Oxidative stress is involved in LLLT mechanism of action on skin healing in rats.

The skin injury healing process involves the main phases of homoeostasis, inflammation, proliferation, and remodeling. The present study aimed to analyze the effects of low-level laser therapy (LLLT) on hematological dynamics, oxidative stress markers, and its relation with tissue healing following skin injury. Wistar rats were divided into control, sham, skin injury, and skin injury LLLT. The biochemical and morphological analyses were performed in the inflammatory (1 and 3 days) and regenerative phases (7, 14, and 21 days) following injury. The skin injury was performed in the dorsal region, between the intrascapular lines, using a surgical punch. LLLT (Al-Ga-In-P, λ=660 nm, energy density of 20 J/cm2, 30 mW power, and a time of 40 s) was applied at the area immediately after injury and on every following day according to the experimental subgroups. LLLT maintained hematocrit and hemoglobin levels until the 3rd day of treatment. Surprisingly, LLLT increased total leukocytes levels compared to control until the 3rd day. The effects of LLLT on mitochondrial activity were demonstrated by the significant increase in MTT levels in both inflammatory and regenerative phases (from the 1st to the 7th day), but only when associated with skin injury. The results indicated that LLLT modulated the inflammatory response intensity and accelerated skin tissue healing by a mechanism that involved oxidative damage reduction mostly at early stages of skin healing (inflammatory phase).

Oxidative stress is involved in LLLT mechanism of action on skin healing in rats

The skin injury healing process involves the main phases of homoeostasis, inflammation, proliferation, and remodeling. The present study aimed to analyze the effects of low-level laser therapy (LLLT) on hematological dynamics, oxidative stress markers, and its relation with tissue healing following skin injury. Wistar rats were divided into control, sham, skin injury, and skin injury LLLT. The biochemical and morphological analyses were performed in the inflammatory (1 and 3 days) and regenerative phases (7, 14, and 21 days) following injury. The skin injury was performed in the dorsal region, between the intrascapular lines, using a surgical punch. LLLT (Al-Ga-In-P, λ=660 nm, energy density of 20 J/cm2, 30 mW power, and a time of 40 s) was applied at the area immediately after injury and on every following day according to the experimental subgroups. LLLT maintained hematocrit and hemoglobin levels until the 3rd day of treatment. Surprisingly, LLLT increased total leukocytes levels compared to control until the 3rd day. The effects of LLLT on mitochondrial activity were demonstrated by the significant increase in MTT levels in both inflammatory and regenerative phases (from the 1st to the 7th day), but only when associated with skin injury. The results indicated that LLLT modulated the inflammatory response intensity and accelerated skin tissue healing by a mechanism that involved oxidative damage reduction mostly at early stages of skin healing (inflammatory phase).

Anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide in mice.

Several isatin derivatives have shown important biological activities, which have attracted interest from researchers. For this reason, the present study aimed to evaluate the anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide (COPHCT) in mice. Three doses of this compound were tested: 1.0, 2.5, and 5.0 mg/kg. The anti-inflammatory activity was assessed using the carrageenan-induced paw edema model and the zymosan-induced air pouch model. The evaluation of the antinociceptive effect was performed through the formalin test and the acetic acid-induced abdominal writhing test. The paw edema assay demonstrated that all doses of the compound showed a significant reduction of the edema in the second hour evaluated, but a better response was observed in the fourth hour. The zymosan-induced air pouch model indicated that the compound, in all doses, significantly reduced leukocyte migration and total protein concentration levels. In the formalin test, the doses 1.0, 2.5, and 5.0 mg/kg of COPHCT showed activity only in the second phase, with reduction in paw pain time of 73.61, 79.46, and 73.85%, respectively. The number of abdominal writhings decreased with the increasing dose, but only 5.0 mg/kg COPHCT exhibited a significant response, with a reduction of 24.88%. These results demonstrated the ability of this compound to interfere in the anti-inflammatory activity of edema, vascular permeability, and cell migration. In addition, its possible antinociceptive effect may be related to the dose used.

Anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide in mice

Several isatin derivatives have shown important biological activities, which have attracted interest from researchers. For this reason, the present study aimed to evaluate the anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide (COPHCT) in mice. Three doses of this compound were tested: 1.0, 2.5, and 5.0 mg/kg. The anti-inflammatory activity was assessed using the carrageenan-induced paw edema model and the zymosan-induced air pouch model. The evaluation of the antinociceptive effect was performed through the formalin test and the acetic acid-induced abdominal writhing test. The paw edema assay demonstrated that all doses of the compound showed a significant reduction of the edema in the second hour evaluated, but a better response was observed in the fourth hour. The zymosan-induced air pouch model indicated that the compound, in all doses, significantly reduced leukocyte migration and total protein concentration levels. In the formalin test, the doses 1.0, 2.5, and 5.0 mg/kg of COPHCT showed activity only in the second phase, with reduction in paw pain time of 73.61, 79.46, and 73.85%, respectively. The number of abdominal writhings decreased with the increasing dose, but only 5.0 mg/kg COPHCT exhibited a significant response, with a reduction of 24.88%. These results demonstrated the ability of this compound to interfere in the anti-inflammatory activity of edema, vascular permeability, and cell migration. In addition, its possible antinociceptive effect may be related to the dose used.

Rhabdias glaurungi sp. nov. (Nematoda: Rhabdiasidae), parasite of Scinax gr. ruber (Laurenti, 1768) (Anura: Hylidae), from the Brazilian Amazon.

The genus Rhabdias Stiles & Hassal, 1905 includes about 83 species of nematodes parasitic in amphibians and reptiles worldwide. Herein, we describe Rhabdias glaurungi sp. nov. from the hylid frog Scinax gr. ruber (Laurenti, 1768) in the Gunma Ecological Park, Santa Bárbara municipality, state of Pará, Brazil. This species has six small lips, an inflated cuticle along the entire body and a cup-shaped buccal capsule with smooth internal surface of its anterior part and irregularly folded internal surface of its posterior part in apical view. From the 17 valid species recognized in the Neotropical realm, the new species can be distinguished by the number of lips, the morphology and size of its buccal capsule, as well as the extent and shape of its cuticular inflation; in addition, there are molecular differences. Sequences of the mitochondrial Cytochrome c Oxidase subunit I gene strongly support the status of this form as a separate species. Molecular phylogenetic analysis shows R. glaurungi sp. nov. nested within the R. pseudosphaerocephala Kuzmin, Tkach & Brooks, 2007 species complex. Rhabdias glaurungi sp. nov. is the second species of the genus described from hosts of the family Hylidae in the Neotropical realm. We conclude that the diversity of Rhabdias within the Neotropics is likely largely underestimated.

Co-infection with filarial nematodes in Sapajus macrocephalus and Cebus albifrons (Primates: Cebidae) from the Peruvian Amazon.

Dipetalonema caudispina (Molin, 1858) and D. gracile (Rudolphi, 1809) (Filarioidea: Onchocercidae) are two of six known species of filarial nematodes that parasitize Neotropical non-human primates. Adult filariae were collected from the thoracic and abdominal cavities of 38 of 44 specimens of Sapajus macrocephalus (Spix, 1823) and nine of ten specimens of Cebus albifrons (Humboldt, 1812) (Primates: Cebidae), distributed in the Yavarí-Mirín river basin and used locally for human consumption. Co-occurrence of D. caudispina and D. gracile is reported for the first time, with a prevalence of 18.5% (10 of 54 hosts examined). Our finding of D. caudispina and D. gracile in cebids from the Peruvian Amazon constitutes a new geographical record for both filariae, two new host records for D. caudispina, and the first report of D. gracile in S. macrocephalus. In addition, we provide morphometric data for D. caudispina, complementing the original description, as well as scanning electron microscopy details on the structure of the area rugosa and number of caudal papillae in males.

Cryotherapy: biochemical alterations involved in reduction of damage induced by exhaustive exercise.

When exercises are done in intense or exhaustive modes, several acute biochemical mechanisms are triggered. The use of cryotherapy as cold-water immersion is largely used to accelerate the process of muscular recovery based on its anti-inflammatory and analgesic properties. The present study aimed to study the biochemical effects of cold-water immersion treatment in mice submitted to exercise-induced exhaustion. Swiss albino mice were divided into 4 treatment groups: control, cold-water immersion (CWI), swimming exhaustive protocol (SEP), and SEP+CWI. Treatment groups were subdivided into times of analysis: 0, 1, 3, and 5 days. Exhaustion groups were submitted to one SEP session, and the CWI groups submitted to one immersion session (12 min at 12°C) every 24 h. Reactive species production, inflammatory, cell viability, and antioxidant status were assessed. The SEP+CWI group showed a decrease in inflammatory damage biomarkers, and reactive species production, and presented increased cell viability compared to the SEP group. Furthermore, CWI increased acetylcholinesterase activity in the first two sessions. The present study showed that CWI was an effective treatment after exercise-induced muscle damage. It enhanced anti-inflammatory response, decreased reactive species production, increased cell viability, and promoted redox balance, which could decrease the time for the recovery process.

Efeito do tipo de alimentação de cães saudáveis sobre análises clínicas e aspectos comportamentais / Effect of the diet of healthy dogs on clinical analysis and behavioral aspects

Objetivou-se estudar o efeito da alimentação caseira, da ração comercial a granel e da ração comercial fechada para cães adultos sobre análises clínicas e aspectos comportamentais de ingestão das dietas. O delineamento experimental foi inteiramente ao acaso, com três tratamentos, sendo o tratamento 1 constituído de dieta caseira, o tratamento 2 por ração a granel tipo premium, e o tratamento 3 por ração comercial fechada do tipo premium. Foram utilizados três cães em cada um dos tratamentos, totalizando nove animais. As variáveis analisadas foram: teste de glicemia, odor fecal, escore fecal, volume fecal, análise química da urina (pH urinário, presença de proteína, sangue, corpos cetônicos, bilirrubina e glicose), avaliação visual do pelo e consumo voluntário da dieta. Os cães submetidos à dieta caseira apresentaram menor resultado de glicemia sanguínea (83,90). A urina apresentou pH mais alcalino para cães que receberam a ração a granel (8,06). Cães da dieta caseira apresentaram o mais alto escore para a qualidade do pelo. Houve um maior volume das fezes de cães que receberam a dieta de ração a granel. Cães que receberam a dieta de alimentação caseira ou a ração comercial fechada apresentaram um consumo total do alimento em menos de 10 minutos. A maioria dos resultados foi semelhante entre a dieta caseira e a ração fechada do tipo premium. A ração a granel apresentou os piores resultados.(AU)

The objective of this study was to study the effect of homemade feed, commercial feed in bulk, and closed commercial feed for adult dogs on clinical analysis and behavioral aspects of diet intake. The experimental design was completely randomized with three treatments: homemade diet, bulk premium type feed, and closed commercial feed of the premium type. Three dogs were observed in each of the treatments, totaling nine animals. The variables analyzed were: blood glycemia, fecal odor, fecal score, fecal volume, chemical urine analysis (urinary pH, presence of protein, blood, ketone bodies, bilirubin and glucose), visual evaluation of hair and voluntary dietary intake. Dogs submitted to a home diet showed lower blood glucose results (83.90). Urine was more alkaline pH for dogs that received the bulk feed (8.06). Dogs from the homemade feed presented the highest score for hair quality. There was a greater volume of feces from dogs that received the diet of bulk feed. Dogs that received the diet of home feed or closed commercial feed showed a total consumption of food in less than ten minutes. Most of the results were similar between the homemade feed and the closed-type premium diet. The dogs bulk feed presented the worst results.(AU)

Helmet retention system types as a risk factor to properly attach the chin strap.

OBJECTIVES: This study aimed to determine the factors associated with the incorrect use of a helmet retention system (loose or open) and how often this happens. METHODS: This was an observational transversal study conducted in Curitiba, Brazil. Trained observers positioned at traffic lights collected information about the helmet fixation mode, the helmet model (full-face, open-face, modular, half), and the helmet retention system model (micrometric, double-D, fast-release). Additional data including position on the motorcycle, gender, and function of the motorcycle (as a work vehicle) were collected. The observers, collection site, and periods were randomly selected by lots. RESULTS: From a total of 3,050 motorcyclists, 1,807 (59.2%) had their helmets fastened correctly, 907 (29.7%) had the retention system fastened loosely, and in 336 (11.0%), the retention system was completely open. Increased odds of incorrect use were observed for the fast-release and double-D buckles compared to the micrometrics buckles, with a fixed odds ratio (OR) of 4.62 (95% confidence interval [CI], 3.89-5.51) and 3.54 (95% CI, 2.46-5.09), respectively (P <.0001). Full-face helmets had a higher chance of incorrect use (P <.0001), and passengers had a higher incidence of incorrect use of the helmet than drivers (P <.0001). CONCLUSION: An important risk factor related to the incorrect use of the helmet was the type of retention system. The helmet model and being a passenger had a secondary influence on incorrect use of helmets.

Efeito do tipo de alimentação de cães saudáveis sobre análises clínicas e aspectos comportamentais / Effect of the diet of healthy dogs on clinical analysis and behavioral aspects

Objetivou-se estudar o efeito da alimentação caseira, da ração comercial a granel e da ração comercial fechada para cães adultos sobre análises clínicas e aspectos comportamentais de ingestão das dietas. O delineamento experimental foi inteiramente ao acaso, com três tratamentos, sendo o tratamento 1 constituído de dieta caseira, o tratamento 2 por ração a granel tipo premium, e o tratamento 3 por ração comercial fechada do tipo premium. Foram utilizados três cães em cada um dos tratamentos, totalizando nove animais. As variáveis analisadas foram: teste de glicemia, odor fecal, escore fecal, volume fecal, análise química da urina (pH urinário, presença de proteína, sangue, corpos cetônicos, bilirrubina e glicose), avaliação visual do pelo e consumo voluntário da dieta. Os cães submetidos à dieta caseira apresentaram menor resultado de glicemia sanguínea (83,90). A urina apresentou pH mais alcalino para cães que receberam a ração a granel (8,06). Cães da dieta caseira apresentaram o mais alto escore para a qualidade do pelo. Houve um maior volume das fezes de cães que receberam a dieta de ração a granel. Cães que receberam a dieta de alimentação caseira ou a ração comercial fechada apresentaram um consumo total do alimento em menos de 10 minutos. A maioria dos resultados foi semelhante entre a dieta caseira e a ração fechada do tipo premium. A ração a granel apresentou os piores resultados.(AU)

The objective of this study was to study the effect of homemade feed, commercial feed in bulk, and closed commercial feed for adult dogs on clinical analysis and behavioral aspects of diet intake. The experimental design was completely randomized with three treatments: homemade diet, bulk premium type feed, and closed commercial feed of the premium type. Three dogs were observed in each of the treatments, totaling nine animals. The variables analyzed were: blood glycemia, fecal odor, fecal score, fecal volume, chemical urine analysis (urinary pH, presence of protein, blood, ketone bodies, bilirubin and glucose), visual evaluation of hair and voluntary dietary intake. Dogs submitted to a home diet showed lower blood glucose results (83.90). Urine was more alkaline pH for dogs that received the bulk feed (8.06). Dogs from the homemade feed presented the highest score for hair quality. There was a greater volume of feces from dogs that received the diet of bulk feed. Dogs that received the diet of home feed or closed commercial feed showed a total consumption of food in less than ten minutes. Most of the results were similar between the homemade feed and the closed-type premium diet. The dogs bulk feed presented the worst results.(AU)

Cryotherapy: biochemical alterations involved in reduction of damage induced by exhaustive exercise

When exercises are done in intense or exhaustive modes, several acute biochemical mechanisms are triggered. The use of cryotherapy as cold-water immersion is largely used to accelerate the process of muscular recovery based on its anti-inflammatory and analgesic properties. The present study aimed to study the biochemical effects of cold-water immersion treatment in mice submitted to exercise-induced exhaustion. Swiss albino mice were divided into 4 treatment groups: control, cold-water immersion (CWI), swimming exhaustive protocol (SEP), and SEP+CWI. Treatment groups were subdivided into times of analysis: 0, 1, 3, and 5 days. Exhaustion groups were submitted to one SEP session, and the CWI groups submitted to one immersion session (12 min at 12°C) every 24 h. Reactive species production, inflammatory, cell viability, and antioxidant status were assessed. The SEP+CWI group showed a decrease in inflammatory damage biomarkers, and reactive species production, and presented increased cell viability compared to the SEP group. Furthermore, CWI increased acetylcholinesterase activity in the first two sessions. The present study showed that CWI was an effective treatment after exercise-induced muscle damage. It enhanced anti-inflammatory response, decreased reactive species production, increased cell viability, and promoted redox balance, which could decrease the time for the recovery process.

Effects of cryolipolysis on abdominal adiposity of women.

　　BACKGROUND: The cryolipolysis is on the spotlight as a non-invasive method which reduces fat layer thickness with no damage to surrounding tissues. OBJECTIVE: This study aims to verify the effectiveness of cryolipolysis in the reduction of localized adiposity in women. MATERIALS AND METHODS: This is an experimental study, without a control group, with pre- and post- treatment evaluation through a single application on the lower abdominal area. SETTING: Research conducted in the period from July to December 2015 at the University Potiguar. PARTICIPANTS: A group of 15 women, age between 25-50 years. The cryolipolysis was used in the following parameters: temperature (-7 degree C); suction power (30 kPa), and application time (60 min). MEASUREMENTS: After the cryolipolysis was performed, a follow-up of 2 months was conducted to verify the changes related to weight, body circumference, fat layer thickness, which were evaluated by ultrasonography and photogrammetry. RESULTS: From data analysis, the reductions observed on perimeter (p=0.03) and ultrasonography (p=0.03) showed significant results, considering p <0.05. As of body weight results (p=0.57), the average value varied during the study; however, at the end of the research, no significant weight increase or decrease was reported, as it is known that this method does not interfere with this variable. Additionally, quantitative data were satisfactory. The photogrammetry analysis showed that cryolipolysis positively affected subjects' results. CONCLUSION: A change in body contouring, especially in individuals with lower body mass, reinforces the idea that the parameters must be suitable for individual needs.

Rohdella amazonica n. sp. (Aspidogastrea: Aspidogastridae) from the Amazoninan banded puffer fish Colomesus psittacus (Bloch & Schneider, 1801).

Aspidogastreans are commonly found infecting freshwater and marine molluscs, teleosts fishes and freshwater turtles. The subclass comprises four families - Rugogastridae Schell 1973, Stichocotylidae Faust & Tang 1936, Multicalycidae Gibson & Chinabut 1984 and Aspidogastridae Poche 1907 - and it is characterized by the presence of a ventral adhesive disc divided into rows of alveoli. In the current work, using light and scanning electron microscopy and molecular approaches, a new species of Aspidogastridae of the genus Rohdella Gibson & Chinabut, 1984, is described as a parasite of Colomesus psittacus in Brazil. The new taxon is distinguishable by the presence of oesophageal glands, teguments covered by ciliated papillae, and the position and shape of the hermaphroditic duct. The present work describes the third species of the genus Rohdella, thereby adding new morphological and molecular data regarding Aspidogastridae.

Caracterização anatômica, histoquímica e de classes de metabólitos secundários de folhas de Guzmania lingulata (L. ) Mez / Anatomical, histochemical and class characterization of secondary metabolites in leaves of Guzmania lingulata (L. ) Mez

Na medicina natural pouco se sabe sobre a importância curativa da maior parte das espécies pertencentes à Bromeliaceae. Tal fato evidencia a necessidade da realização de investigações para a comprovação de possíveis potenciais fitoterápicos em espécies dessa família. Guzmania lingulata é caracterizada como sendo uma das espécies de bromélia mais comercializada e conhecida popularmente, a qual é amplamente propagada no Brasil. O presente trabalho objetivou a caracterização anatômica e a identificação de classes de metabólitos secundários presentes em G. lingulata, por meio de análises anatômicas, histoquímicas e químicas. As folhas foram fixadas em FAA 50 e transferidas para etanol 50%. Para o estudo anatômico, secções transversais da porção mediana das lâminas foliares foram obtidas à mão livre e submetidas ao processo de dupla coloração com safranina e azul de astra, sendo, logo após, montadas em gelatina glicerinada. Para a triagem das classes de metabólitos secundários foram coletadas 10 folhas frescas que passaram pelo processo de maceração em solução hidroalcoólica 70% por sete dias, à temperatura ambiente. O filtrado resultante da filtração a vácuo foi evaporado sob vácuo a temperatura de 50ºC até a remoção total do etanol, sendo obtido desta forma o extrato bruto hidroalcoólico a frio (EBHF). Os resultados das secções histológicas permitiram a caracterização anatômico-foliar baseada na qualidade das secções manuais obtidas. Os testes em relação às classes de metabólitos secundários revelaram a presença de antocianinas e taninos e ausência de flavonóides e saponinas. Esses resultados estimulam a realização de estudos mais aprofundados para identificar e quantificar os compostos dessas e de outras classes de metabólitos.

In natural medicine, little is known about the healing importance of most of the species belonging to the family Bromeliaceae. Such a fact points tothe need to perform investigations toprovethe possible phytotherapic potentials in species of the family. Guzmania lingulata is characterized as one of the most popular well-known and commercialized species of bromeliads, being widely spread in Brazil. This work aimed at the anatomic characterization and identification of classes of the secondary metabolites present in G. lingulata, by anatomic, histochemical and chemical analysis of the leaves. The leaves were fixed in FAA 50 and, afterwards, transferred to 50% ethanol. For the anatomic study, transverse sections of the middleproportion of the leafblades were obtained by hand and exposed to the process of double coloration with safranin and astra blue, being, right after that, mounted on glycerin jelly. For the sorting of the classes of secondary metabolites, 10 fresh leaves were collected and then put through the maceration process in 70% hydroalcoholic solution for 7 days, at room temperature. The resulting filtered solution from the vacuum filtration was evaporated under vacuum at 40ºC until the complete removal of the ethanol, obtaining, this way, the cold crude hydroalcoholic extract (EBHF). The results ofhistological sectionsallowed the anatomical characterization based onthe quality ofthe manual sectionsobtained. The testsin relationto the classes of secondary metabolitesrevealed the presence ofanthocyanins andtannins andabsence of flavonoids and saponins. These results encourage the completionoffurther studies to identify and quantify the compounds of these and other classes of metabolites.

Isolation and characterization of canine parvovirus type 2c (CPV-2c) from symptomatic puppies

Canine parvovirus type 2 (CPV-2) is a leading cause of diarrhea in puppies in several parts of the world. In this study CPV-2 was detected and recovered from puppies showing clinical disease from Montevideo, Uruguay. Samples were processed and used to infect CRFK and MDCK cells in order to isolate the virus. Out of twelve, two samples were positive for CPV-2. A genomic region of 583 bp was amplified and the molecular characterization was performed by sequencing, phylogenetic analysis and Restriction Fragment Length Polymorphism (RFLP). Two isolated viruses (UY1 and UY2) were CPV-2c-like viruses. The comparison between the cytophatic effect (CPE) of CPV-2 (vaccinal virus) and CPV-2c (isolated virus) on primary canine cells cultures and on CRFK line cells, demonstrated that CPV-2c is less citopathogenic in CRFK than in primary cultures. Our study represents the first report on isolation and characterization of canine parvovirus type 2c (CPV-2c) in cell cultures from South American dogs.

Isolation and characterization of canine parvovirus type 2C (CPV-2C) from symptomatic puppies.

Canine parvovirus type 2 (CPV-2) is a leading cause of diarrhea in puppies in several parts of the world. In this study CPV-2 was detected and recovered from puppies showing clinical disease from Montevideo, Uruguay. Samples were processed and used to infect CRFK and MDCK cells in order to isolate the virus. Out of twelve, two samples were positive for CPV-2. A genomic region of 583 bp was amplified and the molecular characterization was performed by sequencing, phylogenetic analysis and Restriction Fragment Length Polymorphism (RFLP). Two isolated viruses (UY1 and UY2) were CPV-2c-like viruses. The comparison between the cytophatic effect (CPE) of CPV-2 (vaccinal virus) and CPV-2c (isolated virus) on primary canine cells cultures and on CRFK line cells, demonstrated that CPV-2c is less citopathogenic in CRFK than in primary cultures. Our study represents the first report on isolation and characterization of canine parvovirus type 2c (CPV-2c) in cell cultures from South American dogs.

Isolation and characterization of canine parvovirus type 2c (CPV-2c) from symptomatic puppies

Canine parvovirus type 2 (CPV-2) is a leading cause of diarrhea in puppies in several parts of the world. In this study CPV-2 was detected and recovered from puppies showing clinical disease from Montevideo, Uruguay. Samples were processed and used to infect CRFK and MDCK cells in order to isolate the virus. Out of twelve, two samples were positive for CPV-2. A genomic region of 583 bp was amplified and the molecular characterization was performed by sequencing, phylogenetic analysis and Restriction Fragment Length Polymorphism (RFLP). Two isolated viruses (UY1 and UY2) were CPV-2c-like viruses. The comparison between the cytophatic effect (CPE) of CPV-2 (vaccinal virus) and CPV-2c (isolated virus) on primary canine cells cultures and on CRFK line cells, demonstrated that CPV-2c is less citopathogenic in CRFK than in primary cultures. Our study represents the first report on isolation and characterization of canine parvovirus type 2c (CPV-2c) in cell cultures from South American dogs.

Patient-based dengue virus surveillance in Aedes aegypti from Recife, Brazil.

BACKGROUND & OBJECTIVES: Dengue is currently one of the most important arthropod-borne diseases and may be caused by four different dengue virus serotypes (DENV-1 to DENV-4), transmitted mainly by Aedes aegypti (Diptera: Culicidae) mosquitoes. With the lack of a dengue vaccine, vector control strategies constitute a crucial mode to prevent or reduce disease transmission. In this context, DENV detection in natural Ae. aegypti populations may serve as a potential additional tool for early prediction systems of dengue outbreaks, leading to an intensification of vector control measures, aimed at reducing disease transmission. In Brazil, this type of surveillance has been performed sporadically by a few groups and has not been incorporated as a routine activity in control programs. This study aimed at detecting DENV in natural Ae. aegypti from Recife, Pernambuco, to check the circulating serotypes and the occurrence of transovarial transmission in local mosquito populations. METHODS: From January 2005 to June 2006, mosquitoes (adults and eggs) were collected in houses where people with clinical suspicion of dengue infection lived at. RNA was extracted from pooled mosquitoes and RT-PCR was performed in these samples for detection of the four DENV serotypes. RESULTS & CONCLUSION: Out of 83 pools of adult mosquitoes collected in the field, nine were positive for DENV: five for DENV-1, two for DENV-2 and two for DENV-3. From 139 pools of adult mosquitoes reared from collected eggs, there were 17 positive pools: three for DENV-1, 10 for DENV-2, and four for DENV-3. These results are discussed in the paper in regard to the local dengue epidemiological data. The conclusions clearly point to the informative power and sensitivity of DENV entomological surveillance and to the importance of including mosquito immature forms in this strategy.

Resistance to the organophosphate temephos: mechanisms, evolution and reversion in an Aedes aegypti laboratory strain from Brazil.

Insecticide resistance is one of the main problems in vector control programs. Because insects have developed resistance to all classes of available chemical insecticides, a proper surveillance and management of resistance in areas where these compounds are being utilized is crucial for the success of control programs. Since the mechanisms and molecular bases of resistance are various, they must be characterized to allow efficient monitoring strategies. Here we report the establishment of an Aedes aegypti strain resistant to temephos, named RecR, selected under laboratory conditions. The parental A. aegypti population was obtained from eggs collected in an area where temephos had been used for 8 years, and presented a baseline resistance ratio (RR) of 7. After 17 generations under selective pressure, the RR has increased to 180. Biochemical assays indicate that metabolic mechanisms are involved on temephos resistance in the selected strain. These experiments showed that, compared to the susceptible colony Rockefeller, RecR present higher activity of glutathione S-transferases (GSTs), alpha- and beta-esterases, and, to a lesser degree, mixed function oxidases (MFO). At the 14th or 17th generations, there was no cross resistance of these insects to deltamethrin, cypermethrin and malathion, while a low resistance level (RR=3) was observed for pyriproxyfen, a juvenile hormone analogue. Experiments on resistance reversal, performed through three different field simulated schemes using the resistant strain, showed that temephos susceptibility can be recovered. The establishment of an A. aegypti colony resistant to temephos is extremely valuable for a deeper understanding of resistance mechanisms and thus for further improvements in control strategies against this vector. With the urgent need on improving methodologies to monitor resistance, molecular studies such as microarrays, and resistant colonies such as RecR will certainly hasten such studies.

Phagocytosis of latex beads and bacteria by hemocytes of the triatomine bug Rhodnius prolixus (Hemiptera: Reduvidae).

Insect circulating hemocytes are primarily responsible for the immune defense against parasites and pathogens. Here, we have analyzed phagocytosis of both biotic (bacteria) and abiotic (latex) particles by circulating hemocytes of 5th-instar nymphs of the triatomine bug Rhodnius prolixus. The following hemocyte types were identified: prohemocytes, plasmatocytes, granulocytes, oenocytoids and adipohemocytes. There was a considerable change in the relative percentage of plasmatocytes and prohemocytes in the hemolymph after challenge with both latex beads and bacteria. Granulocytes and oenocytoids also change their relative percentage in response to latex bead and Staphylococcus aureus, respectively. No significant change was observed in adipohemocytes at any time or treatment. Our data demonstrated that plasmatocytes were the only cell type involved in phagocytosis of foreign particles. As in mammal cells, phagocytosis by both zipper and trigger mechanisms were observed for the uptake of latex beads and bacteria. Neither melanization nor micro-aggregation was observed towards latex particles or Escherichia coli. On the other hand, R. prolixus produced a strong melanization reaction against S. aureus, thus showing that differences exist in the responses to E. coli and to S. aureus. Ultrastructural changes observed in plasmatocytes, adipohemocytes and oenocytoids suggest that these hemocyte types are directly involved in the immune defense of R. prolixus against foreign particles.