A phase Ib/II and pharmacokinetic study of EP0057 (formerly CRLX101) in combination with weekly paclitaxel in patients with recurrent or persistent epithelial ovarian, fallopian tube, or primary peritoneal cancer.

BACKGROUND: EP0057 (formerly CRLX101) is an investigational nanoparticle-drug conjugate (NDC) of a cyclodextrin-based polymer backbone plus camptothecin, a topoisomerase-1 inhibitor. Prior studies showed efficacy in recurrent or persistent, epithelial ovarian, fallopian tube or primary peritoneal cancer (EOC). METHODS: This phase Ib/2 trial assessed safety and efficacy of EP0057 Q2W plus weekly paclitaxel in patients with EOC. The recommended phase 2 dose (RP2D) was identified using a 3+3 design. The single-arm phase 2 assessed overall response (ORR) per RECIST 1.1 in patients previously treated with bevacizumab. Secondary objectives included progression free survival (PFS) and duration of response. RESULTS: The RP2D was established as 15 mg/m2 EP0057 Q2W plus 80 mg/m2 paclitaxel administered 3 weeks on/1 week off. Nine patients enrolled on phase 1b, with no DLTs; 21 additional patients enrolled on phase 2. All completed >1 cycle. Median age was 62 (44-76) years, 57% ≥3 prior therapies. For the primary analysis, 6/19 patients with prior bevacizumab had confirmed responses (ORR=31.6% (95% CI: 15.4% to 54.0%)) including one complete response (CR). Median PFS was 5.4 months. Most common grade 3/4 adverse events attributed to treatment were decreased neutrophil count (13, 43%) and anemia (3, 10%). CONCLUSIONS: Although the observed ORR was not statistically better than the historical control rate, EP0057 remains an interesting option for treatment of recurrent EOC. EP0057 exhibits high plasma drug retention, slow clearance, and controlled slow release of CPT from the polymer when administered alone and with paclitaxel. (NCT02389985) 242 words.

Haemoglobin A1c levels and subsequent cardiovascular disease in persons without diabetes: a meta-analysis of prospective cohorts.

AIMS/HYPOTHESIS: The aim of this meta-analysis was to determine the relationship between HbA(1c) levels and subsequent cardiovascular outcomes in individuals without diabetes. METHODS: We searched Medline, Embase and Scopus from initiation of the study until the end of 2009. One reviewer searched and another verified findings. Data were extracted by one reviewer and verified by another. We accepted prospective studies in any language reporting three or more quartiles for HbA(1c) levels. Within quartiles, authors must have presented both numbers of patient-years at risk and cardiovascular outcomes. Outcomes per person-time at risk were regressed on average HbA(1c) values using Poisson regression. We pooled ß coefficients using Cochran's semi-weighted (inverse variance) random-effects model. Study quality was assessed using the Downs-Black scale. RESULTS: We investigated 16 datasets (nine for total cardiovascular events and seven for death) from five papers with 44,158 patients (44% men) over 404,899 patient-years of follow-up. There were 1,366 cardiovascular deaths (3.1%; 3.37/1,000 person-years) and 2,142 cardiovascular events (4.9%; 5.29/1,000 person-years). The overall meta-analytic ß coefficients were 0.720 (95% CI 0.307-1.133) and 0.757 (95% CI 0.382-1.132) for cardiac death and events, respectively. Compared with the baseline value of 0.0427, an HbA(1c) level of 0.05 was associated with a relative risk for cardiovascular death of 1.13 (95% CI 1.05-1.21), a 0.06 value with 1.34 (95% CI 1.13-1.58), and a 0.07 HbA(1c) with relative risk 1.58 (95% CI 1.22-2.06). Results for total cardiovascular events were similar. The average study quality was 0.7 (70%). CONCLUSIONS/INTERPRETATION: We conclude that HbA(1c) was significantly associated with cardiovascular events and deaths in persons without diabetes.

Determination of water quality variables, endotoxin concentration, and Enterobacteriaceae concentration and identification in southern High Plains dairy lagoons.

The objectives of this study were to determine the concentration of endotoxin, determine 20 water quality variables, and identify and enumerate fungal and bacterial pathogens from United States southern High Plains dairy lagoons and control lakes during summer and winter. Water samples were collected in triplicate from the north, south, east, and west quadrants of each body of water. The mean (+/- SEM) winter dairy lagoon endotoxin concentration was significantly higher (9,678+/-1,834 ng/mL) than the summer concentration (3,220+/-810 ng/mL). The mean endotoxin concentration of the 2 control lakes (summer: 58.1+/-8.8 ng/mL; winter: 38.6+/-4.2 ng/mL) was significantly less than that of the dairy lagoons. Two hundred-one Salmonella enterica spp. isolates were identified, 7 serovars were recovered from the dairy lagoons, and 259 Salmonella ssp. were identified from 5 other dairy locations (milk barn, ditch effluent, settling basin, feed alley pad flush, and center pivots). Twenty-eight Salmonella spp. were identified from center pivot water. Escherichia coli O157:H7 pathogens were isolated from other dairy locations but not from lagoons. Neither Salmonella spp. nor E. coli O157:H7 were identified from control lakes. Enterobacteriaceae opportunistic pathogens were isolated from both dairies and control lakes. Important mesophilic and thermophilic catabolic (to manure biosolids) fungal isolates were identified from dairy effluent locations, but no thermophilic fungal isolates were cultured from the control lakes. Adequate curing of green forage following center pivot irrigation is important to kill lagoon water enteric pathogens, even though the lagoon water is mixed with fresh water. Recirculating lagoon water to flush the feed alley pad, where cows stand while eating, to remove manure and using lagoon water to abate dairy dust in loafing pens and unimproved dairy roads is inconsistent with good environmental practice management.

Ambient and indoor particulate aerosols generated by dairies in the southern High Plains.

The objectives were to quantify and size ambient aerosolized dust in and around the facilities of 4 southern High Plains dairies of New Mexico and to determine where health of workers might be vulnerable to particulate aerosols, based on aerosol concentrations that exceed national air quality standards. Ambient dust air samples were collected upwind (background) and downwind of 3 dairy location sites (loafing pen boundary, commodity, and compost field). The indoor milking parlor, a fourth site, was monitored immediately upwind and downwind. Aerosolized particulate samples were collected using high-volume sequential reference air samplers, laser aerosol monitors, and cyclone air samplers. The overall (main effects and estimable interactions) statistical general linear model statement for particulate matter (PM(10); particulate matter with an aerodynamic diameter of up to 10 microm) and PM(2.5) resulted in a greater mean concentration of dust in the winter (PM(10) = 97.4 +/- 4.4 microg/m(3); PM(2.5) = 32.6 +/- 2.6 microg/m(3)) compared with the summer (PM(10) = 71.9 +/- 5.0 microg/m(3); PM(2.5) = 18.1 +/- 1.2 microg/m(3)). The upwind and downwind boundary PM(10) concentrations were significantly higher in the winter (upwind = 64.3 +/- 9.5 microg/m(3); downwind = 119.8 +/- 13.0 microg/m(3)) compared with the summer (upwind = 35.2 +/- 7.5 microg/m(3); downwind = 66.8 +/- 11.8 microg/m(3)). The milking parlor PM(10) and PM(2.5) concentration data were significantly higher in the winter (PM(10) = 119.5 +/- 5.8 microg/m(3); PM(2.5) = 55.3 +/- 5.8microg/m(3)) compared with the summer (PM(10) = 88.6.0 +/- 5.8 microg/m(3); PM(2.5) = 21.0 +/- 2.1 microg/m(3)). Personnel should be protected from high aerosol concentrations found at the commodity barn, compost field, and milking parlor during the winter.

Terfenadine pharmacokinetics in breast milk in lactating women.

The excretion of terfenadine into breast milk has not been reported previously. Disposition of terfenadine was prospectively studied in four healthy lactating mothers (age, 33 +/- 4 years). Subjects received 60 mg terfenadine every 12 hours over a period of 48 hours to achieve steady-state milk and plasma concentrations. Milk and plasma samples were collected at 1/2, 1, 1 1/2, 2, 3, 4, 6, 8, 12, 24, and 30 hours after the last dose. Terfenadine and its active metabolite milk and plasma concentrations were quantitated by HPLC. Terfenadine was not detected in milk or plasma. Mean +/- SD active metabolite data for milk and plasma are as follows: Cmax (ng/ml), 41.0 +/- 16.4 for milk, 309.0 +/- 120.5 for plasma; tmax (hours), 4.3 +/- 2.4 for milk, 3.9 +/- 3.0 for plasma; t1/2 beta (hours), 14.2 +/- 5.4 for milk, 11.7 +/- 6.4 for plasma; AUC(0-12) (ng.hr/ml) 320.4 +/- 99.8 for milk, 1590.0 +/- 300.4 for plasma. Metabolite milk/plasmaAUC(0-12) ratios ranged from 0.12 to 0.28 (mean, 0.21 +/- 0.07). Newborn dosage estimates based on the highest measured concentration of terfenadine metabolite in milk suggests the maximum level of newborn exposure would not exceed 0.45% of the recommended maternal weight-corrected dose. Estimated amounts consumed by the neonate after the mother is given the recommended dose of the drug are not likely to result in plasma levels producing untoward effects.

The role of the physiotherapist in the management of repetitive strain injury.

A review of current literature concerning the aetiology, diagnosis, role and involvement of physiotherapists in the treatment of repetitive strain injury (R.S.I.) is presented as a basis for investigation. To determine the modes of treatment used by physiotherapists in the management of R.S.I. and to analyse their efficacy, a questionnaire was designed. Forty centres were surveyed and the results are presented. Information concerning the most commonly encountered conditions, treatment given, and physiotherapists' opinions on prevention, patient education and further training in RSI management was also sought. An attempt is made to define the role of the physiotherapist in the recognition, treatment and education aspects of over-use injury, and recommendations for further research and physiotherapy involvement are presented.

Influence of fasting and post-fast diet energy level on feed intake, feeding pattern and blood variables of lambs.

Trials were conducted to determine the influence of feed and water deprivation on feed intake, plasma glucose, free fatty acids (FFA), urea-N (PUN), serum insulin and growth hormone (GH) in lambs. In Trial 1, 12 Hampshire X Suffolk lambs (avg wt 30.5 kg) were deprived of feed and water for 0, 24, 48 or 72 h. During the first 8 d of realimentation, feed intake was depressed more (P less than .05) by longer periods of deprivation. In Trial 2, 12 crossbred lambs (avg wt 50 kg) were deprived of feed and water for 0 or 72 h. During the first 4 d of realimentation, feed intake was lower (P less than .05) in deprived than in fed lambs. The depressed feed intake could be detected within 30 min of feeding. In Trial 3, 12 crossbred lambs (avg wt 40 kg) were fasted for 0 or 72 h, and blood samples were obtained at -5, 30, 60, 120 and 180 min postprandial. On the 1st d of realimentation, lambs previously fasted had abnormal serum hormone patterns compared with nonfasted controls. On d 2 of realimentation, lambs previously fasted had higher (P less than .05) insulin, glucose and FFA than controls. On d 4 of realimentation, lambs previously fasted had higher insulin:GH ratios and lower PUN than controls. Results of these trials suggest that depriving lambs of feed and water for 72 h reduces subsequent feed intake for 4 d or more. Postprandial blood metabolite patterns are abnormal for a similar length of time.

Influence of yeast culture on feeder calves and lambs.

Four experiments were conducted to determine the influence of yeast culture on 1) the health and performance of feeder calves, 2) the response of calves to an infectious bovine rhinotracheitis virus (IBRV) infection, and 3) nutrient utilization in lambs fasted for 3 d. In Exp. 1, 108 feeder calves were transported from Tennessee to Texas (1,600 km) and fed receiving diets containing 0 or .75% yeast culture and .35 or .69% P in a 2 x 2 factorial arrangement of treatments. In Exp. 2, 101 calves were transported 950 kg from Austin, TX to Bushland, TX and fed receiving diets containing 0, .75, 1.125, or 1.5% yeast culture. Yeast culture did not significantly affect the health or performance of calves in either experiment, although morbid calves fed yeast culture required fewer (P less than .05) days of antibiotic therapy in Exp. 2. In Exp. 3, feeder steers were fed diets containing 0 or .75% yeast culture and challenged intranasally with IBRV. Calves fed yeast culture tended to maintain heavier weights and higher DMI during IBRV infection than did steers fed the control diet. In Exp. 4, feeder lambs were fasted for 3 d and refed diets containing 0, .75, 1.125, or 1.5% yeast culture during a N and mineral balance trial. Lambs fed yeast culture had greater (P less than .08) N balance and tended to have greater Zn and Fe balance than control lambs. Results of these studies are interpreted to suggest that supplementation of morbid calves with yeast culture can have beneficial effects (fewer sick days, higher feed intakes) and that these effects may be mediated by improved N, Zn, and Fe metabolism.

Effect of simulated ambient particulate matter exposure on performance, rectal temperature, and leukocytosis of young Spanish goats with or without tilmicosin phosphate.

Dust is an environmental stressor and can become extensive in agricultural production systems. Thirty-six female, Spanish goats (average BW 21.1 kg, SEM = 1.31; age = 4 mo) were randomly assigned to simulated dust events or no dust, with or without tilmicosin phosphate treatment in a 2 x 2 factorial arrangement of treatments to determine effects on performance, rectal temperature, and leukocyte changes. All goats were fed a standard growing diet (13.6% CP) consisting of 37% roughage and 63% concentrate (DM basis). Feed intake was measured daily, and BW (unshrunk) measured individually every 7 d. The tilmicosin-treated group received tilmicosin phosphate (10 mg/kg BW s.c.) before starting the study. Goats exposed to dust were enclosed as a group inside a canvass tent for 4 h each day and ground feed yard manure dust (mean particle size 100 microm) was aerosolized inside the tent to simulate a dust event. There was one single dust event (Phase I) followed by rectal temperature measurement, and heparinized blood collection for complete cell counts at 0 (pretrial), 4, 12, 20, 44, 68, and 210 h after dust exposure. This was followed by 21 d of chronic dust events (Phase II). The sampling procedures for Phase II were exactly the same as in Phase I, except that samples were obtained daily at 0 (before dust application), 4, 8, and 12 h after each dust event. Dust treatment had no effect (P > 0.05) on feed intake or ADG, but the gain:feed (G:F) ratio was lower (P < 0.05) in the control goats than the dust exposed group. Tilmicosin phosphate-treated goats had a higher (P < 0.05) G:F ratio than untreated goats. Dust exposure increased (P < 0.002), but tilmicosin treatment decreased (P < 0.05) rectal temperature at 4 and 8 h. Dust exposure increased (P < 0.02) blood lymphocyte counts compared with controls. These results suggest that simulated dust events altered rectal temperature and leukocyte counts of goats.

Influence of triiodothyronine injections on calf immune response to an infectious bovine rhinotracheitis virus challenge and nitrogen balance of lambs.

Three experiments were conducted to determine the influence of triiodothyronine (T3) or propylthiouracil (PTU) on the humoral immune response of calves challenged with infectious bovine rhinotracheitis virus (IBRV) and on nitrogen depletion and repletion of lambs deprived of feed and water for 3 d. In Exp. 1, 18 steer calves (BW 284 +/- 6 kg) challenged with IBRV were limit-fed (1.5% BW) a 60% concentrate diet and injected (s.c.) daily with alkaline saline, .4 mg of T3, or .8 mg of T3. Injections of T3 did not affect serum antibody titers to IBRV, blood leukocyte counts, or plasma free fatty acid, ceruloplasmin, and cholesterol concentrations but increased (P < .05) plasma glucose concentrations and decreased (P < .05) plasma urea N concentrations. In Exp. 2, 36 IBRV-challenged steers (BW 266 +/- 8 kg) were given ad libitum access to a 60% concentrate diet and injected (s.c.) daily with alkaline saline, .2 mg of T3, or .4 mg of T3. In contrast to Exp. 1, injections of T3 did not affect plasma glucose or urea N concentrations and reduced (P < .05) serum antibody titers to IBRV. In Exp. 3, eight wether lambs were limit-fed (600 g/d) a 36% concentrate pelleted diet and assigned to one of four treatments in a replicated 4 x 4 Latin square designed nutrient balance experiment involving periods of nutrient depletion and repletion. Treatments were as follows: 1) alkaline saline injection (s.c.), 2) 4 mg of PTU/kg BW in water, 3) .15 mg of T3 s.c. daily for 15 d, and 4) .15 mg of T3 s.c. daily for 7 d after the 3-d feed and water deprivation period. Thyroid status affected (P < .05) predeprivation N balance but did not affect N losses during the feed and water deprivation period. Retention of N during realimentation was affected (P < .05) by T3 treatment. Results of these experiments indicate that there is a complex interrelationship among stress, nutrient status, and thyroidal status and that the effects of T3 injections on immune and metabolic responses may be dependent on the nutritional status of the animal.(ABSTRACT TRUNCATED AT 250 WORDS)

Bovine viral diarrhea viral infections in feeder calves with respiratory disease: interactions with Pasteurella spp., parainfluenza-3 virus, and bovine respiratory syncytial virus.

The prevalence of bovine viral diarrhea virus (BVDV) infections was determined in a group of stocker calves suffering from acute respiratory disease. The calves were assembled after purchase from Tennessee auctions and transported to western Texas. Of the 120 calves, 105 (87.5%) were treated for respiratory disease. Sixteen calves died during the study (13.3%). The calves received a modified live virus BHV-1 vaccine on day 0 of the study. During the study, approximately 5 wk in duration, sera from the cattle, collected at weekly intervals, were tested for BVDV by cell culture. Sera were also tested for neutralizing antibodies to BVDV types 1 and 2, bovine herpesvirus-1 (BHV-1), parainfluenza-3 virus (PI-3V), and bovine respiratory syncytial virus (BRSV). The lungs from the 16 calves that died during the study were collected and examined by histopathology, and lung homogenates were inoculated onto cell cultures for virus isolation. There were no calves persistently infected with BVDV detected in the study, as no animals were viremic on day 0, nor were any animals viremic at the 2 subsequent serum collections. There were, however, 4 animals with BVDV type 1 noncytopathic (NCP) strains in the sera from subsequent collections. Viruses were isolated from 9 lungs: 7 with PI-3V, 1 with NCP BVDV type 1, and 1 with both BVHV-1 and BVDV. The predominant bacterial species isolated from these lungs was Pasteurella haemolytica serotype 1. There was serologic evidence of infection with BVDV types 1 and 2, PI-3V, and BRSV, as noted by seroconversion (> or = 4-fold rise in antibody titer) in day 0 to day 34 samples collected from the 104 survivors: 40/104 (38.5%) to BVDV type 1; 29/104 (27.9%) to BVDV type 2; 71/104 (68.3%) to PI-3V; and 81/104 (77.9%) to BRSV. In several cases, the BVDV type 2 antibody titers may have been due to crossreacting BVDV type 1 antibodies; however, in 7 calves the BVDV type 2 antibodies were higher, indicating BVDV type 2 infection. At the outset of the study, the 120 calves were at risk (susceptible to viral infections) on day 0 because they were seronegative to the viruses: 98/120 (81.7%), < 1:4 to BVDV type 1; 104/120 (86.7%) < 1:4 to BVDV type 2; 86/120 (71.7%) < 1:4 to PI-3V; 87/120 (72.5%) < 1:4 to BRSV; and 111/120 (92.5%) < 1:10 to BHV-1. The results of this study indicate that BVDV types 1 and 2 are involved in acute respiratory disease of calves with pneumonic pasteurellosis. The BVDV may be detected by virus isolation from sera and/or lung tissues and by serology. The BVDV infections occurred in conjunction with infections by other viruses associated with respiratory disease, namely, PI-3V and BRSV. These other viruses may occur singly or in combination with each other. Also, the study indicates that purchased calves may be highly susceptible, after weaning, to infections by BHV-1, BVDV types 1 and 2, PI-3V, and BRSV early in the marketing channel.

Bovine viral diarrhea virus (BVDV) 1b: predominant BVDV subtype in calves with respiratory disease.

The prevalence of bovine viral diarrhea virus (BVDV) infections was determined in 2 groups of stocker calves with acute respiratory disease. Both studies used calves assembled after purchase from auction markets by an order buyer and transported to feedyards, where they were held for approximately 30 d. In 1 study, the calves were mixed with fresh ranch calves from a single ranch. During the studies, at day 0 and at weekly intervals, blood was collected for viral antibody testing and virus isolation from peripheral blood leukocytes (PBLs), and nasal swabs were taken for virus isolation. Samples from sick calves were also collected. Serum was tested for antibodies to bovine herpesvirus-1 (BHV-1), BVDV1a, 1b, and 2, parainfluenza 3 virus (PI3V), and bovine respiratory syncytial virus (BRSV). The lungs from the calves that died during the studies were examined histopathologically, and viral and bacterial isolation was performed on lung homogenates. BVDV was isolated from calves in both studies; the predominant biotype was noncytopathic (NCP). Differential polymerase chain reaction (PCR) and nucleic acid sequencing showed the predominant subtype to be BVDV1b in both studies. In 1999, NCP BVDV1b was detected in numerous samples over time from 1 persistently infected calf; the calf did not seroconvert to BVDV1a or BVDV2. In both studies, BVDV was isolated from the serum, PBLs, and nasal swabs of the calves, and in the 1999 study, it was isolated from lung tissue at necropsy. BVDV was demonstrated serologically and by virus isolation to be a contributing factor in respiratory disease. It was isolated more frequently from sick calves than healthy calves, by both pen and total number of calves. BVDV1a and BVDV2 seroconversions were related to sickness in selected pens and total number of calves. In the 1999 study, BVDV-infected calves were treated longer than noninfected calves (5.643 vs 4.639 d; P = 0.0902). There was a limited number of BVDV1a isolates and, with BVDV1b used in the virus neutralization test for antibodies in seroconverting calves' serum, BVDV1b titers were higher than BVDV1a titers. This study indicates that BVDV1 strains are involved in acute respiratory disease of calves with pneumonic Mannheimia haemolytica and Pasteurella multocida disease. The BVDV2 antibodies may be due to cross-reactions, as typing of the BVDV strains revealed BVDV1b or la but not BVDV2. The BVDV1b subtype has considerable implications, as, with 1 exception, all vaccines licensed in the United States contain BVDV1a, a strain with different antigenic properties. BVDV1b potentially could infect BVDV1a-vaccinated calves.

Comparison of four immune variables and pulmonary lesions of goats with intrapulmonary exposure and subsequent intrathoracic challenge exposure with Pasteurella haemolytica.

A comparison of immune variables following lung sensitization with live Pasteurella haemolytica serotype 1 (Ph1)-impregnated agar beads was done in 2 separate trials. The Ph1 immune variables studied were blood bactericidal activity, serum bacteriolysis, total classical complement, and indirect hemagglutination antibody. Each trial had 16 male weanling goats: 6 controls and 10 principals. In trial 1, each goat was surgically catheterized through the trachea, then the material was deposited in a bronchus. The controls received only agar beads and the principals received agar beads impregnated with live Ph1. These goats were studied for 32 days, euthanatized, and necropsied. In trial 2, the controls were each transthoracically injected with agar beads into the left lung and the principals were similarly injected with agar beads impregnated with live Ph1. These goats were studied for 35 days, then challenge exposed transthoracically by injection of Ph1 in saline solution (1.2 x 10(7) CFU/ml) into the right lung. Four days later, they were euthanatized and necropsied. The volume of lung consolidated tissue was an excellent measure of Ph1 immunity. Principal goats generated solid protective immunity to subsequent challenge exposure because minimal or no lung consolidation was observed, whereas large volumes of lung consolidation were seen in the controls. The principal goats in trial 1 gave a weak serum indirect hemagglutination Ph1 antibody response, which was attributed to the bronchial method of depositing the Ph1. The corresponding response of the control group remained negative. The Ph1 agar beads (1 x 10(6) CFU in 0.5 ml) protected the bacteria from immediate phagocytosis and lysis as indicated by the induced pneumonic deaths of 2 principals 5 days later.(ABSTRACT TRUNCATED AT 250 WORDS)

Efficacy of Pasteurella haemolytica subunit antigens in a goat model of pasteurellosis.

The effectiveness of Pasteurella haemolytica biovar A, serovar 1 (Ph1) subunit vaccines was tested in goats, using challenge exposure by transthoracic injection. Twenty-two weanling male Spanish goats were randomly allotted to 4 groups. Six goats were given 2 transthoracic injections into the lung 18 days apart with live Ph1 impregnated in agar beads (positive controls). Six goats were not given injections (negative controls). Five goats were given 2 transthoracic injections into the lung 18 days apart with 4.6 mg of cytotoxin in agar beads. The remaining 5 goats were given 2 IM injections, 18 days apart, into the thigh with 4.6 mg of cytotoxin emulsified in incomplete Freund's adjuvant. Twenty-four days after the second injection, all goats were challenge-exposed to live Ph1 by transthoracic injection into the lung, and 4 days later, all goats were euthanatized and necropsied. Serum neutralizing anticytotoxin titer was measured throughout the experiment. Mean volume of consolidated lung tissue was 0.38 cm3 for the positive control group, 32 cm3 for the negative control group; 19 cm3 for the cytotoxin-lung group; and 88 cm3 for the cytotoxin-adjuvant-IM group. Only the positive control group was protected from Ph1 challenge exposure. The Ph1 cytotoxin subunit vaccine alone appeared to be ineffective, and the anticytotoxin titer was not correlated with protection. In a separate trial, 32 weanling male Spanish goats were randomly allotted to 5 groups. Each was given 2 transthoracic injections into the lung 22 days apart. Six goats were given Ph1 cytotoxin impregnated into agar beads; 6 were given Ph1 lipopolysaccharide impregnated in agar beads; 6 were given Ph1 capsule impregnated in agar beads. Six goats were given agar beads only (negative controls), and 6 were given live Ph1 impregnated into agar beads (positive controls). Twenty days after the second injection, all goats were challenge-exposed to live Ph1 by transthoracic injection into the lung, and 4 days later, all goats were euthanatized and necropsied. Mean volume of consolidated lung tissue was 0.14 cm3 for the positive control group, 7.59 cm3 for the negative control group, 11.21 cm3 for the cytotoxin group, 10.19 cm3 for the lipopolysaccharide group, and 1.6 cm3 for the capsule group. Again, only injection of live Ph1 (positive controls) induced solid protection; however, the capsule subunit vaccine induced partial protection against challenge exposure in this trial. Lipopolysaccharide and cytotoxin subunit vaccines were ineffective in protecting goats against challenge exposure with live Ph1.

Effects of marketing stress on fecal excretion of Salmonella spp in feeder calves.

Fecal samples were collected from 200 feeder-calves on farms in Tennessee, after assembly at a Tennessee auction market, and after transport to a Texas feedyard. A final fecal sample was collected from each calf after 30 days of feedyard confinement. The fecal samples were cultured for the presence of Salmonella spp. Salmonella isolates were serotyped and antimicrobial drug-resistance patterns determined. The number of calves fecal culture-positive for Salmonella spp increased from 0 on the Tennessee farms and auction market to 3/200 (1.5%) at entry into the Texas feedyard, and 16/200 (8%) after 30 days of feedyard confinement. Salmonella serotypes isolated and the number of isolates of each serotype were S reading (8), S cerro (4), S newbrunswick (3), S anatum (2), and S typhimurium (copenhagen; 2). All Salmonella isolates were resistant to 5 or more of 13 antimicrobial drugs tested. Salmonella reading isolates were resistant to 10 or 11 of 13 antimicrobial drugs. The results indicated that the calves could have been infected with Salmonella spp prior to or during the course of the study, and that marketing stress as they moved from farm through feedyard may have induced fecal excretion of salmonellae. In addition, the pattern of antimicrobial drug resistance in the Salmonella isolates was broad.

Market stress-associated changes in serum complement activity in feeder calves.

Classical hemolytic complement (C) of calves was analyzed during a protocol designed to imitate the usual market handling of feeder calves from the southeastern United States. Serum C concentrations of the calves (n = 100 x 4 years) were evaluated on their farm of origin, on arrival at an auction market, on arrival at a feedyard, and during their first 4 weeks in the feedyard. Complement concentrations (measured in CH50 units) were typically lowest at the farm of origin and highest when the calves entered the auction market 28 to 133 days later. Serum C concentrations decreased after the calves encountered the severe stresses of being in the auction market for 7 days, 24-hour truck transport (1,932 km) to the feedyard, and the first 7 days in the feedyard. The C concentrations recovered after 21 to 28 days in the feedyard. Steers had significantly (P less than or equal to 0.05) lower C concentrations than did heifers in 3 of 4 years at the farm of origin, and in 2 of 4 years at the auction market. Morbid calves had significantly (P less than or equal to 0.05) lower C values than did healthy calves on day 7 in the feedyard in 3 of 4 years. There were significant differences in C concentrations of calves from different farms of origin in each of the 4 years. There was no significant difference in C concentrations of calves that were vaccinated vs those not vaccinated with Pasteurella haemolytica.

Efficacy of a subcutaneously administered, ultraviolet light-killed Pasteurella multocida A:3-containing bacterin against transthoracic challenge exposure in goats.

OBJECTIVE: To determine the effectiveness of Pasteurella multocida biovar A, serovar 3 (Pm A:3) killed by exposure to UV light and incorporated with a polyacrylate bead carrier as a vaccine. ANIMALS: 18 weanling male Spanish goats. PROCEDURE: Prospective, randomized controlled study with 3 treatment groups: positive-control (PC), negative-control (NC), and principal Pm A:3 bacterin (PA) groups. Six PC goats each received live Pm A:3 and polyacrylate beads twice, 22 days apart, by transthoracic injection into the left lung. Six NC goats each received only PA beads twice, 22 days apart, by transthoracic injection. Six principal goats each received Pm A:3 vaccine SC twice, 22 days apart. Fourteen days after the second vaccination, all goats were challenge exposed with live Pm A:3 by transthoracic injection into the right lung, and 4 days later they were euthanatized and necropsied. RESULTS: Mean volume of consolidated lung tissue at the challenge site was 1.75 cm3 for the PC group, 15.18 cm3 for the NC group, and 3.9 cm3 for the PA vaccine group. The NC group had a significantly (P < or = 0.002) larger mean volume of consolidated lung tissue than did the PC and PA groups after challenge exposure. CONCLUSIONS: The PA bacterin and the PC groups developed protective immunity against live Pm A:3 challenge exposure. An SC administered, UV light-killed, Pm A:3 bacterin induced protective immunity similar to that induced by virulent live Pm A:3 injected into the target organ, the lung.

Blood bactericidal assay (Pasteurella haemolytica) comparison of morbidity in marketed feeder calves.

An in vitro bactericidal assay that used bovine heparinized blood was investigated for its usefulness in detecting differences in the bactericidal immunity of calves against Pasteurella haemolytica serotype 1 (Ph1). Greater than 90% of killing occurred within 30 minutes. The substitution of fetal calf serum for autologous calf plasma caused loss of bactericidal activity of the blood. Decomplemented calf serum also was low in bactericidal activity. The blood bactericidal assay appears to be opsonin antibody-dependent and complement-dependent. The coefficient of variation (CV) that can be expected with this assay was established by use of a group of 8 calves; within-day CV maximum was 0.9, and between-day CV maximum was 2.1. The blood bactericidal assay was used to evaluate 30 calves under typical market stress from 4 farms in eastern Tennessee. All calves had decreased bactericidal activity, as they moved into a feedyard in Texas. The bactericidal activity was reduced among sick calves, based on the severity of clinical signs. Morbidity was highest during the first 14 days in the feedlot. During this period, healthy calves had a decreased bactericidal index (BI) of 4 points, and calves with clinical signs of bovine respiratory tract disease for 3 days had a decreased BI of 8 points. The average reduction in the BI of calves with clinical signs of bovine respiratory tract disease for 6 or more days was 14 points.

Equine rhinopneumonitis virus (herpesvirus type 1): attenuation in stable monkey cell line.

An isolate of virulent equine herpesvirus (EHV) type 1 was adapted to Vero stable cell line by 13 serial passages at 37 C and 50 serial passages at 26 C. Characteristics of the attenuated EHV-1 were found to be avirulent, but immunogenic in horses if injected intramuscularly. The attenuated virus was regularly isolated from peripheral leukocytes in inoculated horses, but was not recovered from nasal turbinate tissues. A mild leukopenia was noticed. The attenuated virus produced characteristic large syncytia on primary isolation in rabbit kidney (RK13) or Vero cells at 37 C in contrast to cell rounding observed with virulent EHV-1. The syncytial marker was stable through 20 serial passages in Vero cells at 37 C. New application of double immunodiffusion test for distinguishing between EHV-1 and EHV-2 also is described.

Immune response to pulmonary injection of Pasteurella haemolytica-impregnated agar beads followed by transthoracic challenge exposure in goats.

A method of inducing Pasteurella haemolytica serotype 1 (Ph1) lung infection in goats, using low numbers of bacteria and without impairing host immunity, was developed. Two trials were conducted. Results of trial 1, using 10 principals (Ph1 agar beads) and 6 controls (agar beads alone), indicated that Ph1 organisms imbedded in agar beads could survive host lung defenses for 32 days. Results of trial 2 indicated that lung immunity in the inoculated goats (principals) was high and they were more protected than controls against a transthoracic challenge of Ph1 (1.18 x 10(7) colony-forming units) injected into a lung of each goat on posttreatment day 35. When comparing challenge-exposed principals with controls, the controls developed rectal temperatures above normal for a longer time, duration of anorexia was longer, and signs of depression were seen. The controls developed large areas of consolidated lung tissue, more Ph1 isolates were recovered from nasal turbinates and lung tissue, and higher Ph1 concentrations were found in the lungs. The serum Ph1 indirect hemagglutination antibody titers in the principals of both trials increased, compared with titers in controls. Principal goats in trial 2 had higher Ph1 indirect hemagglutination antibody titers after injection of Ph1-impregnated agar beads and less severe lung lesions after challenge exposure than did controls. The small pneumonic consolidated lesions in the principals, compared with extensive lesions in controls after Ph1 challenge exposure, indicated a high degree of immunity after exposure to Ph1 organisms imbedded in agar beads.