Relationship of fetal age at conjunctival exposure of pregnant heifers and Brucella abortus isolation

Pregnant heifers were exposed by a conjunctival inoculation with 1 X 10(7) colony-forming units of Brucella abortus strain 2308. At parturition, milk and uterine samples from dams plus samples from dead calves were cultured bacteriologically for Brucella. The logistic regression probability of B abortus isolation increased from 0.22 to 0.90, as fetal age at exposure of heifers increased from 60 to 150 gestation days. Strain 2308 was recovered at parturition from 14 (64%) of 22, 17 (71%) of 24, and all 28 (100%) heifers that were at gestation days less than 127, 127 to 157, and greater than 157, respectively, at time of exposure. The number of infected heifers and the number of samples positive for B abortus were significantly increased as fetal age at exposure of heifers increased from gestation days less than 127 to greater than 157 (chi 2 greater than 10, P less than 0.005).     

Attenuation of a Brucella abortus mutant lacking a major 25 kDa outer membrane protein in cattle

OBJECTIVE: To determine the virulence of a Brucella abortus mutant, BA25, lacking a major 25 kd outer membrane protein (Omp25) in cattle. ANIMALS: 20 mixed-breed heifers in late gestation. PROCEDURE: 10 heifers were inoculated with 1 x 10(7) colony-forming units of the Omp25 mutant via the conjunctival sac, and an equal number were infected with the virulent parental strain B. abortus 2308. The delivery status of the dams was recorded, and colonization was assessed following necropsy. The ability of BA25 to replicate inside bovine phagocytes and chorionic trophoblasts was also evaluated in vitro because of the propensity of virulent brucellae to replicate inside these cells in vivo. RESULTS: The parental strain induced abortions in 5 of 10 inoculated cattle, whereas only 1 of 10 dams exposed to BA25 aborted. Brucella abortus strain 2308 colonized all of the cow-calf pairs and induced Brucella-specific antibodies in 100% of the dams. In contrast, BA25 was isolated by bacteriologic cultural technique from 30% of the calves and 50% of the inoculated dams (n = 10). Of the 10 heifers inoculated with BA25, 4 did not develop Brucella-specific antibodies nor were they colonized by the mutant strain. In bovine macrophages and chorionic trophoblasts, BA25 replicated in significantly lower numbers than the virulent parental strain (n = 3). CONCLUSIONS AND CLINICAL RELEVANCE: The 25 kd outer membrane protein may be an important virulence factor for B. abortus in cattle. The attenuation of the Omp25 mutant in cattle may involve the inability of BA25 to replicate efficiently in bovine phagocytes and chorionic trophoblasts.     

Attenuation and immunogenicity of a Brucella abortus htrA cycL double mutant in cattle

PHE1 is a htrA cycL double gene deletion mutant of virulent Brucella abortus strain 2308 (S2308) which has previously been evaluated in the murine and caprine models of bovine brucellosis. This report describes the results of studies conducted with this mutant in the natural bovine host. Six sexually mature, non-gravid heifers were inoculated via the conjunctival sac with 1 x 10(10) colony forming units (CFU) of either the parental S2308 or the htrA cycL gene deletion mutant, PHE1. At 4, 7 and 11 days post-inoculation, PHE1 was found to colonize the bovine host at lower levels than S2308. In a second experiment, eight heifers in mid-gestation were infected with 1 x 10(7) CFU of either strain via the conjunctival sac. The virulent S2308 caused abortions or weak calves in 4/4 cows, while all four cows infected with PHE1 had healthy calves. Furthermore, PHE1 exhibited decreased resistance to killing by cultured bovine neutrophils and macrophages compared to the parental strain. These studies demonstrate that the B. abortus htrA cycL gene deletion mutant PHE1 is highly attenuated in the bovine host when compared to the virulent parental S2308.     

Prevention of persistent infection in calves by vaccination of dams with noncytopathic type-1 modified-live bovine viral diarrhea virus prior to breeding

OBJECTIVE: To determine the ability of a modified-live virus (MLV) bovine viral diarrhea virus (BVDV) type 1 (BVDV1) vaccine administered to heifers prior to breeding to stimulate protective immunity that would block transmission of virulent heterologous BVDV during gestation, thus preventing persistent infection of a fetus. ANIMAL: 40 crossbred Angus heifers that were 15 to 18 months old and seronegative for BVDV and 36 calves born to those heifers. PROCEDURE: Heifers were randomly assigned to control (n = 13) or vaccinated (27) groups. The control group was administered a multivalent vaccine where-in the BVDV component had been omitted. The vaccinated heifers were administered a single dose of vaccine (IM or SC) containing MLV BVDV1 (WRL strain). All vaccinated and control heifers were maintained in pastures and exposed to BVDV-negative bulls 21 days later. Thirty-five heifers were confirmed pregnant and were challenge exposed at 55 to 100 days of gestation by IV administration of virulent BVDV1 (7443 strain). RESULTS: All control heifers were viremic following challenge exposure, and calves born to control heifers were persistently infected with BVDV. Viremia was not detected in the vaccinated heifers, and 92% of calves born to vaccinated heifers were not persistently infected with BVDV. CONCLUSIONS AND CLINICAL RELEVANCE: These results document that vaccination with BVDV1 strain WRL protects fetuses from infection with heterologous virulent BVDV1.     

Fetal protection following exposure to calves persistently infected with bovine viral diarrhea virus type 2 sixteen months after primary vaccination of the dams

This study demonstrated that the bovine viral diarrhea virus (BVDV; types 1 and 2) fractions of a multivalent vaccine protected pregnant heifers and their fetuses at 149 to 217 days of gestation against exposure to calves persistently infected with BVDV type 2a. Eighty percent (eight of 10) of the control heifers were viremic at least 1 day following challenge, whereas all (20 of 20) BVDV-vaccinated heifers were virus isolation-negative on all postchallenge assessment days. Ninety percent (nine of 10) of the calves born to control heifers but only 5% (one of 20) of calves born to BVDV-vaccinated heifers seroconverted to BVDV type 2 before ingesting colostrum. One calf born to a control heifer was persistently infected. No calves from BVDV-vaccinated heifers were persistently infected.     

Effect of dose of Brucella abortus strain 19 in yearling heifers on the relative risk of developing brucellosis from challenge exposure with strain 2308

Yearling heifers were given SC injections of 10(8) (n = 40), 10(9) (n = 44), or 10(10) (n = 44) colony-forming units of Brucella abortus strain 19 (S19). The proportion of heifers with positive serologic test results at 1 month following vaccination increased as the dose of S19 increased. These proportions decreased with time, and all heifers had negative card, rivanol, and complement fixation test results within 4 months. Positive ELISA results persisted beyond 4 months in all three S19 dose groups; however, all heifers were ELISA-negative within 9 months after vaccination. Comparable lymphocyte transformation activity was stimulated by S19 dose of 10(9) or 10(10) and approximately half of the heifers in both groups had a positive stimulation index at 9 months. Immunity of the pregnant heifers was challenged 9 months after vaccination with 10(7) B abortus strain 2308 as follows: diluent controls (n = 69); 10(8) B abortus S19 (n = 40); 10(9) B abortus S19 (n = 39); and 10(10) B abortus S19 (n = 39). Tissue specimens from heifers were obtained at parturition and necropsy for culturing of B abortus. The proportion of heifers that developed brucellosis, ie, had positive culture results, increased as gestation days at challenge exposure increased. The effect of gestational age was controlled in the analysis using logistic regression. The relative risk of brucellosis was reduced to 0.38, 0.15, and 0.06 for B abortus S19 doses of 10(8), 10(9), and 10(10), respectively, compared with diluent controls at 1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of stage of gestation on efficacy of Brucella abortus strain-19 vaccination in cattle.

Seventy-nine cattle in all stages of gestation were inoculated with a low dose (2.5 x 10(8) colony-forming units) of Brucella abortus strain 19, then challenge exposed with pathogenic B abortus strain 2308 during the subsequent gestation. A brucellosis case was defined by isolation of strain 2308 from dam or calf samples. Cumulative incidence of brucellosis cases was 48, 33, 25, or 47% for cattle that were, respectively, not pregnant, or 19 to 87, 100 to 167, or 190 to 253 days in gestation at vaccination. The cumulative incidence was 56% in 27 nonvaccinated controls. The 95% confidence intervals for risk ratios included 1 in all cattle, except those that were 100 to 167 days in gestation at vaccination (ie, second trimester); the confidence interval for this group was 0.21 to 0.97. The prevented fraction (1-risk ratio) attributed to strain 19, in ascending order, was 0.14, 0.16, 0.4, or 0.55, respectively, for cattle that were not pregnant, or were 190 to 253, 19 to 87, or 100 to 167 days in gestation at vaccination. Potential confounders of breed, pen effect, and gestation days at challenge exposure did not significantly affect results. Results supported the hypothesis that stage of gestation at vaccination will affect the prevented fraction of brucellosis, or efficacy of strain 19, in cattle vaccinated with a low dose and, therefore, is one factor that may explain variation in strain 19-induced protection.     

Efficacy of calfhood vaccination with Brucella abortus strain RB51 in protecting bison against brucellosis

In the studies reported here, protection induced by calfhood vaccination of bison with 1.2-6.1 x 10(10)CFU of Brucella abortus strain RB51 (SRB51) against a virulent strain of B. abortus was evaluated. Non-vaccinated and SRB51-vaccinated bison were intraconjunctivally challenged during midgestation with 3 x 10(7)CFU of virulent B. abortus strain 2308 (S2308). Maternal and fetal tissues were obtained within 24hour after abortion or parturition. Incidence of abortion was greater (P<0.05) in non-vaccinated as compared to SRB51-vaccinated bison (62% and 15%, respectively), with abortions occurring between 5 and 8 weeks after experimental challenge. Calves from bison vaccinated with SRB51 had a reduced (P<0.05) prevalence of fetal infection with S2308 as compared to calves from non-vaccinated bison (19% and 62%, respectively). Although the ability to recover the 2308 challenge strain from maternal tissues did not differ (P>0.05) between nonvaccinates and vaccinates (100% and 78%, respectively), calfhood vaccination with SRB51 reduced (P<0.05) recovery of S2308 from uterine or mammary gland tissues. In bison which did not abort, S2308 was routinely recovered in low numbers from maternal lymphatic tissues; particularly the parotid, bronchial, supramammary, and mandibular lymph nodes. The RB51 vaccine strain was not recovered at any time from maternal or fetal samples obtained at necropsy. Histological lesions associated with Brucella-induced abortions were suppurative placentitis, fetal broncho-interstitial pneumonia, and fetal histiocytic splenitis. The results of our studies suggest that calfhood vaccination of bison with SRB51 is efficacious in protecting against intramammary, intrauterine, and fetal infection following exposure to a virulent strain of B. abortus during pregnancy. As brucellosis is transmitted horizontally through fluids associated with the birth or abortion of an infected fetus, or vertically to the calf through the ingestion of milk containing B. abortus, our data suggest that calfhood vaccination with SRB51 will be beneficial in preventing transmission of brucellosis in bison.     

Protective effects against abortion and fetal infection following exposure to bovine viral diarrhea virus and bovine herpesvirus 1 during pregnancy in beef heifers that received two doses of a multivalent modified-live virus vaccine prior to breeding

Objective-To determine whether administration of 2 doses of a multivalent, modified-live virus vaccine prior to breeding of heifers would provide protection against abortion and fetal infection following exposure of pregnant heifers to cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV) and cattle with acute bovine herpesvirus 1 (BHV1) infection. Design-Randomized controlled clinical trial. Animals-33 crossbred beef heifers, 3 steers, 6 bulls, and 25 calves. Procedures-20 of 22 vaccinated and 10 of 11 unvaccinated heifers became pregnant and were commingled with 3 steers PI with BVDV type 1a, 1b, or 2 for 56 days beginning 102 days after the second vaccination (administered 30 days after the first vaccination). Eighty days following removal of BVDV-PI steers, heifers were commingled with 3 bulls with acute BHV1 infection for 14 days. Results-After BVDV exposure, 1 fetus (not evaluated) was aborted by a vaccinated heifer; BVDV was detected in 0 of 19 calves from vaccinated heifers and in all 4 fetuses (aborted after BHV1 exposure) and 6 calves from unvaccinated heifers. Bovine herpesvirus 1 was not detected in any fetus or calf and associated fetal membranes in either treatment group. Vaccinated heifers had longer gestation periods and calves with greater birth weights, weaning weights, average daily gains, and market value at weaning, compared with those for calves born to unvaccinated heifers. Conclusions and Clinical Relevance-Prebreeding administration of a modified-live virus vaccine to heifers resulted in fewer abortions and BVDV-PI offspring and improved growth and increased market value of weaned calves.     

Gestation nutrition, tissue exchange and maintenance requirements of heifers

Forty-six pregnant, crossbred, 2-yr-old heifers of large or small mature size were individually fed a 70% cottonseed hull diet during gestation. Heifers were fed at either 1.0% (nutritionally restricted) or 1.5% (nonrestricted) of body weight from 90 d gestation through parturition. Live weight, from 90 d gestation to parturition, was reduced by 20.5% and 1.0% for restricted and nonrestricted heifers, respectively. Whereas nonrestricted heifers gained maternal protein (3.2 kg) from d 90 through parturition, restricted heifers lost (P less than .05) protein (-5.4 kg) and mobilized twice (P less than .05) as much fat (49 vs 25 kg). Percentage of empty body protein increased 13.1 and 9.3% in restricted and nonrestricted heifers, respectively, whereas fat decreased (P less than .05) 31.7% (restricted) and 23.5% (nonrestricted) from 90 d to parturition. Daily metabolizable energy for maintenance (MEm) was greater for large than for small mature size heifers (169 vs 158 kcal/kg.75). Efficiency of ME use for gain was greater for small than for large mature size heifers (36.5 vs 31.2%). Efficiency in early gestation (45.4%) was greater than in late gestation (29.4%) and averaged 33.8% for 90 d gestation to parturition. Maintenance ME requirements increased 25% and efficiency of ME use decreased 35% with advancing stage of gestation. Nutritional restriction of heifers reduced (P less than .05) calf birth weight (27.3 vs 30.2 kg) and decreased gestation lengths (275 vs 282 d) compared with nonrestricted heifers. This research indicates that nutritional restriction of beef heifers alters birth weight, repartitions maternal tissues and changes rate of tissue mobilization.     

Effects of feeding beef females supplemental fat during gestation on cold tolerance in newborn calves.

Effects of prepartum fat supplementation of the dam on cold tolerance of calves were determined in two studies. In Exp. 1, 22 F1, crossbred heifers gestating F2 calves received diets containing either 1.7 or 4.7% dietary fat starting at d 230+/-2d of gestation. Safflower seeds (Carthamus tinctorius) containing 37% oil with 79% linoleic acid were the supplemental fat source in isocaloric-isonitrogenous diets. Calves were separated from their dams at birth, fed pooled dairy-cow colostrum, muzzled to prevent sucking, and returned to their dams in a heated (22 degrees C) barn for 3.5 h. At 4 h of age, a jugular catheter was inserted. At 5 h of age, calves were placed in a 0 degrees C room for 140 min and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for glucose, cortisol, and cholesterol. In Exp. 2, 18 multiparous, crossbred beef cows bred to Murray Grey sires were randomly assigned to receive diets containing either 1.7 or 3.1% dietary fat starting at 235+/-2 d gestation. Safflower seeds were used as the supplemental fat source in isocaloric-isonitrogenous diets. At d 260 of gestation, premature parturition was induced in one-half of the cows from each diet group by feeding Ponderosa pine (Pinus ponderosa) needles. Experimental protocols were the same as in Exp. 1, except that cold exposure was at 9 degrees C for 200 min. Rectal temperatures were affected in Exp. 1 by time and diet x time (both P < .01) and diet x calf sex (P < .05) and in Exp. 2 by calf age (P < .05), time, and calf age x time (both P < .01). Plasma cortisol concentrations were affected by time (P < .01) and calf sex x time (P < .05) in Exp. 1 and by time ( P < .01) in Exp. 2. Cholesterol concentrations in Exp. 1 were affected by diet x time (P < .05) and in Exp. 2 by time (P < .05). Plasma glucose concentrations were affected in Exp. 1 by diet (P < .05) and in Exp. 2 by calf age, time, and calf age x time (all P < .01). We conclude from Exp. 1 that feeding heifers supplemental fat during late gestation increased glucose concentrations in the newborn calf, resulting in favorable responses in body temperature in the cold-stressed newborns. This increase in substrate availability suggests a potential positive effect on heat generation in newborns during sustained periods of cold stress. In Exp. 2, premature calves had compromised cold tolerance possibly due to impaired shivering or brown adipose tissue thermogenesis.     

The effects of late gestation nutrient restriction of dams on beef heifer intake,metabolites and hormones during an ad libitum feeding trial

This study's objective was to determine if nutrient restriction during late gestation affected beef heifer feed intake, body weight (BW) gain and endocrine regulation during a 10‐week feeding trial. During the last 100 days of gestation, control (CON) dams were fed to increase body condition score (BCS). Whereas, nutrient‐restricted dams (NR) and NR dams protein supplemented 3 days/week (NRS) were fed to decrease BCS by 1.2. After parturition, all cow‐calf pairs were moved to a common pasture and fed in excess of requirements until weaning. At 15 months of age, heifers were randomly sorted into two pens and adjusted to a commercial total mixed ration over a 2‐week period. Blood samples and BW were taken at the initiation of feeding and on a biweekly basis for the duration of the feeding trial. Feed intake was monitored for 10 weeks using a GrowSafe System. After 10 weeks, an intravenous glucose tolerance test (IVGTT) was performed on 21 randomly subsampled heifers. During the feeding trial, NR heifers consumed more feed than CON and NRS heifers. Heifers from NR dams tended to increase BW compared to NRS and CON heifers when adjusted for initial BW. Heifers from NR and NRS dams had a greater increase in BCS compared to heifers from CON dams. Plasma glucose and insulin concentrations during the feeding trial increased in NR heifers compared to the other groups beginning at 2 and 4 weeks respectively. Plasma leptin concentrations were increased in the NR and NRS heifers compared to the CON heifers beginning at week 4 of feeding. During the IVGTT at the conclusion of the feeding challenge, plasma glucose and insulin were increased in NR heifers compared to other treatment groups. These results show that nutrient restriction during late gestation alters appetite and endocrine regulation in heifer offspring.     

Bacterial survival, lymph node changes, and immunologic responses of cattle vaccinated with standard and mutant strains of Brucella abortus.

Forty-eight cattle were used in 4 experiments; 6-week-old calves in experiments 1-3 (n = 24) and 10-month-old heifers in experiment 4 (n = 24). In experiments 1-3, 7 groups of 3 calves each were inoculated SC with 5 strains of Brucella abortus: virulent strain 2308 (2 groups), vaccine strain 19 (2 groups), and mutant strains RB51. 19 delta 31K, and 19 delta SOD. Sera and lymph node tissues were examined at 2-week intervals for evidence of infection. At postinoculation (PI) week 12, 2 calves in each group were given dexamethasone for 5 days. Calves were then euthanatized and lymphoid tissue, spleen, liver, and bone marrow were examined for evidence of B abortus. Calves given strain 2308 had large numbers of bacteria in their lymph nodes, marked granulomatous lymphadenitis in the deep cortex, and loss of lymphoid cells in superficial cortical areas. In addition, they had high serum antibody titers at PI week 16. Calves given strain 19, or genetic mutants derived from strain 19, cleared bacteria from lymph nodes more rapidly, had less lymphoid destruction, and developed antibody titers that did not persist for 16 weeks. The RB51 strain (rough) was cleared most rapidly from lymphoid tissues and induced serum antibody responses only to the core of the lipopolysaccharide molecule. Treatment of calves with dexamethasone did not cause B abortus to reappear in tissues of any calves, nor did serum antibody titers increase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of porcine relaxin on induced parturition in beef heifers

Two-year-old crossbred beef heifers were used to test the effects of porcine relaxin (pRelaxin) alone, or in combination with dexamethasone, on the induction of parturition, the incidence of dystocia, and retained placentas. Effects of treatment on pelvic area, postpartum interval, milk production, colostrum quality, calf birth weight, calf vigor, and calf performance were also evaluated. On Day 275 of gestation, heifers from two fetal-sire groups were randomly assigned to one of four groups in a 2 × 2 factorial design and received: no treatment (controls, N = 19), 20 mg of dexamethasone intramuscularly (im) (n = 22), 5 mg of pRelaxin (3,000 U/mg) im (n = 19), or 20 mg of dexamethasone plus 5 mg of pRelaxin (n = 17). Length of gestation (in days) was less (P < 0.05) in heifers treated with dexamethasone (279.8 ± 1.0) than in controls (286.6 ± 0.9), but was not influenced (P > 0.05) by treatment with pRelaxin. The incidence of retained placentas in heifers treated only with dexamethasone (27.3%) was not reduced by concomitant treatment with pRelaxin (35.3%). Retained placentas were not observed in any control heifers and in only one heifer (5.2%) treated solely with pRelaxin. Ease of calving (1 = unassisted, 5 = abnormal presentation) was not influenced by treatment (P > 0.05), even though birth weights (in kilograms) of calves from heifers treated with dexamethasone (36.4 ± 0.8) were less (P < .01) than those of calves from nondexamethasone-treated heifers (39.2 ± 0.8). Dexamethasone tended to reduce (P < 0.07) calf vigor (1 = healthy and strong, 5 = dead on arrival; 1.48 ± 0.11 vs. 1.18 ± 0.11), but was not (P > 0.05) influenced by pRelaxin. The duration of the postpartum anestrous interval (73.1 ± 1.8 d across groups) and pelvic areas following treatment and parturition were not influenced (P > 0.05) by dexamethasone or pRelaxin. Although determinants of colostrum quality (P < 0.01) and quantity (P < 0.08) of milk produced were influenced by dexamethasone, adjusted 205-d weights of calves did not differ (P > 0.05) among groups. In conclusion, treatment with pRelaxin alone failed to induce parturition or, when combined with dexamethasone, to reduce the incidence of retained placentas.     

STUDIES ON DYSTOCIA AND BIRTH WEIGHT IN ANGUS HEIFERS CALVING AT TWO YEARS OF AGE

A trial is described in which groups of Angus heifers mated as yearlings were maintained on two widely differing nutritional levels during the last third of gestation in order to study effects on birth weight of calves and dystocia. The different nutritional levels produced a 5 lb difference in birth weight of calves (P < 0.01). As the calving season advanced the mean calf birth weight increased by 1.4 lb every ten days (P < 0.01). Heifers that suffered dystocia had calves 7 lb heavier and pelves 15.6 sq cm smaller than heifers calving normally (P < 0.01). Following observations on the birth process it was concluded that uncomplicated birth is complicated within two hours following the appearance of the amniotic sac.     

Protection by oral administration of brucella abortus strain 19 against an oral challenge exposure with a pathogenic strain of Brucella

Twenty heifers were vaccinated orally with Brucella abortus strain 19. These heifers and 21 control heifers were challenge exposed in midgestation with strain 2308 by the oral route. Ten of 19 pregnant control heifers aborted and 14 were culture positive. Two additional heifers were seropositive at slaughter. Strain 2308 was recovered from 4 vaccinates at slaughter; none of the vaccinates aborted. Titers after oral vaccination persisted for less than 82 days.     

Evaluation of the efficacy of a modified-live combination vaccine against bovine viral diarrhea virus types 1 and 2 challenge exposures in a one-year duration-of-immunity fetal protection study

This study demonstrated that the modified-live bovine viral diarrhea virus (BVDV) type 1 and 2 fractions of a multivalent vaccine protected pregnant heifers and their fetuses against virulent BVDV types 1 and 2 challenge exposures at 370 days after vaccination. All BVDV vaccinated heifers inoculated with either BVDV type 1 or 2 at approximately 62 to 94 days of gestation delivered fetuses or calves that were negative for BVDV by ear-notch immunohistochemistry and virus isolation and serum neutralization on a prenursing serum sample. In comparison, eight of nine and 10 of 10 fetuses or calves from non-BVDV-vaccinated heifers were considered persistently infected following exposure to BVDV type 1 and type 2, respectively.     

Isolation of Mycobacterium paratuberculosis after oral inoculation in uninfected cattle.

Feces from cows naturally infected with Mycobacterium paratuberculosis was given to 6 uninfected heifers by orogastric intubation, to determine whether ingested organisms could be passively excreted and detected by bacteriologic culture of feces (ie, false-positive result). Heifers were paired, and each pair received a different dose of feces on days 1 and 2. Fecal samples were collected from the heifers 3 times daily. Mycobacterium paratuberculosis was detected in fecal samples of all heifers within 18 hours of being given the first dose of feces. The number of colony-forming units peaked on days 3 or 4, and organisms were no longer detected by day 7. The number of colony-forming units in fecal samples from the heifers was approximately proportional to the dose given. On days 15 and 16, the experiment was repeated with feces from a second infected cow. Results were similar to those in the first experiment. All heifers remained seronegative (agar-gel immunodiffusion test and ELISA) and had negative results to the intradermal johnin test throughout the experiment. Lymph node and intestinal tissues were obtained from all 6 heifers at slaughter on day 28. Mycobacterium paratuberculosis was not isolated from mesenteric lymph nodes from the ileocecal valve region, but was isolated from ileal mucosal samples from each heifer.     

Nutritionally altering weight gain patterns of pregnant heifers and young cows changes the time that feed resources are offered without any differences in production

We hypothesized that feed resources could be deferred to a later time in the production cycle without a decrease in fertility or weight of calf produced in heifers and young cows. One-hundred and thirty-one MARC III (four breed composite: (1/4) Hereford, (1/4) Angus, (1/4) Red Poll, and (1/4) Pinzgauer) heifers were divided into three treatments: M-M-M-M (n = 46), L-H-M-M (n = 41), and L-L-L-H (n = 44). The experiment consisted of four feeding periods. Period 1 was 94 to 186 d of gestation, and heifers were fed a moderate (M) or low (L) level of feed. Period 2 was 187 d of gestation to parturition, and heifers were fed moderate, high (H), or low levels of feed. Period 3 was from parturition through 27 d of lactation, and heifers were fed moderate or low levels of feed. Period 4 was from 28 d to approximately 63 d of lactation, and heifers were fed moderate or high levels of feed. Females remained within treatments through their first parity (heifers) and second parity (cows). Feed intake of L-H-M-M and M-M-M-M treatments did not differ from each other either as heifers (P = 0.23) or as second-parity cows (P > 0.59). The L-L-L-H heifers ate less feed than L-H-M-M and M-M-M-M heifers (P < 0.001), and second-parity L-L-L-H cows ate less feed than second-parity L-H-M-M and M-M-M-M cows (P < 0.002). In the first parity, treatments did not differ in the percentage of calves weaned (P = 0.11), weight of calf weaned (P = 0.50), or percentage of cows diagnosed pregnant (P = 0.29) with a second calf. In the second parity, treatments did not differ in the percentage of calves weaned (P = 0.77), weight of calf weaned (P = 0.63), or percentage of cows expressing a corpus luteum at the start of breeding for their third calf (P = 0.21). Our findings suggest that timing nutrient availability to heifers and primiparous cows can be used to change the time that feed resources are used.     

Protection levels in vaccinated heifers with experimental vaccines Brucella abortus M1-luc and INTA 2

Brucella abortus M1-luc is a mutant strain derived from S19 vaccine strain in which most of bp26 sequence has been replaced by the luciferase coding gene. Strain I2 is a double mutant derived from M1-luc in which most of omp19 has been deleted without introduction of any genetic markers. In BALB/c mice, M1-luc presented equivalent performance to S19 regarding persistence, splenomegaly and protection against challenge. Interestingly, I2 was more attenuated than S19, with no reduction of protection against challenge. In order to evaluate the potential for vaccine use of these strains in the natural host, four groups of 15 heifers, 6-month old, were either non-vaccinated or vaccinated with S19, M1-luc or I2. To at reached 17-month old, heifers were synchronized with two doses of PGF2alpha and received natural service during 60 days with two bulls. Pregnant heifers were challenged at approximately six gestation months with virulent B. abortus S2308. Blood samples post-challenge of heifers were collected for serologic test as well as specimens of aborted fetuses and premature calves for bacterial isolation and histopathological analyses. Protection levels against abortion were 78.6% for S19, 81.8% for M1-luc and 45.5% for I2, compared to the 25% that did not abort from the non-vaccinated group. These results indicate that in bovines BP26 had no influence in protective capacity of S19, correlating with the results obtained in mice. However, contrarily to what was previously observed in mice, lack of expression of Omp19 rendered in less protection capacity of S19 in the natural host.