Secretion of multi-protein migratory complex induced by Toxoplasma gondii infection in macrophages involves the uPA/uPAR activation system

Toxoplasmosis is a world wide spread zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite that is able to disseminate into deep tissues and cross biological barriers, reaching immunoprivileged sites such as the brain and retina. The parasite is able to infect macrophages and dendritic cells for dispersal throughout the body. However, the molecular mechanisms or outcomes of the subversion of the host cell are largely unknown. Recently our group established that metalloproteinases are involved in migration of infected macrophages. Herein, we evaluated the recruitment of host invasive machinery components in T. gondii infected murine macrophages. We showed by immunoprecipitation assays that MMP-9, CD44 TIMP-1 and uPAR were secreted as a multi-protein complex by infected macrophages. Zymographic analysis revealed that MMP-9 was present in its pro- and active form. Moreover, inhibition of uPA/uPAR pathway by PAI-1 decreased secretion of MMP-9 active forms, as well those associated to uPAR and TIMP-1, but not to CD44. Data presented here suggest that MMP-9 is secreted as a multiprotein complex by T. gondii infected macrophages, similar to that observed in metastatic cells. We further speculate that uPA/uPAR system is involved in the expression/secretion of complexes containing active MMP-9 forms.     

Evaluation of sodium percarbonate as a bath treatment for amoebic gill disease in Atlantic salmon

Amoebic gill disease (AGD), caused by Neoparamoeba perurans, is a major health challenge for Atlantic salmon aquaculture globally. While freshwater bathing for 2 hr is effective in reducing infection severity, there is need for more rapid and lower cost alternatives. To this end, a combination of sodium percarbonate (SPC) in freshwater was examined for its treatment efficacy. Initial in vitro studies showed a reduction in amoeba viability when exposed for 30 min to freshwater containing >500 mg/L SPC. Subsequently, AGD‐affected salmon were bathed for 30 min in 16°C freshwater containing 100, 500 or 1,000 mg/L SPC, or for 2 hr in 16°C freshwater to mimic industry practice. Treatment at the highest SPC concentration caused extensive gill damage and substantial mortality. Neither occurred to a significant extent at lower SPC concentrations. Gill pathology of surviving fish 10 days post‐treatment (dpt) was comparable to or more severe than pre‐treatment, and significantly (p < .001) more severe than in 2 hr freshwater bathed fish. N. perurans DNA was confirmed by qPCR in all treatment groups at 10 dpt. The data indicate that a 30‐min exposure to SPC in freshwater is not a suitable alternative to existing freshwater treatment of AGD.     

Estimating Population Growth Rate From Capture–Recapture Data in Presence of Capture Heterogeneity

The direct estimation and modeling of population growth rate from capture–recapture data has now seen a number of applications. However, the original model cannot accommodate heterogeneous capture probabilities. While studying a population of small mammals Peromyscus maniculatus, we became concerned that the peak of population size may be estimated too late in the year because of heterogeneous catchability. Hence, we developed a variation of the original model with a finite number of catchability classes. The results obtained with the new model are more in agreement with the known biology of this population. A bibliographic appendix and computer code are available online.     

Haematological and histological findings in Leghorn chickens infected with infectious bursal disease virus strain 73688

In the present study, specific-pathogen-free, 2-week-old Leghorn chickens were experimentally infected with strain 73688 of infectious bursal disease virus (IBDV) in order to evaluate haematological and histological changes that might suggest a pathomechanism for haemorrhages in this disease. At 96 hours post infection (hpi) a significant increase in prothrombin time was detected in the absence of visible lesions in myeloid bone marrow tissue and of significant thrombocytopenia. The aforementioned findings suggest alteration of the secondary coagulation mechanisms and not a direct effect of virus on thrombocytes or its precursors.     

Characterisation and histopathological observations of a selected Brazilian precocious line of Eimeria acervulina

Precocious lines of Eimeria acervulina "Cu" and "I" strains were obtained after 25 passages of oocysts in chickens that showed a shortening of the prepatent period for first oocyst output from 96 h to 81 and 82 h, respectively. Both precocious lines were evaluated for pathogenicity using as criteria weight gain, lesion score and total oocyst production. Infection of the "Cu" precocious line in chickens showed a high weight gain, low lesion score and low oocyst production, when compared to parent strain infected chickens. However, the results did not show a significant difference in relation to the criteria used above for the E. acervulina "I" precocious line when compared to its parent strain. This suggests a low degree of attenuation for the "I" strain but good attenuation for the precocious "Cu" line. The histopathological observations of chickens infected with the E. acervulina "Cu" parent strain and precocious line, comparing life cycle and intestinal lesions, showed: (1) parasite stages only in the border cells of infected chicken intestinal villi, for the precocious line; (2) parasite stages in the border cells of the intestinal villi and submucosa cells near the Lieberkühn glands of the intestine; and (3) high degree of inflammatory cells around the parasites in chickens infected with the parent strain. The "Cu" strain was also characterized for sensitivity against eight anticoccidial drugs. Sensitivity was observed for four anticoccidial drugs and partial resistance for four other drugs, although the strain had never had previous contact with anticoccidial drugs, suggesting the presence of a natural resistance factor. This Brazilian E. acervulina "Cu" precocious line showed attenuation for pathogenicity in chickens, suggesting that it could be a suitable strain for use as a live vaccine in Brazil.     

Assessment of the relationship of bispectral index values, hemodynamic changes, and recovery times associated with sevoflurane or propofol anesthesia in pigs

OBJECTIVE: To evaluate bispectral index (BIS) values in pigs during anesthesia maintained with sevoflurane-fentanyl or propofol-fentanyl as a predictor of changes in hemodynamic parameters and duration of recovery from anesthesia. ANIMALS: 12 pigs. PROCEDURE: Pigs were randomly allocated to undergo 1 of 2 anesthetic regimens. Anesthesia was induced with propofol (2 mg/kg, i.v.); 6 pigs were administered sevoflurane via inhalation (1 minimum alveolar concentration [MAC] at a fresh gas flow rate of 3 L/min; group I), and 6 were administered propofol (11 mg/kg/h, i.v.; group II). All pigs received fentanyl (2.5 mg/kg, i.v., q 30 min). After abdominal surgery, pigs were allowed to recover from anesthesia. Cardiovascular variables and BIS values were recorded at intervals throughout the procedure; duration of recovery from anesthesia was noted. RESULTS: No correlation was established between arterial blood pressure and BIS and between heart rate and BIS. Mean BIS at discontinuation of administration of the anesthetic agent was greater in group-II pigs (65.2 +/- 10.6 minutes) than in group-I pigs (55.8 +/- 2.9 minutes). However, recovery from anesthesia was significantly longer in group II (59.80 +/- 2.52 minutes) than in group I (9.80 +/- 2.35 minutes). CONCLUSIONS AND CLINICAL RELEVANCE: In swine anesthetized with sevoflurane or propofol and undergoing abdominal surgery, the BIS value derived from an electroencephalogram at the end of anesthesia was not useful for predicting the speed of recovery from anesthesia. Moreover, BIS was not useful as a predictor of clinically important changes in arterial blood pressure and heart rate in those anesthetized pigs.     

Bispectral index,spectral edge frequency 95%, and median frequency recorded for various concentrations of isoflurane and sevoflurane in pigs

OBJECTIVE: To evaluate bispectral index (BIS), spectral edge frequency 95% (SEF), and median frequency (MED) in relation to a visual analogue scale (VAS) as indicators of anesthetic depth for various concentrations of sevoflurane and isoflurane in pigs. ANIMALS: 32 pigs. PROCEDURE: Pigs were randomly allocated to 8 groups (4 pigs/group). An electroencephalogram (EEG) was recorded in each conscious pig. Pigs were then anesthetized by use of sevoflurane (n = 16) or isoflurane (16). Agents were administered in oxygen at minimum alveolar concentrations (MACs) of 1, 1.25, 1.5, and 1.75 MAC in a randomized order. End-tidal sevoflurane and isoflurane concentrations were maintained for 30 minutes, after which an EEG was recorded for 5 minutes; BIS, SEF, and MED were then calculated. Anesthetic depth was evaluated by use of the VAS. Cardiovascular and EEG responses to nociceptive stimuli were evaluated for each anesthetic agent. RESULTS: BIS decreased significantly for the various concentrations of each anesthetic. At equivalent MACs, BIS values were significantly higher during sevoflurane-induced anesthesia than during isoflurane-induced anesthesia. Values of MED and SEF decreased significantly from basal values to 1 MAC of sevoflurane and isoflurane. For both agents, there was good correlation between VAS scores and BIS values and between VAS scores and SEF values. CONCLUSIONS AND CLINICAL RELEVANCE: BIS was useful for predicting changes in anesthetic depth at clinical dosages of inhalant anesthetics. Values of BIS, SEF, and MED were significantly higher during anesthesia induced by administration of sevoflurane than during anesthesia induced by administration of isoflurance at equivalent MACs.     

Western blot analysis of the humoral response of dogs experimentally infected with Angiostrongylus vasorum (Baillet, 1866)

Seven cross-bred dogs were inoculated with Angiostrongylus vasorum and serum samples were analyzed using the enzyme-linked immunosorbent assay (ELISA) and Western blot (WB). ELISA detected specific antibodies anti-A. vasorum, from 14 to 28 days after inoculation (DAI) and persisted throughout the experiment. Using WB, the main antigens detected had molecular weight of approximately 115, 102, 86, 76, 69, 56, 41, 32, 28, 20-22 and 10kDa.     

Integrated control of apple postharvest pathogens and survival of biocontrol yeasts in semi-commercial conditions

The biocontrol yeast isolates Rhodotorula glutinis LS11, Cryptococcus laurentii LS28 and Aureobasidium pullulans LS30 were tested against Botrytis cinerea and Penicillium expansum on apples artificially inoculated and stored at 3 and 20 °C. Isolates LS28 and LS30 were most effective, consistently resulting in high reductions of fungal decay, while isolate LS11 was effective only on apples stored at 3 °C. The yeasts showed good in vitro resistance to dicarboximides and copper fungicides, while they were inhibited by triazoles. Isolate LS11, in contrast to LS28 and LS30, was also inhibited by benzimidazoles. The yeasts were tested on naturally-infected apples in semi-commercial conditions for 2 years. They were applied twice: soon after harvesting and 20 days later, at the beginning of the cold storage. The antagonists significantly reduced fungal decay when combined with a low dosage of benomyl showing an activity comparable to that exerted by the fungicide alone at full dosage. Periodical monitoring of the epiphytic biocontrol yeast populations in both the field and cold room showed a good rate of survival of the antagonists on the skin of treated apples. Specific fingerprints relying on amplified restriction length polymorphism (AFLP) were used to integrate the morphology-based monitoring of the yeasts.