Effect of three feeding levels on the pathogenesis and establishment of Haemonchus contortus in parasite-naïve Pelibuey hair sheep lambs during their first infection.

This study evaluated the effect of three feeding levels on the pathogenesis and establishment of H. contortus upon the first infection of parasite-naïve Pelibuey hair sheep lambs. Forty-two 6-month-old hair sheep lambs (24 ± 4 kg) raised parasite free from birth were used. The lambs were assigned to 3 groups (n = 14), and each was fed a diet designed for different daily weight gain (DWG): 75 g/d (Diet 1), 125 g/d (Diet 2) and 200 g/d (Diet 3). After four weeks of diet adaptation, 10 lambs/group were infected with 450 L3H. contortus/kg BW (infected), and 4 lambs/group were kept parasite-free (NInf). DWG, hematocrit (Ht), hemoglobin (Hb), peripheral eosinophils (EOS), IgG concentration against H. contortus, and eggs per gram (EPG) of feces were measured in each lamb from day 14 before infection until day 29 postinfection (PI). On day 29 PI, the lambs were slaughtered to determine the total number of adult parasites (TAW), the length of the female worms, and the number of eggs in utero (EIU). Each group reached the expected DWG (P = 0.001), and there was no effect of infection or the diet × infection interaction. Ht was lower in infected lambs than in NInf lambs, and this difference was significant for animals on Diets 1 and 2 (P = 0.044). From day 14 PI onward, Hb was lower in the infected lambs than in the NInf lambs (P = 0.001). Furthermore, compared with NInf lambs, the infected lambs had higher EOS from day 7 PI and higher IgG from day 14 PI. Neither EOS nor IgG were affected by diet. Lambs on Diet 3 had lower EPG during patency than those fed Diets 1 or 2 (days 25 and 28 PI; P = 0.002). Furthermore, lambs fed Diet 3 had lower TAW (Diet 1 vs 3 P = 0.037; Diet 2 vs 3 P = 0.049) and EIU (P = 0.004) than lambs fed Diet 1 or 2. Lambs were resilient to infection regardless of diet. Although EOS and IgG were higher in all infected animals than in Ninf animals, EPG, TAW and EIU decreased only in lambs fed Diet 3. Thus, a diet targeting a DWG of 200 g/d can significantly limit the establishment of H. contortus in Pelibuey lambs infected for the first time.

Synthesis and Antioxidant/Anti-Inflammatory Activity of 3-Arylphthalides.

Phthalides are a group of compounds with relevant biological activities in different areas such as cytotoxicity, anti-stroke activity, neuroprotection, and inflammation, among others. In this study we designed and synthesized a series of 3-arylphthalide derivatives in order to identify their antioxidant and anti-inflammatory activities. The synthetic methodology was established in terms of atom and step economy through a dehydrative coupling reaction between 3-hydroxyphthalide and different properly functionalized arene rings. The evaluation of the antioxidant activity was performed by the ABTS assay and for the anti-inflammatory activity the inhibition of LPS-induced nitric oxide (NO) production in microglial cells Bv.2 and macrophage cells RAW 264.7 was measured. The synthesized compound 3-(2,4-dihydroxyphenyl)phthalide (5a) showed better antioxidant activity than the Trolox standard and caused strong inhibition of NO production in LPS-stimulated Bv.2 and RAW 264.7 cells. In addition, compound 5a reduced the expression of the pro-inflammatory cytokines Il1b and Il6 in RAW 264.7 cells. These results, which are the first account of the anti-inflammatory activity of 3-arylphthalides, suggest that compound 5a could be a promising candidate for more advanced anti-inflammatory studies.

Diterpenoids from the Brown Alga Rugulopteryx okamurae and Their Anti-Inflammatory Activity.

Brown algae of the Family Dictyotaceae produce an array of structurally diverse terpenoids, whose biomedical potential in the anti-inflammatory area has been scarcely explored. Herein, the chemical study of the alga Rugulopteryx okamurae has led to the isolation of ten new diterpenoids: rugukadiol A (1), rugukamurals A-C (2-4), and ruguloptones A-F (6-10). The structures of the new compounds were established by spectroscopic means. Compound 1 exhibits an unprecedented diterpenoid skeleton featuring a bridged tricyclic undecane system. Compounds 2-10 belong to the secospatane class of diterpenoids and differ by the oxygenated functions that they contain. In anti-inflammatory assays, the new diterpenoid 1 and the secospatanes 5 and 10 significantly inhibited the production of the inflammatory mediator NO in LPS-stimulated microglial cells Bv.2 and macrophage cells RAW 264.7. Moreover, compounds 1 and 5 were found to strongly inhibit the expression of Nos2 and the pro-inflammatory cytokine Il1b in both immune cell lines.

Effect of timing of challenge following short-term natural exposure to bovine viral diarrhea virus type 1b on animal performance and immune response in beef steers.

Bovine respiratory disease (BRD) is the most common and economically detrimental disease of beef cattle during the postweaning period, causing the majority of morbidity and mortality in feedlots. The pathogenesis of this disease often includes an initial viral infection, which can predispose cattle to a secondary bacterial infection. The objective of this experiment was to determine the effects of timing of an intratracheal (MH) challenge relative to 72 h of natural exposure to bovine viral diarrhea virus (BVDV) type 1b persistently infected (PI) calves on performance, serum antibody production, total and differential white blood cell (WBC) count, rectal temperature, clinical severity score (CS), and haptoglobin (Hp). Steers ( = 24; 276 ± 31 kg initial BW) were randomly allocated to 1 of 3 treatments (8 steers/treatment) in a randomized complete block design. Treatments were steers not exposed to calves PI with BVDV 1b and not challenged with MH (CON), steers intratracheally challenged with MH 84 h after being exposed to calves PI with BVDV 1b for 72 h (LateCh), and steers intratracheally challenged with MH 12 h after being exposed to calves PI with BVDV 1b for 72 h (EarlyCh). Performance (ADG, DMI, and G:F) was decreased ( < 0.001) for both EarlyCh and LateCh from d 0 to 4. From d 5 to 17, LateCh appeared to compensate for this lost performance and demonstrated increased ADG ( = 0.01) and G:F ( = 0.01) compared with EarlyCh. Both EarlyCh and LateCh had decreased platelet counts ( < 0.001) compared with CON. Antibody concentrations of BVDV and MH were higher ( < 0.05) for both EarlyCh and LateCh compared with CON. Rectal temperature, CS, and Hp increased ( < 0.001) across time from h 4 to 48, h 4 to 36, and h 8 to 168, respectively. Within 24 h of MH challenge, WBC and neutrophil concentrations within the blood increased whereas lymphocyte concentrations decreased. The timing of BVDV exposure relative to a MH challenge appears to influence the CS and acute phase response associated with BRD. As typical beef cattle marketing channels allow for variation in the timing of respiratory pathogen exposure, understanding the physiological changes in morbid cattle will lead to improved management of BRD.

First Report of Pestalotiopsis clavispora Causing Dieback on Blueberry in Uruguay.

During the last 10 years, blueberry (Vaccinium corymbosum) production in Uruguay has increased to more than 850 ha. From 2005, symptoms of dieback characterized by the death of twigs and branches have been frequently observed in blueberry plants cv. O'Neal in orchards located in Uruguay. Symptomatic 4-year-old plants (cv. O'Neal) were collected and small pieces of necrotic tissues were surface disinfected and plated onto potato dextrose agar (PDA) with 0.2 g liter-1 of streptomycin sulfate. Plates were incubated at 25°C in the dark. All affected tissues consistently developed colonies with white and cottony mycelium, turning slightly yellow after 7 to 10 days. Black acervuli distributed in concentric circles were observed after 10 days. Conidia were fusiform, straight, and had five cells. Basal and apical cells were colorless while the three median cells were dark brown. Conidia (n = 50) had an average of 22.1 (16.5 to 28.2) × 6.6 (5.6 to 7.7) µm. All conidia had one basal appendage of 6.1 (3.9 to 14.3) µm and two to four (usually three) apical appendages of 22.8 (17.4 to 42.9) µm. According to colony and conidia morphology, the isolates were initially identified as Pestalotiopsis clavispora (G.F. Atk.) Steyaert (1). To identify, the internal transcribed spacers (ITS1, 5.8S, ITS2) region of rDNA of a representative isolate (Ara-1) was amplified with ITS1/ITS4 primers (4), sequenced, and compared with those deposited in GenBank. The isolate Ara-1 (Accession No. JQ008944) had 100% sequence identity with P. clavispora (Accession Nos. FJ517545 and EU342214). To confirm pathogenicity, isolate Ara-1 was inoculated onto asymptomatic 1-year-old blueberry plants (cv. O'Neal). Mycelial plugs (4 mm in diameter) from an actively growing colony on PDA were applied to same-size bark wounds made with a cork borer in the center of the stems previously disinfected with 70% ethanol and covered with Parafilm. Control plants were inoculated with sterile PDA plugs. Inoculated plants (five per treatment) were randomly distributed in a greenhouse and watered as needed. After 2 weeks, all stems inoculated with P. clavispora showed brown necrotic lesions 2 to 3 cm in length and 1 to 2 mm deep. White mycelium was observed over lesions. Control plants remained symptomless. The pathogen was reisolated from all necrotic lesions, thus fulfilling Koch's postulates. P. clavispora has been reported as associated with blueberry in Hawaii (3) and Chile (2). To our knowledge, this is the first report of P. clavispora causing dieback disease on blueberry in Uruguay. References: (1) E. F. Guba, Monograph of Pestalotia and Monocheatia. Harvard University Press, Cambridge, MA, 1961. (2) J. G. Espinoza et al. Plant Dis. 92:1407, 2008. (3) L. M. Keith et al. Plant Dis. 90:16, 2006. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and Mannheimia haemolytica challenge on animal performance, nitrogen balance, and visceral organ mass in beef steers.

Bovine viral diarrhea viruses (BVDV) have been isolated alone or in combination with other viral and bacterial pathogens in animals diagnosed with bovine respiratory disease (BRD), a disease causing major economic loss to the feedlot industry. The objective of this experiment was to determine the effects of Mannheimia haemolytica challenge after short-term exposure (72 h) to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on performance, N balance, and organ mass in finishing cattle. Treatments (6 steers/treatment; initial BW = 314 +/- 31 kg) were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV1b (BVD) for 72 h; 3) steers intratracheally challenged with M. haemolytica (MH); or 4) steers exposed to 2 steers PI with BVDV1b for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Steers were housed in metabolism stanchions during the first 5 d after the M. haemolytica challenge and on d 7 to 11, 28 to 32, and for 5 d before slaughter (average 119 d on feed) to determine N balance and were weighed every 28 d. At slaughter, carcass and organ mass data were collected. Data were analyzed as a randomized complete block design with a 2 x 2 factorial arrangement of treatments, and steer was used as the experimental unit. From d -3 (beginning of PI steer exposure) to 4, steers challenged with M. haemolytica had less (P = 0.04) ADG than steers not challenged with M. haemolytica. In addition, steers exposed to steers PI with BVDV tended (P = 0.09) to have less ADG and G:F across the entire finishing period than steers not exposed to BVDV. Before slaughter, retained N expressed as grams per day (P = 0.03) and as a percentage of N intake (P = 0.04) was less in BVD steers compared with steers not exposed to BVDV. There were no effects (P > 0.10) of BVDV exposure or M. haemolytica challenge on empty BW (EBW) or carcass characteristics. Expressed as a percentage of EBW, HCW was less (P = 0.02) and total offal weight was greater (P = 0.02) for steers challenged with M. haemolytica compared with steers not challenged. Results are in agreement with those reported in larger scale finishing studies and suggest that acute exposure to BRD-related pathogens can have long-term effects on animal performance.

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and subsequent infection with Mannheima haemolytica on clinical signs and immune variables: model for bovine respiratory disease via viral and bacterial interaction.

The objective was to determine effects of an intratracheal Mannheimia haemolytica challenge after 72-h exposure to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on serum antibody production, white blood cell count (WBC), cytokine concentrations, and blood gases in feedlot steers. Twenty-four steers (initial BW = 314 +/- 31 kg) were randomly allocated to 1 of 4 treatments (6 steers/treatment) arranged as a 2 x 2 factorial. Treatments were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV for 72 h (BVD); 3) steers intratracheally challenged with M. haemolytica (MH); and 4) steers exposed to 2 steers PI with BVDV for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Rectal temperature was increased (P < 0.001) for MH and BVD+MH during the initial 24 h after the M. haemolytica challenge. For MH and BVD+MH, total WBC count was increased (P < 0.01) at 36 h post M. haemolytica challenge compared with CON, whereas in BVD steers, WBC count was decreased (P < 0.01). Total lymphocyte count was increased (P = 0.004) during the initial 72 h post BVDV exposure for the BVD and BVD+MH groups compared with MH and CON, and this difference remained at 96 h post M. haemolytica challenge. An increased (P < 0.001) total neutrophil count was observed during the initial 36 h for the MH group and at 72 h for the BVD+MH challenge group. Interleukin 1beta, IL-6, and tumor necrosis factor alpha (TNFalpha) concentrations were greater (P

Intranasal vaccination of calves with Mannheimia haemolytica chimeric protein containing the major surface epitope of outer membrane lipoprotein PlpE, the neutralizing epitope of leukotoxin, and cholera toxin subunit B.

This study was done to determine if intranasal vaccination of weaned beef calves with a chimeric protein containing the immunodominant surface epitope of Mannheimia haemolytica PlpE (R2) and the neutralizing epitope of leukotoxin (NLKT) covalently linked to truncated cholera toxin (CT) subunit B (CTB) could stimulate secretory and systemic antibodies against M. haemolytica while enhancing resistance of cattle against M. haemolytica intrabronchial challenge. Sixteen weaned beef calves were intranasally vaccinated with CTB-R2-NLKT chimeric (SAC102) or with R2-NLKT-R2-NLKT chimeric (SAC89) protein with or without native CT on days 0 and 14 and were challenged intrabronchially on day 28. In vitro, SAC102 bound the CT receptor molecule, GM(1)-ganglioside. Mean IgA antibodies to M. haemolytica whole cells (WC) and to LKT were high on day 0. A small, yet significant increase (p<0.05) was found in mean nasal antibodies to M. haemolytica WC for the SAC89+CT and SAC102 vaccinates after the second vaccination. SAC102 stimulated significant (p<0.05) mean serum antibody responses to all three antigens by day 28. Following challenge, mean antibodies to WC and LKT significantly increased (p<0.05) for the SAC102, SAC89 and SAC89+CT groups with the mean antibody responses to rPlpE stimulated by SAC102 vaccination being significantly higher (p<0.05) than for the other vaccinated and control groups. On day 1 after challenge, mean clinical score for the control group was significantly higher (p<0.05) than for the SAC102 and SAC89+CT vaccinates, and by day 2 after challenge, clinical score for the control group was significantly higher (p<0.05) than for all three chimeric vaccinated groups. Therefore, intranasal vaccination with CTB-R2-NLKT (SAC102) and R2-NLKT-R2-NLKT (SAC89) chimeric proteins enhanced resistance against intrabronchial challenge with the bacterium as well as stimulating antibody responses to M. haemolytica antigens.

Immunity of cattle following vaccination with a Mannheimia haemolytica chimeric PlpE-LKT (SAC89) protein.

UNLABELLED: We developed several chimeric PlpE-leukotoxin (LKT) constructs containing the major epitope of Mannheimia haemolytica outer membrane lipoprotein PlpE (epitope R2) and the neutralizing epitope of M. haemolytica LKT (NLKT) [Ayalew et al. Mannheimia haemolytica chimeric protein vaccine composed of the major surface-exposed epitope of outer membrane lipoprotein PlpE and the neutralizing epitope of leukotoxin. Vaccine 2008;26(38):4955-61]. Vaccination of mice with these PlpE-LKT chimeric proteins stimulated anti-PlpE antibodies that caused complement-mediated bacteriolysis of M. haemolytica as well as neutralizing anti-LKT antibodies. Chimeric protein SAC89, which contains two copies of R2 and two copies of NLKT, generally stimulated the best overall responses in mice. The objectives of the current study were: (1) to determine through a dose titration study if vaccination of cattle with SAC89 stimulated antibodies to both PlpE and LKT and (2) evaluate SAC89-induced immunity against experimental M. haemolytica challenge of cattle. In the dose titration study, vaccine doses ranged from 100 to 400 microg. SAC89 significant anti-M. haemolytica surface and LKT antibodies were detected following vaccination with each dose. The vaccination/challenge study was conducted with 30 weaned beef cattle distributed among four groups: Control (no vaccine), 100 microg SAC89, M. haemolytica Bacterin, and SAC89+M. haemolytica bacterin. On day 42 after two vaccinations, cattle were challenged transthoracically with M. haemolytica. There was significant reduction (p<0.05) in lesion scores for the SAC89+bacterin-vaccinated group (74.6% reduction compared to control lesion scores) when compared to the other groups (34.7% and 35.6% reduction compared to control lesion scores). Evaluation of antibody responses demonstrated that the control group failed to develop antibody responses to M. haemolytica surface antigens or to LKT. Bacterin-vaccinated cattle developed anti-M. haemolytica antibodies after the second vaccination. SAC89- and SAC89+bacterin-vaccinated groups developed significant antibody responses 14 days after the first vaccination and further significant increases in antibodies after the second vaccination. CONCLUSIONS: Vaccination with the chimeric protein SAC89 in conjunction with a M. haemolytica bacterin stimulated significant protection against a severe transthoracic challenge with the bacterium.

Effects of commingling beef calves from different sources and weaning protocols during a forty-two-day receiving period on performance and bovine respiratory disease.

The study objective was to determine health and performance of ranch calves from different preconditioning strategies during a 42-d receiving period when commingled with calves of unknown health histories from multiple sources. Steer calves from a single source ranch (RANCH) were weaned and immediately shipped to a feedlot (WEAN, initial BW = 247 +/- 29 kg); weaned on the ranch for 45 d before shipping, but did not receive any vaccinations (WEAN45, initial BW = 231 +/- 26 kg); or weaned, vaccinated with modified live viral vaccine, and held on the ranch for 45 d before shipping (WEANVAC45, initial BW = 274 +/- 21 kg). Multiple-source steers were purchased through auction markets (MARKET, initial BW = 238 +/- 13 kg), and upon receiving, a portion of ranch-origin steers from each weaning group was commingled with a portion of MARKET cattle (COMM). The experimental design was completely randomized with a 2 x 3 +1 factorial arrangement of treatments. Factors were RANCH vs. COMM and weaning management (WEAN vs. WEAN45 vs. WEANVAC45) as the factors; MARKET cattle served as the control. Calves of WEAN, WEAN45, and MARKET were vaccinated on arrival at the feedlot. Ranch-origin calves tended (P = 0.06) to have greater ADG than COMM or MARKET calves, although ADG was not affected (P = 0.46) by weaning management. Across the 42-d receiving period, DMI was not affected (P = 0.85) by cattle origin. However, MARKET, WEAN45, and WEANVAC45 calves consumed more (P < 0.001) DM than WEAN calves. Gain efficiency was not affected (P > or = 0.11) by treatment. Ranch-origin calves were less (P < 0.001) likely to be treated for bovine respiratory disease than MARKET calves; COMM calves were intermediate. Calves that were retained on the ranch after weaning (WEAN45 and WEANVAC45) were also less likely to be treated (P = 0.001) than MARKET or WEAN calves. As expected, differences in morbidity related to differences in health costs. Calves of WEAN45 and WEANVAC45 had less (P < 0.001) health costs than MARKET and WEAN calves. On arrival, serum haptoglobin concentrations were greater (P < 0.001) in MARKET and WEAN compared with WEAN45 and WEANVAC45 calves. Calves from a single source that are retained on the ranch for 45 d after weaning exhibit less morbidity and less health costs during the receiving period at the feedyard than when cattle are commingled or trucked to the feedyard immediately after weaning.

Serum antibodies in mares and foals to Actinobacillus equuli whole cells, outer membrane proteins, and Aqx toxin.

Actinobacillus equuli is carried in the alimentary tract of mares and can cause severe septicemia of neonatal foals. A hemolytic subspecies, A. equuli subsp. haemolyticus, and a non-hemolytic subspecies, A. equuli subsp. equuli, have been identified. Hemolytic strains produce the RTX toxin Aqx. The purpose of this study was to demonstrate sequentially in two sets of mare-foal pairs antibodies to A. equuli whole bacterial cells, outer membrane proteins, and recombinant Aqx and to compare the transfer of antibodies to these antigens between mares and their foals. Two mare/foal sets of sera were evaluated. Cohort A consisted of 18 mare-foal pairs obtained in the spring of 2005. Cohort B consisted of 10 mare-foal pairs obtained in the spring of 2006. For both sets, mare and foal sera were obtained immediately after foaling and prior to nursing (time 0) as well as at 12 and 24h and daily thereafter for 7 days. For Cohort B, sera were also obtained 30 days after birth. At parturition all mares had detectable antibodies to A. equuli whole cells and outer membranes; however, of those mares, two in Cohort A had undetectable antibodies to Aqx and their foals likewise had undetectable anti-Aqx antibodies. Antibodies against whole cells, outer membrane proteins, and Aqx were readily transferred from mares to foals. In most cases, there were significant correlations (p<0.05) between antibodies against whole cells, outer membrane proteins, and Aqx in mares' sera at the time of parturition and foal sera 24 after birth. Antibodies against the three antigen preparations had declined insignificantly (p>0.05) by day 30.

[Estimation of vitamin supplements and minerals consumption dispensed through pharmacy offices in the province of Las Palmas]. / Estimación del consumo de suplementos vitamínicos y minerales dispensados a través de las oficinas de farmacia de la provincia de Las Palmas.

BACKGROUND: Vitamin supplements and minerals consumption (SPM) is increasing in occidental societies due to the growing concern about health by the population. OBJECTIVES: To have a initial approaching to the to SPM consumption in the Province of Las Palmas through 2000 and 2001. To identify SPM proportions that are dispensed as pharmaceutical specialities and those who are sold as parapharmacy products. Finally, to describe the evolution of this consumption throughout a year. METHOD: The information was obtained through the list of the whole pharmaceutical specialities and parapharmacy products through 2000 and 2001 who have at least a vitamin in its composition and/or a mineral. RESULTS: Usually, 297 pharmaceutical specialities and 216 parapharmacy products are currently being sold. Pharmaceutical specialities comprised 65.6% of the whole products sold and within them, vitamins were the most dispensed (41.5%). Regarding parapharmacy products, vitamins and minerals compounds were the products more sold (34.6%). Comparing to 2000, during 2001 there was a statistically significant increase in the consumption of parapharmacy products, remaining without changes the use of pharmaceutical specialities. CONCLUSION: On the basis of sold XX SPM consumption seems to be due mainly by pharmaceutical specialities rather than parapharmacy products. Nevertheless through 2001 there was an increase only in the parapharmacy products.

Effects of dietary energy and starch concentrations for newly received feedlot calves: I. Growth performance and health.

Crossbred calves (n = 572; initial BW = 186 +/- 27 kg) purchased from northern Texas, Arkansas, and southeast Oklahoma auction markets were delivered to the Willard Sparks Beef Research Center, Stillwater, OK, and used to study the effects of dietary energy and starch concentrations on performance and health of newly received feedlot calves during a 42-d receiving period. On arrival, calves were assigned randomly to one of two dietary energy levels (0.85 or 1.07 Mcal NEg/kg DM) and one of two dietary starch levels (34 or 48% of ME from starch) in a 2 x 2 factorial arrangement of treatments. Cattle were weighed and serum samples were collected on d 0, 7, 14, 28, and 42. Individual animal records of morbidity were kept for all cases of respiratory and other disease. Nasal swabs were collected from each morbid animal and cultured for upper-respiratory pathogens. There were no energy x starch level interactions for performance or health response variables. Daily gain (1.14 kg/d) and gain efficiency (ADG:DMI = 0.179) were not affected by increasing dietary energy or starch concentrations. Calves fed low-energy diets consumed (P < 0.05) more DM. No difference (P = 0.54) was detected in morbidity for calves fed high-energy (62.4% calves treated) compared with low-energy (65.8% calves treated) diets; however, calves fed the high-starch diets had numerically (P = 0.11) greater morbidity than calves fed low-starch diets (68.8 vs. 59.4% calves treated, respectively). There were no energy or starch effects on Mannheimia haemolytica or Pasteurella multocida antibody titers; however, day effects (P < 0.02) occurred. On d 7, 14, and 28, calves had antibody titers for P. multocida that were greater (P < 0.05) than titers on d 0. In addition, calves had greater antibody titers to M. haemolytica on d 7 and 14 than on d 0. Nasal swabs revealed that calves fed the high-energy diets tended (P = 0.06) to have a lower percentage of morbid calves with P. multocida during the first antimicrobial treatment and a lower percentage of Haemophilus somnus isolates during the first (P = 0.01) and second (P = 0.06) antimicrobial treatments than calves fed the low-energy diets. Although animal performance was not influenced, the present data suggest that feeding the high-energy diet decreased the percentage of P. multocida and H. somnus pathogens in calves that received one or more antimicrobial treatments.

Effects of dietary energy and starch concentrations for newly received feedlot calves: II. Acute-phase protein response.

Two hundred forty five market-stressed bull and steer calves (205 +/- 14 kg) were received in January and September 2001 to determine the response of acute-phase proteins to dietary energy and starch concentrations and to determine whether acute-phase proteins could be used as a diagnostic or prognostic tool for calves affected by bovine respiratory disease. On arrival, calves were randomly assigned to one of two dietary energy levels (0.85 or 1.07 Mcal of NEg/kg of DM) and one of two dietary starch levels (34 or 48% of dietary ME from starch; n = 5 pens/treatment). All calves were weighed, and plasma and serum samples were collected from a subset of animals (n = 6 calves/pen; 30 calves/treatment) on d 0, 7, 14, 28, and 42 of the receiving period. This subset of calves (n = 120) was used for all subsequent analyses. Concentrations of fibrinogen (Fb), haptoglobin (Hp), and serum amyloid-A (SaA) were determined. In addition, samples were collected from the subset of calves when they received medical treatment and 7 d following treatment to measure serum concentrations of Hp and SaA. Serum concentrations of Fb, Hp, and SaA did not differ among dietary treatments, but decreased (P < 0.03) as day of the receiving period increased. Fibrinogen (P < 0.001) and the ratio of Fb:total blood protein were greater (P < 0.003) in calves treated multiple times than in calves never treated or treated once for bovine respiratory disease. In addition, on d 0 and 7, Hp concentration increased (antimicrobial treatment x day interaction, P < 0.03) as the number of antimicrobial treatments increased, and was greater on d 14 and 28 in calves treated multiple times than in calves never treated or treated once. Haptoglobin concentration was greater (P < 0.05) in calves on medical treatment days compared with recovery days (7 d after medical treatment). Although diet seemed to have little effect on acute-phase protein response, these results suggest that haptoglobin may be useful as a diagnostic tool to make management decisions regarding treatment protocols for calves with bovine respiratory disease.

Intranasal vaccination of rabbits with Pasteurella multocida A:3 outer membranes that express iron-regulated proteins.

OBJECTIVE: To determine efficacy of intranasal vaccination of rabbits with Pasteurella multocida A:3 outer membrane proteins (OMP) expressing iron-regulated OMP (IROMP) in conferring protection against experimental challenge exposure. ANIMALS: 52 male New Zealand White rabbits. PROCEDURE: Rabbits were vaccinated intranasally on days 0, 7, and 14; some vaccines included cholera toxin (CT) as an adjuvant. Concentrations of intranasal IgA and serum IgG antibodies against P multocida OMP were determined. In experiment A, rabbits were vaccinated with either phospate-buffered saline solution (PBSS), PBSS-CT, OMP-CT, or IROMP-CT, challenge-exposed intranasally on day 16, and euthanatized and necropsied on day 28. Rabbits were also vaccinated with OMP or IROMP without CT and were not challenge-exposed. In experiment B, rabbits were vaccinated with PBSS, PBSS-CT, IROMP, or IROMP-CT. On day 17, rabbits were challenge-exposed intranasally. Nasal bacteria and antibodies were determined on day 24. RESULTS: In experiment A, OMP-CT vaccination stimulated mucosal and systemic antibody responses to the bacterium and enhanced resistance against challenge exposure. Intranasal bacterial counts were not significantly reduced. Vaccination with IROMP-CT stimulated mucosal and systemic antibodies, enhanced resistance to challenge exposure, and significantly reduced nasal bacterial counts. In experiment B, natural infection was detected in several rabbits at challenge exposure; however, IROMP-CT-vaccinated rabbits had significantly higher serum and nasal antibody responses, compared with other rabbits IROMP-CT-vaccinated rabbits had significantly lower nasal bacterial counts compared to control rabbits. CONCLUSIONS AND CLINICAL RELEVANCE: Intranasal vaccination of rabbits with P multocida outer membranes containing IROMP and CT stimulated immunity against experimental pneumonic pasteurellosis.

Characterization of rabbit Pasteurella multocida isolates by use of whole-cell, outer-membrane, and polymerase chain reaction typing.

PURPOSE: To characterize Pasteurella multocida isolates from laboratory rabbits using serotyping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins (WCPs) and outer-membrane proteins (OMPs), and polymerase chain reaction (PCR) fingerprinting. METHODS: Fifty isolates were obtained from five sources: ATCC (1), Oklahoma (4), Michigan (9), Minnesota (7), and Texas (29). The PCR fingerprinting was conducted using two minisatellite probes for M13 and a modified M13 core sequence and two microsatellite probes--(GTG)5 and (GACA)4. RESULTS: Forty-five isolates were serogroup A, and five were serogroup D. Ten WCP patterns (W1-W10) with one variation (W1a) and 10 OMP (OM1-OM10) patterns were found. Primers M13 phage, modified M13 phage, (GTG)5, and (GACA)4 generated 7, 9, 5, and 9 fingerprint types, respectively. Combination of WCP, OMP, and PCR fingerprint results yielded 39 groups with a discrimination index of 0.98. The PCR fingerprint results generally indicated clonal association among isolates within geographic locations except for the isolates from Texas, which varied markedly in PCR fingerprint types. CONCLUSION: Single primer PCR fingerprinting provided a simple and rapid means of typing P. multocida isolates from laboratory rabbits. Combinations of conventional and molecular typing enhanced differentiation among P. multocida isolated from rabbits with pasteurellosis.

Genomic DNA restriction site heterogeneity in bovine Pasteurella multocida serogroup A isolates detected with an rRNA probe.

A total of 81 Pasteurella multocida isolates from healthy and diseased dairy and beef cattle originating from various geographical locations was examined by rRNA gene restriction site polymorphism analysis (ribotyping), restriction endonuclease analysis (REA), SDS-PAGE analysis of whole-cell (WCP) and outer-membrane (OMP) proteins, and capsule and somatic serotyping. Bacterial strains were isolated from nose, lung and in one case testicle, of Holstein and cross-bred beef cattle. The isolates represented for the most part serogroup A3 (88%). Ribotyping was performed on DNA digested with HaeII, electrophoresed and then hybridised with 32P-labelled 16S-23S rRNA from Escherichia coli. Six ribotypes (R1-R6) and 10 REA types were found among the 81 isolates with similar discrimination index (DI) of c. 0.60. Protein profiles revealed reproducibility and high levels of polymorphisms among lung isolates. Isolates were compared according to their geographical habitat, their isolation from dairy or from beef cattle and from nasal cavities or lungs. No correlation was apparent between geographical locations and ribotypes. Overall, isolates obtained from dairy cattle were predominantly R1, whereas those obtained from beef cattle were equally distributed between R1 and R2. R1 was more representative of lung isolates. For some strains, particularly the single isolate ribotypes, good correlation was achieved between WCP analysis, REA types and ribotypes. For others, REA to some extent and WCP profiles were able to discriminate among isolates within ribotypes. The data suggest that a combination of ribotyping, REA and WCP analysis is useful for investigating the epidemiology of bovine P. multocida serogroup A.

Serum antibody responses of cattle vaccinated with partially purified native Pasteurella haemolytica leukotoxin.

The objective of these experiments was to study the serum antibody responses of cattle to partially purified, native Pasteurella haemolytica A1 leukotoxin (LKT) formulated with a commercial aluminum hydroxide-DDA-bromide adjuvant. In two experiments, calves received two intramuscular injections 21 days apart and sera were obtained periodically. Serum antibody responses to P. haemolytica outer membrane proteins (OMPs), formalinized P. haemolytica, and LKT were determined. In Experiment A, Holstein calves (140 kg each) were vaccinated with either 10, 1.0 or 0.1 micrograms of LKT, 10(9) c.f.u. of live P. haemolytica, or adjuvanted diluent. In Experiment B, mixed-breed beef calves (200 kg each) were vaccinated with either 100, 50 or 10 micrograms of LKT, 10(9) c.f.u. live P. haemolytica, or adjuvanted diluent. Vaccination of dairy calves with 10 micrograms of partially purified LKT stimulated LKT neutralizing antibody responses similar to those stimulated by vaccination of one calf with live P. haemolytica. In Experiment B, which used larger and different breeds of cattle, two vaccinations 3 weeks apart with 50 micrograms LKT stimulated LKT neutralizing responses equivalent to or greater than those stimulated by vaccination with live P. haemolytica. In both experiments, LKT vaccines stimulated only low antibody responses to formalinized P. haemolytica or to OMPs.