In situ neutral detergent insoluble nitrogen as a method for measuring forage protein degradability.

A method of estimating the undegraded intake protein (UIP) concentration of forages was developed and validated with a series of in situ experiments. The hypothesis was that UIP calculated from in situ neutral detergent insoluble N (NDIN) is equal to total in situ N minus the microbial N that is estimated from purines (MN). The in situ disappearance rates of total in situ N (TN), MN, and NDIN were measured for six hay samples and two range masticate samples. Hypothetical rates of passage (2 or 5%/h) were used to calculate UIP (% of DM) for each N pool. Estimates of UIP from TN were higher (P = .0001) than those from either MN or NDIN, and MN estimates of UIP were similar (P = .48) to NDIN estimates. A low-N fiber source (solka floc) was incubated in situ for 8 h. Analysis of the residue detected purines before, but not after, neutral detergent extraction. Several in situ incubation (i.e., Dacron bag size and number of Dacron bags in a mesh bag) and neutral detergent extraction conditions were tested. None of the factors tested affected in situ NDIN disappearance (P > .05). The hypothesis that NDIN is completely digestible in the rumen was tested. Estimates of the extent of NDIN digestion were made using 96-h in situ incubations, and UIP was recalculated for the test samples. Mean in situ UIP concentration decreased upon recalculation (P = .05). In situ NDIN provides estimates of forage UIP that are equal to estimates from MN. Forage UIP estimates are less when extent of N degradation is estimated and included in the calculation.     

Effects of alpha-tocopheryl acetate supplementation and salt addition on the oxidative stability (TBARS) and warmed-over flavour (WOF) of cooked turkey meat.

1. Day-old turkey poults (n = 14) were randomly divided into 2 groups (n = 7) and fed diets containing 20 (E20) and 600 (E600) mg all-rac-alpha-tocopheryl acetate/kg food for 21 weeks prior to slaughter. Following slaughter, breast and leg meat was removed and 4 batches of patties were produced from each. Two of the batches were formed from E20 meat (E20) and E20 plus 1% salt (E20S). Two similar batches were formed from E600 meat (E600) and E600 plus 1% salt (E600S). 2. Patties were fried, cooled and overwrapped with high oxygen-permeable film. Overwrapped patties were displayed in a 4 microC cabinet under fluorescent light (616 lux). Lipid oxidation (TBARS numbers) was determined on d 0, 2, 4, 6, 8 and 10, while taste panels to assess warmed-over flavour (WOF) were carried out on d 0, 2 and 4 of refrigerated (4 degrees C) display. 3. In the case of both leg and breast meat, E600 patties were the least susceptible of the 4 treatment batches to lipid oxidation. Salt had the effect of promoting lipid oxidation, with E20S and E600S patties having higher TBARS numbers than the corresponding patty batches where salt was absent. 4. Taste panel results showed that leg and breast patties formed from the meat of turkeys given alpha-tocopheryl acetate enriched diets developed significantly (P < 0.05) less WOF than those formed from control turkey meat on d 2 and 4 of refrigerated (4 degrees C) display. Patties containing 1% salt generally exhibited a greater degree of WOF than patties without salt. 5. A linear relationship was observed between TBARS numbers and WOF percentages for all batches of leg and breast patties.     

Heligmosomoides polygyrus and Trypanosoma congolense infections in mice: effect of immunisation by abbreviated larval infection.

Concurrent African trypanosome and gastrointestinal helminth infections are prevalent in sub-humid savannah where they are endemic. However, acquired resistance in animals varies with their responder status and exposure. As a guide to study in the definitive hosts, the effects of Trypanosoma congolense infection on the development and maintenance of homologous Heligmosomoides polygyrus resistance were investigated in outbred TO mice. These mice were immunised by abbreviation of larval infection. Immune or naive mice were either infected with 500 infective larvae (L3) of H. polygyrus and/or 10(4) bloodstream forms of T. congolense or were not infected. The outcome of infection was monitored by routine parasitological and immunological techniques for 30 days after the day of the T. congolense infection. Significantly more immune mice concurrently infected with both parasites survived than did immune mice in which H. polygyrus was superimposed on a 10-day-old T. congolense infection. Although all the mice in this latter group died before the end of the experiment, larval immunisation prolonged their survival, relative to similarly treated naive mice. The antibody titres to H. polygyrus in the sera of immune mice challenged with H. polygyrus alone were significantly higher than those of immune mice concurrently infected with both parasites but the levels of protection obtained were comparable. It is concluded that T. congolense may not completely block the strong acquired resistance induced by abbreviated H. polygyrus larval infection in TO mice but is capable of interfering with protective responses, especially if the trypanosome infection occurs prior to H. polygyrus challenge infection.     

Effect of timing and size of photoperiod change on plasma FSH concentration and the correlation between FSH and age at first egg in pullets.

1. ISA Brown pullets were transferred at 6, 9, 12, 15, 18 or 20.3 weeks of age from an 8 h photoperiod to an 8, 10, 13 or 16 h photoperiod. Plasma follicle stimulating hormone (FSH) concentration was measured at transfer at 7 and 14 d afterwards, and age at first egg (AFE) was recorded. 2. Plasma FSH concentration in pullets reared on constant 8 h photoperiods generally increased with age but with a trough at 12 weeks. Plasma FSH increased during the first 14 d of photostimulation to a significantly higher concentration, compared with constant 8 h controls, when the photoperiod was increased to 13 or 16 h at 9, 12 or 15 weeks; but for the increase from 8 h to 10 h photoperiods FSH was only significantly higher than controls when the change was made at 12 weeks. 3. The change in plasma FSH concentration 14 d after photostimulation was significantly correlated with mean AFE (reported in Lewis et al., 1997) and appears to be a better predictor of gonadal development than concurrent changes in plasma LH concentration previously reported (Lewis et al., 1994).     

Regulation of DNA synthesis in Leydig cells obtained from neonatal pig testes.

Three distinct waves of Leydig cell development are found in the pig testes, which occur during fetal, perinatal, and prepubertal periods. Proliferation of Leydig cells is primarily regulated by luteinizing hormone (LH); however, effects of LH on proliferation of immature rat Leydig cells are mediated by specific growth factors and cytokines such as transforming growth factor-alpha (TGFalpha), insulin-like growth factor-1 (IGF-1), interleukin-1beta (IL-1beta), steroidogenesis-inducing protein (SIP), and TGFbeta. The objective of the present study was to identify growth factors that could possibly be involved in the proliferation of Leydig cells in the neonatal pig testis. Leydig cells were isolated from 3- to 5-d-old pig testes, cultured for 48 hr in serum-free media, washed, and treated with hCG and/or IGF-1, epidermal growth factor (EGF), IL-1beta, SIP, and TGFbeta for 18 hr. Tritiated thymidine incorporation into DNA was assessed over a subsequent 4-hr period. Incorporation of [3H]-thymidine was stimulated by hCG treatment with a 2.3-fold increase over control cultures. SIP also induced a significant increase (P < 0.0001) in the incorporation of [3H]thymidine into Leydig cell DNA. Similarly, EGF and IGF-1 also increased DNA synthesis in neonatal porcine Leydig cells, whereas IL-1beta had no effect. TGFbeta had very little, if any, effect on DNA synthesis when added alone, but inhibited the stimulatory effects of other mitogens (SIP, hCG, EGF/TGFalpha, and IGF-1). Our results indicate that these growth factors may play a role in the LH/hCG-dependent proliferation of Leydig cells during the perinatal period of development.     

Persistent activity of doramectin injectable formulation against experimental challenge with Haemonchus placei in cattle.

Two studies were conducted in North America to evaluate the persistent activity of doramectin injectable formulation against experimental challenge with Haemonchus placei. In both studies, calves were randomly assigned to 1 of 4 treatment groups (n = 10 per group) or a larval viability group (n = 2). Calves were treated subcutaneously in the lateral midline of the neck with saline (1 ml/50 kg) on Day 0, or with doramectin (200 mg/kg = 1 ml/50 kg) on Day 0, 7, or 14. Animals used to assess larval viability did not receive any treatment. Beginning on Day 14 and continuing through Day 28, each of the 40 treated calves were given approximately 300 infective larvae of H. placei per os. The two larval viability animals received approximately 10,000 larvae as a single dose on Day 28. Approximately two weeks later, all animals were slaughtered and the abomasum from each calf processed for nematode recovery. A 2% aliquot of abomasal contents plus wash was examined for enumeration and identification of nematodes. Geometric mean H. placei counts were calculated from the log (H. placei count +1) and used to estimate percentage reduction. Overall, doramectin was > or =96.9% efficacious in reducing infection with H. placei when challenged daily 14-28 days after treatment.     

Ultrastructure of porcine circovirus in persistently infected PK-15 cells.

The ultrastructure of porcine circovirus was examined in persistently infected porcine kidney (PK)-15 cells. Virus-infected PK-15 cells had large numbers of intracytoplasmic inclusions, and a few cells had intranuclear inclusions. Intracytoplasmic inclusions were dispersed throughout the cytoplasm but were most numerous in the perinuclear cytoplasm. Inclusion were of various sizes, round to oval, and electron dense and were of two general types. Inclusions of the first type were small (0.1-0.5 microm diameter), not surrounded by trilaminar membranes, and granular with indistinct margins that blended with surrounding cytoplasm. Some contained 12+/-2-nm-diameter icosahedral virions in loose aggregates or rarely forming paracrystalline arrays. Small inclusions could be sites of viral assembly or maturation. Intracytoplasmic inclusions of the second type were larger (0.5-5.0 microm diameter) and more numerous and had abrupt margins surrounded by trilaminar membranes. They were more electron dense than small inclusions and were heterogeneous, containing various proportions of aggregated virions, electron-dense crystalline lamellae of 5 nm periodicity, and/or whorls of myelinoid membranes. Virions usually formed paracrystalline arrays and occasionally were loosely aggregated. Larger inclusions were typical of autophagolysosomes. Intranuclear inclusions were not membrane bound and were often associated with reticulated nucleoli or aggregates of heterochromatin. Some inclusions were irregularly shaped aggregates of indistinct, circular 10-12-nm-diameter viruslike particles. Others were 0.1-1.0 microm in diameter, round or ring shaped, dense, and finely granular, with sharply demarcated margins.     

Survival of Salmonella in the crop contents of market-age broilers during feed withdrawal.

Recent studies have indicated that crop contamination increases during preslaughter feed withdrawal and that contaminated crop contents may serve as an important source of Salmonella entry into poultry processing plants. During the present study, we evaluated the effect of preslaughter feed withdrawal on crop pH and Salmonella crop contamination in broilers from three commercial broiler flocks. The effect of experimental feed withdrawal on crop pH, lactic acid concentration, and Salmonella crop contamination was also evaluated in market-age broilers challenged experimentally with Salmonella typhimurium. Crop pH increased significantly (P < 0.05) from 3.64 +/- 0.25 before feed removal to 5.14 +/- 0.72 after 8 hr of feed withdrawal in broilers from commercial flocks. The incidence of Salmonella crop contamination in the commercial broilers increased (P < 0.05) from 3.3% before feed removal to 12.6% after 8 hr of feed withdrawal. Similarly, crop pH increased (P < 0.05) by a magnitude of approximately 1 unit in broilers after 8 hr of experimental feed withdrawal. The population of S. typhimurium in the crops of the experimentally challenged broilers increased (P < 0.05) by approximately 1 log unit during the 8-hr experimental feed withdrawal. The concentration of lactic acid in the crop of the broilers during experimental feed withdrawal decreased (P < 0.01) from a range of 119-135 mumol/ml before feed removal to a range of 22-32 mumol/ml after 8 hr of feed withdrawal. The results indicated that feed withdrawal resulted in a decrease in lactic acid in the crop, accompanied by an increase in crop pH, and an increase in Salmonella crop contamination.     

Interbreed variation of DLA-DRB1, DQA1 alleles and haplotypes in the dog.

Although 36 DLA-DRB1 and 10 DLA-DQA1 allele sequences have been published to date, no data on individual allele frequencies exists, either for specific breeds or cross breeds, and the full extent of the polymorphism at each of these loci is still not known. We have used sequence-specific oligonucleotide probing (SSOP) to characterise a series of 367 dogs for their DRB1 and DQA1 alleles. These included individual animals from over 60 different breeds, with numbers per breed ranging from 1 to 39. DLA types were generated from 218 dogs for DRB1 and from 330 dogs for DQA1, while 181 dogs were characterised for both these loci. The frequency of individual DRB1 and DQA1 alleles showed considerable interbreed variation, e.g. 83% of West Highland White Terriers were DRB1*01 as opposed to 9% of Collies. No breed had >9 of the 22 DRB1 types defined in this study; several breeds had only two DRB1 types. DLA-DQA1 showed less variation in allele numbers per breed, but also showed considerable interbreed frequency variation. Haplotype analysis revealed over 44 different DRB1/DQA1 combinations. Of these, 25 were in a number of animals, and also in an animal that was homozygous for one or both of these loci. Some DRB1 alleles could be found in combination with several different DQA1 alleles, while others were only present in one haplotypic combination. DLA allele frequency data in normal dogs will be critical for disease association studies. It may also be possible to use haplotype data to establish the genetic relationships between different dog breeds.     

Evaluation of iodophor skin preparation techniques and factors influencing drainage from ventral midline incisions in horses.

OBJECTIVE: To document natural bacterial flora on the ventral aspect of the equine abdomen, to compare 2 preparation techniques, and to identify potential risk factors that may contribute to incisional drainage. DESIGN: Prospective study. ANIMALS: 53 horses undergoing exploratory celiotomy. PROCEDURE: Group-1 horses (n = 26) were prepared with povidone-iodine and alcohol. Group-2 horses (27) were prepared with a film-forming iodophor complex. Numbers of bacterial colony-forming units (CFU) were measured before and after surgical scrub, following skin closure, and after recovery from general anesthesia. Swab specimens to identify normal skin bacterial flora and potential pathogens were obtained by swabbing a 4 x 4-cm area. Variables that might affect incisional drainage were also investigated. RESULTS: For both techniques, there was a significant reduction in bacterial numbers after skin preparation. Incisional drainage was observed in 14 (26%) horses (8 group-1 and 6 group-2 horses). Preexisting dermatitis, poor intraoperative drape adherence, high number of bacterial CFU obtained after recovery from anesthesia, and high number of CFU obtained from the surgery room environment were the main risk factors associated with subsequent incisional drainage. Bacillus spp, nonhemolytic Staphylococcus spp, Micrococcus spp, Corynebacterium spp, Streptomyces spp, other nonenteric genera, and nonhemolytic Streptococcus spp were the most common isolates obtained before surgical scrub. CONCLUSIONS AND CLINICAL RELEVANCE: Both skin preparation techniques were equally effective in reducing numbers of bacterial CFU by 99%, and a significant difference was not found in incisional drainage rate between groups. Protection of the wound during recovery from anesthesia and the immediate postoperative period may reduce incisional drainage after abdominal surgery in horses.