Reservoirs of Staphylococcus aureus in meat sheep and dairy cattle

The objective of the study was to investigate reservoirs and transmission of S. aureus in ewes and lambs in 3 meat sheep flocks. Repeated sampling of milk, teat skin, nasal- and vaginal mucous membranes was performed and samples were analysed for S. aureus. For comparison, samples were also collected from cows and young heifers in 3 dairy cattle herds. Selected isolates were compared by pulsed-field gel electrophoresis (PFGE). S. aureus was detected in 8 (1.5%) of 520 milk samples from ewes and in 38 (6.4%) of 588 milk samples from cows. From body site swabs, S. aureus was found in 394 (32.6%) of 1208 samples from sheep and in 67 (16.0%) of 420 samples from cattle. The proportion of S. aureus-positive nasal swabs from ewes and cows were 56.7% and 13.9%, respectively. From lambs, 58.2% of the nasal swabs were S. aureus-positive. In each flock, one S. aureus pulsotype predominated. Identical S. aureus pulsotypes were found in milk and from body sites. Paired S. aureus isolates from the nasal cavity of (i) ewes and their lambs, (ii) twins and (iii) from repeated swabs of individual ewes were compared by PFGE, and in the majority of cases the two isolates were identical. The results contribute new knowledge indicating frequent transmission of S. aureus between the dam and her lambs and within animals in a flock. In contrast to cattle, S. aureus is frequently present in the nose of sheep which may represent the primary reservoir of S. aureus in sheep flocks.     

Factors associated with ELISA scores for paratuberculosis in an Angus-Brahman multibreed herd of beef cattle

Cow and calf genetic and environmental factors were evaluated for their association with ELISA scores for paratuberculosis in a multibreed population of beef cattle. The ELISA scores are a measure of the presence or absence of antibodies against Mycobacterium avium subsp. paratuberculosis in bovine serum. The linear mixed-model analysis used 352 ELISA scores from 238 cows: 51 Angus (A); 34 Brahman (B); 41 (3/4 A 1/4 B); 45 (1/2 A 1/2 B); 34 (1/4 A 3/4 B); and 33 Brangus (5/8 A 3/8 B). Cows were assumed to be unrelated. Year affected (P < 0.001) ELISA scores, but age of cow did not, which was expected to be significant because of the chronic progressive nature of this disease. Important regressions on fixed effects associated with cows were 1) a positive estimate of cow B breed effect (0.59 +/- 0.24; P < 0.017), indicating an upward trend of ELISA scores toward 100% B cows; 2) a negative estimate for weight change from before calving (late November) to the date of the blood sample in May (-0.0062 +/- 0.0019 score/kg; P < 0.002), indicating that poorer maintenance of cow weights was associated with higher ELISA scores; and 3) a positive estimate for days in lactation of cow on the date of the blood sample (0.0086 +/- 0.0034 score/d; P < 0.021), indicating the production of larger amounts of antibodies against Mycobacterium avium subsp. paratuberculosis as lactation progressed. Relevant regressions on fixed effects associated with calves were 1) calf birth weight (-0.022 +/- 0.010 score/kg; P < 0.035), and 2) calf gain from birth to the date of the cow blood sample (-0.0092 +/- 0.0027 score/kg; P < 0.001). These estimates indicate that cows that produced lighter calves at birth and/or calves with slower preweaning growth tended to have greater ELISA scores. Although the sensitivity (percentage of infected animals detected) of ELISA was only 50%, these results suggest that subclinical paratuberculosis may be negatively affecting cows and their offspring. Factors identified as associated with ELISA scores could help producers with culling decisions related to paratuberculosis control and eradication in beef cattle.     

Effects of sheep and cattle alternate grazing on sheep parasitism and production

Production of sheep (nursing ewes) grazing alternately with cattle (growing weaned heifers) was compared to the production of sheep or cattle grazing alone (controls). Pasture production and sheep parasitism were also monitored. The herbage allowance was higher for the control heifers than for the alternate heifers, but the leaf to green material ratio (LGMR) was lower, and no difference on heifer growth was revealed (443 vs. 431g.d^-1, P = 0.54). The LGMR was higher for the alternate sheep (+3 points) than for the control sheep, except during the dry season, when the herbage density was lower. The effects of parasitism on the packed cell volume of alternate ewes and lambs were lower than those of control ewes and lambs. However, the infection of sheep by Cooperia sp. (better adapted to cattle) was significantly higher for the alternate sheep than for the controls, and some indication of cattle infection by Haemonchus contortus was suggested. The 70-day lamb weight was higher in the alternate grazing system than in the control (+0.76,+1.11 and+0.61kg for the dry, intermediate and rainy seasons, respectively), and the average 70-day lamb production per ewe exposed was 21.42kg in the alternate grazing system vs. 18.59kg in the control (P = 0.003).     

An evaluation of selected screening tests for bovine paratuberculosis.

The objective of this study was to evaluate the performance of the lipoarabinomannan antigen enzyme-linked immunosorbent assay (LAM-ELISA), carbohydrate antigen complement fixation (CH-CFT), and protein D antigen agar gel immunodiffusion (D-AGID) tests for bovine paratuberculosis, relative to histopathology, and to culture and isolation of Mycobacterium paratuberculosis from tissues and feces. Samples for test evaluation were collected from four sources including blood and tissues from 400 cull cows at three abattoirs in Ontario, blood and feces from a paratuberculosis survey of cattle from 120 dairy farms in Ontario, a serum bank containing samples from cattle from Ontario and Québec, and a bank of sera from cattle from Pennsylvania and the northeastern United States. The data were analyzed using receiver operator characteristic curves, estimates of relative sensitivity and specificity, and kappa statistics of agreement between tests. The LAM-ELISA performed significantly better than both the CH-CFT and the D-AGID tests. The LAM-ELISA was better at predicting fecal shedding status than tissue infection. However, the LAM-ELISA also had limitations. When interpreted as positive or negative (+/-), at a critical optical density of 0.675, its sensitivity and specificity relative to bacteriology were 49% and 87% respectively. Although the serological tests examined in this study provided some information, they did not predict well the infection status of individual animals.     

Serum haptoglobin concentration in cattle

To obtain a basal concentration of serum Haptoglobin (Hp) in cattle in Taiwan, Hp concentrations were measured from serum samples collected from 10 healthy heifers, every week for one year. The values were also compared with those collected from 15 cows diagnosed with postpartum metritis. The heifers were successfully impregnated by artificial insemination six months after the tests. Hp concentrations were also measured in the serum collected from 11 other cows within 3 weeks after parturition. The Hp assay developed in this study gave a good correlation (r=0.893)with Western blotting. The Hp concentration of 454 serum samples from the 10 heifers had a mean value of 83.6 +/- 34.1 mg/l, and there was no significant difference among individual heifers. The basal value of Hp in heifers was calculated as less than 73.6 mg/l. No significant difference in Hp concentration was observed among the 10 heifers during cold and warm seasons (19.8 +/- 2.2 degrees C vs 27.3 +/- 1.4 degrees C), or before and after pregnancy. The mean serum Hp concentration from cows suffering from postpartum reproductive disorders was 1133.5 +/- 627.1 mg/l, which was significantly greater than the serum of healthy heifers and postpartum cows (104.6 +/- 61.0 mg/l) (P<0.05). These results demonstrate that Hp concentration may be a useful indicator for cows with postpartum reproductive disorders.     

Familial associations with paratuberculosis ELISA results in Texas Longhorn cattle

The objective of this cross-sectional study was to estimate familial associations with paratuberculosis ELISA status in beef cattle. Texas Longhorn cattle (n=715) greater than 2years of age were sampled for paratuberculosis testing using ELISA and fecal culture. Diagnostic test results were indicative of substantial numbers of false-positive serological reactions consistent with environmental exposure to non-MAP Mycobacterium spp. Associations between ancestors and paratuberculosis ELISA status of offspring were assessed using conditional logistic regression. The association between ELISA status of the dam and her offspring was assessed using linear mixed-effect models. Significant associations were identified between some ancestors and offspring ELISA status. The odds of being classified as "suspect" or greater based on ELISA results were 4.6 times greater for offspring of dams with similarly increased S:P ratios. A significant positive linear association was also observed between dam and offspring log-transformed S:P ratios. Results indicate that there is familial aggregation of paratuberculosis ELISA results in beef cattle and suggest that genetic selection based on paratuberculosis ELISA status may decrease seroprevalence. However, genetic selection may have minimal effect on paratuberculosis control in herds with exposure to non-MAP Mycobacterium spp.     

Agreement between three ELISAs for Mycobacterium avium subsp. paratuberculosis in dairy cattle

During a 10-month period in 1999, 994 serum and tissue samples were collected from dairy cows at slaughter in eastern Canada. The sources of these cattle were from all four Atlantic Canadian provinces along with some cows from the state of Maine. The sera were used to assess the agreement of three commercially available ELISAs for Mycobacterium avium subsp. paratuberculosis. Two ELISAs were indirect absorbed ELISAs licensed for use in North America, the third was an indirect non-absorbed ELISA licensed for use in Europe. Overall, there was poor agreement between the three ELISAs. The highest and lowest kappa values were 0.33 and 0.18, which is fair and poor agreement, respectively. However, when only tissue culture-positive cattle were compared, the ELISAs had better agreement (kappa=0.37-0.51). The proportions of positive tests, however, were significantly different among the three ELISAs. The poor agreement among the three ELISAs is as concerning as the fact that these tests have low sensitivity. The implications are greatest when the tests are used at the cow level to make individual animal decisions, which is not the recommended method on the product labels. At the cow level, if the result obtained from one ELISA is positive, using a different ELISA in a pre-clinical animal has a high likelihood of giving a different result due to low predictive values of positive test results.     

Efficacy of an intravaginal progesterone insert and an injection of PGF2alpha for synchronizing estrus and shortening the interval to pregnancy in postpartum beef cows, peripubertal beef heifers, and dairy heifers

The objective was to test the efficacy of an intravaginal progesterone insert and injection of PGF2alpha for synchronizing estrus and shortening the interval to pregnancy in cattle. Cattle were assigned to one of three treatments before a 31-d breeding period that employed artificial insemination. Control cattle were not treated, and treated cattle were administered PGF2alpha or an intravaginal progesterone-releasing insert (CIDR) for 7 d and treated with PGF2alpha on d 6. The treatments were applied in one of three experiments that involved postpartum beef cows (Exp. 1; n = 851; 56+/-0.6 d postpartum), beef heifers (Exp. 2; n = 724; 442.5+/-2.8 d of age), and dairy heifers (Exp. 3; n = 260; 443.2+/-4.5 d of age). Luteal activity before treatment was determined for individual cattle based on blood progesterone concentrations. In Exp. 1, there was a greater incidence of estrus during the first 3 d of the breeding period in CIDR+PGF2alpha-treated cows compared with PGF2alpha-treated or control cows (15, 33, and 59% for control, PGF2alpha, and CIDR+PGF2alpha, respectively; P < 0.001). The improved estrous response led to an increase in pregnancy rate during the 3-d period (7, 22, and 36% for control, PGF2alpha, and CIDR+PGF2alpha, respectively; P < 0.001) and tended to improve pregnancy rate for the 31-d breeding period for cows treated with CIDR+PGF2alpha, (50, 55, and 58% for control, PGF2alpha, and CIDR+PGF2alpha, respectively, P = 0.10). Improvements in rates of estrus and pregnancy after CIDR+PGF2alpha, were also observed in beef heifers. Presence of luteal activity before the treatment period affected synchronization and pregnancy rates because anestrous cows (Exp. 1) or prepubertal heifers (Exp. 2) had lesser synchronization rates and pregnancy rates during the first 3 d of the breeding period as well as during the entire 31-d breeding period. The PGF2alpha, and CIDR+PGF2alpha but not the control treatments were evaluated in dairy heifers (Exp. 3). The CIDR+PGF2alpha-treated heifers had a greater incidence of estrus (84%) during the first 3 d of the breeding period compared with the PGF2alpha-treated heifers (57%), but pregnancy rates during the first 3 d or during the 31-d breeding period were not improved for CIDR+PGF2alpha compared with PGF2alpha-treated heifers. In summary, the concurrent treatment of CIDR and PGF2alpha improved synchronization rates relative to PGF2alpha alone or control. Improved estrus synchrony led to greater pregnancy rates for beef cows and beef heifers but failed to improve pregnancy rates for dairy heifers.     

Serotypes and analysis of distribution of Shiga toxin producing Escherichia coli from cattle and sheep in the lower North Island, New Zealand

AIMS: To serotype a subset of Shiga toxin-producing Escherichia coli (STEC) isolates from cattle and sheep to determine whether any corresponding serotypes have been implicated in human diarrhoeal disease, both in New Zealand and worldwide, and to examine the distribution of STEC and enteropathogenic Escherichia coli (EPEC) amongst cattle (calves, heifers and dairy) and sheep (lambs, rams and ewes), to assess whether carriage of identified bacterial genotypes may be associated with a particular age of animal. METHODS: Recto-anal mucosal swabs (RAMS) were taken from 91 calves, 24 heifers and 72 dairy cattle, and 46 lambs, 50 ewes and 36 rams, from four sites in the Manawatu and Rangitikei regions of New Zealand. Strains of E. coli selected from primary isolation plates were subjected to a multiplex polymerase chain reaction (PCR), to determine the presence of Shiga toxin genes (stx1 and stx2) and the E. coli attaching and effacing gene (eae). RESULTS: Overall, 186/319 (58.3%) animals sampled were positive for stx1, stx2, or eae isolates. More sheep (43.9%) were stx1-positive than cattle (2.7%; p = 0.036), and amongst sheep more lambs and ewes were stx1-positive than rams (p = 0.036). Amongst cattle, more calves and heifers were eae-positive than dairy cows (p = 0.030). Two or more different STEC were isolated from at least 28 (9%) animals (three cattle and 25 sheep), based on their stx/eae genotype. Enterohaemolysin genes were found in 39/51 (76%) isolates serotyped. Twenty-one different serotypes were detected, including O5:H-, O9:H51, O26:H11, O84:H-/H2 and O149:H8 from cattle, and O26:H11, O65:H-, O75:H8, O84:H-, O91:H-, O128:H2 and O174:H8 from sheep; O84:H-, O26:H11, O5:H-, O91:H- and O128:H2 serotypes have been associated with human disease. CONCLUSIONS: If nationally representative, this study confirms that cattle and sheep in New Zealand may be a major reservoir of STEC serotypes that have been recognised as causative agents of diarrhoeal disease in humans. Distribution of STEC and EPEC in cattle and sheep indicates that direct contact with, in particular, calves or their faeces, or exposure to environments cross contaminated with ruminant faeces, may represent an increased risk factor for human disease in New Zealand.     

Relation of growth hormone response to growth hormone-releasing hormone to estimation of milk production via deuterium oxide dilution in beef cattle

Current methods of estimating milk production in beef cows can be time-consuming, labor-intensive, and subject to high variability. The weigh-suckle-weigh (WSW) method requires repeated separation of offspring from their dams. Machine milking requires that animals be acclimated to the equipment prior to the estimation. The objective of Exp. 1 was to validate a deuterium oxide (D2O) dilution method of estimating milk production in cattle. In Exp. 1, Holstein calves (n = 5) averaging 29+/-2 d of age and 52.6+/-2.5 kg (+/- SE) were used as the model. Blood was collected for baseline D2O measurements followed by an injection of 300 mg D2O/kg BW. Syringes were weighed before and after the injection to gravimetrically determine the dose. Another blood sample was collected after D2O was allowed to equilibrate with body water for 2 h, and on each of the next five consecutive days, prior to feeding. Actual milk intake was measured by disappearance (i.e., amount of milk replacer offered to the calf minus the amount refused). Deuterium oxide in plasma was measured by mass spectrometry and milk intake was computed from the disappearance curve of D2O in blood plasma for each calf. Accumulated milk intake estimated by D2O dilution was highly correlated (y = 0.9x + 0.6; R2 = 0.99; P < 0.001) with actual milk intake. The objectives of Exp. 2 were to determine whether 1) D2O dilution was comparable to a standard measure of milk production in beef heifers and 2) growth hormone (GH) response to GH-releasing hormone (GHRH) in heifers at weaning is predictive of subsequent milk production. Deuterium oxide dilution and WSW were compared using 14 first-calf Angus heifers and their calves. Deuterium oxide dilution was used to estimate milk production of 40 first-calf Angus heifers that had been challenged with GHRH at weaning. Results indicate that the D2O dilution method is correlated (R2 = 0.89; P = 0.04) to the WSW estimation of milk production. Growth hormone response to GHRH in weanling heifers is positively related (R2 = 0.22; P = 0.03) to their subsequent milk production. Deuterium oxide dilution in calves offers an additional approach to the estimation of milk production of the dam in typical beef cattle production settings.     

Border Disease Virus Transmitted to Sheep and Cattle by a Persistently Infected Ewe: Epidemiology and Control

In a Swedish sheep flock comprising 202 ewes and 13 rams, a pair of twin lambs born in the spring of 1990 demonstrated signs of border disease (BD) and were persistently infected (PI) with border disease virus (BDV). Investigation showed that BDV had been introduced in the preceding tupping period 5-6 months earlier by a bought-in ewe which, on the basis of immunoperoxidase- and polymerase chain reaction techniques, was shown to be PI with BDV. Only 7 of the ewes, all of which had been in close contact with the PI ewe, seroconverted during the subsequent gestation. Apart from the PI twin lambs the losses caused by BDV were restricted to 2 barren ewes. The twin lambs, the PI ewe and lambs from the other 4 ewes that seroconverted were removed from the flock. The flock was thereafter free from an ongoing infection with BDV as shown by the absence of seroconversion.In addition, 5 heifers in late pregnancy most probably seroconverted to bovine virus diarrhoea virus (B VDV) when kept in close contact with the same PI ewe during the winter of 1989-90. When these heifers were reintroduced to the BVDV-free dairy herd from which they originated, their serum antibody titres ranged between 1:250 and 1:1250. Neither these heifers - nor their calves–caused any spread of the infection in the herd, as indicated by the absence of seroconversion in 70 cows. The present investigation shows that in the control of both BDV in sheep and BVDV in cattle, it is important to ensure that the risk of transmission of pestivirus between the 2 species is minimized.     

Insulin-like growth factor-I concentration in Holstein female cattle: variations with age, stage of lactation and growth hormone-releasing factor administration

Plasma insulin-like growth factor-I (IGF-I) concentrations were monitored in Holstein females through different periods of their growth, lactation and after acute or chronic growth hormone-releasing factor (GRF) administration. Plasma samples were radioimmunoassayed using a human IGF-I antibody after a 24 hr incubation in a HCl(.1N)-glycine(.2M) buffer (pH 2). In a first study, IGF-I concentrations were measured in Holstein females of different ages and(or) stages of lactation (n = 6 per group). The IGF-I concentrations in newborn calves (102.0 +/- 11.3 ng/ml) markedly decreased (P less than .01) in 1 mo old animals (50.2 +/- 7.1 ng/ml), then increased (P less than .01) to 137.0 +/- 5.1 and 137.4 +/- 11.0 ng/ml in 6 and 10 mo old heifers, respectively. In dairy cows, IGF-I concentrations were low 24 hr post-partum (44.7 +/- 7.6 ng/ml) and then increased (P less than .05) to remain stable throughout lactation (91.3 +/- 4.9, 92.8 +/- 12.9, 96.1 +/- 7.6, 90.7 +/- 8.8 ng/ml at 2, 3, 6 and 9 mo of lactation, respectively). There was a further increase (P less than .05) to 113.7 +/- 3.1 ng/ml during the dry period. In a second trial, blood samples were collected from lactating dairy cows every 2 hr for 24 hr following a sc injection of saline (n = 4) or human (h) GRF (1-29)NH2 (10 micrograms/kg BW, n = 4). The IGF-I peak concentration was reached on average 10 hr after the GRF injection and was higher (P less than .01) in treated cows than in control cows (135.4 vs 86.9 +/- 16.2 ng/ml). In the last trial, daily sc injections of 10 micrograms of hGRF(1-29)NH2 per kg BW to dairy cows (252 days of lactation) for 57 days, which increased milk production by 14% (2 kg/day), also increased (P less than .01) IGF-I concentration: 127.1 +/- 5.3 and 118.0 +/- 1.6 vs 90.7 +/- 4.7 and 96.0 +/- 5.0 ng/ml on days 29 and 57 of treatment for treated (n = 9) and control (n = 8) cows, respectively. Thus, the IGF-I concentration in dairy cattle varies with age and stage of lactation, and is increased by GRF administration in lactating dairy cows.     

Factors influencing the isolation of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples.

A modified procedure was used for culture of Mycobacterium paratuberculosis (Mptb) from bovine feces. Bovine fecal samples were decontaminated with NaOH, exposed to a mixture of oxalic acid and malachite green, incubated in a mixture of neomycin and amphotericin B. Decontaminated specimens were inoculated onto modified L?wenstein-Jensen medium. Specimens processed by high-speed centrifugation showed growth earlier than specimens prepared by low-speed centrifugation. However, the overall number of positive cultures at 16 weeks was not different for the 2 methods. When infected dairy herds were sampled 4 times at 6-month intervals and culture-positive cows were culled, the prevalence of infected cattle declined over time. After selective culling, the cattle left in the herds shed low numbers of Mptb, which explains why it took longer for cultures to become positive. No heifers younger than 11 months were culture positive, but heifers 13-14 months of age were more frequently culture positive than were heifers of any other age. The 16-week culture period is needed with this method to detect cattle shedding low numbers of Mptb. High-speed centrifugation of samples does not increase the efficiency of identification of animals shedding Mptb.     

Experimental selection for calving ease and postnatal growth in seven cattle populations. II. Phenotypic differences

Effects of selection for 2-yr-old heifer calving ease (reduced calving difficulty score) on phenotypic differences between select and control lines of cattle for birth, growth, yearling hip height, and pelvic measurements were estimated. The selection objective was to decrease calving difficulty score in 2-yr-old heifers, while either maintaining or increasing yearling weight. The control line objective was to maintain or increase yearling weight by the same amount as the select lines and to maintain or proportionally increase birth weight. Select and control lines were formed in 4 purebred and 3 composite populations. Selection began in 1992 and select (n = 6,926) and control (n = 2,043) line calves were born from 1993 through 1999. Selection was based on EBV calculated from a 4-trait BLUP with observations on 2-yr-old calving difficulty scores, birth weight, weaning weight, and postweaning gain. Calving difficulty was scored on a scale from 1 (unassisted) to 7 (caesarean). All birth traits in select lines differed significantly from control lines. Averaged over 7 yr, select lines calved 3.0 +/- 0.5 d earlier, had 1.8 +/- 0.5 d shorter gestations, were 2.99 +/- 0.32 kg lighter at birth, had 5.6 +/- 1.5% fewer calves assisted at birth (averaged across dam ages), and 2-yr-old heifers had 0.80 +/- 0.08 lower calving difficulty score. Select lines averaged 19.8% lower 2-yr-old heifer calving assistance, but there was no difference in calving assistance of older cows, resulting in a highly significant interaction of selection and dam classification. Preweaning ADG was increased 15 +/- 9 g/d (1.7%) in select lines. Increased preweaning gain offset decreased birth weights in select lines, resulting in weaning weights that did not differ (P = 0.71). Postweaning ADG (P = 0.16) and yearling weight (P = 0.41) also did not differ. Increased preweaning ADG in select lines was not maintained after weaning. Select line hip heights were 0.70 +/- 0.21 cm shorter when measured as yearlings. Pelvic height, width, and area of select heifers measured 25 to 74 d after yearling weights were not significantly different. The differences between select and control lines significantly changed over the course of the experiment for some traits. In the final 2 yr of the experiment, select lines had 3.9 kg lower birth weight and 1.3 cm shorter hip heights. Selection can be used effectively to reduce 2-yr-old calving difficulty and calving assistance while maintaining or increasing yearling weight.     

Effects of feed deprivation on behavioral reactivity and physiological status in Holstein cattle

The present study evaluated whether feed deprivation can increase reactivity to stressful events, such as those that can occur at slaughter. Therefore, effects of 30 h of feed deprivation on behavior, including reactions to psychological stressors, and physiological status in cattle were determined. Sixteen Holstein cows (Exp. 1) and 32 Holstein heifers (Exp. 2) were either fed (FE) or 30-h feed deprived (FD). Throughout the first day of feed deprivation and during evening feed distribution to control animals, FD heifers and cows were more active than controls (P < 0.05). In Exp. 1, during a feeding test, in response to a sudden air blast arising from the bucket from which the cow was feeding, FD cows showed a longer latency to return to feed (P = 0.0002), spent less time in the bucket air blast zone (P = 0.008) and less time motionless (P = 0.03), and tended to withdraw over a longer distance (P = 0.07) than FE cows. In Exp. 2, during a reactivity test, FD heifers spent more (P = 0.0001) time motionless in response to social isolation than FE heifers. In Exp. 2, one-half of the FE and FD heifers were subjected to an additional physical and psychological stressor just before the reactivity test by driving them for 5 min through a labyrinth. Within heifers subjected to the additional stressor, FD heifers were less accepting of being detained (P = 0.05) and stroked (P = 0.003) by a familiar stockperson in a corner of the test arena. Compared with FE animals, FD heifers and FD cows had greater plasma cortisol concentrations (P < 0.05). Feed-deprived cows also had reduced β-hydroxybutyrate concentrations (P = 0.02) compared with FE cows. Thus, in cattle, FD influenced some of the classical indicators of energy metabolism and exacerbated reactivity to sudden events. In addition, when additional stressors were applied, FD cattle were more reluctant to accept handling. Results indicate that a multifactorial origin of stressors during the slaughter period may synergistically increase psychological stress of cattle.     

Allergic and serologic diagnosis of paratuberculosis in cattle

The use of allergenodiagnosis in cattle herds where paratuberculosis is to be eradicated depends on the allergenogenic properties of the causative agent of this disease. An evaluation of the reliability of an intradermal allergic test performed to diagnose paratuberculosis in cattle in the given area revealed that this method is important especially if the result of this test is explicitly negative. A cultivation proof of the causative agent of paratuberculosis is of greatest importance among bacteriological methods for infection diagnosis. The agreement of the bacteriological and serological examination which reached 80% in young cattle pointed out the high specificity of an indirect haemagglutination test. In cows, unlike calves and heifers, the specificity of diagnostic tests was considerably lower. It is recommended to use an elimination method mainly in young cattle: regular examinations of calves from their age of three months will help to form a group of heifers free of infection, observing the basic infection controlling measures and providing conventient diet.     

Detection of antibodies against Anaplasma marginale in milk using a recombinant MSP5 indirect ELISA

An indirect enzyme linked immunosorbent assay (iELISA) for diagnosis of anaplasmosis using undiluted individual milk samples from dairy cows was developed. The recombinant 19 kDa major surface protein 5 (rMSP5) of Anaplasma marginale was used as antigen. A monoclonal antibody against bovine IgG1 conjugated with peroxidase and the chromogen 3,5,3',5'-tetramethylbenzidine were used in the test. Strong and weak, positive and negative milk samples were set up as reference controls. Results were expressed as percentage of positivity (PP) contrasting with the strongest positive control. The test was evaluated in two groups (G1 and G2) of lactating dairy cows from herds located in A. marginale non-endemic areas of Argentina. The infection status of both groups, G1 (n=128) sampled after anaplasmosis outbreak, and G2 (n=216) free of anaplasmosis was established by polymerase chain reaction (PCR). Serum samples of cows from G1 and G2 were analyzed by card agglutination test (CAT) and competitive ELISA (cELISA), while the novel iELISA was evaluated in their corresponding milk samples. At a cutoff of 42 PP, the ELISA has 98% sensitivity and 95% specificity. A significant difference (P<0.0001) was found between the mean PP value of negative samples from G1 (17.4+/-14.9), and G2 (8.6+/-7.1). The agreement and kappa (kappa) value between iELISA and PCR was 96%, kappa=0.919; between iELISA and CAT was 97%, kappa=0.880; and between iELISA and cELISA was 97%, kappa=0.899. These results strongly support the usefulness of iELISA to detect A. marginale antibodies in milk. Additional studies are necessary to define the ability of the milk iELISA to detect not only acutely infected, but also carrier cattle.     

Multivariate regression analysis of epidural pressure in cattle

OBJECTIVE: To evaluate the effects of growth, maturity, and pregnancy on epidural pressure in cattle. ANIMALS: 50 healthy Holstein cattle (18 heifers, 23 lactating cows, and 9 pregnant nonlactating cows). PROCEDURE: Each of the cattle was restrained in a standing position. Height of the second lumbar vertebra's transverse process (2LTP) and humeral tuberosity (HT) on the right side as well as abdominal girth (AG) were measured in each animal, and body condition score (BCS) was ascertained. Skin caudal to the first lumbar spinous process was aseptically prepared, and anesthetic was injected. After inserting a 16-gauge 120-mm Tuohy needle in the ligamentum flavum, a calibrated pressure transducer was connected to the needle. Then, the needle was introduced into the epidural space, and epidural pressure was recorded. RESULTS: Mean +/- SD residual epidural pressure of heifers (-9.3+/-3.3 mm Hg) was significantly higher than that of lactating (-174+/-5.5 mm Hg) or nonlactating (-14.5+/-2.4 mm Hg) cows. Stepwise regression of 5 variables revealed that only the difference in height between 2LTP and HT (2LTP - HT) in heifers and only BCS in lactating cows were significantly correlated with residual epidural pressure. For all cattle, the optimal equation (R2 = 0.47) describing the relationship was y = -12.7 + 6.3x, - 0.4x2 - 0.1x3, where y is epidural pressure, x1 is BCS, x2 is 2LTP - HT, and x3 is age. CONCLUSIONS AND CLINICAL RELEVANCE: Negative epidural pressure was detected in standing cattle. Growth, maturity, and pregnancy affect epidural pressure in cattle.     

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.     

Performance of a Johne's disease enzyme-linked immunosorbent assay adapted for milk samples from goats.

Antibody detection-based tests for paratuberculosis offer speed and economy, 2 diagnostic test attributes important to animal industries with narrow profit margins. Application of such tests to individual milk samples instead of serum samples can further improve testing efficiency and decrease testing cost. Accuracy of a commercial bovine paratuberculosis enzyme-linked immunosorbent assay (ELISA) adapted for use on goat serum and milk samples was determined. Fecal, blood, and milk samples were collected from 159 goats belonging to 2 Wisconsin goat herds with a prior history of paratuberculosis and 1 herd of 50 goats from a paratuberculosis-free Wisconsin herd. Fecal samples were cultured using the modified BACTEC 12B media. Sera were tested according to the manufacturer's instructions for bovine samples. Milk samples were centrifuged and mixed with the ELISA kit's Mycobacterium phlei-containing diluent at a ratio of 1:2. Using fecal culture as the "gold standard," the sensitivity of the ELISA on goat serum was 64% and the sensitivity of the ELISA on goat milk was 48%. The milk ELISA had higher agreement with fecal culture results (kappa = 0.525) than the serum ELISA (kappa = 0.425). ELISA specificity was 100% on both serum and milk. Regression analysis also showed good correlation between serum and milk S/P values (r2 = 0.67). Although less sensitive, the ELISA on goat milk samples appears to offer a useful, low-cost alternative for detection of goats with paratuberculosis that have progressed to the stage of shedding M. paratuberculosis in their feces.