The immune response to experimental Pasteurella haemolytica infection in calves

Four male dairy calves, ages 1-9 months, were inoculated intratracheally (IT), with log dilutions (1.5 X 10(3)-1.5 X 10(6)) of an isolate of P. haemolytica A-1. Doses of bacteria varied according to ages of the calves, older calves receiving the larger doses. All four calves became severely ill within 24 h after inoculation and antibiotic treatment was considered essential. Two months later the four calves remained healthy after IT injection of P. haemolytica, again given in log dilution (2.8 X 10(2)-2.8 X 10(5)). The control calf, given a dilution of only 28 viable P. haemolytica (plate count), developed severe respiratory infection 9 days post inoculation. Antibiotic treatment was given to this calf for 7 days, at which time recovery was evident. All five calves developed direct bacterial agglutination titers to P. haemolytica. Persistent leukocyte migration inhibition indexes of all calves were decreased by greater than or equal to 20% compared to their controls. Although the initial doses administered were low, the calves became ill. Most reports refer to massive doses necessary to produce primary disease and significant agglutination titers.     

The pulmonary clearance of Pasteurella hemolytica in calves infected with bovine parainfluenza-3 virus.

The purpose of this study was to determine if parainfluenza-3 virus in calves interfered with normal pulmonary bacterial clearance. Three groups (A, B and C) of four calves each were exposed to an aerosol of parainfluenza-3 virus. Three days later the four hour pulmonary clearance of Pasteurella hemolytica was determined on the first group (A), seven days later on the second (B) and 11 days later on the third (C). Group A had a mean pulmonary bacterial retention of 3.6 +/- 3.5%. Group B was 83.1 +/- 35.9% and Group C was 41.2 +/- 30.9%. The results demonstrate that parainfluenza-3 virus interfered with the pulmonary bacterial clearance of Pasteurella hemolytica particularly on day 7 and also on day 11 but not on day 3. This inhibition of pulmonary clearance caused by the virus may be a key factor in the pathogenesis of pneumonic pasteurellosis. Histological examination of the lungs did not demonstrate a correlation between pulmonary retention of bacteria and the development of pathological changes.     

牛源溶血性巴氏杆菌的分离鉴定

乌鲁木齐某肉牛养殖场育肥牛发生了以体温升高、呼吸困难、咳嗽及快速死亡为主要特征的疾病,每日死亡率超过5%。从发病死亡牛肝脏和肺脏等组织中分离到1株细菌,通过菌落形态、染色特性、培养特性及生化试验等一系列鉴定,确定为溶血性巴氏杆菌。给小白鼠腹腔注射该菌悬液,8 h内全部死亡,结合该批次牛刚从伊犁长途运输而来的流行病学特点,诊断该病是以溶血性巴氏杆菌为主要病原的牛运输热。药敏试验结果表明沙拉沙星和强力霉素最敏感,将它们用于全群预防和治疗后效果显著。     

Dual oxidase 2 and glutathione peroxidase gene expression are elevated in hyperimmunized sheep challenged with Haemonchus contortus

A sequential biopsy sampling method was used to investigate oxidant and antioxidant gene responses in resistant sheep challenged with Haemonchus contortus larvae or a sham saline challenge. The expression of key sheep oxidant and antioxidant producing genes were measured in sequential samples removed from the abomasums at days 0, 1, 2, 3, 5, 7 and 28 post challenge. Gene expression levels at each time point were compared to expression at day 0, and levels of the various genes were also correlated to other markers of infection including immune cell counts and cytokine gene expression. The early response to larval challenge infection in resistant animals was marked by a divergence of two groups of host oxidant producing genes: the dual oxidase group (DUOX2/DUOXA2) showing increases in expression to day 7, while members of the phagocytic NADPH oxidase (PHOX) group showed significant decreases in expression. The change in DUOX2 expression between days zero and seven, when host resistance to infection is mediated, was negatively correlated to final worm burden suggesting NADPH oxidase expression may play a role in parasite expulsion. Expression of the DUOX group oxidants was positively correlated to expression of the Th2 cytokine IL4. Changes in host antioxidant pathways between different members of the glutathione peroxidase family (intestinal and plasma GPX) and genes involved in glutathione metabolism were also observed. This first study of the putative roles of oxidant production by the dual oxidase group, antioxidant glutathione pathways, immune cell populations, and cytokine profiles, in the development of resistance to infection by hyperimmune sheep are discussed.     

Selenium and glutathione peroxidase levels in lambs receiving feed supplemented with sodium selenite or selenomethionine

Twenty-one 6 months old female lambs were divided into 7 groups and fed a basal diet containing 0.13 mg Se/kg. The basal diet was further supplemented with 0, 0.1, 0.5 or 1.0 mg Se/kg either as sodium selenite or as selenomethionine, and was fed for 10 weeks. Both feed additives produced an increase in the selenium concentration in the tissues analysed. Significant correlations were found between the concentrations of selenomethionine or sodium selenite added to the feed and the subsequent tissue levels. However, the selenium levels seemed to plateau at approximately 0.5 mg Se/kg of supplemented sodium selenite. The total glutathione peroxidase (GSH-Px) activity of the tissues increased when the selenium supplementation increased from 0 to 0.1 mg/kg for both selenium compounds. With further increase in selenium supplementation the GSH-Px activity in the tissues plateaued except in the blood where the activity continued to rise with increasing selenomethionine supplementation. The selenium dependent GSH-Px activity in the liver rose with increasing selenomethionine supplementation, but approached a plateau when 0.1 mg Se/kg as sodium selenite was added to the feed. The selenium concentration in whole blood responded more rapidly to the selenium supplementation than did GSH-Px activity. The experiment indicates that the optimal selenium concentration in the feed is considerably higher than 0.1 mg Se/kg, and that selenium levels of 1.0 mg/kg in the feed do not result in any risk for the animals or the consumers of the products.Key words: dietary selenium, lambs, selenium concentrations, glutathione peroxidase activities, tissues     

Blood cellular immune response in pigs immunized and challenged with Haemophilus parasuis

The cellular immune response to an experimental infection by Haemophilus parasuis, the etiological agent of Glässer’s disease in pigs, was characterized studying changes in peripheral blood mononuclear cells (PBMC) in colostrum-deprived pigs. Five groups were studied, four of those were previously immunized with different formulations and the fifth was maintained as non-immunized control. All groups were challenged with 5 × 10⁹ CFU of H. parasuis serotype 5. The non-commercial bacterin conferred a complete protection, while the OMP-vaccine and the exposure to a subletal dose of 10⁵ CFU of H. parasuis protected only partially, and the recombinant Tbp B-vaccine induced no protection. PBMC were analyzed using monoclonal antibodies against porcine CD45⁺, CD3⁺, CD4⁺, CD8α⁺, CD25⁺, CD4⁺ naïve, αIgM⁺ and SWC3⁺ cells in single-colour fluorescence, and CD4⁺/CD8α⁺ and CD8α⁺/CD8β⁺ combinations in two-colour fluorescence. The different groups showed no significant changes in PBMC subsets following vaccination, and only minor changes were encountered after challenge, consisting mainly of significant increases (P < 0.05) in the relative proportions of monocytes and granulocytes (SWC3⁺) and B cells (αIgM⁺), as well as a significant reduction in CD3⁺ cells (P < 0.05). These changes were similar for the five groups compared, except for the significant increase of CD25⁺ cells, which was only observed for the bacterin-vaccinated group. These results suggest an increase of trafficking of inflammatory cells and the onset of the adaptive antibody response against H. parasuis infection; in addition, the blood cellular response developed by the different groups was not relevant to protection.     

Recovery of Pasteurella hemolytica from aerosols at differing temperature and humidity.

A Pasteurella hemolytica suspension with fetal calf serum was aerosolized in a standard system with ambient temperature of 30 or 2 degrees C and relative humidity conditions of 90 or 60%. The number of organisms sprayed in five minutes and the number recovered from one third of the aerosol during these five minutes was determined. Recoveries were influenced by temperature difference between aerosol and collecting fluid. Recoveries ranged between 0.059--0.94%.     

Studies on serum tocopherol, selenium levels and blood glutathione peroxidase activities in calves with white muscle disease

For the purpose of clarifying the cause of white muscle disease (WMD) in calves, tocopherol and selenium levels and blood glutathione peroxidase (GSH-Px) activity were measured on 10 calves with WMD and nine of their dams. The main clinical symptoms of the 10 calves with WMD were motor disturbances including recumbency and stiffness. Serum enzyme activities (GOT, GPT, CPK, LDH) in calves with WMD increased markedly, and this increase was also observed in some of their dams. Serum tocopherol levels of calves with WMD were low, 70% of which showing deficient levels of less than 70 micrograms/100 ml. Serum selenium levels of all the calves were lower than 35 ppb, indicating a deficiency, and were accompanied by low blood GSH-Px activity. alpha-Tocopherol and selenium concentrations in organs were very low. Dams of calves with WMD showed low serum tocopherol levels, 22% of which indicating deficient levels below 150 micrograms/100 ml. Serum selenium levels in dams showed a marked decrease to under 20 ppb, and also low blood GSH-Px activity. Feedstuffs supplied in the farms to affected calves indicated very low alpha-tocopherol contents (below 3 mg/100g DM) and low selenium concentrations below 50 ppb in DM. It was concluded that WMD in calves was attributable to nutritional muscular dystrophy caused by deficiencies in tocopherol and selenium in feedstuffs supplied to their dams.     

Isolation of Pasteurella haemolytica and correlation with serum antibody response in clinically normal beef calves

Bacteria from the nasal cavity and trachea were cultured, and serum antibody titers determined for Pasteurella haemolytica serotype 1 in 164 beef calves obtained from a closed herd on range pasture. At the first sampling, P. haemolytica serotype 1 was cultured from 16.4% of the calves. Antibody titers were determined by a quantitative fluorimetric method and the mean titer was 9.5 +/- 5.8. Fifty-seven randomly selected calves were used to study the correlation of serum antibody response and positive culture of P. haemolytica under natural conditions. Clinical signs of respiratory disease were not observed in those calves. During the observation periods, there was a two-fold increase in the percentage of calves that were culture positive. There was no significant difference between mean serum antibody titers or frequency distribution of antibody titers from the two samplings. Comparisons between serum antibody titers, rise in titers, and P. haemolytica isolation failed to reveal any significant correlation. Of the 9 calves that had a decline in antibody titer to P. haemolytica, none was culture positive. Seroconversion to respiratory viruses did not correlate with P. haemolytica related variables.     

Effect of breed (Angus vs Simmental) on immune function and response to a disease challenge in stressed steers and preweaned calves

Two experiments were conducted with feeder steer calves and preweaned calves to determine the effects of breed on immune response. In Exp. 1, newly weaned Angus (n = 24) and Simmental (n = 24) steer calves were blocked by weight within breed and randomly assigned to 12 pens with four calves per pen. The basal diet consisted of 87% corn silage (DM basis) and 13% of a soybean meal-mineral-vitamin supplement. Steers were allowed ad libitum access to feed throughout the study. On d 2 following weaning, calves received an intranasal inoculation of infectious bovine rhinotraecheitis virus (IBRV; 2.7 x 10(8) CCID50). Rectal temperatures in response to the IBRV were higher (P < .05) in Angus calves. On d 9, calves were injected i.m. with 10 mL of a 25% pig red blood cell (PRBC) suspension. Total immunoglobulin (Ig) and IgM titers against PRBC were higher (P < .05) for the Angus calves. Breed did affect cell-mediated immune response to phytohemagglutinin (PHA). In Exp. 2, preweaned (16 Angus and 16 Simmental) calves were selected based on breed, body weight, and sex. On 0 d, all selected calves were injected i.m. with 10 mL of a 25% PRBC suspension. Total Ig and IgG titers against PRBC were higher (P < .05) for Angus calves. On d 28, lymphocytes were isolated from peripheral blood obtained from eight calves per breed. Peripheral lymphocytes from the Angus calves had a greater (P < .07) blastogenic response to 6.25 microg/mL of PHA than lymphocytes from Simmental calves. Results indicate that the immune response of Angus and Simmental calves may differ.     

Dynamics of the humoral immune response of calves infected and re-infected with Cooperia punctata

The dynamics of the humoral immune response of calves were analysed after primary infection and re-infection with the intestinal nematode Cooperia punctata. 12 male 5 month-old Holstein-Friesian calves were randomly divided into two groups A and B. At the beginning of the experiment Group A animals were each infected experimentally with a single oral dose of 130,000 infective third stage larvae (L3) of C. punctata. The animals of Group B were kept as non-infected controls. The two calves from Group A with the highest infections died of cooperiosis at 32 and 44 days after infection (DAI), respectively. On DAI 100 the calves were treated with the recommended dose of oxfendazole. On DAI 180 the remaining four calves of Group A and three animals of Group B (B1) were infected with 260,000 L3 of C. punctata, while the other three calves of Group B (B2) served as non-infected controls. Monitoring of the humoral immune response predominantly demonstrated an IgG1 response against both adult and L3 antigen of C. punctata. Moreover, re-infections increased the levels of these immunoglobulins. IgA levels were less increased than IgG1 and no significant increase was observed in IgG2 and IgM levels. Immunoblotting analysis showed that total IgG present in the serum of the primary infected animals mainly reacted against adult proteins of 12-14 and 17-20 kDa and against L3 proteins of 33 and 43 kDa. After re-infection total IgG reacted with the same adult proteins but also with an adult 29 kDa protein.     

Colonization of the tonsils of calves with Pasteurella haemolytica.

Tonsils of 10 calves were inoculated with Pasteurella haemolytica (PH) and the degree of colonization was followed by collecting sequential tonsil. wash specimens. Tonsils were colonized for at least 3 weeks after instillation of PH into the tonsillar sinus. Calves with colonized tonsils responded with serum and nasal secretion antibody responses to PH and to leukotoxin. Pasteurella haemolytica was detected in nasal mucus specimens of 2 calves during the week after inoculation of the tonsils, but all other specimens were culture-negative. Infectious bovine rhinotracheitis virus-induced respiratory tract disease 25 days later did not elicit a population increase of PH in the tonsils, and did not elicit shedding of PH in nasal mucus.     

Hemodynamic effects of acute pneumonia experimentally induced in newborn calves inoculated with Pasteurella haemolytica

Hemodynamic responses to acute pneumonia and to hypoxia were investigated in 10 newborn calves. Experiments were performed on heparinized, anesthesized and ventilated calves. Control calves were inoculated intratracheally with bovine fetal serum. Pneumonia was induced in treated calves by intratracheal inoculation with P haemolytica suspended in bovine fetal serum. Before inoculation (base line), at the time of inoculation (T = 0), and at 30-minute intervals for 3 hours, pulmonary arterial pressure (Ppa), systemic arterial pressure, cardiac output (CO), arterial blood gases, pulmonary vascular resistance (PVR), and systemic vascular resistance were determined. At T = 0, calves in both groups became hypoxemic, alveolar-arterial O2 difference, PVR, and Ppa increased, and CO and systemic vascular resistance remained unchanged. At subsequent measurement intervals, all values returned to base-line in control calves, whereas treated calves had progressive hypoxemia associated with a decrease in Ppa and PVR, with no change in CO. Three hours after inoculation and after inhalation of 10% O2 in N2, PVR increased significantly in the control calves. In the treated group, hypoxia did not increase the resistance, compared with base-line and 3-hour values. The data indicate decreased Ppa during pneumonic pasteurellosis is because of a decrease in PVR and that pneumonia may attenuate the normal pulmonary hypoxic vasoconstrictor response.     

浅谈猪伪狂犬病的诊断要点及防治措施

猪伪狂犬病又名传染性延髓麻痹病、奇氧症,本病是由伪狂犬病毒(PRV)引起的多种家畜和野生动物发热,奇氧(猪除外)及脑脊髓炎为主要症状的一种疾病。     

Effect of ascorbic acid supplementation on the immune response of chickens vaccinated and challenged with infectious bursal disease virus

One-day-old chickens were divided into two groups and reared under similar conditions. One group was fed a diet supplemented with 1000ppm ascorbic acid and the other group was fed an identical diet, but not supplemented with ascorbic acid. Both groups were vaccinated against infectious bursal disease (IBD) at 7 days of age and challenged orally with 4x10(5) of 50% embryo-lethal-dose IBDV 14 days later. The number of anti-IBDV antibody secreting cells, production of interleukin-2 (IL-2) by splenocytes, number of CD4(+), CD8(+) and IgM(+) cells in spleen and IgM(+) cells in bursa of Fabricius were compared between the two groups at 7 days (prior to vaccination), 21 days (14 days post-vaccination and prior to challenge) and 31 days (10 days post-challenge) of age. The number of CD8(+) in spleen at 7 days of age and IgM(+) cells in bursa at 7, 21 and 31 days of age were significantly higher in ascorbic acid supplemented group (P<0.05). Production of IL-2 by splenocytes was higher as indicated by higher stimulation indices in ascorbic acid supplemented group. The number of anti-IBDV IgG antibody secreting cells in spleen at 21 and 31 days of age were significantly higher in ascorbic acid supplemented group (P<0.05). Dietary supplementation of ascorbic acid may ameliorate the immunosuppression caused by IBDV vaccination and improve humoral and cellular immune responses.