Isolation, identification and cyfluthrin-degrading potential of a novel Lysinibacillus sphaericus strain, FLQ-11-1.

Strain FLQ-11-1, isolated from sewage sludge, was able to degrade cyfluthrin and was identified as Lysinibacillus sphaericus based on its morphology, 16S rRNA sequence and fatty acid methyl ester (FAME) analyses. This strain could use cyfluthrin as its carbon or nitrogen source. Response surface methodology (RSM) analysis showed that the optimum conditions for degradation were at pH 7.0 and 35 °C, using an inoculum amount with an OD600nm value of 1.6. Under these conditions, approximately 80.4% of cyfluthrin (50 mgl(-1)) was degraded within five days (d) of incubation. Four metabolic compounds were detected during cyfluthrin degradation and identified as methyl-3-(2,2-dichlorovinyl)-2,2-dimethyl-(1-cyclopropane)-carboxylate, 4-fluoro-3-phenoxy-benzoic acid methyl ester, methyl-3-phenoxybenzoate, 3-phenoxy-benzaldehyde by gas chromatography-mass spectrometry (GC-MS) and tandem mass spectrum (MS/MS) analysis and no cyfluthrin was detected after seven days of incubation. A possible degradation pathway was proposed, and our data showed that cyfluthrin could be efficiently degraded by FLQ-11-1, indicating that this strain could potentially be used to eliminate the contamination of pyrethroid herbicides.

Improved early postresuscitation EEG activity for animals treated with hypothermia predicted 96 hr neurological outcome and survival in a rat model of cardiac arrest.

PURPOSE: To investigate the effect of hypothermia on 96 hr neurological outcome and survival by quantitatively characterizing early postresuscitation EEG in a rat model of cardiac arrest. MATERIALS AND METHODS: In twenty male Sprague-Dawley rats, cardiac arrest was induced through high frequency transesophageal cardiac pacing. Cardiopulmonary resuscitation was initiated after 5 mins untreated arrest. Immediately after resuscitation, animals were randomized to either 2 hrs of hypothermia (N = 10) or normothermia (N = 10). EEG, ECG, aortic pressure, and core temperature were continuously recorded for 6 hrs. Neurological outcome was evaluated daily during the 96 hrs postresuscitation period. RESULTS: No differences in the baseline measurements and resuscitation outcome were observed between groups. However, 96 hr neurological deficit score (204 ± 255 versus 500 ± 0, P = 0.005) and survival (6/10 versus 0/10, P = 0.011) were significantly better in the hypothermic group. Quantitative analysis of early postresuscitation EEG revealed that burst frequency and spectrum entropy were greatly improved in the hypothermic group and correlated with 96 hr neurological outcome and survival. CONCLUSION: The improved burst frequency during burst suppression period and preserved spectrum entropy after restoration of continuous background EEG activity for animals treated with hypothermia predicted favorable neurological outcome and survival in this rat model of cardiac arrest.

Effect of therapeutic hypothermia vs δ-opioid receptor agonist on post resuscitation myocardial function in a rat model of CPR.

AIM: This study is to compare the effect of the δ-opioid receptor agonist, D-Ala(2)-D-Leu(5) enkephalin (DADLE) with normothermic control and therapeutic hypothermia on post resuscitation myocardial function and 72-h survival in a rat model of cardiac arrest and resuscitation. METHODS: Ventricular fibrillation (VF) was induced in 15 male Sprague-Dawley rats. After 8 min of untreated VF, cardiopulmonary resuscitation was performed for 8 min before defibrillation. Animals were randomized to three groups of five: (a) normothermia; (b) hypothermia (32 °C); and (c) normothermia with DADLE intravenous infusion (1 mg/kg h(-1)). Hypothermia and drug infusion were started after successful defibrillation. Myocardial functions, including cardiac output (CO), left ventricular ejection fraction (LVEF), and myocardial performance index (MPI) were measured echocardiographically together with duration of survival. RESULTS: The 72-h survival was significantly greater in the hypothermic group than in both DADLE and normothermic group (p = 0.02). However, the survival time of the DADLE treated animals was significantly longer than that of the normothermia group (51.8 ± 18.9 vs 18.8 ± 10.1h, p < 0.01). DADLE group showed significantly better CO (PR 60 min, p = 0.049), better LVEF (PR 60 min, p = 0.044; PR 240 min, p < 0.001) and lower MPI (PR 60 min, p = 0.043; PR 240 min, p = 0.045) than normothermic group. Hypothermia group also showed significantly better CO (PR 60m in, p = 0.044; PR 240 min, p = 0.007), better LVEF (PR 60 min, p = 0.001; PR 240 min, p < 0.001) and lower MPI (PR 60 min, p = 0.003; PR 240 min, p = 0.012) than the normothermic group. CONCLUSIONS: DADLE attenuated post resuscitation myocardial dysfunction and increased short term survival time. However, the 72-h survival in the DADLE group was less than that in the hypothermia group.

What is the optimal dose of epinephrine during cardiopulmonary resuscitation in a rat model?

OBJECTIVE: Because different species may require different doses of drug to produce the same physiologic response, we were provoked to evaluate the dose-response of epinephrine during cardiopulmonary resuscitation (CPR) and identify what is the optimal dose of epinephrine in a rat cardiac arrest model. METHODS: Rat cardiac arrest was induced via asphyxia, and then the effects of different doses of epinephrine (0.04, 0.2, and 0.4 mg/kg IV, respectively) and saline on the outcome of CPR were compared (n = 10/each group). The primary outcome measure was restoration of spontaneous circulation (ROSC), and the secondary was the change of spontaneous respiration and hemodynamics after ROSC. RESULTS: Rates of ROSC were 9 of 10, 8 of 10, 7 of 10, and 1 of 10 in the low-dose, medium-dose, and high-dose epinephrine groups and saline group, respectively. The rates of withdrawal from the ventilator within 60 minutes in the low-dose (7 of 9) and medium-dose epinephrine groups (7 of 8) were higher than in the high-dose epinephrine group (1 of 7, P < .05). Mean arterial pressures were comparable, but the heart rate in the high-dose epinephrine group was the lowest among epinephrine groups after ROSC. These differences in part of time points reached statistical significance (P < .05). CONCLUSION: Different doses of epinephrine produced the similar rate of ROSC, but high-dose epinephrine inhibited the recovery of spontaneous ventilation and caused relative bradycardia after CPR in an asphyxial rat model. Therefore, low and medium doses of epinephrine were more optimal for CPR in a rat asphyxial cardiac arrest model.

Dose-response of vasopressin in a rat model of asphyxial cardiac arrest.

The advantage of vasopressin over epinephrine in the treatment of cardiac arrest (CA) is still being debated, and it is not clear whether a high dose of vasopressin is beneficial or detrimental during or after cardiopulmonary resuscitation (CPR) in a rat model of CA. In this study, asphyxial CA was induced in 40 male Sprague-Dawley rats. After 10 minutes of asphyxia, CPR was initiated; and the effects of different doses of vasopressin (low dose, 0.4 U/kg; medium dose, 0.8 U/kg; and high dose, 2.4 U/kg; intravenous; n = 10 in each group) and a saline control (isotonic sodium chloride solution, 1 mL, intravenous) were compared. Outcome measures included the rate of restoration of spontaneous circulation (ROSC) and changes of hemodynamic and respiratory variables after ROSC. The rates of ROSC were 1 of 10 in the saline group and 8 of 10 in each of the 3 vasopressin groups. There were no differences in mean aortic pressure or changes of respiratory function after CPR among the vasopressin groups. However, the heart rate was lower in the high-dose vasopressin group than in the low- and medium-dose groups. These findings indicate that different doses of vasopressin result in a similar outcome of CPR, with no additional benefits afforded by a high dose of vasopressin during or after CPR, in a rat model of asphyxial CA. The mechanism and physiologic significance of the relative bradycardia that occurred in the high-dose vasopressin group are currently unknown and require further investigation.

Combination of cardiac pacing and epinephrine does not always improve outcome of cardiopulmonary resuscitation.

We hypothesized that the combination of cardiac pacing and epinephrine would yield a better efficacy for cardiopulmonary resuscitation (CPR) and the combination of 2 therapies at different opportunity would achieve the same results of CPR. Cardiac arrest was induced by clamping the tracheal tubes in 60 Sprague-Dawley rats. At 10 minutes of asphyxia, the animals were prospectively randomized into 5 groups (n = 12/group), and received saline (Sal-gro, 1 mL, intravenous [i.v.]), epinephrine (Epi-gro, 0.4 mg/kg, i.v.), pacing (Pac-gro, transesophageal cardiac pacing combined with saline 1 mL, i.v.), pacing + epinephrine group 1 (PE-gro1, transesophageal cardiac pacing combined with epinephrine 0.4 mg/kg, i.v.), or pacing + epinephrine group 2 (PE-gro2, transesophageal cardiac pacing combined with epinephrine 0.4 mg/kg, i.v., 4 minutes after the transesophageal cardiac pacing initiating and failing to resuscitate the animals), followed by initiation of CPR. Restoration of spontaneous circulation in Sal-gro was lower than in Epi-gro, Pac-gro, PE-gro1, and PE-gro2 (16.67% vs 66.67%, 66.67%, 100%, and 100%; P < .05 or P < .001, respectively). The proportions of withdrawing ventilator and 2-hour survival proportions in Pac-gro and PE-gro2 were higher than in Epi-gro and PE-gro1 (8/8, 10/12 vs 1/8, 2/12, respectively, P < .01, and 7/8, 8/12 vs 1/8, 2/12, respectively, P < .05 or P < .01). Mean survival time in Pac-gro and PE-gro2 were longer than in Epi-gro and PE-gro1 (P < .05 or P < .01). Therefore, the combination of 2 therapies does not always improve outcome of CPR. It is obvious that the combination of transesophageal cardiac pacing with delayed administration of epinephrine yields a better outcome compared to the combination of 2 therapies at the same time during CPR in a rat asphyxia cardiac arrest model.

A simpler cardiac arrest model in rats.

Two disadvantages of electrical induction of cardiac arrest used currently are that it is a technically complicated procedure and the consequent thermal injury, which prompts us to search for a simpler method with less adverse effect to induce ventricular fibrillation (VF) in rats. Different potential (18, 24, 30, and 36 V) of alternating current (AC) were administered to elicit VF in 15 rats via pacing electrode placed in esophagus. Four minutes after onset of VF, conventional cardiopulmonary resuscitation (CPR) was initiated. Restoration of spontaneous circulation was defined as the return of supraventricular rhythm with a mean aortic pressure of 20 mm Hg or greater for a minimum of 5 minute. Ventricular fibrillation was achieved by short interval of AC stimulation in all of the rats. After the termination of prolonged AC stimulation, electrocardiogram indicated VF occurred in 6 of 15 rats, asystole in 3 of 15 rats and pulseless electrical activity in 6 of 15 rats. Before CPR, however, electrocardiogram indicated that only 2 of 15 and 4 of 15 animals remained in VF and pulseless electrical activity, respectively, whereas 9 of 15 animals presented as asystole. After CPR, 11 of 15 animals were resuscitated. Necropsy showed that there was no gross evidence of thermal injury on the surface layer of the heart. Therefore, development of a rat cardiac arrest model by transesophageal AC stimulation is simpler and less adverse effect, which may have practical significance for facilitating experimental investigation on cardiac arrest and CPR.

A simpler cardiac arrest model in the mouse.

OBJECTIVE: Delivering alternating currency (AC) to right ventricular endocardium to induce ventricular fibrillation (VF) in mice is complicated. We tried to validate whether transoesophageal AC stimulation could induce VF and how long AC stimulation had to be sustained to prevent the spontaneous cardioversion of VF in mice. METHODS: A pacing electrode was inserted orally into the oesophagus and AC was delivered to esophagus through the pacing electrode to stimulate the heart and induce VF in 15 mice. The incidence of VF and time of AC stimulation were recorded 4min after onset of VF cardiopulmonary resuscitation (CPR) was started. RESULTS: VF was induced by short AC stimulation in all 15 mice. With the prolongation of AC stimulation, the incidences of spontaneous cardioversion of VF decreased whereas the incidence of pulseless electrical activity (PEA) increased accordingly. Following the termination of prolonged AC stimulation, VF occurred only in 1 of 15 mice, but PEA in 14 of 15 mice. Before CPR 1 of 15 and 12 of 15 animals remained in VF and in PEA, respectively, while 2 of 15 animals developed into asystole. After CPR, 11 of 15 animals were successfully resuscitated. CONCLUSION: VF can be induced by a short period of transoesophageal AC stimulation in mice. However, prolonged AC stimulation is prone to induce PEA other than VF. Nonetheless, the development of a mouse CA model in this manner is simpler and easier, which may have practical significance for facilitating experimental investigation on CA and CPR.

Epinephrine, but not vasopressin, improves survival rates in an adult rabbit model of asphyxia cardiac arrest.

Although vasopressin has been reported to be more effective than epinephrine for cardiopulmonary resuscitation in ventricular fibrillation animal models, its efficacy in asphyxia model remains controversy. The purpose of this study was to investigate the effectiveness of vasopressin vs epinephrine on restoration of spontaneous circulation (ROSC) in a rabbit model of asphyxia cardiac arrest. Cardiac arrest was induced by clamping endotracheal tube. After 5 minutes of basic life-support cardiopulmonary resuscitation, animals who had no ROSC were randomly assigned to receive either epinephrine alone (epinephrine group; 200 microg/kg) or vasopressin alone (vasopressin group; 0.8 U/kg). The coronary perfusion pressure (CPP) was calculated as the difference between the minimal diastolic aortic and simultaneously recorded right atrial pressure. Restoration of spontaneous circulation was defined as an unassisted pulse with a systolic arterial pressure of 60 mm Hg or higher for 5 minutes or longer. We induced arrest in 62 rabbits, 15 of whom had ROSC before drug administration and were excluded from analysis. The remaining 47 rabbits were randomized to epinephrine group (n = 24) and vasopressin group (n = 23). Before and after drug administration, CPP in epinephrine group increased significantly (from -4 +/- 4 to 36 +/- 9 mm Hg at peak value, P = .000), whereas CPP in vasopressin group increased only slightly (from 9 +/- 5 to 18 +/- 6 mm Hg at peak value, P = .20). After drug administration, 13 of 24 epinephrine rabbit had ROSC, and only 2 of 23 vasopressin rabbit had ROSC (P < .01). Consequently, we conclude that epinephrine, but not vasopressin, increases survival rates in this adult rabbit asphyxia model.

Ventricular fibrillation induced by transoesophageal cardiac pacing: a new model of cardiac arrest in rats.

OBJECTIVE: To investigate whether transoesophageal cardiac pacing can induce ventricular fibrillation (VF) and how long the cardiac pacing has to be sustained to prevent the reversion of the VF induced. METHODS: A pacing electrode was inserted orally into the oesophagus and high-frequency ventricular pacing was performed so as to elicit VF in 25 Sprague-Dawley rats. Incidences of VF and time of cardiac pacing were observed and recorded. Four minutes after onset of VF cardiopulmonary resuscitation (CPR) was initiated. RESULTS: A short interval of high-frequency ventricular pacing caused an immediate drop of blood pressure, loss of pulse and increase of right atrial pressure in the same time frame. When the cardiac pacing was terminated, VF was elicited at least once or more than once in all of the 25 rats. However, the VF elicited by the burst stimulation could be defibrillated spontaneously. With the prolongation (120-180 s) of cardiac pacing, the incidence of defibrillation of VF decreased from 100 to 0%. VF persisted in 19 of 25 animals, developed into asystole in 5 of 25 animals and converted into pulseless electrical activity in 1 of 25 animals prior to CPR. Following CPR 22 of 25 animals were resuscitated. CONCLUSIONS: Transoesophageal cardiac pacing can induce VF in rats. However, the cardiac pacing is required for at least 120-180 s to ensure that VF does not spontaneously convert. We can use the technique to establish a new and simpler rat cardiac arrest (CA) model, which may facilitate experimental investigation on CPR.

A comparison of transoesophageal cardiac pacing and epinephrine for cardiopulmonary resuscitation.

The use of cardiac pacing to deal with bradycardia is well established. There is debate as to the benefits during cardiopulmonary resuscitation (CPR). This study was performed to compare the effects of transoesophageal cardiac pacing and high-dose epinephrine on the benefits of cardiopulmonary resuscitation after asphyxial cardiac arrest in rats. Thirty Sprague-Dawley rats of both sexes were randomly selected to a saline group (Sal-gro, treated with normal saline 1 mL IV, n = 10), an epinephrine group (Epi-gro, treated with epinephrine 0.4 mg/kg IV, n = 10), or a pacing group (Pac-gro, treated with normal saline 1 mL IV combined with transoesophageal cardiac pacing, n = 10) in a blinded fashion during resuscitation after 10 minutes of asphyxial cardiac arrest. Manual chest compression was in all cases performed using the same methodology by the same personnel who was blinded to hemodynamic monitor tracings. The rate of restoration of spontaneous circulation was 1 (10%), 7 (70%), and 8 (80%) of 10 in Sal-gro, Epi-gro, and Pac-gro, respectively. The rate of ventilator withdrawal within 60 minutes after resuscitation in Pac-gro was higher than that of Epi-gro (8/8 vs 1/7, respectively; P = .001); the survival rate after 2 hours in Pac-gro was significantly higher than that in Epi-gro (7/8 vs 1/7, respectively; P = .01). The data demonstrate that both epinephrine and transoesophageal cardiac pacing are effective within 10 minutes of asphyxia in rats. It is worth noting that transoesophageal cardiac pacing produced a better outcome with respiration and longer survival time compared with epinephrine after restoration of spontaneous circulation.

Does naloxone alone increase resuscitation rate during cardiopulmonary resuscitation in a rat asphyxia model?

Cardiac arrest was induced with asphyxia to identify if naloxone alone increases resuscitation rate during cardiopulmonary resuscitation in a rat asphyxia model. The animals were randomized into either a saline group (Sal-gro, treated with normal saline 1 ml iv, n = 8), a low-dose naloxone group (treated with naloxone 0.5 mg/kg iv, n = 8), or a high-dose naloxone group (HN-gro, treated with naloxone 1 mg/kg iv, n = 8) in a blinded fashion during resuscitation. At the end of 10 minutes of asphyxia, cardiopulmonary resuscitation was started, and each drug was administered at the same time. The rate of restoration of spontaneous circulation was seen in 1 of 8, 3 of 8, and 7 of 8 animals in the Sal-gro, LN-gro, and HN-gro, respectively. The rate of restoration of spontaneous circulation in HN-gro was significantly higher than that in Sal-gro (P < .05). Naloxone (1 mg/kg) alone can increase resuscitation rate following asphyxial cardiac arrest in rats.

Transoesophageal cardiac pacing is effective for cardiopulmonary resuscitation in a rat of asphyxial model.

OBJECTIVE: To investigate effectiveness of transoesophageal cardiac pacing in a rat model of asphyxial cardiac arrest. METHODS: Ten minutes after the tracheal tube had been clamped, cardiac arrest (CA) occurred in 20 Sprague-Dawley rats, and the rats were assigned randomly to receive cardiopulmonary resuscitation (CPR) in a control group or CPR combined with transoesophageal cardiac pacing in a pacing group. Restoration of spontaneous circulation (ROSC) was defined as an unassisted pulse with a mean arterial pressure (MAP) of >or=20 mmHg for >or=1 min. RESULTS: ROSC was significantly more frequent in the pacing group compared with the control group (7/10 versus 1/10, P<0.05). Faster ROSC and longer survival trend in the pacing group were seen compared with the control group. CONCLUSION: Transoesophageal cardiac pacing is effective for CPR in a rat of asphyxial model. However, the precise mechanism is not clear and further experiments will be necessary.