Septic arthritis: immunopathogenesis, experimental models and therapy

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

Nutritional status and immune response in murine experimental Jorge Lobo's disease

There are no studies investigating the role of nutritional status and immunity associated with Jorge Lobo's disease. The objective of this study was to evaluate the effects of protein-calorie malnutrition on the immune response of BALB/c mice inoculated with Lacazia loboi. In this study,the animals were divided into four groups: G1: inoculated with restricted diet, G2: not inoculated with restricted diet, G3: inoculated with regular diet, G4: not inoculated with regular diet. The animals of groups G1 and G2 were submitted to malnutrition for 20 days and once installed the animals were inoculated intradermally into the footpad. After 4 months, they were euthanised for the isolation of peritoneal lavage cells and removal of the footpad. The production of IL-2, IL-4, IL-10, IL-12, IFN-γ, TNF-α, H2 O2 and nitric oxide (NO) was evaluated in the peritoneal lavage cells. The footpad was evaluated regarding the size of macroscopic lesions, number of fungi and viability index. The results showed that the infection did not exert great influence on the body weight of the mice and previous malnutrition was an unfavourable factor for viability index, number of fungi, macroscopic lesion size in the footpad and production of H2 O2 , NO, IL-12, IL-10 and IFN-γ, suggesting that malnutrition significantly altered fungal activity and peritoneal cells. The results suggest considerable interaction between nutrition and immunity in Jorge Lobo's disease.

Competitive interactions between C. albicans, C. glabrata and C. krusei during biofilm formation and development of experimental Candidiasis

In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis.

Effectiveness of hyperbaric oxygen for experimental treatment of schistosomiasis mansoni using praziquantel-free and encapsulated into liposomes: Assay in adult worms and oviposition

The treatment of schistosomiasis depends on a single drug: praziquantel (PZQ). However, this treatment presents limitations such as low and/or erratic bioavailability that can contribute to cases of tolerance. Improvements to the available drug are urgently needed and studies with a controlled system of drug release, like liposomes, have been gaining prominence. The present study evaluated the activity and synergy between liposomal-praziquantel (lip.PZQ) and hyperbaric oxygen therapy (HBO). Mice received doses of 60 or 100mg/kg PZQ or lip.PZQ, 50 days post-infection, and after the treatment, were exposed to HBO (3 atmosphere absolute - ATA) for 1h. The viability of adult worms and oviposition were analyzed, by necropsy and Kato-Katz examination performed after 15 days of treatment. A concentration of 100mg/kg of lip.PZQ+HBO was more effective (48.0% reduction of worms, 83.3% reduction of eggs/gram of feces) and 100% of the mice had altered of oograms (indicating interruption of oviposition) compared to other treatments and to the Control group (infected and untreated). It is known that PZQ requires participation of the host immune system to complete its antischistosomal activity and that HBO is able to stimulate the immune system. The drug became more available in the body when incorporated into liposomes and, used with HBO, the HBO worked as an adjuvant. This explains the decreases of oviposition and worms recovered form hepatic portal system.

Complexation between darunavir ethanolate and beta cyclodextrin: experimental and theoretical studies

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

Infecção experimental para avaliar a excreção do vírus da diarreia viral bovina por leitões desmamados e transmissão viral por via aerógena

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

Ribavirin has an in vitro antiviral effect in rabies virus infected neuronal cells but fails to provide benefit in experimental rabies in mice

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

Efeito da infecção com Candida albicans no desenvolvimento da Encefalomielite Autoimune Experimental

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

Leishmaniose visceral e o sistema nervoso central: inflamação nas infecções natural canina e experimental em camundongos: Guilherme Dias de Melo. -

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

Humoral Immune Responses of White-Tailed Deer (Odocoileus virginianus) to Mycobacterium bovis BCG Vaccination and Experimental Challenge with M. bovis

Monitoring of the kinetics of production of serum antibodies to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of Mycobacterium bovis infection as well as for the characterization of disease progression and the efficacy of intervention strategies in several species. The humoral immune responses to multiple M. bovis antigens by white-tailed deer vaccinated with BCG orally via a lipid-formulated bait (n = 5), orally in liquid form (n = 5), and subcutaneously (n = 6) were evaluated over time after vaccination and after experimental challenge with virulent M. bovis and were compared to the responses by unvaccinated deer (n = 6). Antibody responses were evaluated by using a rapid test (RT), a multiantigen print immunoassay (MAPIA), a lipoarabinomannan enzyme-linked immunosorbent assay (LAM-ELISA), and immunoblotting to whole-cell sonicate and recombinant antigen MPB83. MAPIA and RT detected minimal to no antibody responses over those at the baseline to multiple M. bovis antigens in vaccinated white-tailed deer after challenge. This was in contrast to the presence of more readily detectable antibody responses in nonvaccinated deer with more advanced disease. The LAM-ELISA results indicated an overall decrease in the level of production of detectable antibodies against lipoarabinomannan-enriched mycobacterial antigen in vaccinated animals compared to that in nonvaccinated animals after challenge. Immunoblot data were inconsistent but did suggest the occurrence of unique antibody responses by certain vaccinated groups to Ag85 and HSP70. These findings support further research toward the improvement and potential use of antibody-based assays, such as MAPIA, RT, and LAM-ELISA, as tools for the antemortem assessment of disease progression in white-tailed deer in both experimental and field vaccine trials.

Development of a Model of Natural Infection with Mycobacterium bovis in White-Tailed Deer

The objective of this study was to develop a suitable experimental model of natural Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus), describe the distribution and character of tuberculous lesions, and to examine possible routes of disease transmission. In October 1997, 10 mature female white-tailed deer were inoculated by intratonsilar instillation of 2 3 103 (low dose) or 2 3 105 (high dose) colony forming units (CFU) of M. bovis. In January 1998, deer were euthanatized, examined, and tissues were collected 84 to 87 days post inoculation. Possible routes of disease transmission were evaluated by culture of nasal, oral, tonsilar, and rectal swabs at various times during the study. Gross and microscopic lesions consistent with tuberculosis were most commonly seen in medial retropharyngeal lymph nodes and lung in both dosage groups. Other tissues containing tuberculous lesions included tonsil, trachea, liver, and kidney as well as lateral retropharyngeal, mandibular, parotid, tracheobronchial, mediastinal, hepatic, mesenteric, superficial cervical, and iliac lymph nodes. Mycobacterium bovis was isolated from tonsilar swabs from 8 of 9 deer from both dosage groups at least once 14 to 87 days after inoculation. Mycobacterium bovis was isolated from oral swabs 63 and 80 days after inoculation from one of three deer in the low dose group and none of four deer in the high dose group. Similarly, M. bovis was isolated from nasal swabs 80 and 85 days after inoculation in one of three deer from the low dose group and 63 and 80 days after inoculation from two of four deer in the high dose group. Intratonsilar inoculation with M. bovis results in lesions similar to those seen in naturally infected white-tailed deer; therefore, it represents a suitable model of natural infection. These results also indicate that M. bovis persists in tonsilar crypts for prolonged periods and can be shed in saliva and nasal secretions. These infected fluids represent a likely route of disease transmission to other animals or humans.

Reo-Like Virus in White Shrimp Penaeus vannamei (Crustacea: Decapoda): Co-occurrence with Baculovirus penaei in Experimental Infections

A virus, tentatively identified as reo-like, occurred concurrently with experimentally-induced Baculovirus penaei (BP) infection in cultured white shrimp larvae Penaeus vannamei. Each shrimp with a reo-like viral infection also had a BP infection, but not all BP-infected shrimp had a reo-like infection. Both viruses occurred in the same tissues and occasionally withln the same cell. The reolike virus developed in epithelial cells of the anterior midgut and in reserve- and fibrillar-cells of the hepatopancreas. The paraspherical and non-enveloped reo-like virions (ca. 50 nm diam.) occurred as unordered aggregates in the cell cytoplasm. Their etiology has not been determined. Reo-like virions may have been introduced along with the BP virus, or, were latent and only manifested due to stress induced by the more pathogenic BP virus.

Experimental Bolbophorus damnificus (Digenea: Bolbophoridae) Infections in Piscivorous Birds

In order to determine potential definitive hosts of the digenetic trematode, Bolbophorus damnificus, two American White Pelicans (Pelecanus erythrorhynchos), two Double-crested Cormorants (Phalacrocorax auritus), two Great Blue Herons (Ardea herodias), and two Great Egrets (Ardea alba) were captured, treated with praziquantel, and fed channel catfish (Ictalurus punctatus) infected with B. damnificus metacercariae. Patent infections of B. damnificus, which developed in both American White Pelicans at 3 days post-infection, were confirmed by the presence of trematode ova in the feces. Mature B. damnificus trematodes were recovered from the intestines of both pelicans at 21 days post-infection, further confirming the establishment of infection. No evidence of B. damnificus infections was observed in the other bird species studied. This study provides further evidence that Double-crested Cormorants, Great Blue Herons, and Great Egrets do not serve as definitive hosts for B. damnificus.

Experimental Inoculation of Coyotes with Mycobacterium bovis Susceptibility and Shedding

Several wildlife species have tested positive for bovine tuberculosis in Michigan and may potentially transmit the disease to other animals. Coyotes have the highest known prevalence in the endemic area and thus, our objective was to investigate the shedding of Mycobacterium bovis by coyotes. Four coyotes were orally inoculated with 1 ml of 1 x 105 CFU/ml of M. bovis. Oral and nasal swabs, and feces were collected regularly and tested by culture. Fecal samples were also tested by exposing guinea pigs to the coyotes' feces. All animals were necropsied to determine if infection occurred. All swabs, feces and tissues were negative on culture. The dosage of M. bovis given to these coyotes was considered biologically relevant, but was insufficient for causing infection. Due to the lack of infection, we still do not know the risk coyotes pose for shedding M. bovis.