A PCR-based epidemiological survey of Hepatozoon canis in dogs in Nigeria

The prevalence of Hepatozoon canis infections in dogs in Nigeria was surveyed using molecular methods. DNA was extracted from blood samples obtained from 400 dogs. A primer set that amplified the Babesia canis 18S rRNA gene, which has high similarity to the H. canis 18S rRNA gene, was used for the PCR. As a result, samples from 81 dogs (20.3%) produced 757 bp bands, which differed from the 698 bp band that corresponded to B. canis infection. The sequence of the PCR products of 10 samples were determined, all of which corresponded with the H. canis sequence.     

Effect of low pH on the susceptibility of isolated cod (Gadus morhua) microsomes to lipid oxidation

During the extraction of muscle to produce protein isolates by acid or alkali solubilization, membranes are exposed to abnormally low or high pH. Low but not high pH treatment induces rapid oxidation of membrane phospholipids in the presence of hemoglobin. The goal of this research work was to study the oxidative stability of microsomes under the conditions met during acid solubilization. Isolated microsomes from cod muscle were used as a model system. At pH 5.3 or lower, 99% of isolated cod membranes sedimented at low centrifugation speeds. Isolated membranes that were exposed to pH 3.0 were less susceptible to hemoglobin-mediated lipid oxidation. Cod hemoglobin exposed to pH 3 was rendered less pro-oxidative than the untreated cod hemoglobin. However, when microsomes and hemoglobin were together exposed to low pH, oxidation was promoted. Citric acid and calcium chloride, as well as press juice isolated from cod muscle, were able to inhibit lipid oxidation of microsomal suspensions.     

Comparative susceptibility of chickens, turkeys and ducks to infectious bursal disease virus using immunohistochemistry

Twenty chicks, 12 turkey poults and 10 ducklings, all 5 weeks old were infected with 2 × 10^3.5 chick LD₅₀ IBD virus to determine the course of the virus in the 3 poultry species. Uninfected control birds were kept separately. Two infected and 2 control birds/species were euthanized at time intervals between 3 and 168 hours post infection (pi). Sections of thymus, bursa of Fabricius, spleen, liver, kidney, proventriculus and ceacal tonsil were stained for the detection of IBD virus antigen using immunoperoxidase technique. IBD virus antigen positive cells stained reddish-brown and the amount of such cells in tissue sections were noted and scored. Stained cells were present in all organs examined for up to 168 hours pi in the 3 poultry species except ceacal tonsils of ducks at 72 and 120 hours pi. Antigen score was highest in chickens and least in ducks as reflected by average of total scores/sampling time of 12, 10.8 and 8 in chickens, turkeys and ducks respectively. Total antigen score/sampling time in infected chickens peaked twice; 24/48 and 144 hours pi, whereas such bi-phasic peaks were absent in turkeys and ducks. Range of total antigen score at different sampling times was 7–17.5 in chickens, 10–13 in turkeys and 7–10 in ducks indicative of marked viral replication in chickens. Presence of IBD viral antigen in organs of all 3 poultry species is indicative of infections. The innate ability of turkeys and ducks to prevent appreciable replication of IBD virus after infection requires further investigation.     

Retention of alkamides in dried Echinacea purpurea

Different drying methods, such as freeze-drying (FD), vacuum microwave drying (VMD), and air-drying (AD), were applied to fresh roots and leaves of Canadian-grown Echinacea purpurea to determine the optimal method for preserving alkamide levels. Using HPLC, six alkamide fractions (alkamides 1, 2, 3, 6a/6, 7, 8/9) were quantitated in dried roots, whereas four alkamide fractions (alkamides 1, 2, 3, 8/9) were measured in dried leaves. Different elution conditions used in HPLC for alkamide analysis did not affect the eluted fractions nor the quantitation of different alkamides. Individual alkamide concentrations in roots and leaves were affected by the drying methods used. To preserve higher levels of total alkamides, FD was found to be the best method, VMD was a superior method for drying roots than AD at 70 degrees C, while AD at 50 degrees C was the preferred method for drying leaves of E. purpurea.     

Partitioning of exogenous delta-tocopherol between the triacylglycerol and membrane lipid fractions of chicken muscle

The partitioning of exogenous delta-tocopherol, added dissolved in ethanol, between the neutral triacylglycerols and membranes of chicken leg muscles was investigated. The two lipid fractions were separated using differential ultracentrifugation techniques. Triacylglycerols were obtained after high-speed centrifugation of the minced muscle at 130000 g for 30 min. Membranes were collected from a muscle-buffer homogenate (pH 7.5) between 10000 g for 20 min and 130000 g for 30 min. The triacylglycerols collected represented from 15 to 80% of the total triacylglycerols of the minced muscle, the yields increasing with increasing muscle triacylglycerol content. The phospholipids in the isolated membrane fraction represented from 20 to 35% of the total phospholipids of the muscle. At low muscle total lipid contents (3-5%), the added delta-tocopherol was present in approximately the same concentration in both muscle lipid fractions. At higher total lipid contents, achieved by adding exogenous triacylglycerols, the delta-tocopherol concentration in the membranes increased relative to that in the triacylglycerols.     

Effect of pH on lipid oxidation using trout hemolysate as a catalyst: a possible role for deoxyhemoglobin

Hemoglobin-mediated lipid oxidation was studied by adding hemolysate to washed cod muscle. Three pH values were examined (pH 7.6, 7.2, and 6.0). The lag time prior to rancidity and thiobarbituric acid reactive substance development decreased greatly as the pH was reduced (p < 0.01). Formation of methemoglobin due to autoxidation of the heme pigment was found to occur more rapidly at reduced pH. Also, the level of deoxyhemoglobin was found to sharply increase with pH reduction in the range of pH 7.6-6.0. This suggested a potential role for deoxyhemoglobin as a catalyst. ATP lowered hemoglobin oxygenation at pH 7.2. Peroxidation of linoleic acid by oxy/deoxyhemoglobin and methemoglobin was investigated at two levels of preformed lipid hydroperoxides. At a reduced level of preformed lipid hydroperoxides, oxy/deoxyhemoglobin stimulated peroxidation of linoleic acid, whereas methemoglobin did not. At the higher level of preformed lipid hydroperoxides, both oxy/deoxyhemoglobin and methemoglobin were active. This investigation suggests that reduced hemoglobins played an important role in lipid oxidation processes.