Cross-immunoreactivity between anti-potato apyrase antibodies and mammalian ATP diphosphohydrolases: potential use of the vegetal protein in experimental schistosomiasis

We have previously showed that Schistosoma mansoni ATP-diphosphohydrolase and Solanum tuberosum potato apyrase share epitopes and the vegetable protein has immunostimulatory properties. Here, it was verified the in situ cross-immunoreactivity between mice NTPDases and anti-potato apyrase antibodies produced in rabbits, using confocal microscopy. Liver samples were taken from Swiss Webster mouse 8 weeks after infection with S. mansoni cercariae, and anti-potato apyrase and TRITC-conjugated anti-rabbit IgG antibody were tested on cryostat sections. The results showed that S. mansoni egg ATP diphosphohydrolase isoforms, developed by anti-potato apyrase, are expressed in miracidial and egg structures, and not in granulomatous cells and hepatic structures (hepatocytes, bile ducts, and blood vessels). Therefore, purified potato apyrase when inoculated in rabbit generates polyclonal sera containing anti-apyrase antibodies that are capable of recognizing specifically S. mansoni ATP diphosphohydrolase epitopes, but not proteins from mammalian tissues, suggesting that autoantibodies are not induced during potato apyrase immunization. A phylogenetic tree obtained for the NTPDase family showed that potato apyrase had lower homology with mammalian NTPDases 1-4, 7, and 8. Further analysis of potato apyrase epitopes could implement their potential use in schistosomiasis experimental models.

The subgenus Migonemyia Galati 1995 (Diptera, Psychodidae, Phlebotominae), with description of a new species Migonemyia vaniae: a review

The capture of a new species of the subgenus Migonemyia Galati, 1995 (Diptera, Psychodidae, Phlebotominae), Migonemyia vaniae sp. nov. in the Ribeira Valley, state of São Paulo, Brazil, together with the other two species: Mg. migonei (França, 1920) and Mg. rabelloi (Galati & Gomes, 1992) lead us to review this subgenus. The new species was described and illustrated. The genitalia of the two other species were also illustrated and some genital characteristics (number of setae on the gonocoxite tuft, ejaculatory ducts and pump and ducts/pump ratio; and number of setae on the tergite VIII of the females) considered important to differentiate the three species, including five populations of Mg. migonei (from Northeastern, Southeastern, and Southern Brazilian regions and of Peru) were submitted to variance analyses. The Mg. migonei population of Northeastern Brazilian region showed distinct smaller values (P < 0.05) than the other Brazilian populations studied as regarding these characteristics. The capture of both sexes of these three species in sympatry confirms the association between the sexes of Mg. rabelloi, recognised as doubtful when this species was originally described. Identification keys for male and female of the three species are presented.

A simple approach improving the performance of urine reagent strips for rapid diagnosis of urinary schistosomiasis in Nigerian schoolchildren

In Nigeria, schistosomiasis, caused predominantly by the species Schistosoma haematobium, is highly endemic in resource-poor communities. We performed a school-based survey in two rural communities in Osun State (Southwestern Nigeria) and assessed macrohaematuria, microhaematuria and proteinuria as indirect indicators for the presence of disease. Urine samples were inspected macroscopically for haematuria and screened for microhaematuria and proteinuria using urine reagent strips. The microscopic examination of schistosome eggs was used as the gold standard for diagnosis. In total, 447 schoolchildren were included in this study and had a 51% prevalence of urinary schistosomiasis. The sensitivity of microhaematuria (68%) and proteinuria (53%) for infection with S. haematobium was relatively low. In patients with a heavy infection (>500 eggs/10 mL), the sensitivity of microhaematuria was high (95%). When the presence of macrohaematuria and the concomitant presence of microhaematuria and proteinuria were combined, it revealed a sensitivity of 63%, a specificity of 93% and a positive predictive value of 91%. Macrohaematuria also showed high specificity (96%) and a positive predictive value of 92%, while sensitivity was < 50%. These data show that combining urine reagent strip tests (presence of proteinuria and microhaematuria) and information on macrohaematuria increased the accuracy of the rapid diagnosis of urinary schistosomiasis in an endemic rural West African setting. This simple approach can be used to increase the quality of monitoring of schistosomiasis in schoolchildren.

Availability of soil water under tillage systems, mulch management and citrus rootstocks

The increased availability of soil water is important for the management of non-irrigated orange orchards. The objective of this study was to evaluate the availability of soil water in a Haplorthox (Rhodic Ferralsol) under different tillage systems used for orchard plantation, mulch management and rootstocks in a "Pêra" orange orchard in northwest Paraná, Brazil. An experiment in a split-split-plot design was established in 2002, in an area cultivated with Brachiaria brizantha grass in which three tillage systems (no tillage, conventional tillage and strip-tillage) were used for orchard plantation. This grass was mowed twice a year between the rows, representing two mulch managements in the split plots (no mulching and mulching in the plant rows). The split-split-plots were represented by two rootstocks ("Rangpur" lime and "Cleopatra" mandarin). The soil water content in the plant rows was evaluated in the 0-20 cm layer in 2007 and at 0-20 and 20-40 cm in 2008-2009. The effect of soil tillage systems prior to implantation of orange orchards on soil water availability was less pronounced than mulching and the rootstocks. The soil water availability was lower when "Pêra" orange trees were grafted on "Cleopatra" mandarin than on "Rangpur" lime rootstocks. Mulching had a positive influence on soil water availability in the sandy surface layer (0-20 cm) and sandy clay loam subsurface (20-40 cm) of the soil in the spring. The production of B. brizantha between the rows and residue disposal in the plant rows as mulch increased water availability to the "Pêra" orange trees.

Soil microbial biomass under different management and tillage systems of permanent intercropped cover species in an orange orchard

To mitigate soil erosion and enhance soil fertility in orange plantations, the permanent protection of the inter-rows by cover species has been suggested. The objective of this study was to evaluate alterations in the microbial biomass, due to different soil tillage systems and intercropped cover species between rows of orange trees. The soil of the experimental area previously used as pasture (Brachiaria humidicola) was an Ultisol (Typic Paleudult) originating from Caiuá sandstone in the northwestern part of the State of Paraná, Brazil. Two soil tillage systems were evaluated: conventional tillage (CT) in the entire area and strip tillage (ST) (strip width 2 m), in combination with different ground cover management systems. The citrus cultivar 'Pera' orange (Citrus sinensis) grafted onto 'Rangpur' lime rootstock was used. Soil samples were collected after five years of treatment from a depth of 0-15 cm, under the tree canopy and in the inter-row, in the following treatments: (1) CT and an annual cover crop with the leguminous species Calopogonium mucunoides; (2) CT and a perennial cover crop with the leguminous peanut Arachis pintoi; (3) CT and an evergreen cover crop with Bahiagrass Paspalum notatum; (4) CT and a cover crop with spontaneous Brachiaria humidicola grass vegetation; and (5) ST and maintenance of the remaining grass (pasture) of Brachiaria humidicola. Soil tillage and the different cover species influenced the microbial biomass, both under the tree canopy and in the inter-row. The cultivation of brachiaria increased C and N in the microbial biomass, while bahiagrass increased P in the microbial biomass. The soil microbial biomass was enriched in N and P by the presence of ground cover species and according to the soil P content. The grass species increased C, N and P in the soil microbial biomass from the inter-row more than leguminous species.

Production of annual winter forage sown before and after soybean harvest under different nitrogen fertilization levels

The objective of this work was to evaluate the effect on forage yield of sowing winter forage species before and after soybean harvest, at different nitrogen application levels. The experiment was set out in a randomized block design with a strip-split plot arrangement, and three replicates. Sowing methods (18 days before soybean harvest and six days after soybean harvest) were allocated in the main plots, and the combination among forage species (Avena strigosa cv. IAPAR 61 + Lolium multiflorum; A. strigosa cv. Comum + L. multiflorum; A. strigosa cv. Comum + L. multiflorum + Vicia villosa; A. strigosa cv. Comum + L. multiflorum + Raphanus sativus; and L. multiflorum) and nitrogen levels (0, 140, 280 and 420 kg ha-1) in the plots and subplots, respectively. Forage sowing before the soybean harvest made it possible to anticipate first grazing by 14 days, with satisfactory establishment of forage species without affecting forage production. This method permitted a longer grazing period, preventing the need for soil disking, besides allowing the use of no-tillage system. The mixture of forage species enables higher forage yield for pasture in relation to single species pastures, with response to nitrogen fertilization up to 360 kg ha-1.

VARIATION OF MICROFIBRIL ANGLE OF Pinus radiata D. Don IN RELATION TO TREE SPACING IN CHILEAN PLANTATIONS

ABSTRACTFour stands of 28-year-old radiata pine (Pinus radiata D. Don) grown in the eighth region (Biobio) of Chile were sampled to determine the effect of tree spacing on the microfibril angle. The samples were taken at two different stem levels of the tree, 2.5 m and 7.5 m, with increment strip taken in the Nothern direction. The four experimental stands were characterized by the following spacing 2x2, 2x3, 3x4 and 4x4. The microfibril angle was measured by X-ray diffraction with the SilviScan technology at the FP-Innovation-Paprican Division in Vancouver, Canada. The results showed a significant effect of tree spacing on the microfibril angle in both juvenile wood and mature wood as well as at the two stem levels considered. The minimum (9.42º) was reached in 2x2 stand at 7.5 m in mature wood, while maximum microfibril angle (24.54º) was obtained in 2x3 stand at 2.5 m in juvenile wood. Regarding the effect of tree spacing, 4x4 stand had the lowest microfibril angle,except in mature wood at 7.5 m where 4x4 had the highest microfibril angle (11°) of the four stands.

Water storage in wetted strips under irrigated coffee trees with different criteria of irrigation management

The increasing demand for water resources accentuates the need to reduce water waste through a more appropriate irrigation management. In the particular case of irrigated coffee planting, which in recent years presented growth with the predominance of drip irrigation, the improvement of drip irrigation management techniques is a necessity. The proper management of drip irrigation depends on the knowledge of the spatial pattern of soil moisture distribution inside the wetted strip formed under the irrigation lines. In this study, grids of 24 tensiometers were used to determine the water storage within the wetted strip formed under drippers, with a 3.78 L h-1 discharge, evenly spaced by 0.4 m, subjected to two different management criteria (fixed irrigation interval and 60 kPa tension). Estimates of storage based on a one-dimensional analysis, that only considers depth variations, were compared with two-dimensional estimates. The results indicate that for high-frequency irrigation the one-dimensional analysis is not appropriate. However, under less frequent irrigation, the two-dimensional analysis is dispensable, being the one-dimensional sufficient for calculating the water volume stored in the wetted strip.

Papillary neoplasias of the biliary tract

The authors conducted a revisional study of intraepithelial papillary lesions of the bile ducts, characterized by being a kind of rare, intraductal growing cholangiocarcinoma. Articles published in the last 10 years were reviewed. The authors considered that the adenoma-carcinoma development is an important feature to warrant prophylactic measures through excisions. The histological type and biomolecular behavior may have relevance in the postoperative course of such lesions, which have a better prognosis when compared with other histological types.

Evaluation of tear production with modified Schirmer tear test-1 during the neonatal period in cats

The study aimed to evaluate tear production by means of modified Schirmer tear tes-1 (mSTT-1) in neonate cats. Likewise, correlation between mSTT-1 and STT-1 was assessed in vitro. Standard SST strips were cut in half and after eye lid opening, tear production of neonates (n=15) was daily measured in both eyes (mSTT-1), until the 7th day, and at day 14, 21, and 28. Animals were daily weighted until 28 days of age. Results were compared statistically (P<0.05). During the first 7 days, the overall mSTT-1 mean was 0.76 wetting/minute. Significant differences between right and left eyes were not observed at any time point (P=1.00). Tear secretion increased significantly, from the 14th to 28th day, in comparison with 7 first days (P<0.05). Positive correlation between maturity parameters and tear secretion was observed (P<0.0001). Distance between slopes of each strip changed significantly (P<0.0001). It was concluded that tear secretion in the neonatal period of cats is very below the reference values described for young and adults of the same species. It is not possible to extrapolate results obtained with mSTT-1 to standard STT-1.

Comparison of two methods of tear sampling for protein quantification by Bradford method

The aim of this study was to compare two methods of tear sampling for protein quantification. Tear samples were collected from 29 healthy dogs (58 eyes) using Schirmer tear test (STT) strip and microcapillary tubes. The samples were frozen at -80ºC and analyzed by the Bradford method. Results were analyzed by Student's t test. The average protein concentration and standard deviation from tears collected with microcapillary tube were 4.45mg/mL ±0.35 and 4,52mg/mL ±0.29 for right and left eyes respectively. The average protein concentration and standard deviation from tears collected with Schirmer Tear Test (STT) strip were and 54.5mg/mL ±0.63 and 54.15mg/mL ±0.65 to right and left eyes respectively. Statistically significant differences (p<0.001) were found between the methods. In the conditions in which this study was conducted, the average protein concentration obtained with the Bradford test from tear samples obtained by Schirmer Tear Test (STT) strip showed values higher than those obtained with microcapillary tube. It is important that concentration of tear protein pattern values should be analyzed according the method used to collect tear samples.

Clinical and pathological aspects of chronic Senecio spp. poisoning in sheep

This paper describes an outbreak of chronic Senecio spp. poisoning in grazing sheep in Rio Grande do Sul, Brazil, causing the death of 10 out of 860 adult sheep. Eight sick ewes were euthanized and necropsied. Cattle from this farm were also affected. Clinical signs included progressive weight loss, apathy and photosensitization. Four out of seven tested sheep had increased gamma-glutamyl transferase serum activity and two of them presented serum elevation of alkaline phosphatase. At necropsy, three out of eight ewes presented slightly irregular toughened livers with multifocal nodules, two out of eight ewes had a whitish liver with thickened fibrotic Glisson's capsule partially adhered to the diaphragm, and three out of eight ewes had smooth and grossly normal livers. Necropsy findings attributed to liver failure included hydropericardium (7/8), ascites (5/8), icterus (2/8), hydrothorax (1/8), and edema of mesentery (1/8). The main hepatic histological findings that allowed the establishment of the diagnosis were megalocytosis, proliferation of bile ducts and fibrosis. Spongy degeneration was observed in the brains of all eight necropsied sheep and was more severe at the cerebellar peduncles, mesencephalon, thalamus, and pons. These are suggested as the portions of election to investigate microscopic lesions of hepatic encephalopathy in sheep with chronic seneciosis. The diagnosis of Senecio spp. poisoning was based on epidemiology, clinical signs, laboratory data, necropsy and histological findings.

Expression of CD14 and toll-like receptors 2 and 4 by milk neutrophils in bovine mammary glands infected with Corynebacterium bovis

This study evaluated the expression of CD14, toll-like receptor (TLR) 2 and TLR4 on the surface of milk neutrophils in bovine mammary glands infected with Corynebacterium bovis. Here, we used 23 culture-negative control quarters with no abnormal secretion on the strip cup test and milk somatic cell count lower than 1x105 cells/mL, and 14 C. bovis infected quarters. The identification of neutrophils, as well as, the percentage of neutrophils that expressed CD14, TLR2 and TLR4 were analyzed by flow cytometry using monoclonal antibodies. The present study encountered no significant difference in the percentages of milk neutrophils that expressed TLR2 and TLR4 or in the expression of TLR4 by milk neutrophils. Conversely, a lower median fluorescence intensity of TLR2 in milk neutrophils was observed in C. bovis-infected quarters. The percentage of neutrophils that expressed CD14 and the median fluorescence intensity of CD14 in milk neutrophils was also lower in C. bovis-infected quarters.

Research of Klebsiella pneumoniae in dairy herds

Klebsiella pneumoniae is a common environmental agent of clinical and subclinical mastitis affecting dairy herds, and may be present in the final product decreasing its quality. Mastitis caused by K. pneumoniae is even more severe due to its poor response to antibiotic therapy, rapid evolution to toxic shock and death of the animal. This paper aimed to study the prevalence of this pathogen among dairy herds in ten farms located in different municipalities of São Paulo State based on size and use of milking technology. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20mL) was collected from all CMT-positive quarters and bulk tanks, whereas swab samples were collected from feces, hind limbs of the animals, bedding and milking parlor. Identification of K. pneumoniae was performed using conventional microbiology culture, biochemical assay and Polimerase Chain Reaction. The primers were designed and tested at the Laboratory of Molecular Biology applied to Zoonoses (FMVZ, Unesp-Botucatu) targeting the 16S rRNA gene. This study included 1067 animals. Six cases of intramammary infection by K. pneumoniae were detected in six different cows in two farms. Moreover, K. pneumoniae was isolated in 77 swabs (34 from bedding in 9 farms, 7 from waiting rooms in 5 farms, 6 from milking parlors in 4 farms, 11 from rectums in six farms, and 19 from hindlimbs in 7 farms. Molecular analysis confirmed the agent was K. pneumoniae. At least one strain of the agent was identified in a certain site in all farms, showing the need of maintaining the hygiene in dairy farms.

Pathological and parasitological aspects of the peacock (Pavo cristatus) infection by Tanaisia(Paratanaisia)bragai

Abstract:Trematodes belonging to the family Eucotylidae, including Tanaisia(Paratanaisia)bragaiSantos, 1934are parasites of the kidney and ureter that affect several species of domestic and wild birds. Tanaisia bragaiis considered a low pathogenic parasite, but high worm burdens may determine clinical complications, including signs of apathy, weight loss, diarrhea and death. This paper describes the first report of infection by T. bragai in peacocks (Pavo cristatus), which constitutes a new host record and offers data on the lesions associated to this parasitism, although the degree of pathogenicity and parasite load may be considered mild. These birds did not exhibit clinical signs of parasitism. The macroscopic exam revealed discreet yellow spots on the liver. In the histological sections of the kidney, specimens of T. bragai were found in the collecting ducts, which were markedly dilated, with a thickened wall. Other findings included a mild inflammatory reaction in the wall of the ducts (but sometimes absent), flattening of lining epithelial cells and small, multifocal points of calcification around the collecting ducts. The microscopic examination of the parasites revealed trematodes with an elongated body, well-developed sub terminal oral sucker, pharynx present, short esophagus, cecum somewhat undulating or not, with blind end, testes symmetrical, equatorial, irregular in shape or slightly lobed, vitelline fields extending in both pre-ovarian and post ovarian fields, uterus very long, intercecal or sometimes overlapping the cecum and containing large quantities of eggs. The present findings suggest the need for further diagnostic studies on the prevalence of this trematode in peacocks as well as pathologic studies for the determination of the potential pathogenicity of this parasite in this species of bird. Moreover, infected peacocks could serve as carriers of T. bragai to be transferred to other bird species, thereby contributing to the dispersion of the parasite.