Antimicrobial TiN-Ag Coatings in Leather Insole for Diabetic Foot.

This work reports on TiN-Ag antimicrobial coatings deposited by d.c. magnetron sputtering on leather used for insoles on the footwear industry, studies involving the antimicrobial properties of Ag-based functionalized leathers by sputtering techniques are shown. The X-ray diffraction (XRD) results suggested the presence of crystalline fcc-TiN phase for the sample without silver, and also a fcc-Ag phase in the samples containing silver. According to the Scanning Electron Microscopy (SEM) analysis, the coatings were homogeneous and dispersed Ag clusters were detected on the surface of samples with silver content above 8 at. %. The Inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis showed that the ionization of silver over time depends on the morphology of the coatings. The samples did not present cytotoxicity and only samples with incorporated silver presented antibacterial and antifungal activity, highlighting the potential of the TiN-Ag insole coatings for diseases such as diabetic foot.

Crassicaudiasis in three geographically and chronologically distant Cuvier's beaked whales (Ziphius cavirostris) stranded off Brazil.

The Cuvier's beaked whale (CBW; Ziphius cavirostris) is a cosmopolitan marine mammal found in deep tropical and temperate waters of all oceans. CBW strandings have been recorded sporadically in Brazil; however, there is lack of information available regarding their causes of stranding and/or death. Herein, we report the epidemiologic, pathologic, morphologic parasitologic features and molecular identification of arterial and renal crassicaudiasis by Crassicauda sp. in three geographically and chronologically distant CBW stranded off Brazil. CBW-1 was an adult male stranded dead in Rio Grande do Sul State. CBW-2 was an adult female that stranded alive in Sergipe State and died shortly after. CBW-3 was and adult male that stranded dead in Santa Catarina State. The most relevant pathologic findings in these three CBW were severe, chronic proliferative mesenteric and caudal aortic endarteritis and chronic granulomatous and fibrosing interstitial nephritis with renicular atrophy and loss, and numerous intralesional Crassicauda sp. nematodes. Furthermore, CBW-1 had concomitant gram-negative bacterial pneumonia and pulmonary and hepatic thromboembolism. Morphologic analysis of renal adult nematodes identified Crassicauda sp. in the three CBW. Molecular analyses targeting the 18S and ITS-2 ribosomal loci of renal nematodes in CBW-2 and CBW-3 identified C. anthonyi. It is believed that severe arterial and renal crassicaudiasis likely resulted or contributed significantly to morbidity and death of these animals. These results expand the known geographical range of occurrence of crassicaudiasis in CBW. Specifically, the present study provides the first accounts of arterial and renal crassicaudiasis in CBW off the southern hemisphere, specifically in CBW off Brazil, and to the authors' knowledge, it is the first record of C. anthonyi in the southern Atlantic Ocean.

Detection of enteric agents into a cats' shelter with cases of chronic diarrhea in Southern Brazil / Detecção de agentes enteropatogênicos associados à diarreia crônica em um gatil no Sul do Brasil

This study carried out a survey about enteropathogenic agents in domestic cats' shelter as a stage of investigation for the intermittent chronic diarrhea. Individual fecal samples from 39 cats with free access to the external environment were submitted to parasitological examination, parvovirus, and coronavirus by PCR, and Cryptosporidium spp., Giardia spp. and Tritrichomonas foetus by real-time PCR. From the cats evaluated, 30 (76.9%) were positive for one or more enteric agents, and coinfections were observed in 11 cats samples (28.2%). Helminth eggs were observed in 48.7% of cats (19/30), 16 (41%) were positive for parvovirus or coronavirus and 25.6% (10/30) were infected by protozoa. From the positives for protozoa, five cats were positive to T. foetus (12.82%). The first finding of this protozoan through PCR was in the southern Brazil, and the second was in the whole country. Chronic diarrhea in cats may be multifactorial in shelter animals where the population density is high and the control of parasitic, and viral infections are deficient. Moreover, it is due to poor hygiene conditions in these shelters. The factors associated with the proliferation of infectious diseases in shelters are correlated with new pathogens infections such as T. foetus.(AU)

Uma pesquisa de agentes enteropatogênicos em gatos domésticos de um abrigo foi realizado como etapa da investigação das causas de diarreias crônicas intermitentes. Amostras fecais individuais de 39 gatos, com livre acesso ao ambiente externo, foram obtidas para pesquisa de helmintos através do exame parasitológico, investigação de parvovírus e coronavírus e de Cryptosporidium spp., Giardia spp. e Tritrichomonas foetus através de PCR em tempo real. Dos gatos avaliados, 30 (76,9%) foram positivos para algum ou mais de um destes agentes entéricos. Desses, 11 (28,2%) apresentaram co-infecções parasitárias. Ovos de helmintos foram observados em 48,7% dos gatos (19/30), 16 felinos (41%) foram positivos para parvovírus ou coronavírus e 25,6% (10/30) estavam infectados por protozoários. Dos positivos para protozoários, cinco apresentaram Tritrichomonas foetus (12,82%), um organismo pouco relatado no Brasil, sendo este o primeiro relato de detecção deste protozoário através de PCR em fezes de gatos no Sul do Brasil e o segundo no país. A diarreia crônica em gatos pode ser multifatorial em animais de abrigo onde a densidade populacional é elevada e os meios de controle parasitário e viral são deficitários, além das condições de higiene precárias. Os fatores associados à proliferação de doenças infecciosas em abrigos promovem o surgimento de infecções por novos patógenos como o Tritrichomonas foetus, até então pouco relatado no Brasil.(AU)

The use of epifluorescence microscopy and fluorescent dyes for visualization of Oxyuris equi eggs.

This study presents a new method for visualization of Oxyuris equi eggs collected by means of a clear adhesive tape applied to the perianal region of horses. The obtained results indicate that this protocol permits a quick, easy, clear and selective visualization of Oxyuris equi eggs even in the presence of feces and other material. Another advantage of this method is that it can be used with fluorescent dyes solubilized in water, which will stain biologic material without dissolving or altering the adhesive tape and is also environmentally safe. Other dyes currently used for staining biologic materials use organic solvents, which may be combined with acids or bases in their formulation, preventing their use with the tape method.

Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets / Estudo caso-controle pareado avaliando a frequência dos principais agentes causadores de diarréia neonatal em suínos no Rio Grande do Sul

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3 percent) and in 16 farms (57.1 percent) more than one agent was found. The main agents diagnosed were Coccidia (42.86 percent) and rotavirus (39.29 percent). The main agents identified in litters with diarrhea were Clostridium difficile (10.6 percent), Clostridium perfringens type A (8.8 percent) and rotavirus (7.5 percent); in control litters, Clostridium difficile (16.6 percent) and Coccidian (8.5 percent). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

Um estudo de caso-controle em leitegadas de um a sete dias de idade foi realizado com o objetivo de identificar os principais agentes infecciosos envolvidos na diarreia neonatal de leitões. As amostras de fezes (n=276) foram coletadas em granjas de suínos no Estado do Rio Grande do Sul, Brasil, no período de maio a setembro de 2007. Leitegadas com diarreia foram consideradas casos (n=147) e leitegadas normais, controles (n=129). As amostras foram examinadas através do teste de aglutinação em látex, PAGE, cultivo, ELISA, PCR e métodos microscópicos para a excreção dos principais agentes de diarreia: virais (rotavirus), bacterianos (Escherichia coli, Clostridium perfringens tipos A e tipo C e Clostridium difficile) e parasitários (coccídeos e Cryptosporidium spp.). Durante o período do estudo não foram observados surtos e a diarréia, quando presente, apresentou-se leve. Pelo menos um agente foi identificado nas amostras fecais de 25 entre 28 granjas (89,3 por cento) analisadas e em 16 granjas (57,1 por cento) mais de um agente foi detectado. Os principais agentes encontrados nas granjas foram coccídeos (42,86 por cento) e rotavírus (39,29 por cento). Os principais agentes detectados nas leitegadas com diarreia foram Clostridium difficile (10,6 por cento), Clostridium perfringens tipo A (8,8 por cento) e rotavírus (7,5 por cento). Por outro lado, nas leitegadas controle os agentes mais prevalentes foram Clostridium difficile (16,6 por cento) e coccídeos (8,5 por cento). E. coli Beta hemolítica e Clostridium perfringens tipo C não foram detectados. O presente estudo de caso-controle demonstrou que nenhum agente infeccioso esteve associado significativamente com diarreia (p>0.05). Esses achados reforçam a necessidade de que haja cuidado na interpretação de resultados de exames laboratoriais em materiais coletados de leitões com diarreia neonatal leve, pois a detecção isolada de um agente infeccioso não indica necessariamente que o mesmo seja a causa do problema.

Capillaria hepatica in Puma concolor: first report in Brazil.

Capillaria hepatica was detected by histopathologic diagnosis in two cougars that were shot in April 2008 in Painel, Santa Catarina, Brazil. Macroscopic analysis of their livers revealed the presence of diffuse granulomas, and the histopathologic analysis indicated the presence of C. hepatica eggs, surrounded by mononuclear inflammatory cell infiltrate, small foci of necrosis, and mild-to-moderate fibrosis. This is the first report of C. hepatica in cougars (Puma concolor) in Brazil.

Presence of Fasciola hepatica in feral nutria (Myocastor coypus) living in a public park in Brazil.

Sixteen nutria (Myocastor coypus) from a protected area in Curitiba, Brazil, were sampled to determine the prevalence of Fasciola hepatica eggs and intestinal parasites and the presence of snails in the habitat used by nutria. The overall prevalence rates were 56.25% (9) for F. hepatica eggs, 87.50% (14) for cestode eggs, 56.25% (9) for ascarid eggs, 50% (8) for coccidian (Eimeriidae) oocysts, and 56.25% (9) for Strongyloidea eggs. Fasciola hepatica eggs had an average size of 138 microm x 72 microm. The following mollusks were found: Physa cubensis, Physa marmorata, and Biomphalaria tenagophyla. These results suggest that the infected nutria could serve as a source of contamination for other animals, humans, and surface water. This is the first report about parasites in nutria in the state of Paraná, Brazil.

Comparação da infecção por protozoários em chinchila (Chinchilla lanigera) de uma criação comercial do município de Viamão-RS, Brasil, e de chinchilas no seu habitat natural, Chile / Comparison of protozoan infection between chinchillas chinchilla lanigera) from a commercial breeding facility in southern Brazil and chinchillas from a natural reserve in Chile

Chinchilla lanígera é um roedor proveniente do Chile e sua criação é com fins comerciais. As doencas parasitarias, principalmente giardíase podem causar problemas clínicos e sanitarios, causando perdas produtivas e económicas. Foram colhidas amostras de fezes de 220 chinchilas de urna criação comercial no sul do Brasil e 35 amostras de chinchilas da Reserva Nacional las Chinchillas no Chile, e submetidas ão método de Faust e colaboradores. O total de amostras positivas para cistos de Giardia sp. foi de 31,37 por cento (80/255); da criação comercial foi de 36,36 por cento(80/220). O número de amostras que apresentaram mais de 5 cistos/campo foi 4,55 por cento(10/220). Todas as amostras dos animáis da Reserva foram negativas. Não houve associação entre a positividade e a faixa etária dos animais analisados.

Chinchilla lanígera is a rodent native to Chile which is bred for commercial purposes. Parasitic diseases, mainly giardiasis, may cause clinical and sanitary problems and lead to production and economic losses. Fecal samples were collected from 220 chinchillas pertaining to a commercial breeding facility in southern Brazil and from 35 chinchillas from Las Chinchillas National Reserve in Chile. All samples were analyzed using the method proposed by Faust et al. Positive samples for Giardia cysts amounted to 31.37 percent (80/255); 36.36 percent (80/220) was recovered from the commercial breeding facility. The rate of samples with over 5 cysts/field was equivalent to 4.55 percent (10/220). All of the samples collected from the National Reserve were negative for Giardia sp. No association was found between positive rates for Giardia sp. and the age of chinchillas.

Detection of Cryptosporidium oocysts by auramine and Ziehl Neelsen staining methods

Cryptosporidium spp is a common intestinal pathogen of animals and humans. It may have an important economic impact on farms and cause potentially zoonotic infections. Fecal specimens were collected from 331 domestic animals (81 beef cattle, 50 sheep, 100 pigs and 100 dogs) and checked for the presence of Cryptosporidium oocysts by way of Ziehl Neelsen and auramine staining methods. An overall positivity rate of 7.5 percent (25/331) was found, with rates of 10 percent (10/100) among the dogs and 18.5 percent (15/81) among the beef cattle. The feces of sheep and pigs tested negative. In beef cattle, 15 and 12 positive samples were detected by the auramine and Ziehl Neelsen staining techniques, respectively, with no statistically significant difference between the two methods. In dogs, the same number of positive samples was found by both techniques.

Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin.

A growing interest in the understanding of the ontogeny and mineralization of fish skeleton has emerged from the recent implementation of fish as a vertebrate model, particularly for skeletal development. Whereas several in vivo studies dealing with the regulation of bone formation in fish have been published, in vitro studies have been hampered because of a complete lack of fish-bone-derived cell systems. We describe here the development and the characterization of two new cell lines, designated VSa13 and VSa16, derived from the vertebra of the gilthead sea bream. Both cell types exhibit a spindle-like phenotype and slow growth when cultured in Leibovitz's L-15 medium and a polygonal phenotype and rapid growth in Dulbecco's modified Eagle medium (D-MEM). Scanning electron microscopy and von Kossa staining have revealed that the VSa13 and VSa16 cells can only mineralize their extracellular matrix when cultured in D-MEM under mineralizing conditions, forming calcium-phosphate crystals similar to hydroxyapatite. We have also demonstrated the involvement of alkaline phosphatase, a marker of bone formation in vivo, and Gla proteins (osteocalcin and matrix Gla protein, MGP) in the process of mineralization. Finally, we have shown that VSa13 and VSa16 cell lines express osteocalcin and MGP in a mutually exclusive manner. Thus, both cell lines are capable of mineralizing in vitro and of expressing genes found in chondrocyte and osteoblast cell lineages, emphasizing the suitability of these new cell lines as valuable tools for analyzing the expression and regulation of cartilage- and bone-specific genes.

Restriction of human immunodeficiency virus type 1 Rev function in murine A9 cells involves the Rev C-terminal domain.

The human immunodeficiency virus type 1 (HIV-1) Rev and human T-cell leukemia virus type 1 (HTLV-1) Rex proteins are essential for the expression of viral structural proteins and productive infection. Both contain a nuclear export signal (NES) in their C-terminal domain and a nuclear localization signal (NLS) in their N-terminal domain. The NES and NLS are necessary for shuttling between nucleus and cytoplasm and are therefore indispensable for the transport of unspliced and singly spliced viral transcripts. HIV-1 Rev function is restricted in A9 cells, a murine fibroblast cell line, whereas HTLV-1 Rex is functional in these cells. Immunofluorescence studies with RevGFP fusion protein demonstrate normal import and export of Rev in A9 cells. To ascertain which domains of Rev are necessary for the restriction of Rev function in A9 cells, we studied a chimeric construct in which the NES domain of Rev was exchanged with Rex C-terminal amino acids 79 to 95, the Rev1-79/Rex79-95 chimera, which restored Rev function in A9 cells. In addition, overexpression of a truncated Rev containing the Rev C-terminal domain in the presence of wild-type Rev, led to restoration of Rev function in A9 cells. These results suggest that the C-terminal domain of HIV-1 Rev plays an important role in restricting Rev function in murine cells.