Flea parasitism in wild mammals in the metropolitan region of Sorocaba, São Paulo, Brazil.

Fleas are eurixenous ectoparasites that can parasitize different species of warm-blooded animals, including humans, with the possibility of causing irritation and blood spoliation. They are vectors responsible for the transmission of numerous pathogens and have a wide geographical distribution, more frequently in warm regions. Domestic animals (dogs and cats) are preferred hosts, but parasitism can also occur in wild hosts, with a greater variety of parasitic species and strong interaction between these siphonapters and their hosts. During the period from January 2021 to June 2023, flea specimens were collected from wild animals coming from the metropolitan region of Sorocaba, São Paulo. Some of these animals were animals from the zoo's own stock, which were diagnosed with parasitism during routine examinations and others were rescued from the natural environment and sent to the zoo. The flea specimens collected were packed in alcohol 700 GL and sent for microscopic diagnosis. Four groups were diagnosed at the specific level: Ctenocephalides felis, Rhopalopsyllus lutzi, R. lugrubis and Tunga penetrans, and two groups as unidentified species, belonging to the genera Rhopalopsyllus and Polygenis. The parasitized animals were all mammals, belonging to eleven different species, distributed in the Orders Carnivora, Didelphimorphia, Rodentia and Cingulata. Studies on the parasitofauna of wild animals contribute to a greater knowledge about the distribution of parasitic agents and their relationships with the host species.

As pulgas são ectoparasitos eurixenos que podem parasitar diferentes espécies de animais de sangue quente, incluindo seres humanos, com possibilidade de causar irritação e espoliação sanguínea. São vetores responsáveis pela transmissão de inúmeros patógenos e apresentam ampla distribuição geográfica, com maior frequência em regiões quentes. Os animais domésticos (cães e gatos) são hospedeiros preferenciais, mas o parasitismo pode ocorrer também em hospedeiros selvagens, com maior variedade de espécies parasitas e forte interação entre esses sifonápteros e seus hospedeiros. Durante o período de janeiro de 2021 a junho de 2023, espécimes de pulgas foram coletados de animais selvagens provenientes da região metropolitana de Sorocaba, São Paulo. Alguns destes animais eram animais do plantel do próprio zoológico, que foram diagnosticados com o parasitismo durante exames de rotina e outros foram resgatados do ambiente natural e encaminhados ao zoológico. Os espécimes de pulgas coletadas foram acondicionados em álcool 700 GL e enviados para diagnóstico microscópico. Foram diagnosticados quatro grupos a nível específico: Ctenocephalides felis, Rhopalopsyllus lutzi, R. lugrubis e Tunga penetrans, e dois grupos como espécies não identificadas, pertencentes aos gêneros Rhopalopsyllus e Polygenis. Os animais parasitados eram todos mamíferos, pertencentes a onze espécies diferentes, distribuídas nas Ordens Carnivora, Didelphimorphia, Rodentia e Cingulata. Estudos sobre a parasitofauna de animais selvagens contribuem para um maior conhecimento sobre a distribuição de agentes parasitários e suas relações com as espécies hospedeiras.

Heterotaxy Syndrome with Increased Nuchal Translucency and Normal Karyotype Associated with Complex Systemic Venous Return. Ultrasound Diagnosis with Autopsy Correlation.

Background: Prenatal ultrasound (US) detection of heterotaxy syndrome can be challenging, especially in identifying cardiovascular and associated anomalies. We present a new case of heterotaxy syndrome with anomalous systemic venous return (ASVR) fully displayed at autopsy. Case report: Left heterotaxy syndrome was diagnosed in a 19 weeks' of gestation fetus with right-sided stomach. The heart showed both ventricles with left morphology, a large ventricular septal defect, persistent left superior vena cava draining into the coronary sinus, ASVR with interrupted inferior vena cava (IVC) and azygous continuation. Autopsy dissection further identified the azygous draining into the left lower pulmonary vein (LLPV). Prenatal a-CGH on villous sampling showed 22q13.1 microduplication inherited from the father, not contributory to the phenotype. Conclusion/discussion: Heterotaxy syndrome requires US accuracy for anomaly identification, as they allow legal termination of pregnancy. Our case is unusual as IVC drained into the azygous vein and then into the LLPV.

408 kb 15q11.2 microduplication by array comparative genomic hybridization in a fetus presenting with exomphalos, micrognathia, tetralogy of Fallot and normal karyotype: a genetic counseling dilemma in paternal carrier status.

Exomphalos may be associated with chromosomal abnormalities and syndromes. Severe exomphalos (herniation of liver, midgut and spleen) associated with increased nuchal translucency was seen at first trimester screening test. Karyotype by chorionic villus sampling showed normal male fetus. Follow up scan at 16 and 18 weeks of gestation confirmed the severe exomphalos and detected micrognathia and tetralogy of Fallot. Array comparative genomic hybridization (a-CGH) further demonstrated a 408 kb 15q11.2 microduplication, with the father-to-be as healthy carrier. This is the first case of an association between 15q11.2 micorduplication and fetal sonographic anomalies. Genetic counseling for estimation of recurrent risk of congenital anomalies is discussed.

POLG1 mutations and stroke like episodes: a distinct clinical entity rather than an atypical MELAS syndrome.

BACKGROUND: POLG1 mutations have been associated with MELAS-like phenotypes. However given several clinical differences it is unknown whether POLG1 mutations are possible causes of MELAS or give raise to a distinct clinical and genetic entity, named POLG1-associated encephalopathy. CASE PRESENTATION: We describe a 74 years old man carrying POLG1 mutations presenting with strokes, myopathy and ragged red fibers with some atypical aspects for MELAS such as late onset, lack of cerebral calcification and presence of frontal and occipital MRI lesions better consistent with the POLG associated-encephalopathy spectrum. CONCLUSION: The lack of available data hampers a definite diagnosis in our patient as well as makes it difficult to compare MELAS, which is a clearly defined clinical syndrome, with POLG1-associated encephalopathy, which is so far a purely molecularly defined syndrome with a quite heterogeneous clinical picture. However, the present report contributes to expand the phenotypic spectrum of POLG1 mutations underlining the importance of searching POLG1 mutations in patients with mitochondrial signs and MELAS like phenotypes but negative for common mtDNA mutations.

Prenatal diagnosis of tricuspid atresia with hypoplastic right ventricle associated with truncus arteriosus communis type II.

PURPOSE: To confirm the central role of antenatal echocardiography and necropsy in the prenatal diagnosis of rare congenital heart defects. METHODS: A 33-year-old woman undergoing second trimester scan using 2D transabdominal and Doppler sonography. RESULTS: The echocardiographic examination showed, at the level of the four-chamber view, a predominant left ventricle with a rudimental right ventricle and a single artery emerging with failed visualization of the pulmonary trunk: a diagnosis of truncus arteriosus communis associated with tricuspid atresia and hypoplastic right heart was made. No other ultrasound-associated anomalies were seen. Fetal karyotype and 22q11.2 microdeletion for Di George syndrome were sought using cordocentesis performed at 21 weeks and both the results were normal. After extensive counselling, the couple opted for termination of pregnancy at 22 weeks gestation. Necroscopy confirmed the prenatal ultrasound diagnosis. Necroscopy of the heart was performed in a manner that resembled the fetal echocardiographic examination and revealed two atria, two atrio-ventricular valves with recognizable mitral and tricuspid morphology, a prevalent ventricle of left-type and a rudimental ventricle of right type and a ventricular septal defect. The common truncus was seen coming out above the ventricular septum, whilst the pulmonary arteries arise separated from the truncus communis. No other structural thoraco-abdominal anomalies were found. CONCLUSIONS: A combined diagnostic strategy based on second trimester fetal echocardiography, genetic analysis and necroscopy has made identification of a rare congenital heart disease possible.

Small size-specific umbilical vein diameter in severe growth restricted fetuses that die in utero.

BACKGROUND: To study changes in umbilical vein (UV) blood flow velocity, diameter and blood flow volume in intrauterine growth retardation (IUGR) fetuses who die in utero (IUD-IUGR). METHODS: Twelve singleton IUGR fetuses who died in utero below 600 g were included. All cases had abnormal uterine and umbilical arteries PI. UV diameter and velocity were measured at the time of diagnosis, and at the last exam, within 24 hours prior to intrauterine death. UV flow was calculated per unit weight (mL/min/kg) and abdominal circumference (AC) (mL/min/cm). UV diameter and velocity were normalized per unit AC. Findings were compared to 14 severe viable-IUGR and 22 normal gestational age-matched fetuses. RESULTS: UV flow (mL/min/kg) was significantly lower in IUD-IUGR compared to viable-IUGR (87 +/- 30 mL/min/kg) and control fetuses (131 +/- 33 mL/min/kg) both at the first (79 +/- 40 mL/min/kg) (P < 0.0001), and at the last exam (54 +/- 29 mL/min/kg) (P < 0.0001). No significant longitudinal flow changes were observed. UV velocity/AC was significantly reduced both in IUD-IUGR and viable-IUGR compared to normal fetuses. UV diameter/AC, was significantly reduced only in IUD-IUGR and not in viable-IUGR compared to normal fetuses. CONCLUSIONS: UV flow (mL/min/kg) was significantly lower in IUD-IUGR fetuses both versus viable-IUGR and normal fetuses. A low flow was due to a decreased UV flow velocity, but also due to a reduced vessel size. This significantly smaller UV size observed in IUGR fetuses with the worst outcome could be considered a severe prognostic sign because of the diagnosis of severe growth restriction.

Genotype-based differentiation of the Cryptococcus neoformans serotypes by combined PCR-RFLP analysis of the capsule-associated genes CAP10 and CAP59.

This report describes an indirect identification method for Cryptococcus neoformans serotypes developed using combined restriction enzyme pattern analysis of two PCR-amplified portions of the capsule-associated genes CAP10 and CAP59. The method relies on the recognition of the sequence conformation of nine serotype-related polymorphic sites by the analysis of four restriction profiles. A 610 nucleotides long trait of the CAP10 gene was digested with the enzymes Sty I or Sal I and a 597 nucleotides long trait of the CAP59 gene was digested with the enzymes Sal I or EcoRV+PstI. The resulting profiles, reported as a string of four numbers, defined for each strain an intrinsically coherent allelic profile closely correlated to the serotype. We analyzed by this method 172 C. neoformans strains obtained from different sources. All the serotype A strains examined and all the strains of the B-C serotypes group were recognized by specific allelic profiles, but serotypes B and C could not be distinguished from each other. Of the serotype D strains, 84% were characterized by a unique allelic pattern, while the remaining 16% were genotypically indistinguishable from the AD serotype organisms among which differences in the ploidy number and evidence of recombination could be recognized.

Descriptive epidemiology of selected birth defects, areas of Lombardy, Italy, 1999.

BACKGROUND: Birth defects are a leading cause of neonatal and infant mortality in Italy, however little is known of the etiology of most defects. Improvements in diagnosis have revealed increasing numbers of clinically insignificant defects, while improvements in treatment have increased the survival of those with more serious and complex defects. For etiological studies, prevention, and management, it is important to have population-based monitoring which provides reliable data on the prevalence at birth of such defects. METHODS: We recently initiated population-based birth defect monitoring in the Provinces of Mantova, Sondrio and Varese of the Region of Lombardy, northern Italy, and report data for the first year of operation (1999). The registry uses all-electronic source files (hospital discharge files, death certificates, regional health files, and pathology reports) and a proven case-generation methodology, which is described. The data were checked manually by consulting clinical records in hospitals. Completeness was checked against birth certificates by capture-recapture. Data on cases were coded according to the four-digit malformation codes of the International Classification of Diseases, Ninth Revision (ICD-9). We present data only on selected defects. RESULTS: We found 246 selected birth defects in 12,008 live births in 1999, 148 among boys and 98 among girls. Congenital heart defects (particularly septal defects) were the most common (90.8/10,000), followed by defects of the genitourinary tract (34.1/10, 000) (particularly hypospadias in boys), digestive system (23.3/10,000) and central nervous system (14.9/10,000), orofacial clefts (10.8/10,000) and Down syndrome (8.3/10,000). Completeness was satisfactory: analysis of birth certificates resulted in the addition of two birth defect cases to the registry. CONCLUSION: This is the first population-based analysis on selected major birth defects in the Region. The high birth prevalences for septal heart defect and hypospadias are probably due to the inclusion of minor defects and lack of coding standardization; the latter problem also seems important for other defects. However the data produced are useful for estimating the demands made on the health system by babies with birth defects.

In vitro antimycotic activity and nail permeation models of a piroctone olamine (octopirox) containing transungual water soluble technology.

Several in vitro studies with a new medical device (Myfungar) containing 0.5% of piroctone olamine (CAS 68890-66-4, octopirox) in a hydroxypropyl chitosan hydroalcoholic solution were performed. The chemical name of the active ingredient is 1-hydroxy-4-methyl-6 (2,4,4-trimethylpentyl)-2(1H)-pyridone; combination with 2-amino-ethanol (1:1). The antimycotic activity was determined in the most common fungi responsible of nail infections such as Candida parapsilosis, Scopulariopsis brevicaulis or Trichophyton rubrum. The minimum inhibitory concentration (MIC), found by means of the broth dilution susceptibility method, ranged between 0.0003% and 0.006% for all pathogens considered. The in vitro permeation study was performed by using bovine hoof membranes inserted in a modified Gummer vertical permeation cell. The experiment showed, at 30 h, a retention of the product in the nail membranes by 11.1% of the applied dose. No piroctone permeation through bovine hoof membranes could be detected by HPLC due to the limit of quantitation of this method. On the other hand, permeation of nail membranes has been demonstrated by a biological assay: the study of in vitro permeation through bovine hoof membranes, performed by biological assay, showed dose-dependent inhibition rings of T. rubrum growth by the tested device, placed either on disks for antibiogram or on nail fragments. The placebo did not show any inhibition. In vitro experimental infection by T. rubrum showed a preventive activity of the tested device on fungal growth as well as a curative activity, as the pathogen was eradicated by the tested solution in previously infected nails.

Three cases with de novo 6q imbalance and variable prenatal phenotype.

We describe two families in which three fetuses had a de novo 6q imbalance and abnormal phenotypes. We determined the boundaries and the parental origin of the chromosomal alterations by segregation analysis using a panel of short tandem repeats (STRs) located on 6q. Cases 1 and 2 (family A) were two sibs with 6q imbalance involving different regions. Case 1 was a female fetus with arthrogryposis, who had a complex rearrangement resulting in two deleted regions (6q22 and 6q25.1-q25.2) and a duplication of 6q23-q25.1. This latter imbalance was reported previously and is associated with joint contractures and short neck, also present in this fetus. The sib (case 2) had intrauterine growth restriction (IUGR) and agenesis of the ductus venosus. This male died shortly after birth; postnatal karyotype and molecular investigations showed a 6q21 de novo deletion. Case 3 (family B) had a prenatally detected deletion of 6q14-q16. Autopsy of the fetus documented minor facial anomalies and contractures of the limbs. All rearrangements were de novo and of paternal origin. Our data and the consistent number of cases of de novo 6q alterations previously reported suggest that chromosome arm 6q could be prone to rearrangements resulting in heterogeneous phenotypes. In family A, chromosome 6q imbalances involving different chromosomal regions were present in two consecutive pregnancies. In such cases counseling should suggest the impossibility of excluding recurrence of a chromosomal imbalance, and should discuss the option of early prenatal diagnosis in subsequent pregnancies.

Few modifications of the Cobas Amplicor HIV Monitor 1.5 test allow reliable quantitation of HIV-1 proviral load in peripheral blood mononuclear cells.

Recent studies have suggested that monitoring the amount of HIV provirus in peripheral blood mononuclear cells (PBMCs) may be a useful end point for HAART where, in combination with plasma viral load, it provides additional information as to the possibility of virus eradication. In the present study, a modified version of the Cobas Amplicor HIV-1 Monitor test (CAHIM), currently used to quantify plasma viremia, have been evaluated to also measure the amount of proviral DNA in PBMCs. The analytical and clinical performance of the modified CAHIM test was assessed by quantifying different amounts of a standard HIV-DNA preparation obtained from the 8E5 cell line and by analysing 165 patients and controls samples. In these experiments, the modified test, that showed a linear dynamic range from 1.7 to 4.7 log10 copies/10(6) cells (r = 0.99) with a maximum CV of 20%, proved able to detect and quantify HIV-DNA in all but one clinical samples, with concentrations varying from 1.3 to 3.8 log10 copies/10(6) cells. During anti-retroviral treatment, the assay revealed different proviral DNA time courses associated with viral load changes and inversely correlated with CD4+ cells count. As expected, HIV-DNA was always detectable even when plasma viremia fell below the CAHIM cut-off. The modified CAHIM test specificity was confirmed by testing 20 HIV-negative samples in triplicates. Taken together, the data showed that the modified CAHIM test can be used to monitor HIV proviral DNA changes during HAART and can help in investigating further the clinical use of this marker.

Simultaneous measurements of umbilical venous, fetal hepatic, and ductus venosus blood flow in growth-restricted human fetuses.

OBJECTIVE: The purpose of this study was to evaluate the changes in the distribution of the umbilical venous blood flow to the liver and to the ductus venosus in intrauterine growth-restricted human fetuses in relationship with dilation of the ductal isthmic diameter. STUDY DESIGN: Umbilical venous flow, ductus venosus blood flow, and blood flow to the fetal liver were measured in 56 severely intrauterine growth-restricted fetuses with an abnormal pulsatility index of the umbilical artery and were compared with 137 normal control fetuses. Percentages of umbilical venous blood flow through the ductus venosus and to the fetal hepatic lobes were calculated. Z-scores for control fetuses and intrauterine growth-restricted fetuses were constructed by the evaluation of the inverted smoothed percentiles. The distributions of the Z-scores was compared with a 1-sample t-test. RESULTS: The ductus venosus blood flow that was corrected for fetal weight was increased significantly in intrauterine growth-restricted fetuses compared with control fetuses (P=0); the median values (interquartile range) for comparable ages of gestation was 41.3 mL/min/kg (range, 26.2-64.0 mL/min/kg) and 30.8 mL/min/kg (range, 19.9-42.8 mL/min/kg), respectively. As a consequence, ductus venosus shunting was increased in intrauterine growth-restricted fetuses compared with control fetuses (P =0). In 23 of 30 intrauterine growth-restricted fetuses, the percentage of umbilical blood flow that was shunted through the ductus was>90th percentile of control fetuses. Ductal diameters were significantly greater in growth-restricted fetuses than in control fetuses (P =.0001). The percentage of blood flow to the right lobe showed a significant reduction (P =.0223), with evidence of reversed blood flow from the right lobe and portal system into the ductus venosus that was provided both by volume blood flow calculations and by direct pulsed Doppler waveform direction. CONCLUSION: In severe intrauterine growth-restricted fetuses, Doppler examination of blood flow volume proved a significant increase in the shunting of umbilical vein blood flow through the ductus venosus that was associated with the dilation of the ductal isthmic diameter. These changes provide a relatively constant blood flow to the heart and brain at the expense of fetal hepatic perfusion.