Use of an immortalized bovine mammary epithelial cell line (MAC-T) to measure the mitogenic activity of extracts from heifer mammary tissue: effects of nutrition and ovariectomy

The objectives of the experiment were (1) to determine whether MAC-T cells would accurately mimic the previously observed proliferative responses of primary mammary epithelial cells (MEC) to mammary tissue extracts from high and low-fed heifers and (2) to determine whether mammary tissue extracts from ovariectomized (OVX) heifers would have lower mitogenic activity than intact controls. Addition of mammary tissue extracts to cell culture media of MAC-T cells plated on plastic or collagen-coated plastic to a range of concentrations between 1 and 8% resulted in dose-dependent increases in cell proliferation. Furthermore, mammary tissue extracts from low-fed prepubertal heifers aged 9 months, stimulated significantly more proliferation of MAC-T cells, as measured by 3H-thymidine incorporation into DNA than mammary tissue extracts from high-fed heifers (40.6 cpm x 10(3) per well versus 21.9+/-1.8 cpm x 10(3) per well). These observations suggested that MAC-T cells would be a suitable alternative to primary MECs for measuring the mitogenic activity of mammary tissue extracts. Conversely, no difference was observed in the mitogenic activity of mammary tissue extracts from OVX or control heifers. Possibly, MAC-T cells provide a good model for nutrition- but not ovarian-induced changes in mammary growth. Alternatively, that reduction of in vivo mammary development following OVX did not result in reduced mitogenic activity of the mammary tissue extracts emphasizes that heifer mammary development is the result of complex interactions between local growth factors and systemic hormones.     

An evaluation of bias in estimated breeding values for weaning weight in Angus beef cattle field records. II. Estimates of bias due to genetic trend

Field records (53,989) from the American Angus Association were used to evaluate the impact of bias associated with genetic trend in estimated breeding values for weaning weight. Annual estimates of breeding values were calculated using the reported weaning weight ratios and ratios adjusted for estimated genetic trend. Genetic gains were assumed to be consistently linear. Bias was estimated as the difference between the two estimates, and herd averages across years for dam's bias ranged from .38 to 3.86 ratio units. The bias was a function of the magnitude of the estimated trend and the generation interval. The impact of this bias was small in young cows, but increased rapidly after diet fourth or fifth calves. When annual genetic trend was less than .75 ratio units, the loss in selection response (based upon selecting the top 10 and 50% of males and females, respectively) did not exceed 2.5%. In herds with trends between .75 and 1.1 units/yr, the loss was 6 to 8%. In the five herds where estimated trend exceeded 1.1 units/yr, the loss ranged from 10.8 to 25.2%).     

Effects of tetracycline and zinc on selection of methicillin-resistant Staphylococcus aureus (MRSA) sequence type 398 in pigs

An in vivo experiment was conducted to evaluate the effects of tetracycline and zinc on pig colonization and transmission of methicillin-resistant Staphylococcus aureus (MRSA) sequence type (ST) 398. Eight piglets naturally colonized with MRSA ST398 and 8 MRSA-negative piglets of the same age and breed were assigned to three groups treated with tetracycline and zinc (Group 1), zinc (Group 2) or tetracycline alone (Group 3) and one non-treated group (Group 4), each containing two MRSA-positive and two MRSA-negative animals. Two additional non-treated control groups composed of only MRSA-positive (Group 5) and MRSA-negative (Group 6) animals were used to check for stability of MRSA carriage status. Nasal swabs and environmental wipes were collected on Days 0, 7, 14, and 21, and the occurrence of MRSA in each sample was quantified by bacteriological counts on Brilliance? MRSA agar. Significantly higher nasal MRSA counts were observed in the zinc-treated (p=0.015) and tetracycline-treated (p=0.008) animals compared to the non-treated animals. Environmental MRSA counts appeared to increase over time in Groups 1 and 2 but such an increase was not statistically significant. MRSA-negative animals housed with MRSA-positive animals became positive in all groups, whereas the carriage status of the animals in Groups 5 and 6 did not change. This study demonstrates that feed supplemented with tetracycline or zinc increases the numbers of MRSA ST398 in the nasal cavity of pigs. Transmission of MRSA from positive to negative animals housed within the same pen was not influenced by exposure to these agents.     

Suckling behavior of calves with dams varying in milk production

Two experiments were performed to test the hypothesis that suckling behavior of calves with similar growth potential varies depending on cows' level of estimated milk production and stage of lactation. Eleven mature cows, which varied in estimated 205-d milk production (996 to 2354 kg/205 d), nursing heifer calves of similar growth potential were used in Exp. 1. Suckling behavior of calves was observed for two 24-h periods at three stages of lactation (average of 52, 104 and 167 d postpartum). Suckling frequency (suckling bouts/24 h) declined as milk production increased at 52 d of lactation (-.00382 bouts/kg milk) but was unrelated to milk production at later stages. Duration of suckling (minutes/suckling bout) increased with estimated level of milk production at all stages of lactation (means = .001556 min/kg milk). Total time suckling tended to increase as estimated level of milk production decreased at 52 d of lactation, but this component of suckling behavior was unaffected by milk level at later stages. Suckling frequency declined from 8.6 bouts/24 h at 52 d of lactation to 4.5 bouts/24 h at 167 d of lactation when averaged across all cows. Total minutes nursed/24 h declined in a similar manner (64 min/24 h to 44 min/24 h) between 52 and 167 d of lactation. Duration of each suckling bout did not change with stage. In the second experiment the relationship of suckling behavior to estimated milk production was evaluated at four early stages (average of 17, 38, 59 and 80 d postpartum) of lactation using 20 mature cows.(ABSTRACT TRUNCATED AT 250 WORDS)     

Binding of bovine IgG2a and IgG2b allotypes to protein A, protein G, and Haemophilus somnus IgBPs.

Immunoglobulin binding proteins (IgBPs) are thought to be virulence factors which enable pathogens to evade the host's immune response. Since bovine IgG2 is important in protection against pyogenic infections, the binding characteristics of Staphylococcus aureus protein A (PrA), streptococcal protein G (PrG), or Haemophilus somnus high molecular weight IgBPs to the two bovine IgG2 allotypes were examined. For PrA or PrG binding of IgG2, guinea pig red blood cells coated with specific IgG2a or IgG2b antibodies were used in a competitive binding inhibition assay with unlabeled and horseradish peroxidase-labeled PrA or PrG. To determine which sizes of H. somnus. IgBPs bind to the two IgG2 allotypes, immunoblots with H. somnus culture supernatant were probed with anti-DNP IgG2a and IgG2b. This detects only Fc binding because anti-DNP does not cross-react with H. somnus antigens. Both IgG2 allotypes bound equally well to PrA and PrG. However, IgG2b but not IgG2a bound to H. somnus high molecular weight IgBPs. The lack of differential binding of bovine IgG2 allotypes to PrA and PrG means that these IgBPs can be considered to be unbiased reagents in assays for detection of bovine IgG2 or for immunoaffinity purification. The differential binding of H. somnus IgBPs to the IgG2 allotypes indicates that animals having one allotype may be more resistant to H. somnus infection than animals having the other allotype.     

Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae

The reference strains of the 12 serotypes of Actinobacillus pleuropneumoniae express one or two of three different RTX exotoxins designated Apx I, Apx II and Apx III. The toxins are important virulence factors. In the present study, ELISAs with purified Apx I, Apx II and Apx III, respectively, as antigen were evaluated as candidates for serological diagnosis of Actinobacillus pleuropneumoniae infection in pigs. The pigs were inoculated with biotype 1, serotypes 1-12, and biotype 2, serotype 14, respectively. A strong humoral antibody response was seen to all the three antigens in most pigs irrespective of the serotype used for inoculation. However, titers to the exotoxins secreted by the serotype used for inoculation were generally highest. The results show that toxin proteins of Actinobacillus pleuropneumoniae are antigenically related and that a correlation between serotype and secretion of exotoxin is not revealed serologically in the ELISA test.     

Fatty acid patterns of dog erythrocyte membranes after feeding of a fish-oil based DHA-rich supplement with a base diet low in n-3 fatty acids versus a diet containing added n-3 fatty acids

Background

In dogs, increasing the tissue n-3 fatty acid (FA) content is associated with potential benefit in some medical conditions, e.g. atopic dermatitis, cancer or heart disease. Therefore effectively and conveniently increasing tissue n-3 FA levels in dogs is of interest. Incorporation of dietary n-3 FA into cell membranes may be studied by FA analysis of erythrocyte membranes (EM), because of the correlation of its FA composition with the FA composition of other cells. Aim of the study was to determine whether an n-3 FA additive added to a control diet is as effective in increasing EM n-3 FA content as feeding an n-3 FA enriched diet. Furthermore the time course of the incorporation of dietary n-3 FA into canine EM was investigated.

Methods

Thirty dogs were randomly divided into three dietary groups with ten dogs per group. CONT got a dry dog food diet which did not contain EPA or DHA. FO got a dry dog food diet with a high EPA and DHA content. ADD got the CONT diet combined with an n-3 FA additive rich in DHA and EPA. After a feeding period of 12 weeks the additive was discontinued in ADD and these dogs were fed CONT diet for another four weeks to observe washout effects. Erythrocyte lipids were extracted from venous blood samples and their FA composition was determined by gas chromatography. The Mann-Whitney-U-test was used to detect significant differences between the different groups and time points.

Results

After one week the proportions of n-3 FA, DHA and EPA were already significantly increased in ADD and FO, apparently reaching a plateau within eight weeks. In our study DHA and not EPA was preferably incorporated into the EM. After discontinuing the administration of the additive in ADD, the n-3 FA values declined slowly without reaching baseline levels within four weeks.

Conclusions

In dogs, an increase of dietary n-3 FA content leads to a rapid inclusion of n-3 FA into EM, regardless of whether the n-3 FA are offered as an enriched diet or as a normal diet supplemented with an n-3 FA additive.     

Short-duration exercise and confinement alters bone mineral content and shape in weanling horses

The hypothesis that short-duration exercise may ameliorate the decrease in bone mass observed with confinement was investigated with 18 quarter horses (nine colts and nine fillies) weaned at 4 mo of age and placed into box stalls. After a 5-wk adjustment period, individuals were grouped by age and weight, and then divided randomly into three treatment groups: 1) group housed; 2) confined with no exercise; and 3) confined with exercise. The confined and exercised groups were housed in 3.7 m x 3.7 m box stalls for the 56-d duration of the trial. The exercised group was sprinted 82 m/d, 5 d/wk, in a fenced grass alleyway. The weanlings were led down an alleyway, turned loose in a small pen, and then released and allowed to run back down the alley. The group horses were housed together in a 992-m2 drylot with free access to exercise. On d 0, 28, and 56, dorsopalmar and lateromedial radiographs of the left third metacarpal bone were taken to estimate changes in bone mineral content and cortical widths. Mean values of medial, lateral, and total radiographic bone aluminum equivalence increased over time (P < 0.05), whereas dorsal and palmar radiographic bone aluminum equivalence did not change significantly. Dorsal, medial, and total radiographic bone aluminum equivalence tended (P = 0.09) to differ by a treatment x day interaction, with values increasing over time only in the exercised group. Normalized medial and total radiographic bone aluminum equivalence tended (P < 0.1) to differ (P < 0.01) with treatment, with exercised horses having greater bone aluminum equivalence than confined horses. Dorsopalmar cortical width in exercised horses was greater than on d 56 (treatment x day; P = 0.07). The dorsopalmar medullary cavity decreased in exercised vs. group-housed horses (P = 0.027), whereas dorsal and medial cortical width tended to increase only in the exercised horses (treatment x day; P < 0.01). This study indicated that a short-duration exercise protocol might be effective in improving bone mass and therefore skeletal strength in horses.