Acute phase proteins: a potential approach for diagnosing chronic infection by Trypanosoma vivax

O objetivo do presente estudo foi verificar possíveis alterações nas proteínas de fase aguda em ovinos infectados experimentalmente com Trypanosoma vivax. Para tanto, foram utilizados oito ovinos machos, sendo quatro usados como controle e quatro infectados com 10(5) tripomastigotas de T. vivax. Colheram-se amostras de sangue em dois tempos antes da infecção e, posteriormente, aos 5, 7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 e 120 dias após a infecção (dpi); após centrifugação e aliquotização das amostras. As proteínas de fase aguda foram separadas por eletroforese em gel de acrilamida, contendo dodecil sulfato de sódio, e suas concentrações foram determinadas através de densitometria computadorizada. A dosagem de proteína total foi realizada pelo método colorimétrico do biureto. A contagem dos tripanossomas foi realizada diariamente, utilizando-se uma alíquota de 5 µL de sangue disperso em lâmina de microscopia, sob lamínula de 22 × 22 mm, contando-se os parasitos em 100 campos microscópicos, com objetiva de 40×, multiplicados pelo fator de correção do microscópio, e o resultado expresso em parasitos por mL de sangue. Para a análise estatística, empregou-se o teste de Wilcoxon a 5% de probabilidade. Foi observada a diminuição de diversas proteínas de fase aguda e aumento de antitripsina e transferrina que podem ser utilizadas para auxiliar no diagnóstico da infecção por T. vivax, principalmente na fase crônica da infecção.

Prevention of Salmonella infection by contact using intestinal flora of adult birds and/or a mixture of organic acids

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

Experimental infection by Salmonella enterica subsp enterica serovar kottbus in day-old broiler chickens

The strain used in this work was a Salmonella enterica subsp enterica serovar Kottbus (6,8:e,h:1,5) isolated from imported day-old ducklings in Laboratório Nacional Agropecuário (LANAGRO/SP) of the Ministry of Agriculture of Brazil (MAPA). In view of the lack of information available about this Salmonella isolate and also because it was detected in day-old imported birds, this study was carried out to investigate the dissemination of S. Kottbus among newly hatched chicks. The birds were placed in three groups: one group of 20 birds received 0.1 mL of S. Kottbus culture containing 1.2 x 10(8) CFU/mL, the second group of 20 birds was inoculated with 1.2 x 10(5) CFU/mL and the third group of 10 birds was untreated (control group). Results were similar for both infected groups. The bacterium was recovered from cloacal swabs collected from the first day following the experimental infection until the end of the trial (42 days post-inoculation). At 15 and 42 days post-inoculation (dpi), half of the birds of each group were killed for bacteriological examination of cecal contents, liver and spleen. At 15 dpi, viable cell counts of S. Kottbus were obtained in all kinds of samples. At 42 dpi, Salmonella was present in the liver and spleen of few birds, but in large amounts in the cecal contents of almost all birds.

Comparison of infection by Brucella spp. in free-ranging and captive wild animals from São Paulo State, Brazil

The aim of the current study was to evaluate the infection rate by Brucella spp. in wild and in captive animals. Serum samples from 121 animals (94 free-ranging and 27 captive) of different mammal species were evaluated. Sera were submitted to rose Bengal test (RBT) for screening and serum agglutination tests (SAT) and 2-mercaptoethanol test (2-ME) for confirmatory results. Nine animals (five free-ranging and four captive) tested positive in RBT, but negative in the confirmatory tests. Several domestic animal diseases that have control programs are not focused on wild reservoirs, such as brucellosis in Brazil. The study of new reservoirs in wildlife is essential to prevent emerging diseases.

Genome-Wide Analysis of Differentially Expressed Genes During the Early Stages of Tomato Infection by a Potyvirus

Plant responses against pathogens cause up-and downward shifts in gene expression. To identify differentially expressed genes in a plant-virus interaction, susceptible tomato plants were inoculated with the potyvirus Pepper yellow mosaic virus (PepYMV) and a subtractive library was constructed from inoculated leaves at 72 h after inoculation. Several genes were identified as upregulated, including genes involved in plant defense responses (e. g., pathogenesis-related protein 5), regulation of the cell cycle (e. g., cytokinin-repressed proteins), signal transduction (e. g., CAX-interacting protein 4, SNF1 kinase), transcriptional regulators (e. g., WRKY and SCARECROW transcription factors), stress response proteins (e. g., Hsp90, DNA-J, 20S proteasome alpha subunit B, translationally controlled tumor protein), ubiquitins (e. g., polyubiquitin, ubiquitin activating enzyme 2), among others. Downregulated genes were also identified, which likewise display identity with genes involved in several metabolic pathways. Differential expression of selected genes was validated by macroarray analysis and quantitative real-time polymerase chain reaction. The possible roles played by some of these genes in the viral infection cycle are discussed.

Occurrence of infection by Platynosomum illiciens (Braun, 1901) in captive neotropical primates

Platynosomum illiciens (Trematoda, Plagiorchida) is a trematode parasite reported in felids and falconiforms. It was identified in the gall bladder of eight captive neotropical necropsied primates from the National Primate Center (CENP), Ananindeua, State of Para, Brazil. This is the first description of Platynosomum illiciens as a parasite of primates.

Prevalence of dog intestinal parasites and risk perception of zoonotic infection by dog owners in São Paulo State, Brazil

Coprological examination was used to estimate the prevalence of gastrointestinal parasites in stray and domiciled dogs from Botucatu, São Paulo State, Brazil. Risk factors for dog infection were assessed in relation to demographic, husbandry and management data. The dog owners completed a questionnaire survey on some aspects of dog parasitism such as parasite species, mechanisms of infection, awareness of zoonotic diseases and history of anthelmintic usage. Parasites were found in the faeces of 138 dogs, with an overall prevalence of 54.3%. Dogs harbouring one parasite were more common (31.4%) than those harbouring two (18.5%), three (3.2%) or four (1.2%). The following parasites and their respective frequencies were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Dipylidium caninum (2.4%), Isospora (3.5%), Cryptosporidium (3.1%) and Sarcocystis (2.7%). Stray dogs were found more likely to be poliparasitized (P < 0.01) and presented higher prevalence of Ancylostoma, T. canis and Giardia (P < 0.01) than domiciled ones. Toxocara canis was detected more frequently in dogs with < 6 months of age (P < 0.05) and no effect of sex or breed could be observed (P > 0.05). Except for Ancylostoma, that showed a significantly higher prevalence in dogs living in a multi-dog household (P < 0.01), parasite prevalences were similar in single- and multi-dog household. The answers of dog owners to the questionnaire showed that the majority does not know the species of dog intestinal parasites, the mechanisms of transmission, the risk factors for zoonotic infections, and specific prophylactic measures. The predominance of zoonotic species in dogs in the studied region, associated with the elevated degree of misinformation of the owners, indicates that the risk of zoonotic infection by canine intestinal parasite may be high, even in one of the most developed regions of Brazil.

Physical, epidemiological, and molecular evaluation of infection by Cryptosporidium galli in Passeriformes

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

CLINICAL STUDY of NATURAL INFECTION BY Trypanosoma cruzi IN DOGS RESIDING IN A RURAL AREA IN MATO GROSSO do SUL STATE, BRAZIL

This study was carried out to describe the clinical characteristics of natural infection caused by Trypanosoma cruzi in dogs that reside in a rural area of Mato Grosso do Sul State, Brazil. Conventional and nonconventional diagnostic methods were used for screening T. cruzi infection in 75 dogs that lived in the area. Cardiovascular tests and biochemical examination of sera were also performed in four confirmed positive dogs. The following techniques were employed: indirect immunofluorescence test (IFAT), enzyme-linked immunosorbent assay with T. cruzi epimastigote antigens (EAE-ELISA) and enzyme-linked immunosorbent assay with T. cruzi excreted-secreted trypomastigote antigens (TESA-ELISA) with antibodies detected in 45.33% (n = 34), 24.0% (n = 18) and 12.0% (n = 9) of the dogs, respectively. The current prevalence of the infection was confirmed as 10.7% (n = 8) by immunoblotting test with T. cruzi excreted-secreted antigens (TESA-blot). The test that showed the best concordance index (Kappa; 0.93), sensitivity (100%) and specificity (98.5%) was TESA-ELISA, that when associated with IFAT had the same results as those obtained by TESA-blot (10.7%). Three out of the four chagasic animals showed enlarged cardiac silhouette on X-ray and an increase of the P-wave duration and QRS complex in electrocardiogram. Two dogs presented conduction disturbances, right bundle branch block in one dog and first-degree atrioventricular block and sinus arrest in another. The ecodopplercardiography presented left-ventricular-wall thickness increased during diastole, decrease of the shortening fraction and inversion in the speed peaks of the E and A waves, indicating the presence of systolic and diastolic disorders. The four animals showed enzymatic activities of creatine kinase (221-404 U/L), MB fraction of creatine kinase (189-304 U/L), elevated total proteins (7.6-10.2 g/dL) and total globulins (4.6-7.7g/dL) and reduction of albumin/globulin ratio, which suggested a myocardial injury and continuous antigenic stimulus.

Infection by Austrodiplostomum compactum metacercariae in fish from the Nova Avanhandava reservoir, Tiete river, São Paulo State, Brazil.

This study aimed to evaluate the infection by the Austrodiplostomum compactum metacercariae in fishes from the Nova Avanhandava Reservoir, low Tiete river, São Paulo State, Brazil. The parasites were collected from eye (aqueous and vitreous humor), fixed in AFA solution and stained with carmine. The morphometric analysis was performed using a computerized system for analysis of images QWin Lite 2.5 (Leica). Prevalence, mean intensity of infection and abundance of infected fish were calculated. of the 22 species of fish registered, five were infected by metacercariae: Hoplias malabaricus, Metynnis maculatus, Plagioscion squamosissimus, Satanoperca pappaterra and Schizodon imparts. of the 627 fish evaluated, 34% were infected. A higher prevalence was observed in P. sqnamosissionis and S. pappaterra. Schizodon nasutus and M. maculatus are new hosts reported for A. compactum metacercariae.

First record of infection by entomopathogenic nematodes of the grass bug Collaria scenica Stal (Hemiptera: Miridae).

NARANJO N, MONTERO DAV, SAENZ APONTE A. 2011. First record of infection by entomopathogenic nematodes of the grass bug Collaria scenica Stal (Hemiptera: Miridae). ENTOMOTROPICA 26(3): 117-125. The study was aimed to test the pathogenicity of Steinernema sp. and Heterorhabditis sp. in Collaria scenica. The effect of different concentrations of infective juveniles (IJ) were tested on nymphs and adults of C. scenica. For this purpose, the bugs were inoculated with 5 000 JI of each nematode species in a factorial design (3x2), and seven concentrations were tested in a JI factorial design (7x2x2). The bugs showed 100% mortality and symptoms of pathogenicity. Infection was found with both species of nematodes and penetration was assumed to be through the spiracles and anus. A higher capacity of pathogenicity was observed with Steinernema sp. Based on the results Heterorhabditis sp. and Steinernema sp. could constitute an efficient tool to control populations of C. scenica in pastures.

Natural infection by Paramphistomoidea Stiles and Goldberger, 1910 trematodes in wild Marsh Deer (Blastocerus dichotomus Illiger, 1815) from Sérgio Mottas's hydroelectric power station flooding area.

Studies on helminthfauna of marsh deer Blastocerus dichotomus Illiger, 1815 are rare, although helminthic diseases are an important cause of mortality in these animals. Fifteen male and female adult marsh deer from Sergio Motta's hydroelectric power station flooding area at Paraná River which died during the capture and quarantine procedures, between 1998 and 1999, were necropsied. Three trematodes species, Paramphistomum cervi, Balanorchis anastrofus and Zygocotyle lunatum, all belonging to superfamily Paramphistomoidea, were identified. The obtained trematodes were identified, counted and their respectives descriptors of infection were determined. All necropsied animals were infected by helminths. Paramphistomum cervi was the most prevalent species, while Zygocotyle lunatum was found in only one animal.

Successful prevention of tunneled central catheter infection by antibiotic lock therapy using cefazolin and gentamicin

Catheter-related bacteremia (CRB) is one of the various complications related to hemodialysis (HD). As a result of this high rate of infection, the antibiotic lock technique (ALT) has been recommended to prevent CRB. However, adverse effects of ALT such as increased emergence of strains resistant to antibiotics and increased mechanical dysfunction catheter were poorly evaluated. We prospectively evaluated the efficacy of catheter-restricted filling using an antibiotic lock solution in preventing CRB. A total of 233 HD patients requiring 325 new tunneled catheters while waiting for placement and maturation of an arteriovenous fistula or graft were enrolled in this study. Patients with a tunneled catheter were assigned to receive either an antibiotic-heparin lock solution (antibiotic group: cefazolin 10 mg/ml, gentamicin 5 mg/ml, heparin 1,000 U/ml) or a heparin lock solution (no-antibiotic group: heparin 1,000 U/ml) as a catheter lock solution during the interdialytic period. The present study aimed to assess the efficacy of ALT using cefazolin and gentamicin in reducing CRB in patients undergoing HD with tunneled central catheter and to identify its adverse effects. CRB developed in 32.4 % of patients in the no-antibiotic group and in 13.1 % of patients in the antibiotic group. CRB rates per 1,000 catheter-days were 0.57 in the antibiotic group versus 1.74 in the no-antibiotic group (p < 0.0001). Kaplan-Meier analysis also showed that mean CRB-free catheter survival was significantly higher in the antibiotic group than in the no-antibiotic group (log-rank statistic 17.62, p < 0.0001). There was statistically significant difference between the two groups in causative organisms of CRB, with predominance of negative culture in both groups, but this prevalence was higher in ALT group (57.9 vs 90.1 %, p < 0.0001), and the two groups also were different in prevalence of gram-positive bacteria as causing organisms (ALT group 21.05 vs = 0 % in control group, p < 0.0001). There was no statistically significant difference between the two groups in drug-resistant germs. There were statistically significant differences between the two groups in the catheter removal causes, with higher rate of infectious cause in control group (12.32 vs 2.22 %, p < 0.0001) and mechanical cause in ALT group (28.26 vs 37.78 %, p < 0.0001). The results suggest that ALT may be a beneficial means of reducing the CRB rate in HD patients with tunneled catheter, without association between ALT and emergence of strains resistant. However, mechanical complications were more prevalent in antibiotic group. Further studies are required to determine the optimal drug regimen, concentrations for ALT, and its adverse effects. © 2012 Springer Science+Business Media Dordrecht.

Comparative proteomic analysis reveals that T3SS, Tfp, and xanthan gum are key factors in initial stages of Citrus sinensis infection by Xanthomonas citri subsp citri

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