A immunosensor for the diagnosis of canine distemper virus infection using SPR and EIS

Canine distemper virus (CDV) is a viral disease that affects dogs and many other carnivores. Clinical diagnosis of CDV is difficult due to the broad spectrum of signs that may be confounded with other respiratory and enteric diseases of dogs. Laboratory analysis is required to diagnose suspected cases. In this study, surface plasmon resonance (SPR) and electrochemical impedance spectroscopy (EIS) methodologies were developed for the detection of canine distemper virus simultaneously. The assay exhibited high specificity, as all the negative controls were not mistakenly detected. The CDV concentration was determined from successive injections into the apparatus, with a linear range from 1.1 to 116.0 ng mL-1. The system exhibited good reproducibility with 4.5% variation between runs after regeneration of the coated surface with a solution of 0.1 M glycine-HCL (pH 3.0). The capacitance and resistance values of the modified interface were calculated from EIS data using an equivalent circuit. It was possible to measure CDV in highly concentrated viruses with good specificity and reproducibility. © 2013 The Royal Society of Chemistry.

Ocorrência de microrganismos periodontopatogênicos e viruses herpéticos na cavidade bucal de pacientes portadores de Síndrome de Down

Pós-graduação em Ciência Odontólogica - FOA

Alterações comportamentais, psicológicas, de higiene bucal e presença de microrganismos superinfectantes e viruses da família herpesviridae em pacientes mantidas em programa de desintoxicação para dependentes químicos

Pós-graduação em Odontologia - FOA

Tropical tanniniferous legumes used as an option to mitigate sheep enteric methane emission

This study presents the first results from Brazil using SF6 tracer technique adapted from cattle to evaluate the capability of condensed tannin (CT) present in three tropical legume forages, Leucaena leucocephala (LEU), Styzolobium aterrimum (STA), and Mimosa caesalpiniaefolia Benth (MIM) to reduce enteric CH4 production in Santa Ins sheep. Twelve male lambs [27.88 +/- 2.85 kg body weight (BW)] were allocated in individual metabolic cages for 20-day adaptation followed by 6 days for measuring dry matter intake (DMI) and CH4 emission. All lambs received water, mineral supplement, and Cynodon dactylon v. coast-cross hay ad libitum. The treatments consisted of soybean meal (710 g/kg) and ground corn (290 g/kg) [control (CON)]; soybean meal (150 g/kg), ground corn (30 g/kg), and Leucaena hay (820 g/kg) (LEU); soybean meal (160 g/kg), ground corn (150 g/kg), and Mucuna hay (690 g/kg) (STA); and soybean meal (280 g/kg), ground corn (190 g/kg), and Mimosa hay (530 g/kg) (MIM); all calculated to provide 40 g/kg CT (except for CON). DMI (in grams of DMI per kilogram BW per day) was lower for LEU (22.0) than CON (29.3), STA (31.2), and MIM (31.6). The LEU group showed emission of 7.8 g CH4/day, significantly lower than CON (10.5 g CH4/day), STA (10.4 g CH4/day), and MIM (11.3 g CH4/day). However, when the CH4 emission per DMI was considered, there were no significant differences among treatments (0.37, 0.36, 0.33, and 0.35 g CH4/g DMI/kg BW/day, respectively, for CON, LEU, STA, and MIM). The sheep receiving STA had shown a tendency (p = 0.15) to reduce methane emission when compared to the CON group. Therefore, it is suggested that tropical tanniniferous legumes may have potential to reduce CH4 emission in sheep, but more research is warranted to confirm these results.

Survey of viruses belonging to different genera and species in noble garlic in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bat coronavirus in brazil related to appalachian ridge and porcine epidemic diarrhea viruses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of supplementation frequency on the ruminal fermentation and enteric methane production of beef cattle grazing in tropical pastures

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resistance to tetracycline and β-lactams and distribution of resistance markers in enteric microorganisms and pseudomonads isolated from the oral cavity

This study evaluated the occurrence of enteric bacteria and pseudomonads resistant to tetracycline and beta-lactams in the oral cavity of patients exhibiting gingivitis (n=89); periodontitis (n=79), periodontally healthy (n=50) and wearing complete dentures (n=41). Microbial identification and presence of resistance markers associated with the production of beta-lactamases and tetracycline resistance were performed by using biochemical tests and PCR. Susceptibility tests were carried out in 201 isolates of enteric cocci and rods. Resistance to ampicillin, amoxicillin/clavulanic acid, imipenem, meropenem and tetracycline was detected in 57.4%, 34.6%, 2.4%, 1.9% and 36.5% of the isolates, respectively. beta-lactamase production was observed in 41.2% of tested microorganisms, while the most commonly found beta-lactamase genetic determinant was gene bla(TEM). Tetracycline resistance was disseminated and a wide scope of tet genes were detected in all studied microbial genus.

Preface to Lesser Known Large dsDNA Viruses. Current Topics in Microbiology and Immunology 328

Several large dsDNA-containing viruses such as poxviruses (smallpox) and herpes viruses are well known among the scientific community, as well as the general populace, because they cause human diseases. The large dsDNA insect-infecting baculoviruses are also well known in the scientific community because they are used both as biological control agents and as protein expression systems. However, there are other large dsDNA-containing viruses, including the giant 1.2-Mb mimivirus, which are less well known even though all of them play important roles in everyday life. Seven of these virus families are reviewed in this book.

Antibodies to Influenza and West Nile Viruses in Horses in Mexico

INFLUENZA A virus (IAV) (family Orthomyxoviridae) is a highly infectious respiratory pathogen of birds and mammals, including human beings and horses (Palese and Shaw 2007). The virus is classified into different subtypes based on the antigenic properties of the haemagglutinin (HA) and neuraminidase (NA) proteins. Sixteen HA subtypes (H1 to H16) and nine NA subtypes (N1 to N9) have been identified (Fouchier and others 2005). Two subtypes, H3N8 and H7N7, have been isolated from horses. The H7N7 subtype was first isolated from a horse in Czechoslovakia in 1956 (Prague/56) (Sovinova and others 1958), and the H3N8 subtype was first isolated from a horse in Miami, USA, in 1963 (Waddell and others 1963). The H7N7 subtype has not been isolated from horses for three decades and is presumed to be extinct (Webster 1993). The H3N8 subtype is currently a common cause of disease in horses worldwide. In horses, influenza is characterized by an abrupt onset of pyrexia, depression, coughing and nasal discharge, and is often complicated by secondary bacteria infections that can lead to pneumonia and death (Hannant and Mumford 1996). Although H3N8 is a major cause of morbidity in horses throughout the world, information on the seroprevalence of IAV in horses and other domestic animals in Mexico is limited.

INFLUENCE OF TYPE 2 BOVINE VIRAL DIARRHEA VIRUS NPRO ON ENHANCEMENT OF BOVINE RESPIRATORY SYNCYTIAL VIRUS REPLICATION MEDIATED BY ANTAGONISM OF HOST CELL INTERFERON TYPE I RESPONSES

Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus, Family Flaviviridae. The virus can infect many species of animals of the order Artiodactyla. The BVDV genome encodes an auto protease, Npro, that degrades interferon regulatory factor-3 (IRF-3) reducing type I interferon (IFN-I) production from host cells. Bovine respiratory syncytial virus (BRSV) is a member of the genus Pneumovirus, Family Paramyxoviridae. Concurrent infection with BVDV and BRSV causes more severe respiratory and enteric disease than infection with either virus alone. Our hypothesis was that Npro modulates the innate immune responses to BVDV infection and enhances replication of BVDV or BRSV co-infection. The noncytopathic BVDV2 viruses NY93/c N- Npro 18 EGFP (a mutant with modified Npro fused with enhanced green fluorescent protein), NY93 infectious clone (NY93/c), wild-type NY93-BVDV2 (NY93-wt), and BRSV were evaluated in this study. The objectives of this study were: (1) to characterize the replication kinetics and IFN-I induction in Madin-Darby bovine kidney (MDBK) cells following infection with each of the BVDV isolates, and (2) to characterize the influence of BVDV-mediated IFN-I antagonism on enhancement of BRSV replication in bovine turbinate (BT) cells. NY93/c N- Npro 18 EGFP replicated 0.4 – 1.6 TCID50 logs lower than NY93-wt in MDBK cells. NY93/c N- Npro 18 EGFP-infected MDBK cells synthesized IFN-I significantly higher than NY93/c- and NY93-wt-infected MDBK cells. BT cells co-infected with NY93/c N- Npro 18 EGFP/BRSV or NY93-wt/BRSV were evaluated to determine the effects of co-infection on BRSV replication and IFN-I induction in BT cells. BRSV RNA levels in NY93-wt/BRSV co-infected BT cells were 2.49, 2.79, and 2.89 copy number logs significantly greater than in NY93/c N- Npro 18 EGFP/BRSV co-infected BT cells on days 5, 7, and 9 post-infection, respectively. BVDV RNA levels in NY93/c N- Npro 18 EGFP-infected BT cells were 1.64 – 4.38 copy number logs lower than in NY93-wt-infected BT cells. NY93/c N- Npro 18 EGFP single and co-infected BT cells synthesized IFN-I significantly higher than NY93-wt single and co-infected BT cells. In summary, these findings suggest: (1) NY93/c N- Npro 18 EGFP BVDV2 induced higher levels of IFN-I than BVDV2-wt and may be useful as a safer, replicating BVDV vaccine, and (2) Enhancement of BRSV infection by BVDV co-infection is mediated by antagonism of IFN-I.

In vitro evaluation, in vivo quantification, and microbial diversity studies of nutritional strategies for reducing enteric methane production

The main objective of the present work was to study nutritive strategies for lessening the CH4 formation associated to ruminant tropical diets. In vitro gas production technique was used for evaluating the effect of tannin-rich plants, essential oils, and biodiesel co-products on CH4 formation in three individual studies and a small chamber system to measure CH4 released by sheep for in vivo studies was developed. Microbial rumen population diversity from in vitro assays was studied using qPCR. In vitro studies with tanniniferous plants, herbal plant essential oils derived from thyme, fennel, ginger, black seed, and Eucalyptus oil (EuO) added to the basal diet and cakes of oleaginous plants (cotton, palm, castor plant, turnip, and lupine), which were included in the basal diet to replace soybean meal, presented significant differences regarding fermentation gas production and CH4 formation. In vivo assays were performed according to the results of the in vitro assays. , when supplemented to a basal diet (Tifton-85 hay sp, corn grain, soybean meal, cotton seed meal, and mineral mixture) fed to adult Santa Ines sheep reduced enteric CH4 emission but the supplementation of the basal diet with EuO did not affect ( > 0.05) methane released. Regarding the microbial studies of rumen population diversity using qPCR with DNA samples collected from the in vitro trials, the results showed shifts in microbial communities of the tannin-rich plants in relation to control plant. This research demonstrated that tannin-rich , essential oil from eucalyptus, and biodiesel co-products either in vitro or in vivo assays showed potential to mitigate CH4 emission in ruminants. The microbial community study suggested that the reduction in CH4 production may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.

High Rates of Detection of Respiratory Viruses in Tonsillar Tissues from Children with Chronic Adenotonsillar Disease

Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR) in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (p = 0.05), and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05) in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.

INFLUENZA VIRUSES IN ADULT DOGS RAISED IN RURAL AND URBAN AREAS IN THE STATE OF SAO PAULO, BRAZIL

In 1970, searching for the interspecies transmission of influenza viruses led to the first study on influenza viruses in domestic animals. Birds and mammals, including human beings, are their natural hosts; however, other animals may also play a role in the virus epidemiology. The objective was to investigate the incidence of influenza viruses in adult dogs raised in rural (9, 19.56%) and urban (37, 80.43%) areas in the state of Sao Paulo, Brazil. Dog serum samples were examined for antibodies to influenza viruses by the hemagglutination inhibition (HI) test using the corresponding antigens from the circulating viruses in Brazil. Dogs from rural areas presented antibodies to influenza A H3N2, and influenza A H7N7 and H3N8. In rural areas, dog sera displayed mean titers as 94.37, 227.88, 168.14, 189.62 HIU/25 mu L for subtypes H1N1, H3N2, H7N7, H3N8, respectively. About 84% and 92% of dogs from urban areas exhibited antibodies to human influenza A H1N1 and H3N2, respectively, with statistical difference at p < 0.05 between the mean titers of antibodies to H1N1 and H3N2. About 92% and 100% were positive for H7N7 and H3N8, respectively. In dogs from urban areas, the mean titers of antibodies against influenza A H1N1, H3N2, H7N7 and H3N8, were 213.96, 179.42, 231.76, 231.35 HIU/25 mu L respectively. The difference among them was not statistically significant at p > 0.05. In conclusion, these dogs were positive for both human and equine influenza viruses. The present study suggests the first evidence that influenza viruses circulate among dogs in Brazil.

Analysis of energetically biased transcripts of viruses and transposable elements

RNA interference (RNAi) is a natural endogenous process by which double-stranded RNA molecules trigger potent and specific gene silencing in eukaryotic cells and is characterized by target RNA cleavage. In mammals, small interfering RNAs (siRNAs) are the trigger molecules of choice and constitute a new class of RNA-based antiviral agents. In an efficient RNAi response, the antisense strand of siRNAs must enter the RNA-induced silencing complex (RISC) in a process mediated by thermodynamic features. In this report, we hypothesize that silent mutations capable of inverting thermodynamic properties can promote resistance to siRNAs. Extensive computational analyses were used to assess whether continuous selective pressure that promotes such mutations could lead to the emergence of viral strains completely resistant to RNAi (i.e., prone to transfer only the sense strands to RISC). Based on our findings, we propose that, although synonymous mutations may produce functional resistance, this strategy cannot be systematically adopted by viruses since the longest RNAi-refractory sequence is only 10 nt long. This finding also suggests that all mRNAs display fluctuating thermodynamic landscapes and that, in terms of thermodynamic features, RNAi is a very efficient antiviral system since there will always be sites susceptible to siRNAs.