was significantly different between patients with right ventricular paced rhythms and normal control subjects (P = .001)., Conclusions: A novel SPECT technique for describing left ventricular mechanical dyssynchrony has been developed and may prove useful in the evaluation of patients for CRT.

Background: Transient ischemic dilation (TID) has been established as an important independent marker of severe and extensive coronary artery disease (CAD) in myocardial perfusion imaging (MPI). The accuracy of the TID index is dependent on a well-determined threshold (normal limits) between normal and abnormal values for each study protocol. To date, the effects of neither gender nor attenuation correction (AC) on TID normal limits have been established. Thus, the objectives of this study were to determine if AC processing changes the normal value of the TID index and if there were gender-related differences in the TID index of normal patients who had undergone rest/exercise-stress technetium-99m sestamibi MPI., Methods and Results: Seventy-five patients (33 women, 42 men; mean age, 57.7 +/- 11.7 y and 55.9 +/- 10.0 y, respectively) with less than a 5% likelihood of CAD, who had undergone low-dose rest/high-dose exercise-stress Tc-99m sestamibi MPI, were studied. All studies were acquired using simultaneous emission/transmission scans and were corrected for attenuation, scatter, and resolution effects using the ExSPECT II method. Both the AC and non-AC studies were analyzed using the Emory Cardiac Toolbox (ECTb; Syntermed, Inc, Atlanta, Ga) quantitative software. The TID index was calculated automatically as the ratio of stress mean left ventricular volumes to rest mean left ventricular volumes by ECTb. Patients were grouped by gender and the TID indices from AC and non-AC studies were compared. Linear regressions of the TID index and body mass index were analyzed to exclude differences in body size between male and female patients as a confounding factor in gender-related differences in TID. The TID index upper normal limits were calculated as the mean value plus 2 standard deviations (SDs). AC processing did not change the TID index significantly whether the genders were combined or separated (AC TID = 0.97 +/- 0.14 vs non-AC TID = 0.98 +/- 0.12 for all patients). Female patients showed higher mean TID indices than male patients in both AC (1.01 +/- 0.15 vs 0.95 +/- 0.12) and non-AC studies (1.00 +/- 0.15 vs. 0.97 +/- 0.10), but this difference was statistically significant only in AC studies (p = .03). TID indices remained constant across the range of body mass index studied. The TID index upper normal limit was 1.31 for female and 1.18 for male patients., Conclusion: TID normal values for rest/exercise-stress Tc-99m sestamibi MPI are gender-dependent and not affected by AC processing. Thus, diagnosticians should take into account these gender-related differences, as compared with the traditional value generated from mostly male populations, to ensure both men and women have the same overall accuracy of using the TID index in the diagnosis and prognosis of CAD.

Brazil officially joined the International Long Term Ecological Research (ILTER) network in January 2000, when nine research sites were created and funded by the Brazilian Council for Science and Technology (CNPq). Two-years later some positive signs already emerge of the scientific, social and political achievements of the Brazilian LTER program. We discuss examples of how ecosystem and biodiversity information gathered within a long-term research approach are currently subsidizing decision-making as regards biodiversity conservation and watershed management at local and regional scales. Success in this respect has often been related to satisfactory communication between scientists, private companies, government and local citizens. Environmental education programs in the LTER sites are playing an important role in social and political integration. Most examples of integration of ecological research to decision-making in Brazil derive from case studies at local or regional scale. Despite the predominance of a bottom-up integrative pathway (from case studies to models; from local to national scale), some top-down initiatives are also in order, such as the construction of a model to estimate the inpact of different macroeconomic policies and growth trajectories on land use. We believe science and society in Brazil will benefit of the coexistence of bottom-up and top-down integrative approaches.

We have investigated the presence of three neurochemical markers, glutamate, calbindin-D28k, and nitric oxide synthase, in spinal neurones that transmit chemical noxious inputs from both the skin and the viscera, by combining retrograde labelling with the fluorescent tracer Fluorogold with dual labelling immunohistochemistry. Neurones projecting to the nucleus of the solitary tract (NTS) that expressed Fos protein in response to cutaneous or visceral noxious stimulation were concentrated in lamina I of the cervical and lumbosacral segments, respectively. Although both labelled neuronal populations were numerous, the spino-solitary cells that transmit visceral nociceptive input predominated over those transmitting cutaneous nociceptive input. Calbindin-D28k-immunoreactivity was observed in neurones of three morphological types (fusiform, flattened, and pyramidal) projecting to the NTS that were activated by somatic or visceral nociceptive neurones. Nitric oxide synthase and glutamate immunoreactivities were present only in viscerally activated nociceptive neurones projecting to the NTS. Glutamate-immunopositive NTS-projecting cells were exclusively of the flattened type, and the nitric oxide synthase-immunolabelled NTS-projecting cells comprised 75%/fusiform cells and 25% flattened cells. These data suggest that the involvement of excitatory spinal lamina I projection neurones in the transmission of peripheral chemical nociceptive inputs to the NTS may be restricted to information of visceral origin.