Development and standardization of an indirect ELISA for the serological diagnosis of classical swine fever

An indirect enzyme linked immunoassay (ELISA-I) was developed and standardized for the serological diagnosis of classical swine fever (CSF). For the comparison, nine hundred and thirty-seven swine serum samples were tested by serum neutralization followed by immunoperoxidase staining (NPLA), considered as the standard. Of these, 223 were positive and 714 negative for neutralizing antibodies to classical swine fever virus (CSFV). In relation to the NPLA, the ELISA-I presented a 98.2% sensitivity; 92.86% specificity, 81.11% positive predictive value, 99.4% negative predictive value and a 94.1% precision. Statistical analysis showed a very strong correlation (r=0,94) between both tests. When compared to a commercially available ELISA kit, the performance of both, in relation to the NPLA, was similar. It was concluded that the ELISA-I is suitable for large scale screening of antibodies to classical swine fever virus, although it does not distinguish antibodies to classical swine fever virus from those induced by other pestiviruses.

An outbreak of malignant catarrhal fever in Murrah buffaloes in Minas Gerais, Brazil

An outbreak of Malignant Catarrhal Fever (MCF) resulted in death of five female buffaloes and one domestic cow from the same farm. Four buffaloes died 10-15 days after the appearance of clinical signs, while the fifth was euthanized in extremis, after similar clinical signs. Histopathological lesions included multifocal histiolymphocytic epicarditis, myocarditis and lymphocytic interstitial pneumonia, which are commonly seen in cases of MCF in buffaloes. Furthermore, lymphocytic vasculitis centered in the adventitia, with occasional fibrinoid necrosis in the muscular layer, was found in the kidneys, liver, spleen, lymph nodes and brain. Nucleotide sequencing of DNA fragments from the central nervous system amplified by PCR revealed 98% similarity with known OHV-2 sequences from Genbank. Additionally, PCR analysis also revealed the presence of OHV-2 DNA in the peripheral mononuclear blood cells of two clinically healthy buffaloes. The diagnosis of MCFwas based on epidemiological, clinical, gross and histopathological findings and on the results of a semi-nested PCR followed by nucleotide sequencing.

Molecular confirmation of ovine herpesvirus 2-induced malignant catarrhal fever lesions in cattle from Rio Grande do Norte, Brazil

Molecular findings that confirmed the participation of ovine herpesvirus 2 (OVH-2) in the lesions that were consistent with those observed in malignant catarrhal fever of cattle are described. Three mixed-breed cattle from Rio Grande do Norte state demonstrated clinical manifestations that included mucopurulent nasal discharge, corneal opacity and motor incoordination. Routine necropsy examination demonstrated ulcerations and hemorrhage of the oral cavity, corneal opacity, and lymph node enlargement. Significant histopathological findings included widespread necrotizing vasculitis, non-suppurative meningoencephalitis, lymphocytic interstitial nephritis and hepatitis, and thrombosis. PCR assay performed on DNA extracted from kidney and mesenteric lymph node of one animal amplified a product of 423 base pairs corresponding to a target sequence within the ovine herpesvirus 2 (OVH-2) tegument protein gene. Direct sequencing of the PCR products, from extracted DNA of the kidney and mesenteric lymph node of one cow, amplified the partial nucleotide sequences (423 base pairs) of OVH-2 tegument protein gene. Blast analysis confirmed that these sequences have 98-100% identity with similar OVH-2 sequences deposited in GenBank. Phylogenetic analyses, based on the deduced amino acid sequences, demonstrated that the strain of OVH-2 circulating in ruminants from the Brazilian states of Rio Grande do Norte and Minas Gerais are similar to that identified in other geographical locations. These findings confirmed the active participation of OVH-2 in the classical manifestations of sheep associated malignant catarrhal fever.

Isospora bocamontensis (Protozoa: Apicomplexa) in captive yellow cardinal Gubernatrix cristata (Passeriformes: Emberezidae)

The yellow cardinal (Gubernatrix cristata) is a passerine found in southern Brazil, especially along the border with Uruguay and Argentina. It is an endangered species and its population is decreasing. Among the parasites that affect passerines, the genus Isospora is the most easily found in both captive and free-living birds. This parasite commonly causes injury to the intestinal tissue and could occasionally affect other organs. In this work we examined the occurrence of coccidiosis in captive yellow cardinals and its association with factors such as sex, use of parasiticides, type of enclosure, contact with feces, type of food and cleaning frequency. We collected fecal samples of 45 yellow cardinals, healthy and kept in captivity, in late afternoon at the end of the reproductive period. The examination showed parasitic infection by Isospora bocamontensis in 44.5% of the birds. This infection is not influenced by the sex of birds, but is significantly affected by the type of enclosure, contact with the feces, use of parasiticides, type of food and cleaning frequency. The results indicate that to keep yellow cardinals captive, these factors must be observed.

Celiotomy by plastrotomy in a yellow-footed tortoise (Geochelone denticulata)

One herein reports a successful case of celiotomy by plastrotomy for removal of foreign bodies in yellow-footed tortoise (Geochelone denticulata). The animal was treated at the Veterinary Hospital of the Federal University of Piaui, with appetite loss, regurgitation, constipation, lethargy, reluctance to walk and slightly reddish ocular mucous membranes. Radiographic examination was performed, confirming the presence of foreign bodies in the stomach. The tortoise underwent celiotomy by plastrotomy for the removal of the foreign bodies. The opening of the plastron was performed through the abdominal shields, with the aid of a circular mini grinding saw. One performed an incision in the midline between the two abdominal veins to access the abdominal cavity. A gastrotomy for removal of the foreign bodies (nails, toothpicks, stones, pieces of plastic, glass and crockery pieces) was performed after the location of the stomach. The surgery was successful and confirmed with radiographic evaluation in the immediate postoperative period. The celiotomy by plastrotomy for removal of foreign bodies in that animal proved to be a viable, very important and safe technique to the survival of chelonians.

Influence of environmental factors on seed germination and seedling emergence of yellow sweet clover (Melilotus officinalis)

Laboratory and greenhouse experiments were conducted to determine the effects of drought and salinity stress, temperature, pH and planting depth on yellow sweet clover (Melilotus officinalis) germination and emergence. Base, optimum and ceiling germination temperatures were estimated as 0, 18.47 and 34.60 ºC, respectively. Seed germination was sensitive to drought stress and completely inhibited at a potential of -1 MPa, but it was tolerant to salinity. Salinity stress up to 90 mM had no effect over the M. officinalis seed germination, but the germination decreased by increasing the salt concentration. The drought and salinity required for 50% inhibition of maximum germination were 207 mM and -0.49 MPa, respectively. High percentage of seed germination (>92%) was observed at pH = 5-6 and decreased to 80% at acidic medium (pH 4) and to 42% at alkaline medium (pH 9) pH. Maximum seedling emergence occurred when the seeds were placed at 2 cm depth and decreased when increasing the depth of planting; no seed emerged from depths of 10 cm.

Initial growth and competitive ability of yellow nutsedge and irrigated rice

Weeds cause significant reduction in the irrigated rice crop yield. Cyperus esculentus (yellow nutsedge) is adapted to irrigate environment. Information on the competitive ability of the weed to the culture, and their environmental adaptation, are scarce. In this study, we sought to determine the initial growth and competitive ability of yellow nutsedge and irrigated rice, as a function of cultivar growth cycle. Initial growth and competition studies were conducted in a randomized complete design in a greenhouse in the agricultural year 2010/11. For the initial growth study, the treatments consisted of a factorial combination of a biotype of yellow nutsedge and two rice cultivars in the function of the vegetative cycle (BRS Querência: early cycle - IRGA 424: intermediate cycle) and six evaluation times (10, 20, 30, 40, 50, and 60 days after emergence). Were evaluated: plant height, leaf area, aboveground dry biomass and root dry biomass. In the competitive ability study in the replacement series, the cultivar BRS Querência (early cycle) and yellow nutsedge were utilized and tested in different proportions of competition (100:0, 75:25, 50:50, 25:75, and 0:100). Were evaluated leaf area and aboveground dry biomass. In general, rice cultivars have an adaptive value equivalent to yellow nutsedge. IRGA 424 cultivar has less height than weed, becoming the weed control more important in this cultivar. For rice crop, intraspecific competition is more important, whereas for the weed, interspecific competition is the most pronounced.

Effects of a neuronal nitric oxide synthase inhibitor on lipopolysaccharide-induced fever

It has been demonstrated that nitric oxide (NO) has a thermoregulatory action, but very little is known about the mechanisms involved. In the present study we determined the effect of neuronal nitric oxide synthase (nNOS) inhibition on thermoregulation. We used 7-nitroindazole (7-NI, 1, 10 and 30 mg/kg body weight), a selective nNOS inhibitor, injected intraperitoneally into normothermic Wistar rats (200-250 g) and rats with fever induced by lipopolysaccharide (LPS) (100 µg/kg body weight) administration. It has been demonstrated that the effects of 30 mg/kg of 7-NI given intraperitoneally may inhibit 60% of nNOS activity in rats. In all experiments the colonic temperature of awake unrestrained rats was measured over a period of 5 h at 15-min intervals after intraperitoneal injection of 7-NI. We observed that the injection of 30 mg/kg of 7-NI induced a 1.5oC drop in body temperature, which was statistically significant 1 h after injection (P<0.02). The coinjection of LPS and 7-NI was followed by a significant (P<0.02) hypothermia about 0.5oC below baseline. These findings show that an nNOS isoform is required for thermoregulation and participates in the production of fever in rats.

Fever induction pathways: evidence from responses to systemic or local cytokine formation

The immune and central nervous systems are functionally connected and interacting. The concept that the immune signaling to the brain which induces fever during infection and inflammation is mediated by circulating cytokines has been traditionally accepted. Administration of bacterial lipopolysaccharide (LPS) induces the appearance of a so-termed "cytokine cascade" in the circulation more or less concomitantly to the developing febrile response. Also, LPS-like fever can be induced by systemic administration of key cytokines (IL-1ß, TNF-alpha, and others). However, anti-cytokine strategies against IL-1ß or TNF-alpha along with systemic injections of LPS frequently lead to attenuation of the later stages of the febrile response but not of the initial phase of fever, indicating that cytokines are rather involved in the maintenance than in the early induction of fever. Within the last years experimental evidence has accumulated indicating the existence of neural transport pathways of immune signals to the brain. Because subdiaphragmatic vagotomy prevents or attenuates fever in response to intraperitoneal or intravenous injections of LPS, a role for vagal afferent nerve fibers in fever induction has been proposed. Also other sensory nerves may participate in the manifestation of febrile responses under certain experimental conditions. Thus, injection of a small dose of LPS into an artificial subcutaneous chamber results in fever and formation of cytokines within the inflamed tissue around the site of injection. This febrile response can be blocked in part by injection of a local anesthetic into the subcutaneous chamber, indicating a participation of cutaneous afferent nerve signals in the manifestation of fever in this model. In conclusion, humoral signals and an inflammatory stimulation of afferent sensory nerves can participate in the generation and maintenance of a febrile response.

Use of inflammatory molecules to predict the occurrence of fever in onco-hematological patients with neutropenia

Febrile neutropenia remains a frequent complication in onco-hematological patients, and changes in the circulating level of inflammatory molecules (IM) may precede the occurrence of fever. The present observational prospective study was carried out to evaluate the behavior of plasma tumor necrosis factor alpha (TNF-α), soluble TNF-α I and II receptors (sTNFRI and sTNFRII), monocyte chemoattractant protein-1 [MCP-1 or chemokine (c-c motif) ligand 2 (CCL2)], macrophage inflammatory protein-1α (MIP-1α or CCL3), eotaxin (CCL11), interleukin-8 (IL-8 or CXCL8), and interferon-inducible protein-10 (IP-10 or CXCL10) in 32 episodes of neutropenia in 26 onco-hematological patients. IM were tested on enrollment and 24-48 h before the onset of fever and within 24 h of the first occurrence of fever. Eight of 32 episodes of neutropenia did not present fever (control group) and the patients underwent IM tests on three different occasions. sTNFRI levels, measured a median of 11 h (1-15) before the onset of fever, were significantly higher in patients presenting fever during follow-up compared to controls (P = 0.02). Similar results were observed for sTNFRI and CCL2 levels (P = 0.04 for both) in non-transplanted patients. A cut-off of 1514 pg/mL for sTNFRI was able to discriminate between neutropenic patients with or without fever during follow-up, with 65% sensitivity, 87% specificity, and 93% positive predictive value. Measurement of the levels of plasma sTNFRI can be used to predict the occurrence of fever in neutropenic patients.

Relationship between genetic mutation variations and acute-phase reactants in the attack-free period of children diagnosed with familial Mediterranean fever

Familial Mediterranean fever (FMF) is a periodic autoinflammatory disease characterized by chronic inflammation. This study investigated the relationship between acute-phase reactants and gene mutations in attack-free periods of childhood FMF. Patients diagnosed with FMF were divided into four groups based on genetic features: no mutation, homozygous, heterozygous, and compound heterozygous. These groups were monitored for 2 years, and blood samples were collected every 6 months during attack-free periods. Erythrocyte sedimentation rate, C-reactive protein, fibrinogen, and white blood cell count were measured. A disease severity score was determined for each patient. Mean values for erythrocyte sedimentation rate and fibrinogen were significantly different in the homozygous group. White blood cell count and C-reactive protein were similar between the groups. Disease severity score was higher in patients with the M694V mutation than in individuals without the mutation, as well as in those with other mutation groups. Periodic follow-up of patients with FMF MEFV mutations in subjects with acute-phase reactants may be useful in the prevention of morbidity.

Postharvest respiratory activity and changes in some chemical constituents during maturation of yellow mombin (Spondias mombin) fruit

Mature fruit from the yellow mombin (Spondias mombin) was monitored for its respiration activity. Mature green fruit from the yellow mombin was stored in closed glass chambers and the concentration of oxygen and carbon dioxide at the end of a six hour respiration period was determined. At the same interval of time, the lid of the chamber was opened for air renewal. The increase in carbon dioxide and decrease in oxygen concentration demonstrated that the fruit was climacteric. The maximum liberation of CO2 54.2 mL Kg-1 h-1 and maximum absorption of O2 49.0 mL Kg-1 h-1 occurred 186 hours after the harvest which, obviously, represented the optimum fruit quality after the senescence process started. The respiratory quotient of fruit at a climacteric maximum was 1.11 representing the oxidation of carbohydrates. Total soluble solids increased from 9.1 °Brix (initial) to 13.7 °Brix (climacteric maximum) during maturation, while the total number of acids in the fruit decreased during maturation i.e. from 1.55% initially to 1.40% at pre-climacteric, 1.0% at climacteric maximum and 0.8% in the post-climacteric stage. A similar behaviour was observed in the case of ascorbic acid. There was a continuous decrease in chlorophyll and a continuous increase in the carotenoid content of fruit during maturation.

Quality of yellow passion fruit stored under refrigeration and controlled atmosphere

In this study, it was evaluated the quality of yellow passion fruits stored under refrigeration and controlled atmospheres of different composition aiming to extend the postharvest life of the fruits. The characteristics of skin color, appearance, mass loss, as well as the chemical quality of the juice of yellow passion fruits stored at: 21% O2 plus 0.03% CO2; 1% O2 plus 0.03% CO2; 5% O2 plus 0.03% CO2; 12% O2 plus 5% CO2; and 5% O2 plus 15% CO2, with 1 control treatment (refrigeration at 13 ºC and 90% UR) were determined. The analyses were performed before and after 30 days of storage and after removing the controlled atmospheres and storage for 9 days under refrigeration at ambient atmosphere. The data were interpreted by simple statistical analysis using the test by confidence intervals with 95% of probability. It was concluded that the application of atmospheres with low oxygen concentration and high carbon dioxide level minimized quality losses. At atmosphere with 5% O2 and 15% CO2, it was observed the lowest color change indexes and mass loss, and also the smallest decrease in acidity, soluble solids content, vitamin C, reducing sugars, and total soluble sugars.

Yield of albedo flour and pectin content in the rind of yellow passion fruit

In this study, it was evaluated the influence of different shapes, sizes, and maturation stages on the yield of albedo flour and pectin content of yellow passion fruit rinds. Random samples of 40 fruits were used, and the data were compared using significance intervals at 5%. Weight, skin color, fruit size and shape, pulp yield, mesocarp thickness, amount of epicarp and mesocarp, moisture content, and pectin yield were determined. The maturation stages were defined according to measurements of the yellow color of the skin. The shape and size patterns were defined according to the length/width ratio (equatorial diameter) of fruits. It was found that the epicarp thickness was not correlated to fruit shape and size, but it was thicker in ripe fruits. The mesocarp was thiner in small ripe fruits, but it did not change with fruit shape. Pulp yield was higher in ripe fruits, and it was not influenced by shape and size of fruits. It was concluded that the content of albedo flour can account for 3.9% of the weight of processed fruits, whereas the amount of pectin powder can account for up to 0.9% of the fruit weight.

Bioactive compounds and antioxidant activity in fruits of clone and ungrafted genotypes of yellow mombin tree

Yellow mombin is a fruit tree that grows spontaneously in the Semi-Arid Northeastern Brazil. Its fruits are still extractively exploited. The pulp of yellow mombin fruit stands out regarding the commercial aspect due to the characteristic flavor and aroma felt when consumed in diverse ways. This study aimed to evaluate the presence of bioactive compounds, total extractable polyphenols, and antioxidant activity of yellow mombin fruits (Spondias mombin, L.), from clone and ungrafted genotypes. The fruits were harvested at commercial maturity from twelve yellow mombin tree genotypes from an experimental orchard located at the municipality of Joao Pessoa, Paraíba, Brazil, and evaluated for chlorophyll, carotenoids, yellow flavonoids, total extractable polyphenols, and antioxidant activity, which was measured by the β-carotene/linoleic acid method. The antioxidant activity showed a percentage of inhibition of oxidation higher than 75% for all genotypes evaluated at the time of 120 minutes. The fruits from clone genotypes showed a higher percentage of antioxidant activity.