Pulsed-field gel electrophoresis and distribution of the genes zag and fnz in isolates of Streptococcus equi

Streptococcus equi subsp. equi and subsp. zooepidemicus are important pathogens of the equine respiratory tract. Isolates of both subspecies were examined by pulsed-field gel electrophoresis (PFGE). With the exception of eight isolates, a unique band pattern was displayed for each of the 48 subsp. zooepidemicus isolates tested. A method to distinguish isolates of the genetically very homogeneous subsp. equi has hitherto not been available, although several methods have been tested. By the use of PFGE, 50 isolates of subsp. equi could be divided into eleven groups, each with a unique pulsotype. In addition, the recently characterised genes encoding the cell-wall proteins ZAG and FNZ of S. equi subsp. zooepidemicus strain ZV were shown by Southern blots to be present in all 98 tested isolates, including the type strains of the two subspecies. Binding assays showed that the expression of the two genes clearly differentiate between the two subspecies.     

Influence of the number of days spent training in an abattoir with access to live cows on the efficiency of do-it-yourself artificial insemination

Data relating to 35,389 inseminations carried out in the first three years after inseminators were trained were collected by postal questionnaire from 94 do-it-yourself (DIY) artificial inseminators in the UK. The mean calving rate from 14,528 inseminations they carried out on 92 farms in the first year after they were trained was 59.4 per cent (range 5 to 92 per cent). In the second year the mean calving rate from 11,515 inseminations by 64 inseminators was 62.3 per cent (range 33 to 88 per cent), and in the third year the mean calving rate from 9346 inseminations by 49 inseminators was 64.6 per cent (range 41 to 92 per cent). There was an increase of 5.2 per cent in the mean calving rate over the three years with an overall mean calving rate of 61.5 per cent. Each trainee carried out an average of 376 inseminations (range 20 to 800) during the three years. Thirteen instructors were responsible for the tuition and the average time the trainees spent in an abattoir with access to live cows was 2.88 days, with a range from none to five days. For the trainees who spent up to three days training in an abattoir there was an increase of 5.9 per cent in the calving rate they achieved in their first year for every day they spent training with access to live cows (P < 0.005), but there were no significant effects on the outcome of their inseminations in their second or third years.     

Metabolic changes concerning the effect of castration on some blood constituents in male rabbits

This experiment was conducted to determine the effects of castration on some metabolic alterations concerning proteins, lipids and minerals metabolism in both immature and mature rabbits. Twenty male New Zealand White rabbits (10 immature and 10 mature) were used. Blood samples were collected from the immature and mature intact animals (control non castrated). Moreover, the castrated animal groups (immature and mature) were sampled at 2, 4, 6 and 8 weeks after castration. The obtained results revealed that, castration caused a significant decrease in serum total protein, albumin, alpha 1-globulins and alpha 2-globulins concentrations. Meanwhile, gamma-globulins level was decreased temporarily at 2 weeks after castration in mature rabbits. beta-globulins level revealed non significant decrease at 2 weeks after castration in immature rabbits followed by a non significant increase. In mature rabbits beta-globulins level non significantly increased. A significant decrease in serum total nucleic acid concentrations was observed after castration throughout the experimental periods, whereas the value of serum uric acid level markedly increased after castration. Serum lipids concentrations (total lipids, total cholesterol, phospholipids and non esterified fatty acids) were increased after castration. The value of serum trace elements including copper, iron, zinc and manganese concentrations showed a significant decrease after castration. The decrease in serum copper and zinc levels were markedly observed in mature castrated rabbits. Also, serum sodium and potassium concentrations were decreased after castration. Testosterone propionate administration in mature castrated rabbits restored most of the investigated serum blood parameters to the levels of intact non castrated mature rabbit. It could be concluded that castration (deficiency of testosterone) induced marked decrease in protein synthesis and increased protein degradation, but serum lipids markedly elevated after castration in both immature and mature animals. Also, castration caused a marked decrease in some serum minerals and trace elements.     

First isolation of Sarcocystis hominis from cattle in Japan.

Sarcocystis hominis was first isolated from slaughtered cattle raised in Japan. Cysts were 1,220-4,460 x 80-384 microns in size and their wall was 3 to 6 microns thick and appeared radially striated in the histopathological sections because of the presence of palisade-like villar protrusions on the surface. The protrusions were 3.1-4.3 x 0.7-1.1 microns in size and had many microtubules in the core. Two cynomolgus monkeys, Macaca fascicularis, fed with the Sarcocystis cysts began to pass sporocysts, which measured a size of 14.3-15 x 9.5-10 microns, in the feces 10 days after ingestion.     

Organ culture of chicken bursa as a model to study the pathogenicity of infectious bursal disease virus isolates.

In vitro study with chicken bursal organ culture was attempted to assess the pathogenicity of locally isolated infectious bursal disease virus (IBDV) initially isolated from the bursa of naturally infected birds. In bursal organ culture, lymphoblastic transformation was noticed as early as 24 hr postinoculation and reached maximum at 72 hr postinoculation. The other microscopic changes were increased number of macrophages and formation of plasma cells. The IBDV antigen was detected 24 hr onward by coagglutination test with antibody coated Staphylococcus aureus strain Cowan I. On the basis of lesion score, the three isolates of IBDV (A, B, and C) were graded as virulent (B isolate) and moderately virulent (A and C isolates). A similar pattern of pathogenicity was also observed in the in vivo pathogenicity studies in chicken based on bursa: body weight ratio and histopathologic lesion score. The bursal organ culture thus provides a useful experimental model to differentiate the IBDV isolates on the basis of their virulence.     

Influence of pig substitution on milk yield, litter weights, and milk composition of machine milked sows.

This study was conducted to 1) determine milk yield of sows that were machine milked up to four times daily; 2) determine the effect of pig substitution on milk yield; 3) assess litter weight changes for sows that are milked; and 4) determine milk composition. Eight sows were milked four times daily to d 51 postpartum. Sows either maintained their own litter or had a week-old replacement litter to replace 25-d-old pigs. Individual gland milk yields were obtained on random days throughout lactation, and different diameter and weighted teat cups were rotated so that all glands received all combinations. Composite milk samples were analyzed for fat, protein, and somatic cells. Milk yields peaked at about 19 d postpartum and declined to 45 d postpartum in sows with their own litter, whereas milk yields peaked earlier and had a more dramatic decline after fostering of a younger litter. Litter weights were 17.1 +/- 1.0 kg at farrowing with 13.6 +/- .6 pigs born alive. Final litter weights were 34.4 +/- 11.7 kg for sows with replacement litters and 74.4 +/- 13.5 kg for sows with their own litters, and numbers of pigs weaned were 6.5 +/- 1.3 and 9.7 +/- 1.5, respectively. Milk fat was influenced by route of oxytocin administration (6.53 +/- .12 for i.v. vs 7.21 +/- .19% for i.m. administration; P < .05). Milk fat percentage was highest on d 2 and declined to 13 d postpartum. Milk protein was influenced by time of day of milking (lowest at the fourth milking, 5.57 +/- .11%) and followed a pattern similar to that for milk fat. Milk protein was affected in a linear manner by milk yield, with highest protein associated with lowest milk yields. Somatic cells in milk were influenced by litter replacement (P < .05) and oxytocin administration (P < .01). There was a linear increase in somatic cells from about 8 x 10(6) cells/mL milk at d 2 to more than 12 x 10(6) cells/mL milk at d 51 postpartum. These results show that pig replacement affects the amount of milk obtained. Moreover, milk composition changes throughout lactation. However, milk removal from sows has a severe impact on litter weight gains, and in systems where sow's milk is needed for commercial purposes, pig supplementation is necessary.     

Effect of the ratio between essential and nonessential amino acids in the diet on utilization of nitrogen and amino acids by growing pigs.

In 36 growing pigs (30 to 60 kg), N balance and amino acid (AA) composition of weight gain were measured to evaluate the interactive effect of the ratio between N from essential amino acids (EAA(N)) to nonessential amino acids (NEAA(N)) and total N level (T(N)) in the diet on N retention and utilization of N, EAA(N), NEAA(N), and AA. Nine diets composed from ordinary feedstuffs and supplemented with crystalline AA were used (three EAA(N):NEAA(N) ratios of 38:62, 50:50, and 62:38 at three T(N) levels of 18.8, 22.9, and 30.0 g/kg). Pigs were fed restrictedly, at a level of 2.8 x energy for maintenance. In all diets, EAA (including arginine) supply was according to or slightly above the recommended ratios to lysine. Measurements were done in four blocks of nine pigs each. In a concomitant slaughter experiment, the AA composition of deposited body protein was determined to estimate AA utilization. The effects of T(N) and EAA(N):NEAA(N) and their interaction for N retention and utilization were significant. Nitrogen retention increased with higher T(N) in the diet. Increasing EAA(N):NEAA(N) from 38:62 to 50:50 improved N retention only at the two lower T(N) levels. Increasing EAA(N): NEAA(N) above 50:50 failed to improve N retention significantly at any of the three T(N) levels. Lowering T(N) improved the utilization of total and digested N and of EAA(N) and NEAA(N). The increase in EAA(N): NEAA(N) consistently resulted in a lower utilization of EAA(N), but this was compensated by a higher utilization of NEAA(N). The utilization of T(N) was improved by increasing EAA(N):NEAA(N) from 38:62 to 50:50 at the two lower T(N) levels and was relatively unaffected by EAA(N):NEAA(N) at the highest T(N). However, a lower utilization of N was observed at a ratio of 62:38 at a T(N) level of 22.9 g/kg. The effects were similar for utilization of individual EAA and NEAA. Utilization of alanine, aspartic acid, and glycine was close to or >100% at the highest EAA(N):NEAA(N), which was expected because all of these AA are synthesized in pigs. Also, the utilization of arginine was >100% in most of the treatments, which confirms the semiessential character of this AA for maintenance. We concluded that the required ratio of EAA(N):NEAA(N) for optimal N retention and utilization is approximately 50:50. The EAA(N):NEAA(N) is more important at lower dietary protein levels. This study indicates that EAA(N): NEAA(N) can be increased up to 70:30 without lowering the utilization of N. Thus, deaminated EAA(N) was efficiently utilized for the synthesis of NEAA(N).     

Case life of seven retail products from beef cattle supplemented with alpha-tocopheryl acetate.

Packages (n = 660) of clod, strip loin, tenderloin, bottom round, eye-of-round, and top round steaks, as well as ground beef, from beef cattle fed diets supplemented with vitamin E (500 IU/d for 100 d preharvest) were compared with packages of products derived from carcasses of beef cattle fed control diets to determine the duration of acceptable muscle color during simulated retail display. In addition, the effect of storage before steak fabrication, simulating distribution time, on the acceptability of each treatment and each cut during retail display was investigated. Dietary supplementation of animals with vitamin E increased muscle alpha-tocopherol concentrations (P < .05) in all cuts. The acceptable retail display time of ground beef from vitamin E-supplemented cattle was longer (P < .05) compared to ground beef from unsupplemented cattle by 10.2, 15.6, and 17.6 h following 7, 14, and 21 d of storage in chub packages prior to final grinding and retail display, respectively. However, length of storage in a vacuum package before retail display did not have a consistent effect on retail case life of steaks. Product drip loss during retail display was not affected by storage time or vitamin E treatment. The trained panel color score during retail display of high vitamin E strip loin, bottom round, inside round, and eye-of-round steaks remained acceptable longer (P < .05) by 11.5, 25.2, 8.4, and 29.4 h, respectively, than that of control steaks. The results demonstrated that vitamin E supplementation of cattle feed can be used as a management tool to reduce the economic losses associated with beef muscle color deterioration during retail display of products.     

In situ disappearance of neutral detergent insoluble nitrogen from alfalfa and eastern gamagrass at three maturities.

In situ digestion kinetics of neutral detergent insoluble nitrogen (NDIN) from alfalfa (Medicago sativa L.) harvested at one-tenth bloom and eastern gamagrass (Tripsacum dactyloides L.) harvested at the boot (GGB), anthesis (GGA), and physiological maturity (GGM) stages of growth were determined with nonlinear regression techniques. Whole-plant tissue and associated leaf and stem fractions were incubated in the ventral rumen simultaneously. On a wholeplant basis, potential extents of degradation were particularly high (> or =904 g/kg NDIN) for GGB and GGA, relative to those of GGM and alfalfa (772 and 658 g/kg NDIN, respectively). For all plant parts, degradation rates of NDIN were faster (P<.05) for alfalfa than for all gamagrass forages. Degradation rate of NDIN did not differ (P>.05) across maturities for any gamagrass tissue type. These results indicate 1) that phenological development and lignification do not limit the rate of NDIN degradation in gamagrass forages but do markedly limit the potential extent of NDIN availability and 2) that most of the NDIN in these forages is potentially available in the rumen and can contribute to the ruminal N supply. Our secondary objective was to compare estimates of N escaping ruminal degradation that were determined on the basis of NDIN degradation kinetics (NDIN method) with those determined traditionally, on the basis of total residual N. The NDIN method mathematically eliminates all neutral detergent soluble N from consideration as part of the pool of dietary N potentially escaping the rumen intact. Estimates of rumen escape nitrogen determined on the basis of degradation rates of NDIN were consistently less than corresponding estimates that were determined on the basis of total residual N. When ruminal escape N that was determined with the NDIN method was regressed on corresponding estimates with the total residual N method, the slopes of the regression lines were .53 and .66 for assumed passage rates of .02 and .06 h(-1), respectively. For the forages evaluated in this study, these results indicate that neutral detergent soluble N may make important contributions to the pool of N escaping ruminal degradation.