Effects of intravenous Escherichia coli dose on the pathophysiological response of colostrum-fed Jersey calves

Objectives of the present study were to characterize the dose dependency of an intravenous Escherichia coli O111:H8 challenge in colostrum-fed Jersey calves and to identify any biochemical markers indicative of septicemia. Eighteen 3-week old colostrum-fed Jersey calves were completely randomized to 1 of 6 doses of E. coli O111:H8. The challenge doses included 0, 1.5 x 10?, 1.5 x 10?, 1.5 x 10?, 1.5 x 10?, and 1.5 x 10? colony-forming units (CFU) given intravenously as a bolus in 5 mL of sterile isotonic saline. Peripheral blood samples were collected at 0, 2, 4, 8, 12, 24, and 48 h relative to the challenge for biochemical, total leukocyte count, and differential analyses. Rectal temperatures were collected via indwelling rectal temperature probes at 5-min intervals, and hourly averages calculated from 2 d prior to the challenge till 2 d after the challenge. All calves survived the 48 h observation period following the challenge. Calves given 1.5 x 10? and 1.5 x 10? CFU displayed sickness behaviors (P < 0.01) beginning 0.5 h after the challenge and returned to that of the control calves by 6 and 32 h for calves challenged with 1.5 x 10? and 1.5 x 10? CFU, respectively. There were treatment x time interactions (P < 0.01) on total leukocyte counts and plasma glucose and zinc concentrations. Calves administered 1.5 x 10? and 1.5 x 10? CFU had leucopenia beginning 2 h after the challenge and returning to counts similar to the control calves within 24 h. Additionally, those calves were hypoglycemic from 4 to 12h after the challenge with the degree of hypoglycemia inversely related to the dose of the E. coli. All calves challenged with E. coli had decreased plasma zinc concentrations, and the magnitude was inversely proportional to the challenge dose. There were treatment x time interactions (P < 0.001) on rectal temperatures following the challenge. All calves challenged with E. coli developed a febrile response, but the intensity and duration of the response were dependent on the challenge dose. These data indicate that calves intravenously challenged with 1.5 x 10? and 1.5 x 10? CFU of the E. coli O111:H8 showed immediate clinical and biochemical signs indicative of septicemia. However, calves administered 1.5 x 10? or less of the E. coli had febrile responses, but did not develop septicemia. Blood glucose and zinc concentrations may be dose responsive indicators that could potentially differentiate between a septicemic versus non-septicemic calf.     

Hemodynamics, plasma eicosanoid concentrations, and plasma biochemical changes in calves given multiple injections of Escherichia coli endotoxin

Twelve male neonatal calves (39 to 50 kg) were allotted to 3 groups of 4 calves each. All calves were anesthetized with halothane, and then Escherichia coli endotoxin was given intravenously (3 times) and intraperitoneally (3 times) during a 6-hour period. Group-1 calves were untreated, group-2 calves were pretreated with a low dose of flunixin meglumine (1.1 mg/kg of body weight), and group-3 calves were pretreated with a high dose of flunixin meglumine (4.4 mg/kg). In calves of group 1, the mean systemic arterial blood pressure (MABP) and cardiac output (CO) decreased, but pulmonary arterial pressure increased after the initial intravenous and intraperitoneal injections of endotoxin. In calves of this group, these changes were accompanied by increased plasma thromboxane B2 (TxB2) concentration. During this period, increased plasma TxB2 concentration or hemodynamic changes were not detected in calves of groups 2 and 3. Only calves of group 1 had altered hemodynamics early in the experiment; however, after 6 hours, calves of all 3 groups had similarly decreased CO and MABP. In calves of the untreated group, plasma 6-keto-prostaglandin (PG)F1 alpha concentration increased steadily from the beginning of the experiment until 3 hours later. The CO and MABP were low at the time when serum 6-keto-PGF1 alpha concentration was high; however, these 2 measurements also were low in treated calves who did not have correspondingly high plasma 6-keto-PGF1 alpha concentration. Regional blood flow analysis did not reveal correlations between prostanoid concentrations and altered blood flow to selected tissues.(ABSTRACT TRUNCATED AT 250 WORDS)     

Virulence factors in Escherichia coli isolated from the blood of bacteremic neonatal calves

Twenty-five Escherichia coli isolates from the blood of critically ill bacteremic calves sampled in two separate studies on a calf-rearing farm housing over 15,000 calves, in the San Joaquin Valley, California were studied.Isolates were characterized for O serogroups and for pathotypes as determined by the presence of specific virulence factors including heat-labile enterotoxin (LT), heat-stable enterotoxins a and b (STa, STb), verotoxins 1 and 2 (VT1, VT2), cytotoxic necrotizing factor (CNF), aerobactin, intimin Eae and P, F17 and CS31A fimbrial adhesins, and resistance to bactericidal effects of serum.These isolates constituted a heterogeneous group. However, isolates were mostly aerobactin positive and often resistant to the bactericidal effects of serum. Isolates of pathotypes O78 (n=6), O119:CS31a (n=3), and P positive but O non-typeable (n=3) were associated with a high mortality rate. The remaining isolates belonged to diverse pathotypes, often possessing the adhesins P, F17, CS31A and Eae but belonging to O serogroups other than O78 and O119, and were less frequently associated with mortality.Although no virulence factor common to all isolates was identified, the capacity to use iron by the presence of aerobactin which is important to the capture of iron was a predominant factor. Moreover, certain pathotypes appear to be associated with primary colisepticemia whereas other pathotypes may cause a bacteremia without necessarily leading to septicemia.     

Estimating leukocyte,platelet, and erythrocyte counts in rats by blood smear examination

Background: The CBC is an essential test for assessing the health of rats used in drug development studies. Because of limited blood volume, estimates of cell counts from a blood smear would be valuable when other analytical methods of enumerating cells are not possible or available. Objective: The purpose of this study was to develop a statistical model to accurately estimate WBC, platelet (PLT), and RBC counts in blood smears from rats. Method: Blood smears and quantitative cell counts were obtained from vehicle‐treated male and female Fischer 344 rats (n=65) involved in a variety of studies. The numbers of WBCs, PLTs, and RBCs were estimated in 10 fields in the monolayer of smears using × 20 (WBC) or × 100 (PLT, RBC) objectives. Using a statistical model and the quantitative cell counts obtained on an ADVIA 120 hematology analyzer, formulas were developed to predict the quantitative counts from the estimates. Results: Data were log‐transformed before analysis. A formula was derived using the slope and intercept of the regression line between cell estimates and ADVIA counts to predict WBC, PLT, and RBC counts based only on estimates. A second formula was developed for situations in which limited quantitative analyses may be available, and resulted in even more accurately predicted counts from smear estimates. Conclusion: The formulas developed in this study can be a valuable tool in estimating cell counts from a blood smear when cell counting instruments are not available or when an instrument cell count needs to be verified. These formulas may be useful in the assessment of rat blood in discovery and lead optimization studies.     

Haematological changes in buffalo calves inoculated with Escherichia coli endotoxin and corticosteroids

Haematological studies were conducted on 10 clinically normal water buffalo calves to determine leucocytic responses to Escherichia coli endotoxin, prednisolone and dexamethasone. Intravenous injection of 10 micrograms endotoxin induced minimal decreases in leucocyte numbers, whereas 20, 50 and 100 micrograms produced a marked leucopenia within one hour. Moderate to marked leucopenia, neutropenia and lymphopenia persisted for three to 14 hours. Significant rebound neutrophilia was evident at six to eight hours after inoculation in calves given only 10 and 20 micrograms. Intramuscular injection of prednisolone (100 mg) and dexamethasone (5 mg) produced increases in total leucocyte counts and neutrophil numbers within two hours. Moderate to marked leucocytosis and neutrophilia persisted for eight to 24 hours. Lymphocyte response was unlike that in other species in that lymphopenia was not a consistent feature of the corticosteroid response. A transient monocytosis was seen following administration of prednisolone but not of dexamethasone, while eosinopenia and basopenia developed in both cases. In conclusion, endotoxin and corticosteroid induced changes in total and differential leucocyte counts in water buffalo were largely similar to those seen in cattle.     

Effect of combined rotavirus and Escherichia coli in neonatal gnotobiotic calves

Gnotobiotic calves (24 hours old) were monoinfected with calf rotavirus (CRV) strain NCDV, an enterotoxigenic Escherichia coli (ETEC) strain B44 (K99+), or a nonenterotoxigenic E coli (NETEC) strain 123 (K99-). Calves also were dually infected with CRV and either ETEC or NETEC. Eighteen calves equally allotted between 6 treatment groups were used in these studies: Noninfected controls--group A; CRV--group B; ETEC--group C; NETEC--group D; CRV + ETEC--group E; and CRV + NETEC--group F. Severe diarrhea and villous atrophy were observed in calves of treatment groups B, C, E, and F. Mortality was present only in treatment groups C and E as result of ETEC infection. There were no significant differences in the clinical responses or enteric lesions between treatments B and F, although a significant increase in the concentrations of NETEC was demonstrated in calves dually infected with CRV + NETEC (group F) as compared with calves monoinfected with NETEC (group D). Calves inoculated with ETEC (group C) had severe villous atrophy, neutrophilic infiltration of intestinal lumen, and moderate enterocyte necrosis. Calves dually inoculated with CRV + ETEC (group E) had the most extensive and severe lesions, similar to those in group C, plus a pronounced necrotic fibrino-hemorrhagic enteritis. Infection of enterocytes by CRV did not affect in any way the adherence of ETEC to the intestinal mucosa. Dual viral and bacterial infections of the same enterocytes were evident.     

Occurrence of enterotoxigenic Escherichia coli in calves with acute neonatal diarrhoea

Enterotoxigenic Escherichia coli (ETEC) were isolated from 16.4% of diarrheic calves of up to 30 days old. Of 1-2-day-old calves nearly one half (45.0%) harboured ETEC, while this was the case with only 4.9% of 8-day-old calves. Among 206 9-30-day-old calves just three were found to harbour ETEC.     

Pathotyping and antimicrobial susceptibility testing of Escherichia coli isolates from neonatal calves

Neonatal calf mortality is a major concern to livestock sector worldwide. Neonatal calf diarrhoea (NCD), an acute severe condition causes morbidity and mortality in calves. Amongst various pathogens involved in NCD, E. coli is considered as one of the major causes. The study was targeted to characterize E. coli isolates from neonatal calves for diarrhoeagenic Escherichia coli (DEC) types (pathotyping), antimicrobial resistance (AMR) profiling and to correlate with epidemiological parameters. From neonates, a total of 113 faecal samples were collected, out of that 308, lactose fermenting colonies were confirmed as E. coli. Pathotypable isolates (12.3%) were represented by STEC (6.1%), EPEC (2.9%), ETEC (1.9%), EAEC (0.9%) and EHEC (0.3%). Occurrence of STEC was more in non-diarrhoeic calves, whereas ETEC was observed more in diarrhoeic calves. EPEC occurrence was observed in both diarrhoeic and non-diarrhoeic calves. Fishers extract test showed no significant association for occurrence of DEC types to type of dairies, health status, species, breed, age and sex of neonatal calves. Two hundred and eighty isolates were tested for antimicrobial susceptibility. The isolates showed maximum resistance towards ampicillin (55.4%) followed by tetracycline (54.3%), while minimum resistance was observed towards meropenem (2.5%). Multidrug resistant E. coli isolates were found to be 139 (49.6%), and Extended-spectrum beta-lactamase (ESBL) producers were 120 (42.9%). DEC pathotypes like STEC, ETEC, EHEC and EAEC that are also multidrug resistant present in neonatal calves have zoonotic potential and hence are of public health significance.

     

芽孢杆菌对大肠杆菌感染断奶仔猪生长性能养分消化率及血液白细胞分类的影响

本试验中F4受体阴性断奶仔猪先提前1周灌服低(3.9×108CFU/头·d)、高(7.8×108CFU/头·d)剂量枯草芽孢杆菌及地衣芽孢杆菌混合制剂,紧接着大肠杆菌F4+ETEC感染,观察芽孢杆菌对仔猪生长性能、养分消化率及白细胞分类的影响。结果显示:高、低剂量芽孢杆菌制剂均显著抑制大肠杆菌引起的仔猪采食量下降。大肠杆菌感染降低了粗蛋白质及总能消化率,而低剂量芽孢杆菌制剂显著抑制F4+ETEC引起的粗蛋白质和总能消化率的下降。低剂量芽孢杆菌制剂显著抑制大肠杆菌引起的杆状核嗜中性粒细胞比例升高。结果表明,芽孢杆菌可以改善断奶仔猪生长性能,低剂量芽孢杆菌可以抑制养分消化率降低,但高剂量芽孢杆菌可能引起免疫失调。     

Antimicrobial alternatives for calf diarrhea: enteric responses to Escherichia coli, deferoxamine, or gallium in neonatal calves

Intestinal malabsorption or transferable resistance in enteric bacteria are potentially serious complications of routine oral administrations of antibiotics. On the basis of reports on antimicrobial effects of host iron sequestration and on synthetic iron chelators or competitors in vitro, 2 iron antagonists were studied for their potential as alternative antimicrobials for Escherichia coli diarrhea. Deferoxamine, a fungal iron chelator used to treat acute iron intoxication, and elemental gallium, a competitive inhibitor of iron activity in metabolic enzyme systems, were examined for their effects on enteric morphology and function in neonatal calves. Twelve male calves were allotted to 4 groups: (1) given nonpathogenic E coli (control); (2) given enterotoxigenic B44 E coli; (3) given deferoxamine (50 mg/kg, twice a day); and (4) given gallium (4 mg/kg, twice a day). Calves were studied for 8 days, including the conduct of oral glucose and lactose tolerance tests on days 1, 3, and 7. By day 7, according to oral glucose and lactose tolerance tests, peak plasma glucose concentrations in all calves of groups 2, 3, and 4 were lower than those values in controls. The frequency of diarrhea was significant in all treated calves, and disease was most severe in the deferoxamine-treated calves. Quantitative cultural examination on day 8 showed significant numerical increases of jejunal and ileal E coli and ileal lactobacilli in deferoxamine-treated calves (group 3) and of ileal streptococci in gallium-treated calves (group 4) and showed jejunal and ileal overgrowths of Saccharomyces yeast in deferoxamine-treated calves.(ABSTRACT TRUNCATED AT 250 WORDS)     

牛膝多糖对脂多糖免疫应激仔猪血液生化指标和白细胞分类计数的影响

试验旨在探讨牛膝多糖(ABPS)对脂多糖(LPS)刺激的断奶仔猪血液生化指标和白细胞分类计数的影响。选用48头(28±3)d、体重(8.45±0.14)kg的杜洛克×长白×大白断奶仔猪,采用2×2因子设计,两个因子分别为不同日粮处理(0或500mg/kgABPS)和免疫应激(注射LPS或生理盐水)。试验期28d。在第14天和第21天,每日粮组的一半猪注射100μg/kgBW的LPS,另一半注射生理盐水作对照。注射后3h,采血测定血液生化指标,并进行白细胞分类计数。结果表明:LPS刺激提高了第14天和第21天血清中碱性磷酸酶、谷草转氨酶(GOT)的活性及谷草转氨酶与谷丙转氨酶的比值(P0.05),并降低了第21天血清总蛋白和白蛋白的含量(P0.05);ABPS提高了第14天血清总蛋白和球蛋白的含量(P0.05)。在第14天,LPS刺激降低了白细胞、淋巴细胞、单核细胞和中性粒细胞的含量及单核细胞比例(P0.001)。在第21天,LPS刺激降低了白细胞、淋巴细胞的含量及比例,而提高了中性粒细胞比例(P0.001)。ABPS提高了白细胞(14d:P0.10)和中性粒细胞(14d:P0.05;21d:P0.05)的含量及中性粒细胞比例(14d:P0.05;21d:P0.10),降低了淋巴细胞比例(14d:P0.05;21d:P0.10)。结果提示,LPS可引起仔猪血液生化指标和白细胞分类计数发生显著变化,ABPS在一定程度可缓解上述现象,表现出一定的改善免疫和缓解炎症的作用。     

Effects of pentoxifylline and polymyxin B on the acute-phase-response to Escherichia coli endotoxin in dwarf goats

The purpose of this study was to assess whether polymyxin B together with pentoxifylline, had beneficial effects on the acute-phase-response to E. coli endotoxin in the dwarf goat ( n  = 6). Polymyxin B partly neutralizes E. coli endotoxin by forming inactive polymyxin B-lipopolysaccharide (LPS) complexes; pentoxifylline has been reported to suppress the LPS-induced production of tumour necrosis factor (TNF-α). E. coli LPS (0.0067 μg/kg/min over 30 min) induced fever, tachycardia, inhibition of rumen motility, a decline in WBC, lymphopenia, and decreases in plasma zinc and iron concentrations. Most of the haematological, blood biochemical and clinical effects of E. coli LPS were significantly reduced by polymyxin B pretreatment (0.1 mg/kg/min over 30 min, i.v.). Pentoxifylline (0.3 mg/kg/min over 30 min, i.v.) did not reduce the clinical and blood biochemical effects of E. coli LPS, however, it modulated the number of circulating neutrophils. No synergistic effects were observed after i.v. infusion of polymyxin B with pentoxifylline. The lack of synergy may be due to the production and release of pro-inflammatory cytokines other than TNF-α.     

Septicaemic Escherichia coli and experimental infection of calves

Three strains of Escherichia coli with a common surface antigen, 31a, capable of adhering to calf enterocytes in vitro were compared to reference strains of septicaemic E. coli (RVC 330 and vir E. coli). The surface antigen 31a was present in the RVC 330 reference strain. E. coli vir had a surface antigen which was not present in E. coli 31a or E. coli RVC 330. The RVC 330 and vir reference strains also adhered to calf enterocytes in vitro. Oral infection of calves not receiving colostrum with E. coli 31a was generally followed by septicaemia and death in less than 48 h. Post-mortem examination revealed pneumonia and oedema of the kidneys and gall bladder. Oral infection of calves receiving colostrum had no effect, but intravenous inoculation produced arthritis within 15 days. The comparison of these results with those previously described by other workers did not lead to the identification of pathognomonic characteristics, which could be clearly correlated with properties specific to E. coli 31a. It is suggested that, like ColV and vir, antigen 31a may be a virulence marker for certain strains of bovine septicaemic E. coli. Furthermore, the 31a antigen appears to be carried on a plasmid.     

The effects of ACTH, prednisolone and Escherichia coli endotoxin on some clinical haematological and blood biochemical parameters in dwarf goats

ACTH (microgram kg-1 i.v.) and prednisolone (1 microgram-1 i.v.) caused a moderate but statistically significant inhibition of rumen contractions, whereas no effects on heart rate and body temperature were observed. Both hormones induced hyperglycaemia and leucocytosis, characterised by moderate lymphopenia and a profound increase in the number of circulating neutrophils. A significant decrease in plasma iron and increase in plasma zinc concentrations were observed. After 3 daily i.m. injections of ACTH (10 micrograms-1 day-1) decreases were seen in both serum Alkaline phosphatase (ALP) activity and plasma trace metal concentrations; heart rate was significantly higher. Intraveneous injection of E. coli endotoxin (0.1 microgram kg-1) caused shivering, fever, inhibition of rumen contractions, changes in heart rate, lymphopenia, neutropenia followed by neutrophilic leucocytosis, hypoferraemia, hypozincaemia, hypoglycaemia and a decline in serum ALP activity. ACTH, given i.m. for 3 days, reduced the febrile responses to E. coli endotoxin, modified the changes in heart rate, intensified the inhibition of rumen contractions, and induced a more marked decrease in the number of circulating neutrophils. ACTH pretreatment did not affect the endotoxin-induced decrease in blood glucose concentrations nor the drop in plasma zinc and iron values. These results suggest that glucocorticosteroids are not primarily involved in the fall in plasma iron and zinc concentrations during E. coli endotoxin-induced fever, the effects of endotoxin released glucocorticosteroids on white blood cells and blood glucose are masked by some other effect(s) of endotoxin, and in dwarf goats, ACTH has antipyretic properties without influencing normal body temperature. This effect is probably not dependent on adrenal cortical activity.