Improvements to the hemagglutination inhibition test for serological assessment of recombinant fowlpox-H5-avian-influenza vaccination in chickens and its use along with an agar gel immunodiffusion test for differentiating infected from noninfected vaccinated animals

In general, avian influenza (AI) vaccines protect chickens from morbidity and mortality and reduce, but do not completely prevent, replication of wild AI viruses in the respiratory and intestinal tracts of vaccinated chickens. Therefore, surveillance programs based on serological testing must be developed to differentiate vaccinated flocks infected with wild strains of AI virus from noninfected vaccinated flocks in order to evaluate the success of vaccination in a control program and allow continuation of national and international commerce of poultry and poultry products. In this study, chickens were immunized with a commercial recombinant fowlpox virus vaccine containing an H5 hemagglutinin gene from A/turkey/Ireland/83 (H5N8) avian influenza (AI) virus (rFP-H5) and evaluated for correlation of immunological response by hemagglutination inhibition (HI) or agar gel immunodiffusion (AGID) tests and determination of protection following challenge with a high pathogenicity AI (HPAI) virus. In two different trials, chickens immunized with the rFP-H5 vaccine did not develop AGID antibodies because the vaccine lacks AI nucleoprotein and matrix genes, but 0%-100% had HI antibodies, depending on the AI virus strain used in the HI test, the HI antigen inactivation procedure, and whether the birds had been preimmunized against fowlpox virus. The most consistent and highest HI titers were observed when using A/turkey/Ireland/83 (H5N8) HPAI virus strain as the beta-propiolactone (BPL)-inactivated HI test antigen, which matched the hemagglutinin gene insert in the rFP-H5 vaccine. In addition, higher HI titers were observed if ether or a combination of ether and BPL-inactivated virus was used in place of the BPL-inactivated virus. The rFP-H5 vaccinated chickens survived HPAI challenge and antibodies were detected by both AGID and HI tests. In conclusion, we demonstrated that the rFP-H5 vaccine allowed easy serological differentiation of infected from noninfected birds in vaccinated populations of chickens when using standard AGID and HI tests.     

Heterogeneous aspects of lipid oxidation in dried microencapsulated oils

This work was aimed at studying lipid oxidation in dried microencapsulated oils (DMOs) during long-term storage. Samples were prepared by freeze-drying of emulsions containing sodium caseinate and lactose as encapsulating components. Evaluation of lipid oxidation was approached by quantitative analysis of nonvolatile lipid oxidation products and tocopherol. Lipid oxidation products were analyzed by separation of polar compounds by adsorption chromatography followed by HPSEC with refraction index detection for quantitation of oxidized triglyceride monomers, dimers, and oligomers. The analytical method applied enabled the detection of different oxidative patterns between the free and encapsulated oil fractions. The free oil fraction of DMOs showed a typical oxidative pattern for oils in continuous phase, which consisted of a clear induction period, in which hydroperoxides (oxidized triglyceride monomers) accumulated, before oxidation accelerated. The end of the induction period was marked by the total loss of tocopherol and the initiation of polymerization. On the contrary, the encapsulated oil showed a pattern characteristic of a mixture of oils with different oxidation status. Thus, high contents of advanced oxidation compounds (polymerization compounds) were detected when the antioxidant (tocopherol) was still present in high amounts. It is concluded that the encapsulated oil was comprised of oil globules with very different oxidation status. The results obtained in this study gave evidence of heterogeneous aspects of lipid oxidation in a dispersed-lipid food system.     

Chemical composition of the Lippia origanoides essential oils and their antigenotoxicity against bleomycin-induced DNA damage

The present work evaluated the chemical composition of the essential oils (EO) obtained from Lippia origanoides and their DNA protective effect against bleomycin-induced genotoxicity. L. origanoides EO chemical composition was determined by gas chromatography–mass spectrometry (GC–MS). The major compounds of the L. origanoides EOs were thymol (34–58%) and carvacrol (26%). The antigenotoxic effects of the EOs, major compounds and standard compound (epigallocatechin gallate) were assayed in co-incubation procedures using the SOS chromotest in Escherichia coli. Both EOs and their major compounds protected bacterial cells against bleomycin-induced genotoxicity indicating that these two compounds were principally responsible for the antigenotoxicity detected in the oils. Thymol and carvacrol antigenotoxicity was lower than those observed with epigallocatechin gallate. The results were discussed in relation to the chemopreventive potential of L. origanoides EOs and their major components, carvacrol and thymol.     

Canine circovirus and Canine adenovirus type 1 and 2 in dogs with parvoviral enteritis

Veterinary Research Communications - Canine parvovirus type 2 (CPV-2) is one of the most relevant pathogens associated with enteritis in dogs and is frequently reported in association with the...     

Cellulose acetate electrophoresis of canine plasma after fibrinogen precipitation by ethanol

Background: In routine canine medicine, anticoagulated blood is often the only sample sent to laboratories for diagnostic purposes. This hampers the interpretation of protein electrophoretic tracings because plasma contains fibrinogen, which migrates in the β–γ region. In human medicine, fibrinogen can be precipitated from plasma using ethanol. Objectives: The purpose of this study was to assess ethanol precipitation as a method for removing fibrinogen from canine plasma so as to facilitate the interpretation of electorphoresis results. Methods: Blood samples collected from 40 dogs were divided into plain tubes and tubes containing EDTA (n=20) or lithium–heparin (n=20). An aliquot of plasma from each sample was incubated with ethanol at a final concentration of 100 mL/L. Cellulose acetate electrophoresis was then performed on serum, plasma, and plasma treated with ethanol. To verify the efficiency of ethanol treatment, fibrinogen was added to 5 canine serum samples at final concentrations of 2.5, 5.0, and 10.0 g/L, and electrophoresis was performed before and after ethanol treatment. Results: Visual analysis of electrophoretograms from ethanol‐treated samples confirmed the disappearance of the fibrinogen peak from the β₂‐globulin region. Treatment with ethanol caused a significant decrease in the percentage of β₂‐globulins and a significant increase in the percentage of α₂‐globulins. Absolute values of most electrophoretic fractions were significantly decreased in ethanol‐treated plasma compared with serum. Conclusions: Ethanol treatment successfully removed fibrinogen from canine plasma and normalized electrophoretic profiles, but probably also precipitated proteins other than fibrinogen. Ethanol treatment is recommended to facilitate visual identification of abnormal monoclonal peaks, but not for determining absolute protein concentrations in electrophoretic fractions.     

Effects of Dietary Fiber on Growth and Gastric Emptying Time of the Freshwater Prawn Macrobrachiurn rosenbergii (De Man, 1879)

Feeding experiments were conducted on large and small adult freshwater prawns Macrobrachium rosenbergii (36.96 ± 1.65 and 13.48 ± 1.65 g in weight) to determine the impact of dietary fiber on growth, digestion, and gastric emptying time. Fiber was incorporated as Q- cellulose into four semi-purified diets at calculated levels of 0, 5, 10 and 15% at the expense of starch and fed for an 8-wk period. The experiment was conducted in a closed, filtered, recirculating freshwater system. The results indicate that the specific growth rates, feed conversion ratios, and protein efficiency ratios improved as the levels of dietary fiber increased from 0.4% to 8%. Gastric emptying times increased as the level of inclusion of cellulose in the diets increased. It was concluded that diets containing up to 10% dietary fiber increases growth rates in adult prawn Macrobrachium rosenbergii by increasing nutrient residence time in the gastrointestinal tract, thereby increasing absorption.     

Color quality of pigments in cochineals (Dactylopius coccus Costa). Geographical origin characterization using multivariate statistical analysis

The commercial value of a cochineal (Dactylopius coccus Costa) sample is associated with its color quality. Because the cochineal is a legal food colorant, its color quality is generally understood as its pigment content. Simply put, the higher this content, the more valuable the sample is to the market. In an effort to devise a way to measure the color quality of a cochineal, the present study evaluates different parameters of color measurement such as chromatic attributes (L*, and a*), percentage of carminic acid, tint determination, and chromatographic profile of pigments. Tint determination did not achieve this objective because this parameter does not correlate with carminic acid content. On the other hand, carminic acid showed a highly significant correlation (r = - 0.922, p = 0.000) with L* values determined from powdered cochineal samples. The combination of the information from the spectrophotometric determination of carminic acid with that of the pigment profile acquired by liquid chromatography (LC) and the composition of the red and yellow pigment groups, also acquired by LC, enables greater accuracy in judging the quality of the final sample. As a result of this study, it was possible to achieve the separation of cochineal samples according to geographical origin using two statistical techniques: cluster analysis and principal component analysis.     

A comparison between the spatial distribution pattern of flagellates and some physicochemical parameters in a waste stabilization pond

Protozoological and physicochemical analyses were performed in a waste stabilization pond at Sto. Tomas Atzingo, a small town near Mexico City, in order to discern the internal dynamics of the pond water through the study of the spatial distribution pattern of flagellates and the parallel water volume distribution pattern of some selected physicochemical determined parameters. The statistical method applied to the data obtained was by cluster analysis. The results showed a slight trend to heterogeneity in the system, when it was evaluated through the physicochemically determined parameters. On the other hand, the spatial distribution pattern of the flagellates pointed out a remarkable heterogeneity in the pond. In systems like the one studied where the main feature is an organic matter overload, such a difference in findings indicates a low correlation between the biological parameters, in this case the flagellates, and the physicochemical variables, that should be considered when evaluating the water quality of such systems, either through the physicochemical, the biological or both scopes.     

A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins

The isolation of broadly neutralizing antibodies against influenza A viruses has been a long-sought goal for therapeutic approaches and vaccine design. Using a single-cell culture method for screening large numbers of human plasma cells, we isolated a neutralizing monoclonal antibody that recognized the hemagglutinin (HA) glycoprotein of all 16 subtypes and neutralized both group 1 and group 2 influenza A viruses. Passive transfer of this antibody conferred protection to mice and ferrets. Complexes with HAs from the group 1 H1 and the group 2 H3 subtypes analyzed by x-ray crystallography showed that the antibody bound to a conserved epitope in the F subdomain. This antibody may be used for passive protection and to inform vaccine design because of its broad specificity and neutralization potency.