Evaluation of modified vaccinia virus Ankara expressing VP2 protein of infectious bursal disease virus as an immunogen in chickens

A recombinant modified vaccinia Ankara (MVA) virus expressing mature viral protein 2 (VP2) of the infectious bursal disease virus (IBDV) was constructed to develop MVA-based vaccines for poultry. We demonstrated that this recombinant virus was able to induce a specific immune response by observing the production of anti-IBDV-seroneutralizing antibodies in specific pathogen-free chickens. Besides, as the epitopes of VP2 responsible to induce IBDV-neutralizing antibodies are discontinuous, our results suggest that VP2 protein expressed from MVA-VP2 maintained the correct conformational structure. To our knowledge, this is the first report on the usefulness of MVA-based vectors for developing recombinant vaccines for poultry.     

Multiple strains of Mycobacterium bovis revealed by molecular typing in a herd of cattle

Mycobacterium bovis isolates from an outbreak of bovine tuberculosis in a herd of cattle in Rio de Janeiro, Brazil, were analysed by spoligotyping and variable-number tandem repeat PCR analysis of the mycobacterial interspersed repetitive unit and exact tandem repeats. Molecular typing revealed a high genetic diversity of strains in the herd. The genetic diversity could be explained by the introduction of infected animals from different sources.     

Ovarian and endocrine responses associated with the treatment of cystic ovarian follicles in dairy cows with gonadotropin releasing hormone and prostaglandin F2alpha, with or without exogenous progesterone

The objectives of this observational study were to document ovarian and endocrine responses associated with the treatment of cystic ovarian follicles (COFs) in dairy cows, using gonadotropin releasing hormone (GnRH) and prostaglandin F2alpha (PGF) with or without exogenous progesterone. A secondary objective was to determine pregnancy establishment following synchronization of ovulation and timed insemination in cows diagnosed with COFs. In trial I, 18 Holstein cows diagnosed with COFs received 2 injections of 100 microg GnRH, 9 d apart, with 25 mg PGF given 7 d after the 1st GnRH. A new follicle developed in all 18 cows after the 1st GnRH, and 83% of cows ovulated following the 2nd GnRH. Cows were inseminated 16 h after the 2nd GnRH. Of the 17 cows available for pregnancy diagnosis, 7 were confirmed pregnant. In trial II, 8 cows with COFs received GnRH and an intravaginal progesterone device (CIDR) concurrently, then PGF 7 d later. The CIDR was removed 2 d after PGF administration. Plasma estradiol concentrations declined following CIDR insertion. In all cows, a new follicle developed following GnRH treatment; estradiol-surge and estrus occurred spontaneously after CIDR-removal. Seven of 8 cows ovulated the new follicle. In dairy cows diagnosed with COFs, treatment with GnRH followed by PGF 7 d later, with or without exogenous progesterone, resulted in the recruitment of a healthy new follicle; synchronization of ovulation and timed insemination resulted in a 41% pregnancy rate.     

Marine-Derived Penicillium purpurogenum Reduces Tumor Size and Ameliorates Inflammation in an Erlich Mice Model

Background: This study addresses the antitumoral properties of Penicillium purpurogenum isolated from a polluted lagoon in Northeastern Brazil. Methods: Ethyl Acetate Extracellular Extract (EAE) was used. The metabolites were studied using direct infusion mass spectrometry. The solid Ehrlich tumor model was used for antitumor activity. Female Swiss mice were divided into groups (n = 10/group) as follows: The negative control (CTL−), treated with a phosphate buffered solution; the positive control (CTL+), treated with cyclophosphamide (25 mg/kg); extract treatments at doses of 4, 20, and 100 mg/kg; animals without tumors or treatments (Sham); and animals without tumors treated with an intermediate dose (EAE20). All treatments were performed intraperitoneally, daily, for 15 days. Subsequently, the animals were euthanized, and the tumor, lymphoid organs, and serum were used for immunological, histological, and biochemical parameter evaluations. Results: The extract was rich in meroterpenoids. All doses significantly reduced tumor size, and the 20 and 100 mg/kg doses reduced tumor-associated inflammation and tumor necrosis. The extract also reduced the cellular infiltration of lymphoid organs and circulating TNF-α levels. The extract did not induce weight loss or renal and hepatic toxic changes. Conclusions: These results indicate that P. purpurogenum exhibits immunomodulatory and antitumor properties in vivo. Thus, fungal fermentation is a valid biotechnological approach to the production of antitumor agents.     

Cercosporiosis resistance in coffee germplasm collection

Cercosporiosis, or brown eye spot, is currently one of the main diseases of the coffee tree. It is caused by Cercospora coffeicola Berk. & Cooke. Nevertheless, genetic resistance to this disease has not yet been explored in any depth. Our objectives (a) were evaluate the response of 124 accessions from the germplasm collection of the Minas Gerais State (GC), Brazil, and eight commercial cultivars of C. arabica to cercosporiosis and (b) determine the best way to perform early progenies selections via controlled greenhouse experiments. Three controlled greenhouse experiments (1–3) were run in different seasons to determine the best way to proceed the selection. The seedlings were inoculated with a four isolates mixture obtained from different regions. The experimental data were analyzed individually (1–3), in a joint analysis, and as repetitions of a randomized complete block design. In each analysis we estimated genetic parameters and E-BLUP (empirical best linear unbiased predictor) genotypic values of the access. There was genetic variability to C. coffeicola resistance among the coffee tree accessions germplasm collection. Therefore, genetic improvements could be obtained by selection. Experimental repetitions in different seasons increase the selection efficiency and reliability of resistant genotypes with low cercosporiosis severity. The genotype Sarchimor MG 8840 showed the highest resistance level followed by Guatenano and the Timor Hybrid UFV 377-34, Timor Hybrid UFV 376-14 BE 5, and Wush–Wush × Timor Hybrid UFV 366-08.     

The cardiopulmonary effects of a peripheral alpha‐2‐adrenoceptor antagonist,MK‐467, in dogs sedated with a combination of medetomidine and butorphanol

ObjectiveTo compare the cardiopulmonary effects of intravenous (IV) and intramuscular (IM) medetomidine and butorphanol with or without MK-467.Study designProspective, randomized experimental cross-over.AnimalsEight purpose–bred beagles (two females, six males), 3–4 years old and weighing 14.5 ±1.6 kg (mean ± SD).MethodsAll dogs received four different treatments as follows: medetomidine 20 μg kg^?1 and butorphanol tartrate 0.1 mg kg^?1 IV and IM (MB), and MB combined with MK-467,500 μg kg^?1 (MBMK) IV and IM. Heart rate (HR), arterial blood pressures (SAP, MAP, DAP), central venous pressure (CVP), cardiac output, respiratory rate (f_R), rectal temperature (RT) were measured and arterial blood samples were obtained for gas analysis at baseline and at 3, 10, 20, 30, 45 and 60 minutes after drug administration. The cardiac index (CI), systemic vascular resistance index (SVRI) and oxygen delivery index (DO₂I) were calculated. After the follow-up period atipamezole 50 μg kg^?1 IM was given to reverse sedation.ResultsHR, CI and DO₂I were significantly higher with MBMK after both IV and IM administration. Similarly, SAP, MAP, DAP, CVP, SVRI and RT were significantly lower after MBMK than with MB. There were no differences in f_R between treatments, but arterial partial pressure of oxygen decreased transiently after all treatments. Recoveries were uneventful following atipamezole administration after all treatments.Conclusions and clinical relevanceMK-467 attenuated the cardiovascular effects of a medetomidine-butorphanol combination after IV and IM administration.     

Phenolic antioxidants and the protection of low density lipoprotein from peroxynitrite-mediated oxidations at physiologic CO2

Dietary phenolic antioxidants have been shown to prevent LDL modifications mediated by several physiologic oxidants including peroxynitrite. However, more recent data demonstrated that CO(2) affected the fate of peroxynitrite in biological fluids and significantly reduced peroxynitrite scavenging by polyphenols, raising doubts concerning their antioxidant activity. We found that the oxidation of LDL lipids mediated by peroxynitrite decreased in the presence of bicarbonate, while Trp oxidation and 3-nitroTyr formation increased, suggesting a redirection of peroxynitrite reactivity toward the protein moiety. We therefore evaluated the protective activity of some phenolic antioxidants (quercetin, oleuropein, resveratrol, (+)-catechin, (-)-epicatechin, tyrosol, alpha- and gamma-tocopherol, ascorbate) on peroxynitrite-mediated oxidation of LDL aromatic residues. Some of these phenols protected LDL Trp from oxidation better than ascorbate or alpha-tocopherol, although protection at 100 microM did not exceed 30-40%. However, the same phenolic antioxidants were more active in inhibiting 3-nitroTyr formation and those with a catechin structure provided significant protection (IC(50%) 40-50 microM). Red wine, a polyphenol-rich beverage, showed a protective effect comparable to that of the most active phenolic antioxidants. Direct EPR studies showed that bicarbonate significantly increased the peroxynitrite-dependent formation of O-semiquinone radicals in red wine, supporting the hypothesis that polyphenols are efficient scavengers of radicals formed by peroxynitrite/CO(2). Ascorbate was a poor inhibitor of peroxynitrite/CO(2)-induced LDL tyrosine nitration, but the simultaneous addition to the most active polyphenols halved their IC(50%). In conclusion, although cooperation with other antioxidants can further decrease the IC(50%) of polyphenolics, as demonstrated for ascorbate, their antioxidant activity appears to occur at concentrations at least 1 order of magnitude higher than their bioavailability.     

Single superphosphate fertilizer distribution among seeder-fertilizer rows

The objective was to verify the uniformity of single superphosphate distribution among seeder-fertilizer rows and its consequences on crops. The experiment was installed in the municipality of Tibagi—PR, involving the following crop rotation in the same area: black oat (Avena strigosa) in 2012, corn (Zea mays) in the 2012–13 season, wheat (Triticum aestivum) in 2013, soybeans (Glycine max) in the 2013–14 season, black oat in 2014 and corn closing the cycle in the 2014–15 season. The experimental design was used in randomized blocks, considering the number of rows of the seeder-fertilizer (ranging from 11 for corn to 27 for wheat) as treatments; with three replicates. During the experimental period, 250 kg ha^?1 of single superphosphate were distributed per crop at sowing, with a mean deviation of up to 14% and variation in the fertilizer mass among the seeder-fertilizer rows in up to 52%. The variables evaluated were initial population and income components. It was concluded that it is not possible to correlate the variation of fertilizer distribution among rows of the seeder-fertilizer with the variables that presented significant differences in the crops through three agricultural seasons. This fact may be related to the alternation of the highest and lowest doses of simple superphosphate. Due to the succession of seeder-fertilizers used over the years to crops seeding, there was a compensation for the random variation of fertilizer among rows and crop spacings along the crop seasons.

     

Natural progression of Bovine Leukemia Virus infection in Argentinean dairy cattle

We describe the progression of Bovine Leukemia Virus (BLV) infection from birth until the first lactation in 61 animals from a typical large dairy herd of Argentina, with more than 85% of prevalence. The purpose was to identify potential points to effectively break the BLV cycle of transmission in our dairy productive system. We detected early infection in 11.47% of newborn calves by nested PCR. From birth to 12 months, no evidence of new infections was observed. After 12 months of age, the detection of new reactors increased slowly with time, from 15.09% at 15 months to 24% at 27 months. After that, the number of reactors increased rapidly up to 40% and 60.76% at 30 and 36 months, respectively. This last 9-month period coincided with parturition and the entry into the milking herd. Real-time PCR showed that more than 75% of adult animals had low peripheral-blood proviral load. Complementary, all infected animals showed low levels of provirus in milk and colostrum. The most important finding was that even when management procedures to prevent BLV iatrogenic transmission were followed, no significant change was observed in the prevalence after three years, strongly suggesting that other way/s of transmission play a key role under natural conditions. This study showed an interesting baseline to draw an alternative approach based on selective segregation according to the peripheral-blood proviral load as a potential indicator of risk transmission, and as an alternative to classical control measures.