Determinants of clinic absenteeism in a chemotherapy service of a cancer center located at Rio de Janeiro, Brazil.

INTRODUCTION: Clinic absenteeism promotes higher waiting lists for medical procedures and public resources waste. OBJECTIVES: The present work aimed to identify the reasons for clinic absenteeism from each cycle of the antineoplastic chemotherapy treatment, as well as to determine the socio-demographic, clinical and treatment profiles of this population. METHODS: This observational prospective work evaluated pediatric and adult patients which missed their chemotherapy cycle between May and October 2023 in a Cancer Center located in Rio de Janeiro, Brazil. Clinic absenteeism rate was calculated, and socio-demographic profile was described. Reasons for absenteeism, treatment protocol and most used drugs were also identified. RESULTS: This work analyzed data from 69 patients, the majority above 60 years old. Approximately 60% were male, 33.3% had little to no education and 63.8% lived outside the center city. Absenteeism average monthly rate was 1.73% for adults and 0.87% for children. The most related non-attendance reasons were patient feeling too ill to attend their chemotherapy session, failure to remember the cycle day and lack of means of transportation. Most prevalent neoplasms were from the digestive tract (46%). Fluorouracil, irinotecan, oxaliplatin and gemcitabine were the most discarded drugs due to absenteeism. CONCLUSIONS: Older patients and the ones residing far away from the Center tend to miss the scheduled chemotherapy cycles. However, most reasons for absenteeism could be avoided by confirmation calls or text messages. These procedures implementation could lead to a lower absenteeism rate and less resource waste.

CPR and DPANN Have an Overlooked Role in Corals' Microbial Community Structure.

Understanding how microbial communities are structured in coral holobionts is important to estimate local and global impacts and provide efficient environment management strategies. Several studies investigated the relationship between corals and their microbial communities, including the environmental drivers of shifts in this relationship, associated with diseases and coral cover loss. However, these studies are often geographically or taxonomically restricted and usually focused on the most abundant microbial groups, neglecting the rare biosphere, including archaea in the group DPANN and the recently discovered bacterial members of the candidate phyla radiation (CPR). Although it is known that rare microbes can play essential roles in several environments, we still lack understanding about which taxa comprise the rare biosphere of corals' microbiome. Here, we investigated the host-related and technical factors influencing coral microbial community structure and the importance of CPR and DPANN in this context by analyzing more than a hundred coral metagenomes from independent studies worldwide. We show that coral genera are the main biotic factor shaping coral microbial communities. We also detected several CPR and DPANN phyla comprising corals' rare biosphere for the first time and showed that they significantly contribute to shaping coral microbial communities.

Oral health-related quality of life determinants throughout adolescence: a cohort study in Brazil.

PURPOSE: Oral health-related quality of life (OHRQoL) is a complex construct that reflects the interaction among well-being, socioeconomic, oral health status, and also contextual factors. This study aimed to verify the association between poorer socioeconomic background and oral health condition on OHRQoL throughout adolescence. METHODS: A cohort study followed a random sample of 1134 participants aged 12 years in 2012 (T1), 770 adolescents with an average age of 14 years in 2014 (T2; retention rate, 67.9%), and 768 adolescents with an average age of 17.5 years in 2018 (T3; retention rate, 67.8%). OHRQoL was collected three times using the Brazilian short version of the Childhood Perception Questionnaire 11-14 (CPQ 11-14). At baseline, the adolescents were clinically assessed by dentists, and their demographic data, socioeconomic status, use of dental services, and toothache were evaluated. The mean income of the neighborhood was used as a proxy of contextual factors. This variable was obtained from an official municipal publication. Poisson multilevel regression analyses were performed using a hierarchical approach to assess the predictors of OHRQoL over time. RESULTS: Adolescents living in a neighborhood with low mean income, low household income, low education levels, girls, and nonwhite individuals reported poor OHRQoL. Clinical variables such as gingival bleeding and malocclusion were also associated with higher overall scores on CPQ11-14, indicating poor OHRQoL. CONCLUSION: Our results showed that the unfavorable contextual and individual socioeconomic environment, and oral disease negatively influenced OHRQoL during adolescence.

Epicardial adipose thickness in youth with type 1 diabetes.

BACKGROUND AND OBJECTIVE: Epicardial adipose thickness (EAT) is increased in adults with type 1 diabetes (T1D) and is thought to contribute to cardiovascular disease (CVD) in this population. Given that CVD risk factors emerge early in life, the purpose of this study was to identify whether EAT is increased in pediatric patients with T1D compared with non-diabetic controls. METHODS: Anthropometric data, blood pressure (BP), and EAT were evaluated in 20 youth with T1D and 20 age, sex, and body mass index (BMI) matched healthy controls between the ages of 5 and 18 years. RESULTS: EAT was 18.5% higher among youth with T1D compared to healthy controls (1.65 ± 0.44 mm vs 1.37 ± 0.27 mm, P = .02). In the entire cohort, EAT was correlated with age (r = 0.71, P < .001), BMI (r = .69, P < .001), waist circumference (r = 0.60, P < .001), systolic BP (r = .34, P = .03), and diastolic BP (r = 0.41, P = .009). Among youth with T1D, there were no significant correlations between EAT and HbA1c (r = -0.16, P = .50), insulin dose (r = .09, P = .71), or duration of disease (r = 0.06, P = .82). CONCLUSIONS: Youth with T1D exhibited significantly higher EAT compared to controls. Increased EAT was associated with adiposity and BP, but not duration of disease, insulin dose, or glycemic control. Increased EAT may represent a pathophysiologic mechanism leading to premature CVD in pediatric patients with T1D.

Influence of the calcium orthophosphate:glass ratio and calcium orthophosphate functionalization on the degree of conversion and mechanical properties of resin-based composites.

The study verified the influence of calcium orthophosphate (CaP):glass ratio on the degree of conversion and mechanical properties of resin-based composites containing either TEGDMA-functionalized dicalcium phosphate anhydrous (DCPA) or non-functionalized DCPA particles. The null hypotheses were that the evaluated variables are not affected by (1) CaP:glass ratio or (2) DCPA functionalization. DCPA particles were synthesized and half of them were functionalized with TEGDMA. Particle characterization included x-ray diffraction, elemental analysis, laser scattering, helium picnometry and scanning electron microscopy. Two series of composites were prepared containing either DCPA-NF (non-functionalized) or DCPA-F (functionalized), with total inorganic content of 50 vol % and DCPA:silanized barium glass (BG) ratios from 10:40 to 50:0. A composite containing 50 vol % BG was tested as control. DC was determined using FTIR spectroscopy. Biaxial flexural strength and modulus were tested after 24 h in water. Data were analyzed using Kruskal-Wallis/Dunn (flexural properties) or analysis of variance/Tukey tests (DC). Materials with similar actual DCPA contents were compared using Student's t test (alpha: 0.05). DC was higher for materials with DCPA-F, except for the 10:40 ratio. DCPA-F resulted in higher strength than DCPA-NF only at 40:10 ratio. Modulus was not affected by functionalization. Materials with similar actual DCPA contents showed differences in DC (F > NF), while no difference in flexural properties was observed between materials with 28%-30% DCPA. Both null hypotheses were rejected.

Compositional boundaries for functional dental composites containing calcium orthophosphate particles.

OBJECTIVES: To investigate the interrelationships among handling, degree of conversion (DC), mechanical behavior and Ca2+ release of composites containing dicalcium phosphate dihydrate (DCPD, CaHPO4.2H2O), as a function of total inorganic content and DCPD: glass ratio. METHODS: Twenty-one formulations (1 BisGMA: 1 TEGDMA, in mols) with inorganic fractions ranging from zero to 50 vol% and different DCPD: glass ratios were evaluated for viscosity (parallel plate rheometer, n = 3), DC (near-FTIR spectroscopy, n = 3), fracture toughness/K1C (single-edge notched beam, n = 7-11) and 14-day Ca2+ release (inductively coupled plasma optical emission spectroscopy, n = 3). Data were analyzed by ANOVA/Tukey test (except viscosity, where Kruskal-Wallis/Dunn tests were used, α: 0.05). RESULTS: Viscosity and DC increased with DCPD: glass ratio among composites with the same inorganic content (p < 0.001). At inorganic fractions of 40 vol% and 50 vol%, keeping DCPD content at a maximum of 30 vol% did not compromise K1C. Ca2+ release showed an exponential relationship with DCPD mass fraction in the formulation (R2 = 0.986). After 14 days, a maximum of 3.8% of the Ca2+ mass in the specimen was released. CONCLUSION: Formulations containing 30 vol% DCPD and 10-20 vol% glass represent the best compromise between viscosity, K1C and Ca2+ release. Materials with 40 vol% DCPD should not be disregarded, bearing in mind that Ca2+ release will be maximized at the expense of K1C.

Effect of the addition of fique bagasse microparticles in obtaining a biobased material based on cassava starch.

The indiscriminate accumulation of plastic waste has prompted research that leads to obtaining biobased materials. The research aim was to evaluate the effect of incorporating fique bagasse microparticles (FBM) in a cassava starch-based foamed material. First, the FBM extraction conditions were established by acid hydrolysis, for which the effect of acid concentration (5, 10 and 15% H2SO4), temperature (70, 80 and 90 °C) and extraction time (3, 5 and 7 h) on particle size, functional groups, color, and thermal properties was evaluated. The addition of FBM to the foamed material was then carried out. To do this, a completely randomized design with five treatments (0, 0.5, 0.75, 1.0 and 1.25% FBM) was evaluated. The response variables were the apparent density, expansion and spring index, compressibility, water absorption, thermal properties and FTIR. The results showed that the acid concentration, temperature and time had an effect on the morphological, chemical and thermal properties of FBM, with 10%, 70 °C and 7 h being the conditions that allowed obtaining the smallest particle size (61.69 ± 12.88 µm2). Moreover, the FBM concentration had a significant effect on the physical and mechanical properties of the foam, unleashing the treatment properties of 0.75%. This indicates that FBM have potential for use in obtaining biobased materials.

Novel Ehrlichia canis genogroup in dogs with canine ehrlichiosis in Cuba.

BACKGROUND: Canine monocytic ehrlichiosis (CME) is caused by the tick-borne pathogen Ehrlichia canis, an obligate intracellular Gram-negative bacterium of the family Anaplasmataceae with tropism for canine monocytes and macrophages. The trp36 gene, which encodes for the major immunoreactive protein TRP36 in E. canis, has been successfully used to characterize the genetic diversity of this pathogen in different regions of the world. Based on trp36 sequence analysis, four E. canis genogroups, United States (US), Taiwan (TWN), Brazil (BR) and Costa Rica (CR), have been identified. The aim of this study was to characterize the genetic diversity of E. canis in Cuba based on the trp36 gene. METHODS: Whole blood samples (n = 8) were collected from dogs found to be infested with the tick vector Rhipicephalus sanguineus sensu lato (s.l.) and/or presenting clinical signs and symptoms of CME. Total DNA was extracted from the blood samples and trp36 fragments were amplified by PCR. Nucleotide and protein sequences were compared using alignments and phylogenetic analysis. RESULTS: Four of the trp36 sequences obtained (n = 8) fall within the phylogenetic cluster grouping the US genogroup E. canis strains. The other E. canis trp36 sequences formed a separate and well-supported clade (94% bootstrap value) that is phylogenetically distant from the other major groups and thus represents a new genogroup, herein designated as the 'Cuba (CUB) genogroup'. Notably, dogs infected with the CUB genogroup presented frequent hemorrhagic lesions. CONCLUSIONS: The results of this study suggest that genetic diversification of E. canis in Cuba is associated with the emergence of E. canis strains with increased virulence.

Susceptibility of the patients infected with Sars-Cov2 to oxidative stress and possible interplay with severity of the disease.

Several recent reviews have suggested a role of oxidative stress in the pathophysiology of COVID-19, but its interplay with disease severity has not been revealed yet. In the present study, we aimed to investigate the association between the severity of COVID-19 and oxidative stress parameters. Clinical data of 77 patients with COVID-19 admitted to the hospital were analyzed and divided into moderate (n = 44) and severe (n = 33) groups based on their clinical condition. Production of oxidant (hydrogen peroxide) and defense antioxidants (total antioxidant capacity, reduced and oxidized glutathione, glutathione s-transferase), and oxidative damage (malondialdehyde, carbonyl, and sulfhydryl) were assessed using the serum samples. The results revealed that severe patients who presented high serum leukocyte count and CRP level stayed for a longer period in the hospital. However, there was no correlation observed between the oxidative stress parameters and degree of COVID-19 severity in the present study. In conclusion, these results indicate that the disease severity may not be a detrimental factor contributing to the changes in the redox profile of hospitalized patients with COVID-19.

A novel evaluation method for Ki-67 immunostaining in paraffin-embedded tissues.

The Ki-67 labeling index is traditionally used to investigate tumor aggressiveness. However, no diagnostic or prognostic value has been associated to the heterogeneous pattern of nuclear positivity. The aims of this study were to develop a classification for the patterns of Ki-67-positive nuclei; to search scientific evidence for the Ki-67 expression and location throughout the cell cycle; and to develop a protocol to apply the classification of patterns of Ki-67-positive nuclei in squamous epithelium with different proliferative activities. Based on empirical observation of paraffin sections submitted to immunohistochemistry for the determination of Ki-67 labeling index and literature review about Ki-67 expression, we created a classification of the patterns of nuclear positivity (NP1, NP2, NP3, NP4, and mitosis). A semi-automatic protocol was developed to identify and quantify the Ki-67 immunostaining patterns in target tissues. Two observers evaluated 7000 nuclei twice to test the intraobserver reliability, and six evaluated 1000 nuclei to the interobserver evaluation. The results showed that the immunohistochemical patterns of Ki-67 are similar in the tumoral and non-tumoral epithelium and were classified without difficulty. There was a high intraobserver reliability (Spearman correlation coefficient > 0.9) and moderate interobserver agreement (k = 0.523). Statistical analysis showed that non-malignant epithelial specimens presented a higher number of NP1 (geographic tongue = 83.8 ± 21.8; no lesion = 107.6 ± 52.7; and mild dysplasia = 86.6 ± 25.8) when compared to carcinoma in Situ (46.8 ± 34.8) and invasive carcinoma (72.6 ± 37.9). The statistical evaluation showed significant difference (p < 0.05). Thus, we propose a new way to evaluate Ki-67, where the pattern of its expression may be associated with the dynamics of the cell cycle. Future proof of this association will validate the use of the classification for its possible impact on cancer prognosis and guidance on personalized therapy.

Histological study and immunohistochemical location of cytoskeletal proteins in the ovaries of the three species of lizards of the family Leiosauridae (Reptilia: Squamata).

The aim of the study is to expand information on the histological aspect, presence, and distribution of cytoskeletal proteins: smooth muscle alpha-actin (α-SMA), desmin and vimentin in ovaries of three lizards Leiosauridae. In all analysed species, the ovaries were paired organs, located inside the coelomic cavity, covered in a simple cubic epithelium. Below the surface was the tunica albuginea. The organs can be divided into two regions: the cortex and the medulla. The pre-vitellogenic follicles consist of an oocyte surrounded by the pellucid zone periodic acid schiff positive, the granulosa layer consisting of three cell types: small, intermediate, and large and the theca layer. The vitellogenic follicles presented only a single layer of cubic granulosa cells. In the three lizards, α-SMA microfilaments (MFs) were verified along the theca layer and in endothelial cells of the blood vessels of the analysed follicles. Researchers have observed desmin intermediate filaments immunostaining exclusively muscle fibers of the albuginea and the ovarian stroma of Enyalius perditus. There was no immunostaining for vimentin in the ovaries of all the lizards studied. Results obtained revealed that the MFs of α-SMA could be responsible for a contractile activity present in the lizards assessed.

Geographic Distribution of Ehrlichia canis TRP Genotypes in Brazil.

Tandem repeat proteins (TRPs) are major immunoreactive proteins of Ehrlichia canis, which have been used in the serological diagnosis of different genotypes of the microorganism. TRP19 is preserved among different E. canis isolates expressed on both reticulate and dense-core cells and observed in the extracellular matrix or associated with the morula membrane. TRP36 is differentially expressed only on the surface of the dense-core form of the bacterium and exhibits more divergence among isolates. The aim of this study was to evaluate the distribution of the American (USTRP36), Brazilian (BrTRP36) and Costa Rican (CRTRP36) genotypes of E. canis in Brazil, using ELISA assays. Serum samples of 814 dogs from 49 municipalities from all over Brazil were analyzed. Our results showed that 33.9% of the samples were reactive to the USTRP36 genotype and 32.6% to the BrTRP36 genotype. The two genotypes appeared to occur equally throughout Brazil, although the frequency of seropositivity was lower in the south than in the country's other regions. Dogs that reacted to at least one of the synthetic peptides (TRP19 and TRP36) were 456 (56%). A few dogs (n = 5; 0.6%) reactive to the E. canis TRP36 genotype (CRTRP36) were also detected in the northeast and southern regions. We concluded that the American and Brazilian genotypes of E. canis are distributed evenly in Brazil, especially in the tropical region, while the temperate region in the south presented the lowest prevalence rates. This study offers the first report of dogs seropositive for the Costa Rican genotype in Brazil.

Canonical correlative analyses among an enzyme-linked immunosorbent assay using synthetic peptides, an indirect fluorescent antibody test, and hematologic measurements in dogs infected with Ehrlichia canis.

BACKGROUND: Immunoreactive tandem repeat proteins (TRPs) in amino acid sequences were identified and employed in the serologic diagnosis of canine monocytic ehrlichiosis (CME). OBJECTIVES: This study evaluated using TRP19 and TRP36 synthetic protein antigens with enzyme-linked immunosorbent assays (ELISAs) and compared the results with an indirect fluorescent antibody test (IFAT) to diagnose CME in the serum of dogs with suspected CME. METHODS: The sera of 243 dogs that exhibited clinical and hematologic signs suggestive of CME had IFATs performed. An ELISA with synthetic TRP19 and TRP36 antigens from two E canis genotypes (USTRP36 and BrTRP36) that circulate in Brazil were also performed. Canonical correlations and chi-square tests were evaluated on the results of these tests. RESULTS: Among the 243 dogs, 179 (73.6%) were reactive at an IFAT ≥ 40, 167 (68.72%) reacted at the cut-off ≥160, and 149 (61.31%) reacted at ≥640. The ELISA revealed that 172 (70.8%) dogs reacted to TRP19 peptide, 163 (67.1%) reacted to USTRP36, and 114 (46.9%) reacted to BrTRP36. The canonical correlation analyses showed that the TRP19 ELISA was the most effective diagnostic method. Serum total protein levels showed strong positive correlations with the USTRP36 peptide. Qualitative analyses revealed an association between the TRP19 peptide and the presence of anemia, thrombocytopenia, and hyperproteinemia. The IFAT results at titers of ≥160 and ≥640 were associated with thrombocytopenia. CONCLUSIONS: The results of this study indicated that the TRP19 peptide was an excellent antigen and that the IFAT was diagnostically important in confirming the diagnosis of CME in serum samples.

Effect of calcium orthophosphate: Reinforcing glass ratio and prolonged water storage on flexural properties of remineralizing composites.

OBJECTIVES: To compare the effects of replacing reinforcing barium glass particles by DCPD (dicalcium phosphate dihydrate), as opposed to simply reducing glass filler content, on composite flexural properties and degree of conversion (DC). On a second set of experiments, composites with different "DCPD: glass" ratios were exposed to prolonged water immersion to verify if the presence of DCPD particles increased hydrolytic degradation. METHODS: Two series of composites were prepared: 1) composites with total inorganic content of 50 vol% and "DCPD: glass" ratios ranging from zero (glass only) to 1.0 (DCPD only), in 0.25 increments, and 2) composites containing only silanized glass (from zero to 50 vol%). Disk-shaped specimens were fractured under biaxial flexural loading after 24 h in water. Another set of specimens of composites with different "DCPD: glass" ratios was stored in water for 24 h, 30, 60, 90 and 120 days and tested in flexure. DC was determined using FTIR spectroscopy. Data were analyzed using Kruskal-Wallis/Dunn test (flexural properties) or ANOVA/Tukey test (DC, alpha: 0.05). RESULTS: For glass-only composites, reducing inorganic content caused a linear decrease in strength. The presence of DCPD did not affect composite strength up until a "DCPD: glass" ratio of 0.5. On the other hand, materials with 0.75 and 1.0 DCPD showed significantly lower strength than the glass-only composite with 12.5 vol% filler and the unfilled resin, respectively (p < 0.001). Except for the 0.25 DCPD composite, the presence of DCPD did not contribute to increase flexural modulus. After water storage, composites containing DCPD showed higher percent reductions in properties than the control, but only in a few cases the effect was statistically significant (strength: 0.5 DCPD, modulus: 0.25 and 1.0 DCPD). DC was only marginally affected by DCPD fraction. SIGNIFICANCE: For composites with "DCPD: glass" of 0.25 and 0.5, reductions in strength were related to the lower glass content, and not due to the presence of DCPD. Flexural modulus was primarily defined by glass content. Overall, composites containing DCPD particles presented higher reductions in properties after water storage, but it remained within limits reported for commercial materials.

Tick-borne pathogens in dogs, wild small mammals and their ectoparasites in the semi-arid Caatinga biome, northeastern Brazil.

Caatinga is a biome exclusive to the semiarid zone of Brazil, where studies on ticks and tick-borne diseases are scarce. Herein, we investigated the occurrence of Rickettsia, Ehrlichia, and Coxiella in wild mammals, domestic dogs and their ectoparasites using molecular and serological techniques. During 2014-2016, blood samples and ectoparasites were collected from 70 small mammals (51 rodents, 18 marsupials, 1 wild canid) and 147 domestic dogs in three areas of the Caatinga. Through serological analyses of domestic dogs of the three areas, 8 to 11 % were seropositive for Rickettsia rickettsii, 9 to 37 % for Rickettsia amblyommatis, 61 to 75 % for Ehrlichia canis, and 0-5% for Coxiella burnetii. All wild mammals were seronegative for Rickettsia spp. and C. burnetii, except for one rodent (Wiedomys pyrrhorhinos) and one marsupial (Didelphis albiventris) that were seroreactive to C. burnetii, one wild canid (Cerdocyon thous) for R. amblyommatis, and two Rattus rattus for Rickettsia spp. Through PCR targeting DNA of Rickettsia, Ehrlichia or Coxiella, all blood samples were negative, except for the presence of Ehrlichia canis DNA in 8.8 % of the domestic dogs, and a recently reported novel agent, Ehrlichia sp. strain Natal, in one marsupial (Gracilinanus agilis). A total of 222 ticks, 84 fleas, and six lice were collected. Ticks were mostly Rhipicephalus sanguineus sensu lato, some Ixodes loricatus, Ornithodoros rietcorreai, Haemaphysalis sp., and Amblyomma spp.; fleas were Ctenocephalides felis felis, Pulex sp. and Polygenis (Polygenis) bohlsi jordani; and lice were Polyplax sp. and Gyropus sp. Through molecular detection of microorganisms, 9% of C. felis felis contained Rickettsia felis, 20 % of A. auricularium contained R. amblyommatis and 13 % of A. parvum contained 'Candidatus Rickettsia andeanae', whereas Ehrlichia canis DNA was detected in at least 6% of the R. sanguineus s.l. from one area. We report a variety of ectoparasites infesting small mammals and domestic dogs in the Caatinga biome, where these ectoparasites probably act as vectors of rickettsiae, ehrlichial agents (E. canis and Ehrlichia sp. strain Natal) and C. burnetii. Our results highlight to the potential risks of human infection by these tick-borne agents in the Caatinga biome.

Uncommon Ehrlichia canis infection associated with morulae in neutrophils from naturally infected dogs in Brazil.

Ehrlichia and Anaplasma species are the most common tick-borne disease (TBD) pathogens in dogs worldwide. Ehrlichia canis, the aetiological agent of the Canine Monocytic Ehrlichiosis (CME), is known to replicate within the cytoplasm of mononuclear cells into clusters of organisms called morulae. However, detection of morulae in neutrophils is commonly observed in dogs infected by Ehrlichia ewingii or Anaplasma phagocytophilum. We report uncommon clinical cases of canine ehrlichiosis presenting morulae compatible with E. ewingii and A. phagocytophilum in dogs from two distinct regions of Brazil. Eight dogs were admitted to two veterinary teaching hospitals from Brazil, showing clinical or haematological signs suggestive of TBD. Blood or peritoneal fluid was withdrawn for haematological and cytologic analysis. All samples were evaluated by PCR assays for Ehrlichia and Anaplasma using genus-specific primers for dsb, 16S rRNA and groEL genes, followed by sequencing. Samples were also evaluated by nested PCR assays for the 16S rRNA gene of E. ewingii and groEL gene of A. phagocytophilum and Anaplasma platys. Seven dogs revealed thrombocytopenia, six dogs had monocytosis and five presented lymphopenia and anaemia. All dogs showed morulae structures compatible with Ehrlichia spp. in neutrophils and were PCR-positive for the dsb and 16S rRNA gene fragments of Ehrlichia, with sequences showing 100% identity with multiple E. canis sequences deposited in the GenBank™. Sequencing of 16S rRNA and groEL gene fragments from one PCR-positive dog showed 100% identity with A. platys. Overall, our data suggest that in endemic regions for E. canis, that is Brazil, the presence of morulae in neutrophils may indicate infection by this bacterium. Herein, morulae were also found in neutrophils present in the peritoneal fluid of a dog. Also, this is the first report of E. canis and Hepatozoon canis co-infection in neutrophils from naturally infected dogs confirmed by DNA sequencing.

Natural pigment extraction optimization from coffee exocarp and its use as a natural dye in French meringue.

This study aimed to optimize the pigment extraction process for coffee exocarp and to evaluate its coloring effect on French meringue. The anthocyanins were determined with the differential pH method and the process was optimized with the response surface methodology. The solvent concentration (SC) and solid solvent ratio (SSR) had a significant effect on the anthocyanin content and surface color of the coffee cherry extracts. The optimal extraction conditions 60% SC and 25% SSR resulted in the highest concentration of anthocyanins (0.145 mg cyanidin 3-glucoside/g of coffee fresh exocarp). For the French meringue, the 3% extract concentration had the smallest total color difference (ΔE), as compared to the control, evidencing the potential of coffee exocarp dyes in place of synthetic dyes in the manufacture of French meringue.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil.

Rangelia vitalii infects erythrocytes, leukocytes and endothelial cells of dogs. The present study aimed to report the molecular detection confirmed by sequencing of R. vitalii in the state of Paraná, as well as describe the clinical, hematological and biochemical alterations of the infected dogs. Three sick dogs from the metropolitan area of Curitiba, PR, Brazil, underwent a physical exam, and laboratory tests included hematology, biochemistry, polymerase chain reaction (PCR), and gene sequencing. Clinical signs included apathy, anorexia, and hemorrhage. Intra-erythrocytic and extracellular piroplasms were found on peripheral blood smears from all three dogs. Blood samples from these animals were positive for Babesia sp. by PCR targeting 18S rRNA. PCR products from all three dogs were sequenced, and BLAST analysis showed that the PCR-generated sequences were highly homologous with those of R. vitalii previously reported. Hematologic findings included severe anemia, shift of neutrophils to the regenerative left, and thrombocytopenia. Serum urea levels were increased in all three dogs, and direct bilirubin levels were elevated in one dog.

Tick-borne pathogens in carthorses from Foz do Iguaçu City, Paraná State, southern Brazil: A tri-border area of Brazil, Paraguay and Argentina.

Tick-borne diseases (TBD) constitute an important group of illness affecting animals and humans worldwide. In Brazil, carthorses are frequently exposed to ticks and tick-borne pathogens, leading to impairment of horse performance and imposing restrictions by the international veterinary authorities for the importation of horses. Accordingly, this study has aimed to i) determine the prevalence of the TBD agents Theileria equi, Babesia caballi, Ehrlichia spp., and hemotropic mycoplasmas in carthorses, ii) identify the tick species parasitizing the animals, and iii) determine factors associated with exposure/infection in Foz do Iguaçu City, Parana state, southern Brazil. A total of 103 carthorses were screened for anti-T. equi and anti-Ehrlichia spp. antibodies by indirect fluorescent antibody assays (IFA). Samples were also tested by PCR assays targeting the 18S rRNA gene of T. equi and B. caballi, and 16S rRNA gene of hemoplasmas. Additionally, PCR assays targeting the 16S rRNA, disulfide bond formation protein (dsb) and tandem repeat proteins 36 (trp36) genes of Ehrlichia spp. were also performed. Antibodies to T. equi and Ehrlichia spp. were detected in 43/103 (41.75%; 95% CI: 32.10-51.88%) and 5/103 (4.85%; 95% CI: 1.59-10.97%) horses by IFA, respectively. DNA of T. equi and B. caballi were found in 25/103 (24.27%; 95% CI: 16.36-33.71%) and 10/103 (9.71%; 95% CI: 4.75-17.13%) carthorses, respectively, and all tested negative for Ehrlichia spp. and hemoplasmas. All sequences showed ≥99% identity with multiple T. equi and B. caballi 18S rRNA gene sequences deposited in GenBank. Overall, 191 Dermacentor nitens ticks were collected from 25/103 (24.27%) animals. Carthorses older than 5 years were more likely to be positive for T. equi (p < 0.05). In conclusion, equine piroplasmosis agents are highly prevalent in carthorses from Foz do Iguaçu City. The low prevalence of Ehrlichia spp. found may be due to the absence of Amblyomma ticks infesting animals, which should be further investigated.

Distribution and sources of organic contaminants in surface sediments of Hooghly river estuary and Sundarban mangrove, eastern coast of India.

This study investigated polycyclic aromatic hydrocarbons (PAHs) and organochlorine compounds such as polychlorinated biphenyls (PCBs) and DDT-related pesticides in surface sediments of Hooghly estuary and the Sundarban mangrove wetlands. Concentrations of ∑17PAH, ∑182PCB and ∑6DDT ranged from 15.4 to 1731, not detected (nd) to 13.5 and nd to 8.97 ng g-1 dry weight, respectively. Low levels of PCBs and low to moderate concentrations of DDTs and PAHs reflected recent development in West Bengal, which was dominated by agriculture and multifarious industries in the past. Diagnostic ratios suggested that major sources of PAHs are combustion processes, DDTs are input by agriculture, antifouling paints and public health campaigns, and organochlorines are predominantly from industrial origin. Heavier PCB congeners suggest local sources and short-range transport of such chemicals. Decision makers may use these findings for managing the Hooghly River watershed in order to promote a sustainable development on the eastern coast of India.