DNA extraction from bovine mummified fetuses and detection of factor XI gene deficiency in the mummies

Genomic DNA extracted from bovine mummified tissue is valuable material for detection of some genes that may contribute to fetal abnormalities. In this study bovine genomic DNA was extracted from the hardened tissue samples of ten bovine mummified fetuses. The amount of genomic DNA extracted from 2 g of the mummified tissues by the phenol/chloroform-ethanol method was low (less than 4 microg/ml) for all samples. The extracted DNA was then amplified by the GenomiPhi DNA amplification system. After amplification, the amount of DNA was increased to more than 100 microg/ml for all samples. This amplification system was shown to be a good tool for amplifying the genomic DNA of the mummified fetuses. The amplified genomic DNA was used for testing the mummies for Factor XI gene deficiency, an autosomal recessive deficiency involved in the early stages of the intrinsic blood coagulation pathway. Exon 12 of the Factor XI gene of the mummies was amplified by PCR. Two of the ten mummified fetuses were heterozygous for the Factor XI gene as indicated by the presence of two amplified DNA fragments of 320 bp and 244 bp. Factor XI deficiency has already been described in Holstein cattle. However, no report is available for bovine fetus. In this study, DNA was extracted and amplified from the bovine mummified fetuses, and the samples were successfully tested for Factor XI gene deficiency in the mummies.     

The Zinc and Copper Content of the Plasma of Sudanese Camels (Camelus dromedarius)

A study was undertaken to investigate the variations in the content of zinc and copper in the plasma of Sudanese camels (Camelus dromedarius). A total of 993 Arabi camels, aged 0.5-8 years, were used to assess the effect of season, age, sex and physiological status on the plasma concentrations of copper and zinc. There was an increase in the concentration of Cu and a decrease in the concentration of Zn in the plasma with age. The concentrations of both Cu and Zn in the plasma were higher in the rainy season than in the dry season. The plasma copper concentrations in pregnant, low-lactating and high-lactating camels were 81.3 +/- 4.7, 59.7 +/- 6.1 and 61.3 +/- 5.5 microg/100 ml, respectively. The corresponding values for zinc were 51.0 +/- 8.9, 53.4 +/- 6.4 and 67.1 +/- 5.5 microg/100 ml, respectively. However, there was no effect of sex on the content of these minerals in the plasma.     

Metabolic profiles and bile acid extraction rate in the liver of cows with fasting-induced hepatic lipidosis

This study was designed to monitor lipid profile in the portal and hepatic blood of cows with fasting-induced hepatic lipidosis, and to compare the results with those in the jugular blood. The work was also carried out to investigate bile acid (BA) in these vessels, and further to investigate BA extraction rate in the liver. Five cows were equipped with catheters in the portal, hepatic and jugular veins (day 0), fasted for 4 days (day 1-day 4) and then refed (day 5-day 11). Before morning feeding, blood was sampled before, during and after fasting from the catheterized vessels. In the portal blood, the concentration of non-esterified fatty acids (NEFA) showed a progressive increase and at day 5 there was an approximate twofold rise. Increased NEFA concentrations were also found similarly in the other two veins. At day 5, beta-hydroxybutyrate (BHBA) in the portal, hepatic and jugular blood rose to 197, 190 and 186% of the pre-fasting value, respectively. However, the concentrations of NEFA and BHBA in the three veins gradually returned to pre-fasting concentration during the refeeding period. Compared with the pre-fasting value at day 0, the content of liver triglyceride (TG) increased significantly at day 5 (P < 0.01). In the liver, the hepatic extraction rate of BA dropped from 3.1 times pre-fasting to 2.2 times during fasting. There were no significant differences in the concentrations of glucose, TG, total cholesterol, cholesterol esters, free cholesterol and phospholipids. The results of the current study show that metabolic alterations occur in the portal, hepatic and jugular veins during induction of hepatic lipidosis in cows, and mostly metabolites, with exception of BA concentration, run parallel. The decreased BA extraction rate in the liver of fasted cows was considered to reflect hepatic cell impairment caused by TG accumulation. Hopefully, the findings, at least in part, contribute to the explanation of the pathophysiology of hepatic lipidosis in dairy cows.     

Impact of browning reactions and bran pigments on color of parboiled rice

Rice color changes from white to amber during parboiling (soaking and steaming). Color parameters indicated that, during soaking, yellow bran pigments leached out in the water. The levels of the Maillard precursors (i.e., reducing sugars (RS) and free alpha-amino nitrogen (FAN)) depended on soaking temperature and time: leaching of RS was compensated by enzymic formation for long soaking times (>60 min), while proteolytic activity was too low to compensate for FAN leaching. Rice soaking under nitrogen, oxygen, or ambient conditions and determination of polyphenol oxidase activity allowed us to conclude that the effect of enzymic color changes on the soaked rice color was rather small. Color measurements of brown and milled mildly, intermediately, and severely parboiled rice samples showed that both brown and milled rice samples were darker and more red and yellow after parboiling and that the effect depended on the severity of parboiling conditions. Furthermore, steaming affected the rice color more and in a way opposite to that observed in soaking. The changes in RS and the loss of FAN during parboiling suggested that Maillard type reactions occur during brown rice steaming. Analyses of furosine levels confirmed Maillard browning of outer bran layers and endosperm during steaming. The level of this Maillard indicator increased with the severity of parboiling conditions in both brown and milled parboiled rice. Measurements of the levels of bran pigments indicated that bran pigments diffuse into the endosperm during parboiling and contribute to the parboiled rice color.     

Bovine hepatocyte growth factor and its receptor c-Met: cDNA cloning and expression analysis in the mammary gland

Hepatocyte growth factor/scatter factor (HGF/SF) is a pleiotropic cytokine that plays a crucial role in the embryonic and postnatal development of various organs including the mammary gland. We cloned bovine HGF and its c-Met receptor cDNAs, and examined their expression during mammary gland development in dairy cows. The 2.5-kbp HGF cDNA clone contained a 2190 bp open reading frame coding a 730 amino acid protein, while the 4.8-kbp c-Met cDNA clone contained a 4152 bp open reading frame coding a 1384 amino acid protein. The bovine HGF and c-Met sequences exhibited more than 87% identity with those of other mammals. RT-PCR analysis revealed ubiquitous expression of both HGF and c-Met mRNAs in various bovine tissues tested. HGF mRNA was detected only in the inactive stage of bovine mammary gland development and not in the developing, lactating, and involuting stages, while c-Met mRNA was detected in the inactive and involuting stages. Immunohistochemical analysis demonstrated that the c-Met protein was found on mammary epithelial cells in the inactive, developing, and involuting stages, and on myoepithelial cells in all stages. These results suggest pivotal roles of HGF and c-Met in the development of bovine mammary gland.     

An immunocytochemical procedure for protein localization in various nematode life stages combined with plant tissues using methylacrylate-embedded specimens

Plant-parasitic nematodes possess a large number of proteins that are secreted in planta, allowing them to be successful parasites of plants. The majority of these proteins are synthesized mainly in the nematode subventral and dorsal glands as well as in other organs. To improve the immunovisualization of these proteins, we adapted a methacrylate embedding method for the localization of proteins inside nematode tissues, and extracellularly when secreted in planta or within plant cells. An important advantage is that the method is applicable for all nematode stages: preparasitic as well as parasitic stages, including large mature females. Herein, the method has been successfully applied for the localization of four nematode secreted proteins, such as Mi-MAP-1, Mi-CBM2-bearing proteins, Mi-PEL3, and Mi-6D4. In addition, we could also localize 14-3-3 proteins, as well as two cytoskeletal proteins, by double-immunolabeling on preparasitic juveniles. Superior preservation of nematode and plant morphology, allowed more accurate protein localization as compared with other methods. Besides excellent epitope preservation, dissolution of methacrylate from tissue sections unmasks target proteins and thereby drastically increases antibody access.     

Induction of defence related enzymes and phenolic compounds in lupin (Lupinus albus L.) and their effects on host resistance against Fusarium wilt

Two different biotic inducers [Pseudomonas fluorescens and Pseudomonas putida] and three different abiotic inducers [copper sulphate, indole butyric acid and potassium chloride] were tested for their efficacy in inducing resistance in lupin plants against Fusarium wilt disease caused by Fusarium oxysporum f. sp. lupini. Application of the biotic and abiotic inducers as seed treatments significantly reduced wilt disease incidence under greenhouse and field conditions. Potassium chloride and Pseudomonas fluorescens were superior. A time course of defence-related enzymes showed substantial increases in enzyme activities in induced infected seedlings compared with untreated healthy plants or infected controls. However, the magnitude of the increase varied among treatments. The maximum increases in chitinase and ??- glucanase activities were recorded at 12 and 8?days after inoculation with the pathogen, respectively. Also, the activity of phenylalanine ammonia lyase increased dramatically 8?days after inoculation. Greater accumulation of phenolic compounds and specific flavonoids upon infection with the pathogen was found in induced and/ or infected seedlings compared with healthy plants. In addition to inducing disease resistance, the treatments were accompanied by significant increases in crop parameters and seed yield compared with untreated controls.     

An unusual outbreak of histomonosis in a commercial turkey flock

In the past histomonosis was very well controlled with Dimetridazole as a treatment and/or Nifursol as feed additive. In the European Union both products were banned in 1995 and 2003, respectively. This was followed by the re-emergence of the disease in the recent years. In the present case a farm with two houses was affected by the disease. In each house 2620 hens and 2620 toms were kept, separated by wire mesh. At the 53rd day of age the toms in house 1 showed general clinical symptoms, accompanied by a slightly raised mortality, which sharply increased in the following days. At necropsy all dead birds showed lesions typical for histomonosis in caeca and liver. Histomonosis was diagnosed by histopathology and PCR. Within five days cumulative mortality was 25.1%.The hens kept at the same house didn't show any symptoms. At day 57 two toms, which were kept in house 2, died and showed similar symptoms and lesions. Within the next three days 48 more birds died. Again the hens in house 2 showed neither clinical signs nor mortality. Treatment trials using herbal products and a change of litter directly after the onset of clinical signs did not reduce the mortality. On day 62 the toms of both houses were euthanized by CO2 in closed containers. The hens were kept until they were slaughtered in week 16 and did not show any evidence of histomonosis.