Congenital cytomegalovirus infection in twin pregnancy.

Cytomegalovirus (CMV) infection is one of the preeminent congenital viral infections, and despite its potential morbidity, uncertainty about its physiopathology, prevention and treatment remains until now. We report a case of a dichorionic and diamniotic twin pregnancy in which only one of the fetus had signs of being affected. The first twin had prenatal diagnosis of intrauterine growth restriction and hyperechogenic bowel, attributable to CMV infection, while there was no evidence of infection of the second one. Prenatal treatment was done with maternal administration of valacyclovir and postnatal treatment of the infected newborn with oral valganciclovir with normal neurodevelopment assessment at 12 months corrected age. In this case, maternal CMV infection was not equally transmitted to both fetuses, suggesting that there may be intrinsic fetal and placental factors influencing both transmission and the clinical features of the infection.

Ticks and Tick-Borne Diseases in Cuba, Half a Century of Scientific Research.

Ticks and the vast array of pathogens they transmit, including bacteria, viruses, protozoa, and helminths, constitute a growing burden for human and animal health worldwide. In Cuba, the major tropical island in the Caribbean, ticks are an important cause of vector-borne diseases affecting livestock production, pet animal health and, to a lesser extent, human health. The higher number of tick species in the country belong to the Argasidae family and, probably less known, is the presence of an autochthonous tick species in the island, Ixodes capromydis. Herein, we provide a comprehensive review of the ticks and tick-borne pathogens (TBPs) affecting animal and human health in Cuba. The review covers research results including ecophysiology of ticks, the epidemiology of TBPs, and the diagnostic tools used currently in the country for the surveillance of TBPs. We also introduce the programs implemented in the country for tick control and the biotechnology research applied to the development of anti-tick vaccines.

Molecular and morphometric identification of Trypanosoma (Megatrypanum) minasense in blood samples of marmosets (Callithrix: Callithrichidae) from the city of Rio de Janeiro, Brazil.

Callithrix jacchus and C. penicillata marmosets are invasive to the state of Rio de Janeiro, Brazil, threatening the native and vulnerable C. aurita. Both invasive species can be hosts of Trypanosoma cruzi, T. minasense, T. rangeli and T. devei. We aim to investigate the occurrence of trypanosomatids in Callithrix sp. from Jardim Botânico do Rio de Janeiro, located in a central and populous area of the city. Fifteen marmosets were captured. Blood samples were collected for light microscopy and molecular genetics analysis. Parasites morphometric values were evaluated for species identification. DNA was extracted from blood samples by phenol-chloroform method, for partial amplification of the 18S rRNA gene. PCR products were sequenced and aligned using BLAST®. A maximum likelihood phylogenetic tree was constructed to analyze the proximity between the observed sequences. By light microscopy, trypomastigotes were detected in five of the fifteen marmosets. Morphometric measurements and size polymorphism corresponded to those previously described for T. minasense. The DNA sequences of approximately 600 base pairs of the 18S rRNA gene were obtained for three samples with 99% identity with T. minasense sequence, forming a cluster in the phylogenetic tree and corroborating morphometric analysis. Trypanosoma minasense is a highly specific parasite to non-human primates considered as non-pathogenic. There is no evidence of infection in humans and these parasite findings from invasive marmosets do not support additional risks for the native species.

Brazilian National Service of Telediagnosis in Electrocardiography.

Access to specialized care remains unsolved in Brazil. The National Telediagnosis Project was created to expand successful telediagnosis experiences nationwide. The Telehealth Network of Minas Gerais (TNMG) was selected as a reference for tele-electrocardiogram (ECG). We aim to describe the experience of TNMG of developing and implementing the Brazilian National Service of Telediagnosis in Electrocardiography. Implementation planning includes discussion of workflows, standard procedures, responsibility definition for stakeholders, and adaptation of TNMG telediagnosis system. Tele-ECG has been implemented in 79 municipalities in 5 states. In a survey with 152 health professionals, 57% noted that ECG was not available in the local public health system before, 63% indicated tele-ECG service utilization ≥3 days per week, 96% considered the service very useful and 89% were very satisfied with it. In conclusion, the service fills a gap in specialized care in the public system and can improve access to a basic exam in remote and underserved regions.

Genetic Diversity of Theileria equi From Horses In Different Regions of Brazil Based On the 18S rRNA Gene.

Equine piroplasmosis stands out among the diseases that affect Equidae in Brazil and the world. It is caused by the protozoa Theileria equi and Babesia caballi. The objective of the present study was to carry out the molecular characterization of T. equi using equine blood samples collected in the 5 geographic regions of Brazil. Samples from all over the country were tested for the presence of T. equi by real-time PCR. The 18S rRNA sequences (∼1,600 bp) obtained from 23 samples taken from naturally infected horses were characterized by sequencing and analyzed to identify the genotypes and the possible sites of genetic variability. Thirteen different T. equi 18S rRNA sequences were identified, and 2 different genotypes were demonstrated to be in circulation in Brazil. Alignment entropy analysis demonstrated the existence of three hypervariable regions (V2, V4, and V8) within the 18S rRNA sequence of T. equi. The V2 region is located between nucleotides 63 and 75, V4 is located between nucleotides 524 and 586, and V8 is located between nucleotides 1,208 and 1,226. The hypervariable region V4 demonstrated the greatest variation within the 18S rRNA sequence of T. equi. Phylogenetic analysis based on the 18S rRNA sequences revealed the formation of 3 distinct clades (A, B, and C). The Brazilian samples belonged to 2 clades (A and C). The present study describes the characterization and heterogeneity of the circulating T. equi 18S rRNA sequences in Brazil. The results confirm that the country is an endemic area for the disease, and they indicate that at least 2 distinct T. equi genotypes are naturally infecting equines in Brazil.

Comparison of heat shock protein 70 kDa and 18S rDNA genes for molecular detection and phylogenetic analysis of Babesia vogeli from whole blood of naturally infected dogs.

A total of 300 blood samples of domiciliated dogs in rural and urban areas of southeast Rio de Janeiro State, Brazil, were used to compare the 18S ribosomal DNA region (18S rDNA) and the heat shock protein 70 kDa (hsp70) gene for molecular detection of Babesia vogeli and to perform a phylogenetic study comparing the two genes for B. vogeli classification. Using conventional polymerase chain reaction (cPCR) of 18S rDNA and hsp70 sequences, we were able to detect B. vogeli with the same sensitivity (96.15%) and specificity (99.63%). However, sequencing revealed one false positive (Rangelia sp.) for 18S rDNA that was not detected by hsp70. This is the first report of an organism closely related to the Rangelia vitalii parasite of dogs in Brazil. In the hsp70-cPCR and hsp70-qPCR comparison, 15.66% of samples were considered positive by quantitative (q)PCR, significantly more than was detected by cPCR (8.66%). In addition to the high conservation of the 18S rDNA, phylogenetic analysis showed that the hsp70 gene can be used to describe phylogenetic relationships between canine piroplasmids with more accuracy than 18S rDNA. According to these findings, the qPCR method has greater sensitivity than cPCR for detection of B. vogeli in naturally infected dogs. The hsp70-qPCR detection limit was 10 copies, with an efficiency of 100.30% and a determination coefficient (R2) of 0.998. The development of this qPCR method provides a highly sensitive approach for B. vogeli molecular detection and a tool that is capable of quantifying parasitemia levels in whole blood samples from dogs. The primers and probes were designed to be specific for B. vogeli, though analytical specificity of the assay has not been tested in vitro with DNA of certain Babesia species that infect dogs. The hsp70 gene is a precise molecular marker for Babesia phylogeny, especially species that infect dogs.

Serological and molecular diagnosis of Ehrlichia canis and associated risk factors in dogs domiciled in western Cuba.

Ehrlichia canis is a rickettsia transmitted by the tick, Rhipicephalus sanguineus, and is the causative agent of canine monocytic ehrlichiosis (CME). In Cuba, the first diagnosis of CME was made in 2001, but few studies have since investigated this disease locally. The objective of this study was to estimate the prevalence of E. canis in dogs domiciled in four municipalities within the western region of Cuba and determine the associated risk factors. Blood was drawn from 378 selected dogs living in four municipalities in two provinces of western Cuba. From the total number of samples, 206 plasma samples were selected to perform an enzyme-linked immunosorbent assay (ELISA) to detect antibodies against E. canis. Using the original 378 samples of extracted DNA, a nested polymerase chain reaction (nPCR) was performed to amplify a specific fragment of the 16S rRNA gene of E. canis. Analysis of the 206 plasma samples revealed a total of 162 animals that were seropositive for E. canis (78.64%) with a density index between 109.5 and 970.7. In contrast, 179 samples were positive based on the nPCR assay (47.35%). As well, there was a high concordance (kappa = 0.7), calculated through the Kappa index, between the animals found to be positive based on nPCR and those determined based on ELISA. The analysis of risk factors showed that residing in the municipality of Boyeros in addition to having a history of infestation by ticks increases the probability of having a positive result based on nPCR.

Molecular epidemiology of Babesia vogeli in dogs from the southeastern region of Rio de Janeiro, Brazil.

Hemoparasitic diseases are prominent in domestic animals, particularly in Brazil, a tropical country with a wide range of vectors. This study investigated the epidemiology of Babesia vogeli in the whole blood of dogs from the southeastern region of Rio de Janeiro, Brazil. Whole blood samples from 390 dogs were screened for the presence of B. vogeli DNA by qPCR using the heat shock protein 70 kDa (hsp70) gene of B. vogeli. Characteristics related to the host and its environment were collected using a questionnaire. Bivariate analysis was used to evaluate each factor individually. A phi correlation test was used to verify collinearity. The variables with p < .1 and a low or moderate correlation with the other variables were selected for the multivariate analysis. Multiple models were created, and the best logistic regression model was chosen using the Akaike Information Criterion (AIC). The final model was used to determine which variables were closely related to B. vogeli infections in dogs. Of the 390 dog blood samples, 15.66% were positive for B. vogeli. The variables cat contact, age, shelter, street or woods access, tick infestation and fur lengthwere included in the final model. Per the logistic regression analysis, three variables explained B. vogeli detection in dogs: age (odds ratio [OR] = 2.12; p-value <.05; confidence interval [CI]: 1.13-3.96), tick infestation (OR = 2.08; p-value <.05; CI: 1.10-3.93) and shelter (OR = 2.22; p-value <.05; CI: 1.16-4.26). These variables were determined to be associated with B. vogeli detection in domiciled dogs in the southeastern region of Rio de Janeiro, Brazil. These data indicate that the age of the animal, the presence of ticks and the lack of shelter directly affect the epidemiology of B. vogeli.

A new quantitative PCR method for the detection of Anaplasma platys in dogs based on the citrate synthase gene.

Anaplasma platys is an obligate intracellular bacterium that primarily affects dogs, but it can also infect humans. Our study aimed to standardize a quantitative real-time (q)PCR method using the citrate synthase gene (gltA) as a specific target for A. platys detection in naturally infected dogs. Primers (gltA84F and gltA84R) and probe (PLATYSp) were designed to amplify an 84-bp fragment based on the gltA gene sequences of A. platys available in GenBank. A total of 186 dog blood samples originating from the Brazilian state of Rio de Janeiro were tested by qPCR. Additionally, the same samples were tested by cytology and a nested (n)PCR that targeted the 16S ribosomal DNA to determine the performance of our qPCR method compared to these existing techniques. Among the samples tested with qPCR, 17.2% were considered positive, significantly more than detected by nPCR (14.0%). Under optical microscopy, inclusions were observed in platelets of 25.3% of the samples, and among these samples, only 33.9% were identified as positive for A. platys using qPCR. The qPCR technique proved to be more specific than cytology and to have superior sensitivity to nPCR for detecting A. platys in dogs. The development of this new qPCR method contributes to the advancement of research involving A. platys Furthermore, it can be used to quantify the presence of this bacterium to evaluate the treatment of infected animals, or even as a more sensitive and specific tool for situations indicating possible clinical disease but with negative cytology.

Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil.

BACKGROUND: Anaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil. METHODS: We tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum. RESULTS: Among the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum infection in dogs. CONCLUSIONS: This is the first report of A. phagocytophilum in ixodid ticks from Brazil. The detection of A. phagocitophylum in A. cajennense, an aggressive feeder on a wide variety of hosts, including humans, is considered a public health concern.

Molecular epidemiology of Theileria equi in horses and their association with possible tick vectors in the state of Rio de Janeiro, Brazil.

The aim of this study was to detect Theileria equi (Laveran 1901) DNA in horses and ticks using real-time PCR and to list the factors associated with infection in animals located in the Seropedica and Petropolis municipalities of the state of Rio de Janeiro. We tested blood samples from 314 horses and samples from 300 ticks, including 191 Amblyomma cajennense, 104 Dermacentor nitens, and 5 Ixodida larvae. Factors inherent to the horse, the ownership, and animal management were obtained from an epidemiological questionnaire and were evaluated in association with the presence of T. equi DNA in the animals. Among the horses in the study, 81 % (n = 253/314) presented T. equi DNA, and the animals of the Seropedica municipality had the highest infection frequency (91 %, n = 128/141, p < 0.001). The factors that had significantly different infection frequencies by chi-squared or Fisher's exact tests (p < 0.2) were included in a logistic regression model using the R programming package. Work and walking activity (odds ratio [OR] = 5.7, CI = 2.3-14.4), reproductive activity (OR = 3.8, CI = 1.3-11.5), and tick infestation (OR = 2.6, CI = 1.1-6.2) were factors that favored the presence of T. equi DNA in the animals (p < 0.05). Among the tick samples, A. cajennense and D. nitens were the identified species. The presence of T. equi DNA was observed in 9.9 % (n = 19/191) of the A. cajennense samples and 3.8 % (n = 4/104) of the D. nitens samples. A multivariate analysis revealed that the presence of A. cajennense on the animals (OR = 4.1, CI = 1.8-9.1) was associated with the presence of T. equi DNA in the horses. In the studied municipalities, activities related to work, walking, and reproduction and the presence of ticks on the horses, particularly an intense infestation of A. cajennense, are factors that lead to infection with T. equi in the horses.

Esquema corporal e nível de atividade física em adultos jovens universitários / Body schema and level of physical activity in university students young adults

O objetivo deste estudo foi analisar o esquema corporal, o nivel de atividade física e a associação existente entre estas variáveis. A amostra foi composta por 52 indivíduos com média de idade de 22,98 ± 4,85 anos, sendo 20 homens (23 ± 7,30 anos) e 32 mulheres (22,97 ± 3,19 anos). Os participantes assinaram um termo de consentimento livre e esclarecido e o trabalho foi aprovado pelo comitê de ética da Universidade São Judas Tadeu. As variáveis analisadas foram o esquema corporal, através do Image Marking Procedure (IMP) e o nível de atividade física, através do IPAq. Utilizou-se a estatística descritiva para caracterizar e conhecer a distribuição da amostra. Em seguida, os dados foram comparados utilizando o teste de análise de variância (ANOVA), seguido do teste de Bonferroni como post-hoc. A associação foi verificada com o teste qui-quadrado e o odds ratio. Os homens apresentaram uma maior porcentagem de indivíduos hipoesquemáticos e com esquema corporal adequado e as mulheres uma maior porcentagem de hiperesquematia. Quanto a simetria, somente a altura do trocânter no sexo feminino apresentou diferenças estatísticas (p = 0,02). A inatividade física foi evidente em todos os domínios para a amostra total, porem no domínio 4 verificou-se uma maior porcentagem de indivíduos fisicamente ativos no sexo masculino. O nível de atividade física geral apresentou uma maior porcentagem de indivíduos classificados como fisicamente ativos. Não houve associação entre o esquema corporal e o nível de atividade física. Nota-se que há uma tendência na população de perceberem suas dimensões corporais maiores, ou seja, tendem a hiperesquematia. O fato do esquema corporal está intimamente relacionado com as aferências cinestésicas e proprioceptivas, não foi confirmado nesta amostra ao comparar o nível de atividade física e o esquema corporal.

The objective of this study was to analyze the body scheme, the level of physical activity and the association between these variables. The sample comprised 52 individuals with mean age of 22.98 ± 4.85 years, 20 men (23 ± 7.30 years) and 32 women (22.97 ± 3.19 years). The participants signed an informed consent and the study was approved by the ethics committee of Universidade São Judas Tadeu. The variables analyzed were body image through the Image Marking Procedure (IMP) and the level of physical activity by IPAQ. We used descriptive statistics to characterize and understand the distribution of the sample. Then the data were compared using analysis of variance test (ANOVA) followed by Bonferroni test as post hoc. The association was found with chi-square and odds ratio. Men had a higher percentage of individuals with body scheme and hipoesquemáticos adequate and women a higher percentage of hiperesquematia. The symmetry, only the height of the trochanter in women showed no statistical differences (p = 0.02). Physical inactivity was evident in all areas for the total sample, however in the field 4 there was a higher percentage of physically active males. The level of overall physical activity had a greater percentage of individuals classified as physically active. There was no association between body scheme and physical activity level. Note that there is a trend in the population perceive their body size larger, that is, tend to hiperesquematia. The fact that body image is closely related to kinesthetic and proprioceptive afferents, was not confirmed in this sample to compare the level of physical activity and body scheme.