Phenotype and function of murine discrete Peyer's patch macrophage derived - dendritic cells

The phenotype and function of peritoneal cavity macrophage-derived dendritic cells (PEC-DC) was previously reported. In this study we have gone further in using our established culture system to generated discrete Peyer's patch dendritic cells (DPP-DC) from murine discrete Peyer's patch macrophages (DPP-M?), following stimulation with granulocyte macrophage colony stimulating factor (GM-CSF) plus interleukin 4 (IL-4) for 7 days. DPP-M? from murine small intestines were obtained by mechanical disruption of discrete Peyer's patches (DPP), followed by metrizamide density gradient centrifugation to remove Peyer's patch resident DC and debri, after which an overnight adherent step in tissue culture medium was carried out for macrophage enrichment. Characterization of the generated DPP-DC was carried out using well-established criteria of morphology, expression of membrane antigens and capacity for antigen presentation. Dendritic cells expressed DEC-205, F4/80 and CD34 at high levels, but exhibited very low CD11c levels. They were shown to present soluble protein antigen to CD3(+) spleen T cells. A comparison of the surface antigen expression in the progenitor DPP-M? population and the generated DPP-DC showed a significant decrease in MHC class II levels and a marked down regulation of the co-stimulatory molecule CD86 (B7-2). High expression of the haemopoietic progenitor marker CD34 indicates that the generated DC, possess a haemopoietic rather than myeloid origin. Taken together, these results may provide a better understanding of the complex network regulating mucosal immune responses.     

Efficacy of extended pirlimycin therapy for treatment of experimentally induced Streptococcus uberis intramammary infections in lactating dairy cattle

Streptococcus uberis is an important cause of mastitis in dairy cows throughout the world, particularly during the dry period, around the time of calving, and during early lactation. Strategies for controlling S. uberis mastitis have not received adequate research attention and are therefore poorly defined and inadequate. Objectives of the present study were to evaluate the efficacy of extended therapy regimens with pirlimycin for treatment of experimentally induced S. uberis intramammary infections in lactating dairy cows during early lactation and to evaluate the usefulness of the S. uberis experimental infection model for evaluating antimicrobial efficacy in dairy cows. The efficacy of extended pirlimycin intramammary therapy regimens was investigated in 103 mammary glands of 68 dairy cows that became infected following experimental challenge with S. uberis during early lactation. Cows infected with S. uberis in one or both experimentally challenged mammary glands were randomly allocated to three groups, representing three different treatment regimens with pirlimycin, including 2-day (n = 21 cows, 31 mammary quarters), 5-day (n = 21 cows, 32 quarters), and 8-day (n = 26 cows, 40 quarters). For all groups, pirlimycin was administered at a rate of 50 mg of pirlimycin hydrochloride via intramammary infusion. A cure was defined as an experimentally infected mammary gland that was treated with pirlimycin and was bacteriologically negative for the presence of S. uberis at 7, 14, 21, and 28 days after treatment. Experimental S. uberis intramammary infections were eliminated in 58.1% of the infected quarters treated with the pirlimycin 2-day regimen, 68.8% for the 5-day regimen, and 80.0% for the 8-day regimen. Significant differences (P <.05) in efficacy were observed between the 2-day and 8-day treatment regimens. The number of somatic cells in milk decreased significantly following therapy in quarters for which treatment was successful in eliminating S. uberis. However, there was no evidence to suggest that extended therapy with pirlimycin resulted in a greater reduction in somatic cell counts in milk than the 2-day treatment. The S. uberis experimental infection model was a rapid and effective means of evaluating antimicrobial efficacy during early lactation at a time when mammary glands are highly susceptible to S. uberis intramammary infection.     

Evaluation of survival of Actinobacillus pleuropneumoniae and Haemophilus parasuis in four liquid media and two swab specimen transport systems

OBJECTIVE: To determine duration and rates of recovery of Actinobacillus pleuropneumoniae and Haemophilus parasuis from 4 liquid media and 2 swab specimen transport systems and compare findings with those of Escherichia coli. SAMPLE POPULATION: One strain each of A pleuropneumoniae (biovar 1, serotype 1), H parasuis (serovar 5), and E coli (serotype O149:K91:H19). PROCEDURE: Strains were incubated in brain heart infusion broth supplemented with horse serum and other nutrients or in horse serum alone, with and without nicotinamide-adenine dinucleotide in both instances, for 150 days at 4 degrees C or room temperature (21 degrees C). Similarly, strains were tested in Stuart and Amies transport systems after storage at room temperature for 8 days. RESULTS: Colony counts greater than those of the initial inoculum were observed after incubation in horse serum for A pleuropneumoniae but not for H parasuis. Overall, incubation at 4 degrees C in the 4 liquid media resulted in longer recovery duration and higher rates than at room temperature. Culture of H parasuis resulted in lower recovery rates and shorter durations of recovery than culture of A pleuropneumoniae, except for culture in horse serum. Haemophilus parasuis survived longer than A pleuropneumoniae in the transport systems, and all organisms survived longer in the Amies system. CONCLUSIONS AND CLINICAL RELEVANCE: Survival of A pleuropneumoniae and H parasuis indicated that horse serum prolongs survivability, which may result in exposure of more animals during a prolonged period. The Amies system might be a good choice for collection of clinical samples from animals, especially for recovery of H parasuis.     

Evaluation of 65% permethrin spot-on for prevention of canine visceral leishmaniasis: effect on disease prevalence and the vectors (Diptera: Psychodidae) in a hyperendemic area

A study was designed to examine the effect of 65% permethrin spot-on on the prevalence of canine visceral leishmaniasis and the abundance of sand flies in two neighborhoods in Corumbá, Mato Grosso do Sul, Brazil known to have a high prevalence of visceral leishmaniasis. An enrollment survey was conducted to determine the prevalence of visceral leishmaniasis. Area residents were provided with information about the project; the study area was defined, and all dogs (160 in Cristo Redentor and 230 in Popular Velha) identified in the study area were enrolled. Three 65% permethrin spot-on treatments (June 15-25, July 13-15, and August 10-12) were administered to 150, 110, and 99 dogs, respectively, in Popular Velha, according to label recommendations. Dogs in Cristo Redentor were untreated controls. Visceral leishmaniasis was diagnosed periodically by indirect immunofluorescence assay. A reduction in canine visceral leishmaniasis prevalence was observed at the Popular Velha site. The infection rate for treated dogs 1 month following the final treatment was approximately 50% reduced from that observed before treatment(19.3% vs 9.6%). Conversely, the infection rate at the control site was more than 80% higher at the September sampling than that observed pretreatment (4.1% vs 7.4%). Similar numbers of sand flies were captured and identified from both sites throughout the study. The results suggest that regular use of 65% permethrin during months of high risk for canine visceral leishmaniasis can be a useful strategy for reducing the prevalence of this disease in hyperendemic areas. It should be stressed, however, that the success of this strategy depends not only on the efficacy of the product itself but also on the adoption of other control measures and on economic variables, considering the low purchasing power of the populations living in higher-risk neighborhoods.     

Taenia solium oncosphere antigens induce immunity in pigs against experimental cysticercosis

Immunity to Taenia solium infection was investigated using an experimental intramuscular oncosphere infection assay (IMOA) model in pigs. Three naturally infected pigs with cysticercosis were treated with oxfendazole (OFZ), a drug demonstrated to kill cysts in porcine muscle. These animals were then challenged with oncospheres but did not develop any cysts while three uninfected pigs that were similarly challenged, did develop intramuscular cysts. In another study, two groups of three pigs each were immunized with crude T. solium oncosphere and metacestode antigens, respectively, and tested with the IMOA. Immunization with crude oncosphere antigens (OAs) induced 100% protection, while metacestode antigens provided only partial protection. Immunoblots showed that pigs with complete immune protection to oncosphere intramuscular challenge had antibodies to two OAs at 31.3 and 22.5 kDa, respectively. Antibody to these two antigens was absent in pigs immunized with metacestodes or in uninfected control pigs. This study demonstrated the presence of two antigens that are unique to the oncosphere. Although, antibody to these two antigens is consistently present in pigs that are protected from an oncosphere intramuscular challenge their role in preventing infection by T. solium larval cysts is still hypothetical.     

Restriction endonuclease and monoclonal antibody analysis of Brazilian isolates of bovine herpesviruses types 1 and 5

Twelve Brazilian isolates and three reference strains of bovine herpesviruses (BHVs) were subjected to restriction endonuclease analysis (REA) and monoclonal antibody (MAb) analysis. Viral DNA was cleaved with BamHI, BstEII, EcoRI, HindIII and PstI. The monoclonal antibody panel allowed the differentiation between types 1 and 5 viruses, while REA with BstEII and HindIII showed the distinction between BHV-1 and -5 subtypes. Typical 1.1 and 1.2a patterns were observed with two isolates from respiratory disease. An isolate from semen of a clinically healthy bull displayed 1.2b profile, whereas another displayed a clear 5a pattern, which was never reported before. Seven out of nine Brazilian type 5 (BHV-5) isolates displayed REA patterns similar to the Australian BHV-5 strain N569 (BHV-5a), and differing from the Argentinean A663 strain (BHV-5b) virus. Another two BHV-5 isolates, which displayed an unusual MAb pattern of reactivity, showed a BstEII profile different from both reference strains of BHV-5. These two viruses were considered BHV-5 "non-a/non-b" subtype.     

Rabies seroprevalence of street dogs from São Paulo City, Brazil

A sample of 602 street dogs captured by the São Paulo zoonosis-control municipal service was analyzed for the presence of serum antibodies against the rabies virus and compared to a control sample of 37 dogs from the kennel of the police of São Paulo. Data were stratified into 4 age intervals and the age from the street-dog sample was estimated by general aspects and dental observation according to standard techniques. We assumed that seropositivity was due to previous vaccination (no sub-clinical infection). From the prevalence data, we estimated the age-dependent incidence-density rate of vaccination by a simple mathematical model, which also allowed the estimation of the period of antibody protection given by the supposed vaccination. Crude seroprevalence in the street-dog sample was 16.5% and in the sample from the police-kennel dogs was 80%. The average age of first vaccination calculated for the street-dog sample was 5 years of age (if we assumed an average period of vaccine protection of 3 years) — but 5 months of age for the police-kennel sample.     

Uncoupling and inhibition properties of 3,4-seco-friedelan-3-oic acid isolated from Maytenus imbricata

3,4-Seco-friedelan-3-oic acid was isolated from Maytenus imbricata (Celastraceae). At low concentrations it inhibited non-cyclic electron transport and ATP synthesis in spinach chloroplasts, i.e., it behaved as a Hill reaction inhibitor, and at high concentrations it acts as an uncoupler by enhancing uncoupled electron transport and Mg²⁺-ATPase activity. 3,4-Seco-friedelan-3-oic acid did not inhibit PSII electron transport from DPC to DCPIP_ox and photosystem I activity, but it enhanced from TMQH₂ to MV, corroborating its action as uncoupler. It inhibits electron flow through PSII from water to sodium silicomolybdate. The whole results indicate that the 3,4-seco-friedelan-3-oic acid target is at the OEC complex enzyme, the donor side of PSII. The fluorescence decay data shows the formation of the K-band, which match this result, acting as inhibitor at the donor side of PSII and it as an uncoupler.     

Susceptibility of citrus species to Alternaria alternata, the causal agent of the Alternaria brown spot

Alternaria alternata, the causal agent of Alternaria brown spot (ABS), causes necrosis on leaves, twigs, and fruit, reducing the productivity and quality of fruits. Tangerines and their hybrids are highly susceptible to the disease. Species, hybrids, and cultivars of Citrus from the germplasm bank of the Estação Experimental de Citricultura de Bebedouro, São Paulo, Brazil, were evaluated in 2004 and 2005 with respect to their resistance to A. alternata, both through natural infection and by inoculation. Detached leaves were also used to demonstrate susceptibility or resistance to the disease. Ten cultivars of Satsumas (Citrus unshiu), and 14 cultivars of Clementine mandarin (C. clementina) did not show any symptoms of the disease in their leaves, either through natural infection or when inoculated in the field. The Burguess SRA-412, Wallent SRA-438, Carvalhais, Ampefy SRA-459, Ananas SRA, and Macaque SRA-426 mandarin hybrids (C. reticulata) did not show symptoms of the disease under natural or artificial infection in the field. Some cultivars of C. deliciosa, C. tangerina, C. erythrosa, and C. temple showed symptoms of the disease, even though no previous record of their susceptibility to Alternaria brown spot had been previously reported. The hybrids Fairchild, Nova, Page, Fortune, and Sunburst were susceptible to the disease. However, Fremont mandarin (a crossing between C. clementina and C. reticulata), Encore (C. nobilis × C. deliciosa), and Fallglo (C. reticulata × C. paradisi) did not show symptoms in field, and few symptoms were verified in detached leaves. These materials are promising for the cultivation of tangerines, and will enable genetic improvement for the development of cultivars resistant to Alternaria brown spot.     

Growth,Feed Utilization,and Glycemic Response in European Sea Bass,Dicentrarchus labrax,Juveniles Fed Carbohydrate of Different Complexities

The effect of dietary carbohydrate complexity on growth, feed utilization, and glycemia was studied in European sea bass juveniles. Four isonitrogenous (50% crude protein) and isolipidic (15% crude lipids) diets were formulated to contain 20% pregelatinized maize starch (PGS diet), dextrin (DEX diet), maltose (MAL diet), or glucose (GLU diet). No effect of dietary carbohydrate complexity on growth was noticed. Feed efficiency and protein efficiency ratio were lower in fish fed the GLU diet than in the other groups, whereas the opposite was observed for feed intake. Plasma glucose peaked 3–4 h after feeding in fish fed the MAL and GLU diets, whereas in fish fed the PGS and DEX diets the peak was reached 5–6 h after feeding. Peak plasma glucose concentration (13 mmol/L) was higher in fish fed the GLU diet than the other diets (9 mmol/L). Shorter hyperglycemia duration was observed in fish fed the MALT and GLU diets (6 h) than the PGS and DEX diets (10 h). Complex carbohydrates delayed plasma glucose concentration peak compared with simple sugars, whereas the opposite was observed for hyperglycemia duration. Overall, dietary maltose, dextrin, and starch were apparently better utilized as energy source than glucose by European sea bass juveniles.