Ox Blood Research Articles

Isolation of naturally infecting Leishmania infantum from canine samples in Novy-MacNeal-Nicolle medium prepared with defibrinated blood from different animal species

The most commonly used culture medium for the in vitro isolation of Leishmania spp. from canine biological samples is biphasic Novy-MacNeal-Nicolle (NNN) medium, whose solid phase is prepared using rabbit blood. Leishmania infantum parasites from natural infections are highly sensitive and demanding for growth in axenic conditions when firstly obtained from the dog’s body. The objective of this study was to evaluate whether NNN medium (NNN-test) prepared with chicken blood (NNN-C), ox blood (NNN-O), horse blood (NNN-H) or sheep blood (NNN-S) was viable for the isolation of parasites from naturally infected dogs, in an endemic area for visceral leishmaniasis caused by L. infantum. Spleen aspirates from six dogs previously diagnosed as infected by parasitological methods were simultaneously inoculated in each NNN-test medium, including the conventional medium prepared with rabbit blood (NNN-R), and the cultures were examined for three weeks under optic microscopy. Spleen samples were also analyzed for parasite loads by quantitative PCR (qPCR). Cultures from three of the six dogs (50%) were positive in at least one of the NNN-test media: one sample presented the highest spleen parasite load by qPCR (1.19 × 104 parasites/mL) and was positive in all test media; the second sample presented parasitic isolation in the first week of culture in all inoculated media, of which the NNN-C medium had the highest mean parasite count (NNN-C = 23.5 × 104/mL vs. NNN-R = 3.25 × 104/mL); the third sample was positive only in the NNN-S medium besides the conventional control NNN-R. Cultures from the three remaining dogs were negative in all NNN media, including the control and test media; of those three dogs, two presented the lowest spleen parasitic loads according to qPCR. Blood from chicken, ox, horse and sheep shown to be viable for the preparation of NNN culture medium for the primary isolation of L. infantum from samples of naturally infected dogs and can be considered as an alternative to rabbit blood when necessary.

Influence of Ox Blood on Water Absorption of and Chloride Penetration into Concrete

Many concrete structures can not reach their designed service life because of their poor durability. Water and aggressive substances such as chloride ions are prerequisite to the deterioration of concrete. In aggressive environments, water repellent treatment is an effective path to delay the repair and improve the durability of concrete structures. Thousands of years ago, people used oils, animal fats or wax to treat the surface of natural stones, bricks or put some animal blood in mortar to make them water repellent. Many historical monuments were protected in these old methods. In this paper, different amounts of ox blood had been added in concrete. Compressive strength, water absorption and chloride penetration tests had been conducted. In this way, the influence of ox blood on water absorption of and chloride penetration into concrete had been investigated. The results indicated that the compressive strength of concrete with addition of ox blood decreased. And water absorption and chloride penetration into concrete were effectively reduced by adding some amounts of ox blood. The concrete with the surface abraded off 5mm, 15mm and 25mm which were mixed with ox blood had higher water absorption than concrete without abrasion.

An in vivo method for the assay of heparin.

Summary The anticoagulant activity of heparin has been assayed in sheep, by giving small consecutive doses intravenously at intervals of 4 hours, and determining the coagulation time 4 minutes after injection. A curve is plotted, using doses of a standard heparin varying in strength by 10 to 20 per cent. This is followed by 1 or 2 injections of the unknown heparin in suitable doses. The figures found agree, with a margin of a few per cent., with the theoretical ones. The fresh whole-blood method, using ox blood in vitro, is found to give figures identical with those obtained with the in vivo method. The unreliability of the methods using artificial systems with citrated plasma or salted whole blood, including the U.S.P. XIV method, is discussed. An in vivo method is devised as a reference technique in assaying heparin.

Complete genome sequence of Jonesia denitrificans type strain (Prevot 55134T)

Jonesia denitrificans (Prevot 1961) Rocourt et al. 1987 is the type species of the genus Jonesia, and is of phylogenetic interest because of its isolated location in the actinobacterial suborder Micrococcineae. J. denitrificans is characterized by a typical coryneform morphology and is able to form irregular nonsporulating rods showing branched and club-like forms. Coccoid cells occur in older cultures. J. denitrificans is classified as a pathogenic organism for animals (vertebrates). The type strain whose genome is described here was originally isolated from cooked ox blood. Here we describe the features of this organism, together with the complete genome sequence and annotation. This is the first completed genome sequence of a member of the genus for which a complete genome sequence is described. The 2,749,646 bp long genome with its 2558 protein-coding and 71 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project.

Organic waste reclamation, recycling and re-use in integrated fish farming in the Niger Delta

The purpose of this paper is to create awareness on the significance of integrated fish farming in organic waste reclamation, recycling and re-use. Example of integrated fish farming practiced at a micro-level in the Niger Delta of Nigeria is crop-snailry-poultry (Chicken) – livestock (pig) – cum-fish production. In this system, fish production remains the most important activity. The organic wastes reclaimed, recycled and re-used in this system are: plant matter, chicken and pig excrements; snail, oyster and periwinkle shells, ox blood and bone, fish wastes, palm kernel cake, groundnut cake, pond silt and aquatic fern, Azolla pinnata . Other reclaimable, recyclable and re-usable materials are also discussed in this paper. Integrating organic waste reclamation, recycling and re-use in integrated fish farming with treatment, incineration, landfill and composting will help to drastically reduce waste in both rural and urban areas. Journal of Applied Sciences and Environmental Management Vol. 10 (3) 2006: 47-53

Structural Basis for Reduced Staphylocoagulase-mediated Bovine Prothrombin Activation

Staphylocoagulase (SC) is a protein secreted by the human pathogen, Staphylococcus aureus, that activates human prothrombin (ProT) by inducing a conformational change. SC-bound ProT efficiently clots fibrinogen, thus bypassing the physiological blood coagulation pathway. The crystal structure of a fully active SC fragment, SC-(1-325), bound to human prethrombin 2 showed that the SC-(1-325) N terminus inserts into the Ile(16) pocket of prethrombin 2, thereby inducing expression of a functional catalytic site in the cognate zymogen without peptide bond cleavage. As shown here, SC-(1-325) binds to bovine and human ProT with similar affinity but activates the bovine zymogen only very poorly. By contrast to the approximately 2-fold difference in chromogenic substrate kinetic constants between human thrombin and the SC-(1-325).human (pro)thrombin complexes, SC-(1-325).bovine ProT shows a 3,500-fold lower k(cat)/K(m) compared with free bovine thrombin, because of a 47-fold increase in K(m) and a 67-fold decrease in k(cat). The SC-(1-325).bovine ProT complex is approximately 5,800-fold less active compared with its human counterpart. Comparison of human and bovine fibrinogen as substrates of human and bovine thrombin and the SC-(1-325).(pro)thrombin complexes indicates that the species specificity of SC-(1-325) cofactor activity is determined primarily by differences in conformational activation of bound ProT. These results suggest that the catalytic site in the SC-(1-325).bovine ProT complex is incompletely formed. The current crystal structure of SC-(1-325).bovine thrombin reveals that SC would dock similarly to the bovine proenzyme, whereas the bovine (pro)thrombin-characteristic residues Arg(144) and Arg(145) would likely interfere with insertion of the SC N terminus, thus explaining the greatly reduced activation of bovine ProT.

Identification of animal species by protein radioimmunoassay of bone fragments and bloodstained stone tools

In forensics and archaeology, it is important to distinguish human from animal remains and to identify animal species from fragmentary bones and bloodstains. We report blind tests in which a protein radioimmunoassay (pRIA) was used to identify the species of six bone fragments lacking morphological specificity and 43 bloodstained lithic tools, knapped experimentally and soaked in blood of known animal and human origin. The submitters of the bone fragments and the bloodstained tools each listed a number of possible species, from which the testers selected the best match with the pRIA results. All six bone fragments were correctly identified: three humans, a deer, a dog, and a cow. Forty-three tools were stained with blood from a wide variety of species including ungulates, carnivores, a fish, and a bird. On 40 of these 43, at least one species (or blood-free control) was identified correctly. Some of the tools were stained with blood of two different species. A mixture of sheep and musk ox blood was correctly identified; in several other mixtures, only a single species was detected. Two tools with human blood and one with human sweat were correctly reported as human. There was a single false positive (one of three controls reported as weakly bovine) and no false negatives. We conclude that the pRIA technique shows a high degree of accuracy in discriminating human from animal bone fragments and bloodstains and in identifying animal species.

Immunolocalization of vesicular stomatitis virus in black flies (Simulium vittatum).

Vesicular stomatitis, a disease of cattle, horses, and swine, is caused by either vesicular stomatitis virus, New Jersey serotype (VSV-NJ), or vesicular stomatitis virus, Indiana serotype, which are related viruses in the genus Vesiculovirus, family Rhabdoviridae. Although recognized for at least 160 years, the epidemiology and pathogenesis of this disease remains undefined. Black flies have been suggested as a vector for VSV-NJ. In this study we infected three- to four-week-old female black flies with VSV-NJ via feeding of virus-spiked ox blood or intrathoracic inoculation, and demonstrated the location of virus by immunohistochemistry. These preliminary findings suggest that VSV-NJ initially infects the gut in the natural situation but that subsequent spread to the salivary gland may be blocked in older flies, decreasing their ability to transmit the virus. The pattern of staining was different in intrathoracic inoculated flies. In these flies, salivary gland involvement was more likely, and extensive staining of eye, brain, and hemolymph suggested a more generalized infection that apparently circumvented the gut. We conclude that intrathoracic inoculation may be an inappropriate method of infection for determining vector competence and that the age of the vector should be considered when conducting competency studies.

Extraction of amino acids from protein hydrolysates by electrodialysis

Protein hydrolysates were obtained by acid hydrolysis from animal or human residues, such as poultry feathers, ox blood and human hair. After neutralization and discolouration with active charcoal, the hydrolysates were treated by successive electrodialysis (ED) in order to extract amino acids into several fractions. The current density and pH were optimized for each ED operation performed with preindustrial pilot scale equipment. The first step was the demineralization of amino acid mixtures using an ED stack with two compartments. The salt removal was achieved with extraction degrees higher than 90% and current efficiencies of about 80%. In the most favourable case, the amino acid losses did not exceed 10%. The second step was the extraction of the charged amino acids using an ED stack with four compartments. Three fractions were obtained, corresponding to the acidic, basic and neutral amino acids. The extraction degrees varied from 80% to 100%. In the third step, the fractionation of basic amino acids on the one hand, and neutral amino acids on the other hand, was carried out with enrichment degrees varying from 50% to 80%. © 1998 SCI

Hemolysis with the F21 Hemopump

The aim of this study was to determine the influence of mechanical factors on free hemoglobin using an F21 hemopump cannula. These factors (flow, rotary speed, reduction of the cross section area of the silicone tube and foreign material on the screw) were tested in a mock circulation with ox blood. Irrelevant increase in free hemoglobin were induced by foreign bodies either in the inlet or on the screw. The hemopump flow was easily affected and almost suppressed by small amounts of foreign bodies on the screw. Other reasons for elevated concentrations of plasma free hemoglobin may imply a general disorder of the pump, anticoagulation, renal function, red cells or a combination of these factors.

Current progress in the development of an intravenous membrane oxygenator.

In vitro testing of an intravenous membrane oxygenator (IMO) consisting of hollow fiber membranes arranged around a centrally positioned balloon is reported. A total of six IMO prototypes were mounted in a specially designed mock circulatory loop and perfused with physiologic saline or fresh abattoir ox blood to investigate their oxygen and carbon dioxide transfer capabilities. One IMO prototype was mounted in the flow loop and perfused with saline for 13 continuous days to test the durability and reliability of the prototype design. It is the authors' hypothesis that the rhythmic inflation and deflation of the balloon increases convective mixing and cross-flow of blood around the fibers, thereby enhancing gas exchange capabilities. The results of these trials support this contention, namely that gas exchange efficiency rose with increasing frequency of balloon pulsation. No significant deterioration in oxygen transfer was observed in the durability test prototype, which was continuously perfused with saline for 13 days.

Kininogenase activity of tonin.

Tonin, known for its specific and direct generation of angiotensin II, was highly purified from rat submaxillary gland and investigated for kininogenase activity. For the substrate, heat-treated plasma from ox blood, and highly purified low-molecular-weight (LMW) and high-molecular-weight (HMW) kininogens, were used. The reaction product formed at pH 8.0 well satisfied the characteristics of kinin, i.e., depressor and oxytocic activities and reactivity with antibradykinin antiserum. Kinin formed by tonin from purified LMW kininogen was identified with bradykinin in high performance liquid chromatography and radioimmunoassay. The results revealed tonin's new capability of forming kinin in addition to the hitherto known pressor angiotensin II, indicating tonin, too, is a member of the "kinin- tensin enzyme system."

Contrasting roles of 5-hydroxytryptamine and noradrenaline in fever in rats.

1. We have investigated the effects of endogenous monoamine depletion on the development of fever in rats. 2. Fever was produced in rats by a single intraperitoneal injection of Salmonella typhosa endotoxin or leucocyte pyrogen manufactured from ox blood. 3. Depletion of brain 5-hydroxytryptamine (5-HT) by pretreatment with p-chlorophenylalanine prevented the rise in rectal temperature produced both by bacterial endotoxin and by leucocyte pyrogen. 4. On the other hand depletion of noradrenaline (NA) by 6-hydroxydopamine pretreatment resulted in a potentiation of the fever produced by bacterial endotoxin. 5. These results suggest that 5-HT is involved in mechanisms responsible for the rise in temperature during fever and that NA is involved in mechanisms which serve to attenuate the rise.

The febrile responses in rabbits and rats to leucocyte pyrogens of different species.

1. We have investigated the effects on body temperature of rats and rabbits of leucocyte pyrogen derived from the blood of rat, rabbit, ox, pig and baboon. 2. In the rabbit intravenous injections (3.5 ml.) of solutions containing leucocyte pyrogen derived from ox, pig and rabbit blood produced fevers with short latencies; no fevers resulted from injections of similar solutions derived from rat or baboon blood. 3. In the rat intraperitoneal injections (2.0 ml.) of solutions containing leucocyte pyrogen derived from ox blood caused a fever, while pig leucocyte pyrogen produced a marked hypothermia. Neither rabbit, baboon, nor rat leucocyte pyrogen had any significant effect on rectal temperature of the rats. 4. Our results show that there is a variability of response in the rat and the rabbit to injection of leucocyte pyrogen of different species; leucocyte pyrogen may be species specific.

Isolation of sodium-complexing polypeptides from mammalian blood and cardiac muscle.

Two distinct polypeptides have been isolated from rat heart and ox blood. They are both found to be effective in forming complexes with sodium ions, and it is suggested that they may have a function in stabilizing sodium ion activity.

Hemolysis in several animal species after rapid freezing of blood.

AbstractThe effect of various freezing rates on the extent of hemolysis in human, bovine and ovine erythrocytes, which are known to have different cell volumes, water contents and permeabilities, was investigated. Blood in stainless steel capillary tubes was frozen at various rates by abrupt immersion of the capillaries into cooling baths at temperatures ranging from −20° to −130°C. Minimum lysis values were obtained at freezing temperatures of −40°, −50° and −70°C with, respectively, human, bovine and ovine blood. The smallest, highly permeable sheep erythrocytes were the least damaged at the highest freezing rates; the largest human cells with the highest water content, suffered the greatest damage; intermediate values were obtained with ox blood. At the lower freezing rates, the largest, human cells were the least damaged; the highest hemolysis values were obtained with the smallest, highly permeable sheep erythrocytes; ox blood again gave intermediate values. These results are in agreement with current views that, (1) very rapid freezing results in the formation of damaging intracellular ice; (2) injury associated with slow freezing is related to the extent of dehydration or to the increase in electrolyte concentration which accompanies ice formation; (3) minimum hemolysis is obtained under those freezing conditions in which osmotic dehydration has been sufficient to prevent the formation of intracellular ice, but has left enough water in the cells to prevent the damaging effects of dehydration and high electrolyte concentrations.

A membrane feeding technique for the infection of Culicoides nubeculosus mg. and Culicoides variipennis sonorensis coq. with Onchocerca cervicalis Rail. and Henry

The transmission of Onchocerca cervicalis Rail. and Henry in the horse is being investigated at Winches Farm Field Station, St. Albans. Laboratory colonies of Culicoides nubeculosus Mg., the natural vector of O. cervicalis in Europe, and of C. variipennis sonorensis Coq., a North American midge, have been established. A technique has been developed to facilitate the artificial infection of these midges with O. cervicalis. Experimental flies imbibed a meal of O. cervicalis microfilariae suspended in defibrinated ox blood at 38°C. A plucked skin from a one-day-old chick was used as a membrane. 232 C. nubeculosus and 117 C. v. sonorensis were fed and 5·5% of the C. nubeculosus and 7·7% of the C. v. sonorensis subsequently developed infective (3rd stage) larvae. The fact that 3rd stage larvae developed in specimens of C. v. sonorensis is a point of special interest because this midge has not yet been identified as a vector of O. cervicalis.

B. The fibrinolytic mechanism. The mechanism of fibrinolysis.

In recent years, stimulated by the production by the pharmaceutical industry of fibrinolytic agents suitable for therapeutic use in man, there has been a great increase in interest in the components and functions in health and disease of the fibrinolytic enzyme system. Development of knowledge of fibrinolysis Spontaneous lytic activity of blood clots It has been known for many years that human blood possesses fibrinolytic activity. Hunter (1794) records that in 'animals killed by lightning or by electricity’ or in animals 'who are run very hard, and killed in such a state’ the blood does not clot. This phenomenon was partly explained by Morawitz (1906) who found that the blood from victims of sudden death contained no fibrinogen and could destroy the fibrinogen and fibrin of normal blood. Denis (1838) observed that the blood clots obtained in wet cupping redissolved in less than 24 h. Green (1887) noted that when fibrin prepared from ox blood had dissolved when incubated in saline it could not be clotted again by thrombin. Dastre (1893), during the course of phlebotomy in dogs, observed a reduction in fibrin yield, which he attributed to destruction of fibrin, a process which he named ‘fibrinolysis’; and Hedin (1904) found spontaneous proteolytic activity in the globulin fraction of ox blood. The main outline of present-day knowledge of spontaneous fibrinolytic activity in the blood was finally achieved when Macfarlane (1937) showed that in man, fibrinolytic activity in the blood could be provoked by surgical operation.

Further Studies on the “Dynia” Clotting Abnormality

Summary1. One patient with multiple bleeding episodes and 4 asymptomatic relatives in 3 generations reveal a coagulation abnormality that cannot be attributed to any known coagulation factor deficiency or to a circulating inhibitor. The abnormality is characterized by deficient generations of intrisic thromboplastin and intermediate product I, and can be corrected by normal plasma and serum.2. The ability of normal blood to correct the coagulation defect in vitro is heat labile; it is not removed from plasma by adsorption with BaSO4 or Al(OH)3. Neonatal blood and blood taken from patients either receiving coumarin drugs or with severe liver disease sustain the ability to correct the described defect. Ox, rabbit and dog blood are also corrective.3. The finding of normal factors XII, XI and IX activities, as well as normal serum thrombosis accelerator (STA) activity, places this abnormality beyond the stage of factor IX activation.4. The data suggest that the familial coagulation abnormality is located in the clotting sequence either in the activation of factor VIII, or in the activation of factor X by factor VIII. Either interpretation is in agreement with the finding of abnormal intermediate product I generation, which reflects primarily the intrinsic activation of factor X.5. The frequency of this defect is unknown. Conceivably some patients considered to have factor XI (PTA) deficiency because of a plasma and serum correctable defect in their thromboplastin generation, may actually have a coagulation defect similar to that described in the Dynia family.6. It is suggested that the name “Dynia defect” be temporarily assigned to this clotting abnormality.

On peptide synthesis. Synthesis of methionyl-lysyl-bradykinin, a kinin from ox blood

The synthesis and the pharmacological actions of methionyl-lysyl-bradykinin, a new kinin from ox blood, are described.