Evolution and divergence of the genetic lineage Desmodus rotundus/Artibeus lituratus of rabies virus in São Paulo State.

The last record of a rabies case caused by the dog-specific rabies virus (RABV) lineage in dogs or cats in São Paulo State was in 1998. From 2002 to 2021, 57 cases of rabies in these animals were reported, and the vast majority (51) were genetically characterized as belonging to the Desmodus rotundus/Artibeus lituratus RABV lineage. However, it is not currently possible to infer which of these bats is the source of infection by genome sequencing of RABV isolates. The aims of this study were (a) to characterize the Desmodus rotundus/Artibeus lituratus lineage to determine the relationships between the RABV lineages and each reservoir, (b) to assess the phylogeny and common ancestors of the RABV lineages found in D. rotundus and A. lituratus, and (c) to further understand the epidemiology and control of rabies. In this study, we genetically analyzed 70 RABV isolates from São Paulo State that were received by the Virology Laboratory of the Pasteur Institute of São Paulo between 2006 and 2015. Of these isolates, 33 were associated with the hematophagous bat D. rotundus and 37 with the fruit bat A. lituratus. A genomic approach using phylogenetic analysis and nucleotide sequence comparisons demonstrated that these isolates belonged to the same genetic lineage of RABV. We also found that, in São Paulo State, the D. rotundus/A. lituratus lineage could be subdivided into at least four phylogenetic sublineages: two associated with D. rotundus and two with A. lituratus. These results are of importance for the epidemiological surveillance of rabies in São Paulo.

Could Fidicina mannifera (Hemiptera: Cicadoidea: Fidicinini) promote a resource pulse in two Brazilian Cerrado vegetation classes? / Fidicina mannifera (Hemiptera: Cicadoidea: Fidicinini) promove pulsos de nutrientes em duas fitofisionomias do Cerrado?

Abstract Cicadas are usually studied regarding their importance in agriculture. However, the possibility of this group to represent a pulse of nutrients can also imply on the success of a given species in the biome. The aim of this study was to assess the level of total soluble proteins and lipids of Fidicina mannifera (Fabricius, 1803), and to determine whether the species can promote a pulse of nutrients in two vegetation classes of the Brazilian Cerrado. To assess the pulse of nutrients, it was concluded the determination of total soluble proteins and lipids from samples of males and adult females of F. mannifera, and the spatial distribution of exuviae of this species was also calculated in two vegetation classes of the Brazilian Cerrado. The amount of protein provided by each individual did not differ between males and females (p = 0.66) but females had 40% more lipids than males (p = 0.05). Regarding F. mannifera the gallery forest offered 11.75 g/ha of protein, 3.91 g/ha of lipids, and the Cerrado stricto sensu offered 4.25 g/ha of protein, and 1.41 g/ha of lipid. The male cicadas have a hollow abdomen, which houses a resonance chamber for sound production in order to attract females to mate, and females store larger amounts of lipids, mainly located in the abdominal cavity, where the body fat is directly linked to the reproductive system for the development of the ovaries and egg production after emergence. The mass occurrence of F. mannifera in the Brazilian Cerrado and the fast availability of proteins and lipids make this species a food resource that can directly impact the diet of secondary consumers and scavengers, although the amount of nutrients available by F. mannifera does not promote a pulse of nutrients in the study site.

Resumo O interesse no estudo de cigarras geralmente é despertado pela sua importância na agricultura. Entretanto, a possibilidade de que este grupo represente um pulso de nutrientes pode também implicar no sucesso de sobrevivência de espécies importantes num determinado bioma. O objetivo deste trabalho foi verificar o nível de proteínas solúveis e lipídios totais de Fidicina mannifera (Fabricius, 1803) e determinar se a espécie pode promover um pulso de nutrientes em duas fitofisionomias do Cerrado. Para avaliar o pulso de nutrientes, foram determinados os níveis de proteínas solúveis e lipídios totais de machos e fêmeas adultos de F. mannifera, e também a distribuição espacial de exúvias desta espécie, em duas fitofisionomias de Cerrado. A quantidade de proteínas disponibilizada por indivíduo não diferiu entre machos e fêmeas (p = 0,66), porém fêmeas apresentaram 40% mais lipídios do que machos (p = 0,05). Para mata de galeria F. mannifera ofertou 11,75 g/ha de proteína, 3,91 g/ha de lipídios, e no cerrado stricto sensu, 4,25 g/ha de proteína, e 1,41 g/ha de lipídio. Machos de cigarras possuem um abdômen oco, que abriga uma câmara de ressonância para a produção de som com o objetivo atração de fêmeas para acasalar, por outro lado as fêmeas armazenam maiores quantidades de lipídios, localizados principalmente na cavidade abdominal, onde a gordura corporal está diretamente ligada ao aparelho reprodutor para o desenvolvimento dos ovários, e produção de ovos após a emergência. A ocorrência em massa de F. mannifera no Cerrado, bem como a disponibilidade de proteínas e lipídios de fácil acesso, faz desta espécie um recurso alimentar que pode impactar diretamente na dieta de consumidores e detritívoros secundários, embora quantidade de nutrientes disponibilizados por F. mannifera não promova um pulso de nutrientes no local estudado.

Could Fidicina mannifera (Hemiptera: Cicadoidea: Fidicinini) promote a resource pulse in two Brazilian Cerrado vegetation classes?

Cicadas are usually studied regarding their importance in agriculture. However, the possibility of this group to represent a pulse of nutrients can also imply on the success of a given species in the biome. The aim of this study was to assess the level of total soluble proteins and lipids of Fidicina mannifera (Fabricius, 1803), and to determine whether the species can promote a pulse of nutrients in two vegetation classes of the Brazilian Cerrado. To assess the pulse of nutrients, it was concluded the determination of total soluble proteins and lipids from samples of males and adult females of F. mannifera, and the spatial distribution of exuviae of this species was also calculated in two vegetation classes of the Brazilian Cerrado. The amount of protein provided by each individual did not differ between males and females (p = 0.66) but females had 40% more lipids than males (p = 0.05). Regarding F. mannifera the gallery forest offered 11.75 g/ha of protein, 3.91 g/ha of lipids, and the Cerrado stricto sensu offered 4.25 g/ha of protein, and 1.41 g/ha of lipid. The male cicadas have a hollow abdomen, which houses a resonance chamber for sound production in order to attract females to mate, and females store larger amounts of lipids, mainly located in the abdominal cavity, where the body fat is directly linked to the reproductive system for the development of the ovaries and egg production after emergence. The mass occurrence of F. mannifera in the Brazilian Cerrado and the fast availability of proteins and lipids make this species a food resource that can directly impact the diet of secondary consumers and scavengers, although the amount of nutrients available by F. mannifera does not promote a pulse of nutrients in the study site.

The Epidemiological Importance of Bats in the Transmission of Rabies to Dogs and Cats in the State of São Paulo, Brazil, Between 2005 and 2014.

In Brazil, rabies control in dogs and cats was pioneered by the state of São Paulo with the adoption of the Pan American Health Organization recommendations for prophylaxis and control, which led to a reduction in rabies cases from 1994 onwards. As a result of these measures, the rabies virus (RABV) genetic lineage associated with dogs has not been found in the state since 1998, and all the cases in domestic animals reported since then have been caused by bat-associated lineages of RABV. In the light of this, this study sought to investigate rabies cases in dogs and cats in the state of São Paulo between 2005 and 2014 and identify the associated transmission cycles by characterizing the RABV lineages responsible for these cases. Nine samples from dogs (n = 5) and from cats (n = 4) were collected between 2005 and 2014. The tenth animal, a rabid cat, was analysed by a different laboratory. The N gene nucleotide sequences obtained were analysed with the neighbor-joining algorithm and Kimura 2-parameter model using the MEGA 6 program. Phylogenetic analysis revealed that the genetic lineages identified in all the samples were those circulating in Brazilian bats. The findings of this study demonstrate that bats play an important role in the transmission of rabies to domestic animals in São Paulo state and that emphasis should be placed on the implementation of public policies to support surveillance of chiropterans for rabies.

Mechanical properties and in vitro characterization of polyvinyl alcohol-nano-silver hydrogel wound dressings.

Polyvinyl alcohol (PVA) hydrogels are materials for potential use in burn healing. Silver nanoparticles can be synthesized within PVA hydrogels giving antimicrobial hydrogels. Hydrogels have to be swollen prior to their application, and the common medium available for that in hospitals is saline solution, but the hydrogel could also take up some of the wound's fluid. This work developed gamma-irradiated PVA/nano-Ag hydrogels for potential use in burn dressing applications. Silver nitrate (AgNO3) was used as nano-Ag precursor agent. Saline solution, phosphate-buffered solution (PBS) pH 7.4 and solution pH 4.0 were used as swelling media. Microstructural evaluation revealed an effect of the nanoparticles on PVA crystallization. The swelling of the PVA-Ag samples in solution pH 4.0 was low, as was their silver delivery, compared with the equivalent samples swollen in the other media. The highest swelling and silver delivery were related to samples prepared with 0.50% AgNO3, and they also presented lower strength in PBS pH 7.4 and solution pH 4.0. Both PVA-Ag samples were also non-toxic and presented antimicrobial activity, confirming that 0.25% AgNO3 concentration is sufficient to establish an antimicrobial effect. Both PVA-Ag samples presented suitable mechanical and swelling properties in all media, representative of potential burn site conditions.

Intercanine distance in the analysis of bite marks: a comparison of human and domestic dog dental arches.

One common parameter considered helpful to identify the origin of bite wounds has been the distance between the canine teeth marks left on the victim. The reliability of this parameter to differentiate the origin of the marks (human or animal) was evaluated using a sample of: a) domestic dogs (n=50) weighting between 4.9 kg and 46 kg of undefined breed and b) human beings (n=50). Dog intercanine distances (ID) were measured directly using calipers, those from the human sample were measured from wax imprints using calipers. It was found that dog bite intercanine distance measurements were overall 2.8% wider for the upper arch and 10.4% wider for the lower arch when compared with the overall result for humans. However, it was observed that the measured values for medium sized dogs (between 9.1 kg and 23.0 kg) are similar to the overall results for humans. Therefore, for this range, the stand alone use of intercanine distance measurements from bite wounds marks are inconclusive with respect of defining if of human origin.

Bovine rotavirus in turkeys with enteritis.

Rotaviruses are the main agents responsible for diarrhea in different animal species and for infantile gastroenteritis. These viruses have been isolated from various avian species and have often been associated with poult enteritis and mortality syndrome. Nevertheless, the knowledge of rotavirus infection in turkeys is scarce. Six group A rotavirus strains obtained from pooled enteric contents of diarrheic turkeys were isolated in MA-104 cell culture and typed as G(6)P(1), a typical bovine rotavirus genotype. Additionally, the electropherotypes showed a migration pattern identical to the Nebraska calf diarrhea virus, and the complete NSP4 gene phylogeny showed that all six strains segregated in the genotype E2. Taken together, these results point toward a cattle-to-turkey rotavirus transmission. As a conclusion, bovine-origin rotavirus can be found in turkeys, and this transmission route must now be considered for the improvement of the health status in turkey farms.

Avaliação hematológica e bioquímica de equinos suplementados com óleo de arroz semirrefinado, rico em gamaorizanol / Hematological and biochemical evaluation of horses supplemented with semi-refined rice oil enriched with gamma orizanol

Avaliou-se o efeito da suplementação com óleo de arroz sobre o peso e perfil hematológico de equinos submetidos a exercício físico moderado. Foram utilizados 14 equinos machos, com peso aproximado de 411kg, distribuídos entre o grupo tratado (GT; n=7), suplementado com óleo de arroz adicionado diariamente à dieta (0,5ml/kg/PV), e o grupo-controle (GC; n=7), tratado com óleo de soja (0,5ml/kg/PV). Foram feitas três avaliações: antes do início e aos 20 e 40 dias após o início do tratamento, as quais consistiram de determinação do peso, exame clínico e coleta de amostras de sangue dos animais, antes e após o exercício, para hematócrito, hemograma, dosagem de glicose, lactato e proteína total. Não houve diferença entre grupos e nem entre avaliações quanto às variáveis peso e proteína total. A glicose aumentou significativamente após o exercício na segunda coleta no GC e na terceira no GT. No GC, o lactato aumentou nas coletas após o exercício, enquanto no GT, os valores foram semelhantes antes e após. A suplementação com óleo de arroz na dieta foi determinante para impedir o aumento de lactato em equinos submetidos a exercício, o que pode ser relevante para aumentar o seu desempenho atlético.

The effect of the supplementation with rice oil was evaluated on the weight and hematologic profile of equines submitted moderate physical exercise. Fourteen male equines, averaging 411kg, were distributed into treated group (GT; n=7), supplemented daily with rice oil added to the diet (0.5ml/kg/BW); and control group (GC; n=7), treated with soybean oil (0.5ml/kg/BW). Three evaluations were made before the treatment, and 20 and 40 days after the beginning of the treatment, consisting of the determination of the weight, the clinical examination, and the collection of blood samples before and after the exercise for hematocrit, hemogram, glucose, lactate, and total protein determinations. There was no difference between the groups, neither between evaluations for weight nor total protein. The glucose increased significantly after the exercise in the second collection in the GC and in the third in the GT. The GC lactate increased significantly in the collections after exercise; while in the GT, the results were similar before and after treatments. The supplementation of the diet with rice oil was determinant to hinder the lactate increase in the animals submitted to exercise, what could be relevant to increase their athletic performance.

Caracterização molecular do vírus da raiva isolado de Desmodus rotundus capturados no Estado do Rio de Janeiro / Molecular characterization of rabies virus isolated from Desmodus rotundus captured in Rio de Janeiro State

Caracterizou-se filogeneticamente o vírus da raiva, isolado de morcegos hematógafos (Demodus rotundus). Cento e noventa e nove D. rotundus foram capturados em cinco abrigos, no Norte e Noroeste do Estado do Rio de Janeiro e sul do Espírito Santo. Sete deles foram positivos para a raiva. Amostras desses vírus foram sequenciadas e comparadas com sequências provenientes de diversos estados brasileiros. As sequências de vírus da raiva isoladas, na região norte do Estado do Rio de Janeiro, mostraram características que as distinguem de amostras de vírus isoladas em outras regiões do país, no entanto foram idênticas às isoladas de bovinos no noroeste do Rio de Janeiro.

Rabies samples isolated from vampire bats captured in the Rio de Janeiro State were phylogenetically analyzed. One hundred and ninety nine vampire bats were captured from five shelters from North and Northwest of Rio de Janeiro and South of Espírito Santo States. Seven of them were positive for rabies. Theses samples were sequenced and compared with rabies virus sequences from several Brazilian states. The sequences of rabies virus, isolated in the present work, from North of Rio de Janeiro State, showed characteristics that differ of the sequences isolated from bats from other Brazilian regions. However, they were identical to samples isolated from cattle in Northwest of Rio de Janeiro state.

X-chromosome genetic variation in São Paulo State (Brazil) population.

As data on X-chromosomal short tandem repeats (X-STRs) for the Brazilian population are scarse, the aim of this study was to determine the allele frequencies of five X-STRs (DXS6854, DXS7424, DXS101, DXS6808 and DXS7132) in the São Paulo State, Brazil. No deviations from the Hardy-Weinberg equilibrium were observed, with the exception of DXS101. The forensic efficiency parameters demonstrated that DXS101 was the most informative marker. Population comparisons revealed that the X-STR profile sampled in the state of São Paulo was more similar to European and African populations than Asiatic populations reported in this work.

Genetic population data of 12 STR loci of the PowerPlex Y system in the state of São Paulo population (Southeast of Brazil).

Allele frequency distributions and population data for 12 Y-chromosomal short tandem repeats (STRs) included in the PowerPlex Y Systems (Promega) were obtained for a sample of 200 healthy unrelated males living in São Paulo State (Southeast of Brazil). A total of 192 haplotypes were identified, of which 184 were unique and 8 were found in 2 individuals. The average gene diversity of the 12 Y-STR was 0.6746 and the haplotype diversity was 0.9996. Pairwise analysis confirmed that our population is more similar with the Italy, North Portugal and Spain, being more distant of the Japan.

Molecular analysis of the N gene of canine distemper virus in dogs in Brazil

Eleven central-nervous-system samples collected from stray dogs between 2000 and 2004 were found positive by RT-PCR, which amplified a 480bp fragment of the N gene of canine distemper virus (CDV). Phylogenetic analysis based on partial N-gene sequences showed four major clusters. All dog strains segregated into cluster I, with a mean nucleotide identity of 95.8 percent and 95.6 percent with the Onderstepoort and Lederle vaccine strains, respectively. Cluster II contained all the raccoon-related strains, cluster III Orient strains and Cluster IV the Onderstepoort and Lederle vaccine strains, with a mean nucleotide identity of 99.7 percent between them. This is the first report of phylogenetic analysis of CDV strains in Brazil.

Onze amostras de sistema nervoso central de cães coletados entre 2000 e 2004 foram positivas pela RT-PCR, a qual amplificou um fragmento de 480pb do gene N do vírus da cinomose canina (VCC). A análise filogenética baseada na seqüência parcial do gene N mostrou quatro principais agrupamentos genéticos. Todas as amostras de cães segregaram no agrupamento I, com identidade média de nucleotídeos de 95,8 por cento e 95,6 por cento com as amostras vacinais Onderstepoort e Lederle, respectivamente. O agrupamento II agregou todas as amostras relacionadas aos guaxinins. O agrupamento III agregou amostras orientais e o agrupamento IV agregou as amostras vacinais Onderstepoort e Lederle, com identidade média de nucleotídeos de 99,7 por cento entre elas. Este é o primeiro relato de análise filogenética de amostras de VCC no Brasil.

Estudo comparativo do óleo essencial de Eugenia punicifolia (HBK) DC. de diferentes localidades de Pernambuco / Comparative study of the essential oil of Eugenia punicifolia (HBK) DC. from diferent places of Pernambuco

O óleo essencial obtido por hidrodestilação das folhas de Eugenia punicifolia, coletados a partir de duas regiões de Matas Serranas do Estado de Pernambuco (Serra Negra e Brejo da Madre de Deus), foi analisado por CG/EM. O monoterpeno oxigenado, linalol (SN: 44,0 por cento, BMD: 61,2 por cento), foi o componente principal do óleo de ambas amostras. Cerca de 66,2 por cento da constituição dos voláteis da amostra proveniente de SN e 74,2 por cento de BMD referem-se aos monoterpenos oxigenados, enquanto 33,6 por cento da amostra de SN e 20,5 por cento de BMD são sesquiterpenos oxigenados, sendo o b-cariofileno o mais abundante (SN: 22,7 por cento, BMD: 16,2 por cento). Com exceção do a-cadinol, presente apenas no óleo proveniente do Brejo da Madre de Deus, a constituição química de ambas as amostras foi à mesma, variando apenas o percentual relativo de seus constituintes.

The essential oils obtained from the leaves of Eugenia punicifolia by hydrodistillation, which were collected from two different locations of Pernambuco (Serra Negra and Brejo da Madre de Deus), have been investigated by GC/MS. Linalol was found to be the main component of this oil from SN (44,0 percent) and BMD (61,2 percent). About 66,2 percent of the oil constituents from SN and 74,2 percent from BMD are oxygenated monoterpenes, with predominance of beta-cariofileno (SN: 22,7 and BMD: 16,2 percent). With exception of a-cadinol, present only in the oils from plants colleted in Brejo da Madre de Deus, the chemical composition of both samples was quite similar, the only difference was the relative content among their components.

Population studies of the Y-chromosome of loci DYS390, DYS391 and DYS393 in Brazilian subjects and its use in human identification.

Human identification can be accomplished by several technical procedures, especially by the comparative analysis of dental documents. Recent advances in molecular biology have now widened the scope for human identification and defined the protocols for the collection of biological material. Allele patterns of a population are particularly useful and they should be verified since they vary in different populations. We have studied the frequency of the alleles in three STR loci of the Y chromosome (DYS390, DYS391 and DYS393) in a group of Brazilian caucasian subjects. Results presented alleles 21, 22, 23, 24, 25 and 26 in locus DYS390, alleles 8, 9, 10, 11 12 and 13 in STR DYS391 and alleles in STR DYS393 were 11, 12, 13 14 and 15. The highest frequencies were 24 (0.46), 11 (0.37) and 13 (0.45). Each of the three STR systems had a PE (power of exclusion) of 0.6764 (DYS390), 0.5988 (DYS391) and 0.6136 (DYS393), The combination of the three STR systems revealed a PE of 0.9498, suggesting that this data can help in human identification.

[Evaluation of 3 methods of DNA extraction from paraffin-embedded material for the amplification of genomic DNA using PCR]. / Avaliação de três métodos de extração de DNA de material parafinado para amplificação de DNA genômico pela técnica da PCR.

There are several protocols reported in the literature for the extraction of genomic DNA from formalin-fixed paraffin-embedded samples. Genomic DNA is utilized in molecular analyses, including PCR. This study compares three different methods for the extraction of genomic DNA from formalin-fixed paraffin-embedded (inflammatory fibrous hyperplasia) and non-formalin-fixed (normal oral mucosa) samples: phenol with enzymatic digestion, and silica with and without enzymatic digestion. The amplification of DNA by means of the PCR technique was carried out with primers for the exon 7 of human keratin type 14. Amplicons were analyzed by means of electrophoresis in an 8% polyacrylamide gel with 5% glycerol, followed by silver-staining visualization. The phenol/enzymatic digestion and the silica/enzymatic digestion methods provided amplicons from both tissue samples. The method described is a potential aid in the establishment of the histopathologic diagnosis and in retrospective studies with archival paraffin-embedded samples.

Postmortem tooth loss in human identification processes.

Teeth provide essential data for human identification. However, they are frequently lost during the process of skeletonization or under manipulation during exhumation. Because of the high frequency of postmortem tooth loss, this phenomenon was examined in three different samples, in an attempt to simulate the actual circumstances experienced in the forensic processes of identification of human remains. The method employed aimed to describe and quantify dental spaces, distinguishing between loss after death or before by extraction. The results showed a high prevalence of postmortem tooth loss: 41.37% in sample A, 56.83% in sample B and 3.96% in sample C. These losses were most frequent in the maxillary incisor group in all three samples analysed. These data underline the need for redoubled care in recovery, transportation and storage of crania, so that teeth are not lost, which could seriously compromise the identification process.