Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.     

Recent Discoveries on Marine Organism Immunomodulatory Activities

Marine organisms have been shown to be a valuable source for biologically active compounds for the prevention and treatment of cancer, inflammation, immune system diseases, and other pathologies. The advantage of studying organisms collected in the marine environment lies in their great biodiversity and in the variety of chemical structures of marine natural products. Various studies have focused on marine organism compounds with potential pharmaceutical applications, for instance, as immunomodulators, to treat cancer and immune-mediated diseases. Modulation of the immune system is defined as any change in the immune response that can result in the induction, expression, amplification, or inhibition of any phase of the immune response. Studies very often focus on the effects of marine-derived compounds on macrophages, as well as lymphocytes, by analyzing the release of mediators (cytokines) by using the immunological assay enzyme-linked immunosorbent assay (ELISA), Western blot, immunofluorescence, and real-time PCR. The main sources are fungi, bacteria, microalgae, macroalgae, sponges, mollusks, corals, and fishes. This review is focused on the marine-derived molecules discovered in the last three years as potential immunomodulatory drugs.     

Integrated production of fish (pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> and red tilapia <Emphasis Type="Italic">Oreochromis</Emphasis> sp.) with two varieties of garnish (scallion and parsley) in aquaponics system

Aquaponics is emerging as an alternative for high-health food production. Being able to identify the technical viability of non-conventional plants and fish species would help to increase the interest and possibilities in aquaponic systems. The goal of the present study was to evaluate the aquaponics production of two garnish species: scallion (S) and parsley (P), using effluents of pacu and red tilapia culture. Two aquaponics devices were used, differing according to the fish species, generating two different effluents. Thus, for plant performance, four treatments were evaluated in a factorial design (plant species and fish effluent as main factors), as followed: Pacu-S, Tilapia-S, Pacu-P, and Tilapia-P, with three replicates each, for 35 days. Fish performance was evaluated using Student’s t test. Each experimental device included a fish tank, filters, and six experimental units for the plants (floating rafts). Results indicated that feed conversion ratio (FCR) was higher in tilapia as compared to pacu (p < 0.05); however, fish productivity and survival were similar between species. Plant performance parameters were similar with no significant differences regardless of the fish effluent (p > 0.05), except for higher number of leaves per plant in scallion cultured using pacu effluent. Plant performance comparing both plant species indicated that scallion performed better as compared to parsley in all parameters. In addition, scallion also performed better related to the plant quality index. The results indicate that pacu presented a viable alternative for the aquaponics production, and regarding to the garnish, scallion performed better results as compared to parsley.     

Serological response of guinea pigs to oily and aqueous inactivated vaccines containing a Brazilian isolate of the Bovine Viral Diarrhea Virus (BVDV)

Bovine Viral Diarrhea Virus (BVDV) is widespread in cattle in Brazil and research shows its large antigenic variability. Available vaccines are produced with virus strains isolated in other countries and may not be effective. In this study, inactivated vaccines containing the Brazilian BVDV-Ib IBSP11 isolate were developed and tested on 6 groups of 10 guinea pigs (Cavia porcellus). Animals in groups A and C received an aqueous vaccine (aluminum hydroxide); B and D groups received an oily vaccine (Montanide ISA50); Group E positive-control animals were given an imported commercial vaccine with BVDV-Ia Singer; Group F animals were sham vaccinated (negative control). Groups A, B and E received two doses, and Groups C and D, three, every 21 days. Twelve blood samples were taken, at 21-day intervals over 231 days, and evaluated for antibody titer through virus-neutralization (VN), using a homologous strain (IBSP11), and a heterologous strain (BVDV-Ia NADL). Most animals, 42 days following the first dose, seroconverted to both strains and, after the second dose, there was a significant increase of titers in all groups. The oily formulation induced greater response after the third administration. This increase was not observed with the aqueous vaccines, regardless of the virus used in the VN. Antibody decline was more rapid in animals that received aqueous vaccines. The results showed the importance of studying the influence of endemic strains of commercial vaccines, to improve the efficacy of BVD vaccination. Use of the endemic strain in vaccine formulation presented promising results, as well as the use of guinea pigs as a laboratory model.     

棉花纤维突变体胚珠发育过程中主要生化物质的积累特征

采用4种不同类型的棉花纤维突变体和一个正常对照TM-1,分析比较了胚珠发育过程中,纤维和种皮内纤维素,种仁内脂肪、可溶性糖和蛋白质等成份的积累特征及诸成份与纤维素含量的相关关系.结果表明,棉花纤维突变体的胚珠不仅生化物质动态变化有着较大的差异,而且其生化物质的组成差异也很大.纤维突变体的纤维素含量低,种仁蛋白质含量低,脂肪含量高.纤维素含量与种仁内脂肪含量呈显著或极显著正相关;与纤维可溶性糖,纤维蛋白质有显著的相关性,呈显著负相关.     

First report of porcine circovirus type 2 infections in Cuba

To obtain information about the porcine circovirus type 2 (PCV2) infection status of pigs in Cuba and the probable association of PCV2 with other porcine viruses, tissue samples collected from ill pigs were evaluated using polymerase chain reaction (PCR). The PCR analysis showed that 67.7% of the samples (23/34) from seven swine herds of six different geographic regions were detected to be positive for PCV2. Ten of the 23 PCV2 positive samples (43.5%) shown a concurrent infection with porcine parvovirus (PPV) and 17 of 23 PCV2 positive samples (73.9%) exhibited a concomitant infection with classical swine fever virus (CSFV). This study is the first report of PCV2 infecting pigs with different clinical conditions in Cuban swine herds and provides evidence of PCV2 co-infection with PPV and CSFV in the field.     

Growth performance of juvenile pacu (Piaractus mesopotamicus) and dourado (Salminus brasiliensis) fed with lyophilized bovine colostrum

Growth performance was evaluated in juvenile pacu (Piaractus mesopotamicus) and dourado (Salminus brasiliensis) fed diets containing 0%, 10% and 20% of lyophilized bovine colostrum for 60 days. The performance variables of weight gain, relative weight gain, feed intake, feed conversion ratio, specific growth rate, total protein intake and protein efficiency ratio were evaluated as well as apparent digestibility coefficients of protein and energy in the diets. Inclusion of bovine colostrum did not induce differences in the performance of pacu and dourado (P > 0.05), indicating that the diets were adequate and met the nutritional needs of the juveniles. The protein digestibility in both species was improved with the inclusion of the bovine lacteal secretion. Based on the variables studied, the diets with lyophilized bovine colostrum inclusion were nutritionally suitable for both pacu and dourado indicating the possibility to use this lacteal secretion as a partial substitute of dietary protein.     

Comparison of slot blot nucleic acid hybridization, immunofluorescence, and virus isolation techniques to detect bluetongue virus in blood mononuclear cells from cattle with experimentally induced infection.

A slot blot hybridization technique was applied for detection of bluetongue virus (BTV) in blood mononuclear cells (BMNC) obtained from cattle with experimentally induced infection. This technique lacked sensitivity to detect the viral nucleic acid directly in clinical specimens. When aliquots of mononuclear cells from these cattle were cultivated in vitro for 10 days to amplify virus titer, only 33.3% of the samples collected during viremia gave a positive signal in the slot blot hybridization format. By contrast, results for 34.3% of noncultured and 63.3% of cultured mononuclear cell samples collected during viremia were positive by immunofluorescence. The average number of infected cells, as detected by immunofluorescence in the noncultured mononuclear cell samples, was 1 to 5/300,000, and was usually > 10/300,000 in the cultured cell samples. Virus was isolated from all postinoculation blood samples obtained from 4 heifers that were seronegative at the time of inoculation, but was not isolated from any of the preinoculation samples, or from any of the postinoculation samples obtained from 2 heifers that were seropositive at the time of inoculation. When virus isolation was attempted from separated mononuclear cells in 2 heifers, 43.7% of the noncultured and 87.5% of the cultured samples had positive results.     

Effect of floor cooling on performance of lactating sows during summer

Forty Landrace × Large White lactating sows were used to evaluate the effects of cooling of the floor when maintained under high temperature conditions during summer on their productive and reproductive performance. The sows were allocated in a completely randomized design with two treatments with 20 replicates according to parity number and body weight, with each animal being considered an experimental unit. The treatments consisted of cooling of the floor under the sow with water circulation at about 17 °C and no cooling. The resulting temperatures of the floor were 27.6 and 35.8 °C for the cooled and the control treatments, respectively. The sows from both treatments were exposed to average maximum and minimum environmental temperatures of 26.9 and 20.8 °C, respectively. Sows maintained on a cooled floor had a higher feed intake (6.47 vs. 5.61 kg/day; P < 0.01). Despite this higher intake, sows maintained on a cooled floor had higher body weight and body protein losses during the lactation period (P < 0.01) in connection with a higher milk yield and subsequent growth of the litter (2280 vs. 1798 g/day; P < 0.01). There was an effect of treatment on rectal temperature, surface temperatures and respiratory rate (P < 0.01) with lower values in sows submitted to floor cooling. It is concluded that floor cooling under the lactating sow improves its productive and reproductive performance, as well as the performance of its litter.