Endostatin inhibits T-type Ca2+ channel current in guinea pig ventricular myocyte

Endostatin, a fragment of collagen XVIII, is known as an endogenous angiogenesis inhibitor, and its serum concentration increases in various cardiovascular diseases. T-type Ca²⁺ channel, low voltage-activated Ca²⁺ channel, is not expressed in adult ventricular myocytes. Re-expression of T-type Ca²⁺ channels in cardiac myocytes is thought to be involved in the development of cardiac hypertrophy. We examined the effects of endostatin on T-type Ca²⁺ channel current by whole-cell patch clamp technique in freshly isolated adult guinea pig ventricular myocytes, which exceptionally express T-type Ca²⁺ channels. Although endostatin 300 ng/ml had no effect on L-type Ca²⁺ current, it significantly inhibited T-type Ca²⁺ current. These data indicate that endostatin can be an endogenous inhibitor of T-type Ca²⁺ channels in the cardiac myocytes.     

Effects of mild cardiac hypertrophy, induced by volume overload in turkeys, on myocardial sarcoplasmic reticulum calcium-pump and calcium-channel activities and on the creatine kinase system.

Recent studies indicate that in animals with marked cardiac hypertrophy, there is depressed function of Ca2+ sequestration by myocardial sarcoplasmic reticulum (SR) because of down regulation of the Ca(2+)-ATPase gene. However, in several animal models we have observed enhancement of myocardial Ca2+ sequestration in response to chronic cardiac stimulation. We tested the hypothesis that in animals with mild cardiac hypertrophy, there is enhanced Ca(2+)-cycling activity by the SR Ca2+ pump and Ca(2+)-release channel. Because creatine kinase activity is consistently decreased in cardiomyopathy, we also determined whether enhanced Ca2+ cycling was accompanied by down regulation or inhibition of the creatine kinase system. Mild cardiac hypertrophy was induced by volume overload; 2% salt was added to the diet of 2-week-old turkey poults for 4 weeks. Compared with age-matched controls, volume overload resulted in 14.3% increase in heart weight and 21.5% increase in heart-to-body weight ratios. The hypertrophied heart had approximately 20% increased activities of the SR Ca2+ pump and the SR Ca2+ channel. Net Ca2+ transport was increased by 16.5%. Compared with controls and in contrast to several other myocardial enzymes, creatine kinase activity was diminished in the hypertrophied hearts by 23% and creatine content was decreased by 8%. Differences between groups were not detected for lactate dehydrogenase, aspartate transaminase, and alanine transaminase. We concluded that an early adaptation of the myocardium undergoing hypertrophy in compensatory response to functional overload is an enhancement of Ca2+ cycling activity by the Ca2+ pump and Ca2+ channel of the SR.(ABSTRACT TRUNCATED AT 250 WORDS)     

Endostatin inhibits bradykinin-induced cardiac contraction

.Endogenous fragments of extracellular matrix are known to possess various biological effects. Levels of endostatin, a fragment of collagen type XVIII, increase in certain cardiac diseases, such as cardiac hypertrophy and myocardial infarction. However, the influence of endostatin on cardiac contraction has not been clarified. In the present study, we investigated the effects of endostatin on bradykinin-induced atrial contraction. Isometric contractile force of mouse isolated left atria induced by electrical current pulse was measured. Voltage-dependent calcium current of guinea pig ventricular myocytes was measured by a whole-cell patch-clamp technique. Endostatin (100–1,000 ng/ml) alone treatment had no influence on left atrial contraction. On the other hand, pretreatment with endostatin (300 ng/ml) significantly inhibited bradykinin (1 µM)-induced contraction and voltage-dependent calcium current. These data suggest that endostatin may decrease bradykinin-induced cardiac contraction perhaps through the inhibition of voltage-dependent calcium channel.     

The effect of three bufadienolide cardiac glycosides on contraction of isolated rat jejunum

Three cardiac glycosides were screened for pharmacological effects on isolated rat jejunum. The contraction of rat jejunum with epoxyscillirosidin, a non-cumulative bufadienolide, and cotyledoside and tyledoside D, both cumulative neurotoxic bufadienolides were compared with methacholine. The results indicate that all three bufadienolides cause contraction of jejunal smooth muscle. When combined with atropine (1 x 10(-6) M) the response of epoxyscillirosidin and tyledoside D decreased, indicating suppression of a cholinergic response caused by the cardiac glycosides.     

Digoxin- and monensin-induced changes of intracellular Ca2+ concentration in isolated guinea-pig ventricular myocyte

This study was undertaken to determine the possible mechanisms of actions of monensin and digoxin by using isolated guinea-pig ventricular myocytes. Since Ca2+ is the major signal for triggering contraction of cardiac muscle, the objective of this study was to determine whether monensin and digoxin affect the [Ca2+]i of cardiac myocytes and if so is this effect due to an increase in [Na+]i. Three different concentrations of digoxin (0.3, 1 and 3 micromol/l) and three different concentrations of monensin (0.3, 1 and 3 micromol/l) were used. Each treatment was monitored for two hours by using computerized fluoroscopy. Both digoxin and monensin increased the [Ca2+]i and accelerated the onset time of [Ca2+]i increase in a dose-dependent manner. Normal myocytes (loaded with fura-2 for 30 min before the treatment) were also compared with 'weakened' myocytes (loaded with fura-2 for 3 h before the treatment to create a 'weakened' condition). It was found that although 0.3 micromol/l monensin and digoxin did not change the [Ca2+]i in normal myocytes, they increased the [Ca2 +]i in 'weakened' myocytes. Finally, a Na+-free medium was used to demonstrate the effect of [Na+]o on both monensin- and digoxin-induced increases in [Ca2+]i. It was found that digoxin did not increase the [Ca2+]i in the Na+-free medium. Although monensin increased the [Ca2+]i in the Na+-free solution, this increase was not as large as in the Na+-containing medium. The results of the study led to the conclusion that the positive inotropic effect of digoxin depends on [Na+]o. However, monensin increases [Ca2+]i in Na+-dependent and -independent ways. An addition conclusion was that 'weakened' myocytes are more sensitive to the monensin and digoxin treatment than normal myocytes.     

Capsaicin-induced relaxation in rabbit coronary artery.

In the present study mechanism of inhibitory effects of capsaicin on the contractility of rabbit coronary artery were studied by measurement of isometric tension and intracellular Ca2+ concentration. Capsaicin (1 microM to 30 microM) relaxed the coronary artery pre-contracted with prostaglandin (PG) F2alpha (1 microM) in a concentration-dependent manner. The PGF2alpha-induced increase in intracellular Ca2+ concentration was also inhibited. The effects of capsaicin were readily reversed by washing capsaicin from the bath. Capsaicin-induced relaxation was not attenuated by pretreatment with capsazepine (1 microM), a blocker of vanilloid receptor or ruthenium red (1 microM), a blocker of non-selective cation channel. Previous exposure to a high concentration of capsaicin (100 microM) or repeated application of capsaicin did not eliminate the relaxation response to subsequent application of capsaicin. Increasing the external K+ concentration to 80 mM significantly attenuated the capsaicin-induced relaxation with simultaneous change in intracellular Ca2+ concentration. Pretreatment with iberiotoxin (100 nM), a blocker of Ca2+-activated K+ channel, only partially inhibited the capsaicin-induced relaxation. However, application of 4-aminopyridine (4-AP, 1 mM), a blocker of delayed rectifier K+ current significantly inhibited the capsaicin-induced relaxation with concomitant attenuation of the effect on intracellular Ca2+ concentration. These results indicate that capsaicin may have a direct relaxing effect on the smooth muscle contractility, and relaxation may be due to activation of the 4-AP-sensitive, delayed rectifier K+ channels in the rabbit coronary artery.     

Anticholinergic effects of artemisinin, an antimalarial drug, in isolated guinea pig heart preparations

Concern has been growing about the cardiac toxicity of antimalarial drugs. Artemisinin, a unique type of antimalarial drug originating from a Chinese medicinal plant, has minimal adverse effects, but it has been reported to inhibit delayed rectifier potassium current, a voltage-gated potassium current. However, no studies have been published concerning the effect of artemisinin on ligand-gated potassium currents. Therefore, in the present study, we examined the influence of artemisinin on the acetylcholine receptor-operated potassium current (IK.ACh), a ligand-gated potassium current, in guinea pig atrial myocytes using a patch clamp technique. Artemisinin (1 to 300 microM) inhibited I(K.ACh) induced by extracellular application of both carbachol (1 microM) and adenosine (10 microM) and that induced by intracellular loading of GTPgammaS (100 microM) in a concentration-dependent manner. Artemisinin inhibited carbachol-induced, adenosine-induced, and GTPgammaS-activated IK.ACh within almost the same concentration range. In left atria, artemisinin (1 to 100 microM) partially reversed the shortening of action potential duration induced by carbachol in a concentration-dependent manner. Carbachol-induced negative inotropic action in left atria was also inhibited by artemisinin (10 to 300 microM). In conclusion, we suggest that the anticholinergic action of artemisinin is mediated through inhibition of IK.ACh via inhibition of the muscarinic potassium channel and/or associated GTP-binding proteins.     

New perspectives in cardiovascular medicine: the calcium channel blocking drugs

Calcium channel blocking drugs reduce the influx of calcium (Ca++) through cell membrane passageways in excitable tissues. This unique pharmacodynamic action represents an important new addition to cardiovascular therapeutics. Clinically available members of this diverse group of compounds are verapamil, nifedipine, and diltiazem. Beneficial responses to these drugs can be explained by vasodilatation and resulting hemodynamic improvement in tissue perfusion-oxygen demand relationships, by suppression of Ca++-dependent arrhythmogenic mechanisms, by direct reduction of pathologic Ca++ overload in ischemic injured cells, or by a combination of these effects. Calcium channel blockade already has become a therapeutic mainstay in human medicine for management of ischemic heart disease and some forms of cardiac dysrhythmias. Relative to veterinary medicine, Ca++ channel blocking drugs may provide a clinical option for controlling supraventricular tachyarrhythmias. Other indications might include obstructive cardiomyopathies, shock-trauma, and congestive heart failure. Importantly, however, essential issues about adverse cardiovascular side effects of Ca++ channel blocking drugs remain unresolved and controversial. Recent studies especially have raised questions about the tendency for Ca++ channel blockade to exacerbate preexisting or occult myocardial contractile failure. Such pharmacodynamic complexities should be assessed judiciously as the Ca++ channel blocking drugs are appraised for entry into veterinary internal medicine.     

Ca2＋对镉胁迫下玉米幼苗生长、光合特征和 PSⅡ功能的影响

为探究钙(Ca~(2+))对玉米镉(Cd)胁迫的缓解作用,采用盆栽试验,研究了根部施加外源Ca~(2+)对Cd胁迫下玉米幼苗生长、光合特征及叶绿素荧光参数等生理指标的影响。结果显示,与对照(CK)相比,100mg/L的Cd处理显著降低了玉米幼苗株高、根、地上部生物量以及玉米叶片叶绿素a、叶绿素b和总叶绿素含量。同时,净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)、PSⅡ最大光化学效率(F_v/F_m)、电子传递速率(ETR)、PSⅡ激发能捕获效率(F_v′/F_m′)、光化学淬灭系数(qP)和PSⅡ实际光化学效率(Φ_(PSⅡ))显著下降,而非光化学淬灭系数(NPQ)和胞间CO_2浓度(C_i)较CK显著上升。外源施加Ca~(2+)可以有效增加镉胁迫下幼苗生物量的积累,明显提高叶绿素a、叶绿素b和叶绿素总量,升高P_n,G_s,T_r,F_v/F_m,ETR,F_v′/F_m′,降低NPQ和C_i,增加幼苗生物量积累,Ca~(2+)浓度为7.5~10.0mmol/L时各指标变化幅度最明显,缓解胁迫的效果最佳。研究结果表明,Cd胁迫使玉米幼苗叶片PSⅡ原初光能转化效率降低,电子传递受到抑制,净光合速率降低。适宜浓度的外源Ca~(2+)能有效缓解Cd对光合机构的伤害,增强幼苗叶片对光的捕获能力,促进光合作用,增加幼苗的生物量,增强玉米幼苗对Cd胁迫的抗性。     

Mechanism of carbon monoxide-induced relaxation in the guinea pig ileal smooth muscle

The mechanism of carbon monoxide (CO)-induced relaxation were investigated in the guinea-pig ileum. CO (10%) inhibited the 40 mM KCl-induced contraction. This effect was antagonized by ODQ (1 microM), a soluble guanylate cyclase inhibitor. In contrast, CO did not inhibit the 40 mM KCl-induced increase in cytosolic Ca2+ level ([Ca2+]i). Cumulative addition of KCl induced a graded increase in both [Ca2+]i and muscle tension. In the presence of CO, the increase in muscle tension was attenuated whereas the increase in [Ca2+]i was only slightly decreased. Thus, the [Ca2+]i-tension relationship constructed by cumulative addition of KCl shifted downwards in the presence of CO. Using the patch clamp, CO was found to have little effect on the peak Ba currents (I(Ba)) when voltage was stepped from -60 mV to 0 mV. From these results, we conclude that CO inhibits contraction of guinea-pig ileum mainly by the decrease in the sensitivity of contractile elements to Ca2+ via a cyclic GMP-dependent pathway but not by the inhibition of L-type Ca2+ channel.     

Homeostasis of intracellular Ca2+ in equine chondrocytes: response to hypotonic shock

REASONS FOR PERFORMING STUDY: Ca2+ homeostasis in articular chondrocytes affects synthesis and degradation of the cartilage matrix, as well as other cellular functions, thereby contributing to joint integrity. Although it will be affected by mechanical loading, the sensitivity of intracellular Ca2+ concentration ([Ca2+]i) in equine articular chondrocytes to many stimuli remains unknown. HYPOTHESIS: An improved understanding of Ca2+ homeostasis in equine articular chondrocytes, and how it is altered during joint loading and pathology, will be important in understanding how joints respond to mechanical loads. METHODS: [Ca2+]i was determined using the fluorophore fura-2. We examined the effects of hypotonic shock, a perturbation experienced in vivo during mechanical loading cycles. We used inhibitors of Ca2+ transporters to ascertain the important factors in Ca2+ homeostasis. RESULTS: Under isotonic conditions, [Ca2+]i was 148 +/- 23 nmol/l, increasing by 216 +/- 66 nmol/l in response to reduction in extracellular osmolality of 50%. Resting [Ca2+]i, and the increase following hypotonic shock, were decreased by Ca2+ removal; they were both elevated when extracellular [Ca2+] ([Ca2+]o) was raised or following Na+ removal. The hypotonicity-induced rise in [Ca2+]i was inhibited by exposure of cells to gadolinium (Gd3+; 10 micromol/l), an inhibitor of mechanosensitive channels. [Ca2+]i was also elevated following treatment of cells with thapsigargin (10 micromol/l), an inhibitor of the Ca2+ pump of intracellular stores. CONCLUSIONS: A model is presented which interprets these findings in relation to Ca2+ homeostasis in equine articular chondrocytes, including the presence of mechanosensitive channels allowing Ca2+ entry, a Na+/Ca2+ exchanger for removal of intracellular Ca2+ and intracellular stores sensitive to thapsigargin. POTENTIAL RELEVANCE: A more complete understanding of Ca2+ homeostasis in equine chondrocytes may allow development of future therapeutic regimes to ameliorate joint disease.     

The heart remembers: observations of cardiac memory in the Dorper sheep heart

Memory is a property common to a diverse range of tissues. Cardiac memory has been demonstrated in the human, dog, rat and rabbit. This is a peculiar phenomenon, reflected in the T wave of the electrocardiogram. The heart is able to remember periods of alterations in the sequence of ventricular activation and once there is a return to a normal sequence of ventricular activation the T waves may manifest memory. Cardiac memory is noted when the T wave during normal ventricular activation retains the vector of the previous abnormal QRS complex, caused by a period of altered ventricular activation. Possible mechanisms of memory in the heart are alterations of the transient outward potassium current (I10) in ventricular myocytes and new protein synthesis inside myocytes. These two mechanisms operate in short- and long-term cardiac memory respectively. Currently, it is unknown whether memory may have adverse structural consequences in the heart. We were able to demonstrate memory in the hearts of Dorper wethers and this is the first report of cardiac memory in Dorper sheep.     

A single injection of periostin decreases cardiac voltage-gated Na+ channel in rat ventricles

Changes in electrophysiological properties, such as ion channel expression and activity, are closely related to arrhythmogenesis during heart failure (HF). However, a causative factor for the electrical remodeling in HF has not been determined. Periostin (POSTN), a matricellular protein, is increased in heart tissues of patients with HF. In the present study, we investigated whether a single injection of POSTN affects the electrophysiological properties in rat ventricles. After male Wistar rats were intravenously injected with recombinant rat POSTN (64 µg/kg, 24 hr), electrocardiogram (ECG) was recorded. Whole-cell patch clamp was performed to measure action potential (AP) and Na⁺ current (I_Na) in isolated ventricular myocytes. Protein expression of cardiac voltage-gated Na⁺ channel (Na_V1.5) in isolated ventricles was examined by Western blotting. In ECG, POSTN-injection significantly increased RS height. POSTN-injection significantly delayed time to peak in AP and decreased I_Na in the isolated ventricular myocytes. POSTN-injection decreased Na_V1.5 expression in the isolated ventricles. It was confirmed that POSTN (1 µg/ml, 24 hr) decreased I_Na and Na_V1.5 protein expression in neonatal rat ventricular myocytes. This study for the first time demonstrated that a single injection of POSTN in rats decreased I_Na by suppressing Na_V1.5 expression in the ventricular myocytes, which was accompanied by a prolongation of time to peak in AP and an increase of RS height in ECG.     

Photodynamic stimulation causes sustained increase in intracellular calcium concentration in cells of small cell lung carcinoma

Photodynamic agents, due to their selective uptake by tumor cells and photon-dependent selective activation, have immense implications for cancer treatment. The present study provided direct evidence that the photon activation of chloro-aluminum phthalocyanine sulphonate (A1PcS4) in the presence of extracellular Ca2+ caused a rapid increase followed by a sustained increase in intracellular concentration of calcium ion ([Ca2+]i) in a small cell lung carcinoma (SCLC) cell line, SBC-3. The [Ca2+]i increase by photodynamic stimulation was completely inhibited by the removal of extracellular Ca2+ and reintroduction of extracellular Ca2+ immediately led to a rapid elevation of [Ca2+]i. However, the increase was not inhibited by application of Ni2+, nifedipine, or SK&F 96365, a receptor-mediated and voltage-dependent Ca2+ entry blocker. The photosensitizer A1PcS4 alone or light alone (4 min) had no effect on [Ca2+]i. Cytotoxicity examination by trypan blue exclusion test, however, suggested photodynamic stimulation-induced cell injury which was observed in both the presence and the absence of extracellular Ca2+. These results indicate that [Ca2+]i increase may not be mandatory for photodynamic stimulation-induced cell injury. Whether [Ca2+]i increase can accelerate, at least in part, cell death under the physiological condition, whether the mechanism(s) of cell death can be different in the presence and the absence of extracellular Ca2+, and whether [Ca2+]i increase can be totally unrelated to cell death await further work.     

肉鸡腹水综合征患鸡右心组织钙沉积和Ca2+-ATPase活性变化

右心肥大衰竭是腹水综合征患鸡发病的重要环节之一,而心肌细胞内Ca^2＋浓度在调节心脏收缩和舒张功能及其生长方面都起着重要作用。本试验应用右心导管法测定AS患鸡右心压力变化情况,采用焦锑酸钾沉淀法、电镜酶细胞化学法研究AS患鸡右心组织Ca^2＋和钙泵（Ca^2＋-ATPase）活性变化及其精确定位。结果显示AS组肉鸡右心室舒张压极显著高于对照组（P〈0.01）,同时,右心室内压最大变化速率也极显著降低（P〈0.01）;对照组肉鸡右心组织偶见少量散在的Ca^2＋沉淀颗粒,低温诱发AS患鸡右心组织发生了明显的钙沉积;对照组肉鸡的右心组织Ca^2＋-ATPase以高电子密度颗粒分布于肌浆网、线粒体膜等处,AS患鸡心脏组织的Ca^2＋-ATPase的电子密度颗粒显著减少或缺失。本研究揭示,在低温条件下AS患鸡具有明显的右心舒张功能障碍,Ca^2＋浓度增高和Ca^2＋-ATPase功能抑制可能在其中起着重要作用。     

Inhibition of adhesion of F18+ Escherichia coli to piglet intestinal villous enterocytes by monoclonal antibody against blood group H-2 antigen

Enterotoxigenic and verotoxigenic F18⁺ Escherichia coli colonising the pig small intestine, adhere to receptors on intestinal villous enterocytes by F18 fimbriae. The aim of the present study was to define the F18R nature. The knowledge on the nature of this receptor could be important for the development of receptor-based treatments against F18⁺ E. coli-induced disease. The adhesion of F18⁺ E. coli to pig intestinal villous enterocytes was analysed in an in vitro assay. The adhesion of F18⁺ E. coli but not of F4ac⁺ E. coli was strongly inhibited by monoclonal antibodies (mAb) with blood group H-2 specificity. Conversely, blood group H-1 specific mAb could not inhibit the adhesion of F18⁺ E. coli nor F4ac⁺ E. coli. Moreover, the blood group H-2 trisaccharide strongly inhibited the adhesion of F18⁺ E. coli, but only partially the adhesion of F4ac⁺ E. coli. These data demonstrate that the F18 receptor contains the blood group antigen H-2 (-fuc-(1-2)-β-Gal-(1-4)-GlcNAc) as major carbohydrate.     

Epigallocatechin-3 gallate prevents cardiac hypertrophy induced by pressure overload in rats

Pressure overload diseases, such as valvular stenosis and systemic hypertension, manifest morphologically in patients as cardiac concentric hypertrophy. Prevention of cardiac remodeling due to increased pressure overload is important to reduce morbidity and mortality. Epigallocatechin-3 gallate (EGCG) is a major bioactive polyphenol present in green tea which has been found to be a nitric oxide-mediated vasorelaxant and to be cardioprotective in myocardial ischemia-reperfusion injury. Therefore, we investigated whether EGCG supplementation could reduce in vivo pressure overload-mediated cardiac hypertrophy. Cardiac hypertrophy was induced by suprarenal transverse abdominal aortic constriction (AC) in rats. Three weeks after AC surgery, heart to body weight ratio increased in the AC group by 34% compared to the sham group. EGCG administration suppressed the load-induced increase in heart weight by 69%. Attenuation of cardiac hypertrophy by EGCG was associated with attenuation of the increase in myocyte cell size and fibrosis induced by aortic constriction. Despite abolition of hypertrophy by EGCG, transstenotic pressure gradients did not change. Echocardiogram revealed that increased left ventricular systolic dimensions and deteriorated systolic function were relieved by EGCG. These results suggest that EGCG prevents the development of left ventricular concentric hypertrophy by pressure overload and may be a useful therapeutic modality to prevent cardiac remodeling in patients with pressure overload myocardial diseases.     

Mechanisms responsible for reduced cardiotoxicity of mitoxantrone compared to doxorubicin examined in isolated guinea-pig heart preparations

We reported previously that doxorubicin, an anticancer agent that has an anthracycline structure, alters Ca2+ releasing and uptake mechanisms in the sarcoplasmic reticulum of myocardial cells. These effects of doxorubicin are apparently related to its cardiotoxicity. Mitoxantrone is a similar anticancer agent with an anthracenedion structure that has been shown to be significantly less cardiotoxic. In the present study, the effects of mitoxantrone on the functions of the sarcoplasmic reticulum were examined in isolated muscle preparations obtained from the guinea-pig heart. In electrically-stimulated left atrial muscle preparations, incubation in vitro for 4 hr with 30 or 100 microM mitoxantrone significantly prolonged the time to the peak of twitch tension, markedly increased the developed tension observed at lower stimulation frequencies, thereby attenuating the slope of positive force-frequency relationships, and increased the postrest contraction observed after a 60-sec quiescent period. In myocytes isolated from ventricular muscles, 30 microM mitoxantrone increased the peak and the size of intracellular Ca2+ concentrations ([Ca2+] i), and prolonged the time to peak [Ca2+]i. In skinned muscle fiber preparations obtained from the left ventricular muscle, 30 muM mitoxantrone significantly increased the caffeine-induced contraction without affecting the Ca2+ sensitivity of contractile proteins. These results suggest that mitoxantrone enhances Ca2+ release from the sarcoplasmic reticulum in isolated atrial muscle preparations obtained from the guinea-pig heart. Apparent enhancement of the sarcoplasmic reticulum functions, in contrast to anthracyclines that has been shown to suppress these functions, seems to explain the relative lack of marked cardiotoxicity of mitoxantrone.     

Identification of common antigens in ribosome-rich extracts from Fusobacterium necrophorum

Ribosome-rich extracts (RRE) were prepared by differential and ultracentrifugation from 25 bovine and 6 ovine isolates of Fusobacterium necrophorum (FN) including both biotypes A and B. A pooled rabbit antiserum was prepared against whole-cell and sonicated whole-cell bacterins of F necrophorum isolate FN 3080, and a 2nd pooled rabbit antiserum was prepared against a RRE of FN 3080. The RRE of the 25 bovine isolates were tested against the FN 3080 whole-cell antiserum, using Ouchterlony double-immunodiffusion procedures. One to 3 precipitin lines were observed with the 25 isolates. The individual bovine isolates were found to have lines of identity with 5 to 21 of the other 24 isolates. The 25 bovine isolates and the 6 ovine isolates were then compared, using the FN 3080 RRE antiserum. One to 3 precipitin lines were observed for the 31 isolates with the RRE antiserum, and lines of identity were observed between all 31 of the isolates. These results indicated that common antigens are present in the RRE from a wide variety of F necrophorum isolates including both A and B biotypes.