一株瘤胃兼性厌氧纤维素酶产生菌的筛选

试验通过选择性培养基初筛、纤维素刚果红培养基鉴别和固体发酵培养复筛,旨在从瘤胃内容物中筛选出一株产纤维素酶活性较高的细菌。结果表明,试验共分离得到19株有纤维降解作用的菌株,其中R2、R3、R4、R5、R7、R14及R197株的降解作用显著,通过刚果红染色及对纤维素酶活、干物质(DM)消失率的测定,最终以R3效果最佳,其透明斑直径/菌落直径为6.27、羧甲基纤维素酶活(CMCase)为5.94 U/g,滤纸糖酶活(FPase)为1.12 U/g、DM消失率达到23.73%。结果提示,筛选出的瘤胃菌中以R3对稻草秸秆的降解作用效果最佳。     

Synovial fluid total protein concentration as a possible marker for canine idiopathic polyarthritis

Idiopathic polyarthritis (IPA) is a very common inflammatory arthropathy in the dog. Canine IPA is diagnosed mainly by detecting increased number of leukocytes in the synovial fluid (SF), which is easily influenced by glucocorticoid therapy. We obtained 31 SF samples from 24 IPA dogs prior to (n=19) and/or after (n=12) 1 to 10 weeks of glucocorticoid therapy. The SF total protein concentrations of IPA dogs were significantly higher than those of dogs with non-arthritis diseases (n=34) and healthy controls (n=10). Our data revealed that the SF total protein concentrations are not influenced by several weeks of glucocorticoid therapy. Hence, the SF total protein concentration is applicable as a diagnostic marker of canine IPA even when the patients are receiving glucocorticoid therapy.     

Controversial aspect of using GFP as a marker for the production of transgenic cattle

The objective of this study was to examine the feasibility of identification and selection of cattle embryos based on green fluorescence (GFP-positive) in order to obtain calves carrying an integrated transgene. The construct used (pbLGTNF-EGFP) contained the human tumor necrosis factor alpha (hTNFalpha) gene fused to the bovine beta-lactoglobulin promoter (bLG) in plasmid vector pCX-EGFP. In four experiments, 76 zygotes were injected; eight of them developed to the morulae/blastocysts stage of which only five were GFP positive (one of them 100%, one-50%, three- 25%). All of the GFP positive embryos were transferred to recipients. Two calves were born: one after transfer of the 100% GFP positive embryo and the other after transfer of one of the 25% GFP positive embryos. Both animals were healthy with normal weight when compared to two control calves. The integration of pbLGTNF-EGFP in the host genome could not be detected in either of the calves, suggesting that GFP is an unreliable marker for preimplantation screening of embryos.     

Evaluation of titanium dioxide as a digestibility marker for cattle

Three studies were conducted to evaluate titanium dioxide (TiO2) as a digestibility marker for cattle. In Exp. 1, eight steers consumed prairie hay ad libitum with or without dietary supplements. Fecal recovery of TiO2 averaged 93% and was not affected (P = 0.47) by supplement. Digestibilities calculated with reference to TiO2 were not different (P = 0.15) from those based on total fecal collections. In Exp. 2, two steers were limit-fed corn-based diets. Fecal recovery of TiO2 averaged 95% and that of chromic oxide (Cr2O3) averaged 113%. Digestibilities calculated with reference to TiO2 were underestimated (P < 0.01) by 1.1 percentage units relative to those based on total fecal collections, and those calculated with reference to Cr2O3 were overestimated (P < 0.01) by 2.0 percentage units. In Exp. 3, eight steers in a replicated 4 x 4 Latin square consumed corn-based diets ad libitum. Fecal recovery of TiO2 averaged 90%, whereas that of Cr2O3 averaged 98%. Digestibilities calculated with reference to TiO2 were underestimated (P < 0.01) by 1.6 to 4.3 percentage units, whereas those calculated with reference to Cr2O3 were not different (P = 0.31) from those based on total fecal collections. Future research is warranted to determine the usefulness of TiO2 in measuring digestibility in cattle.     

Feasibility of using lignin isolated from forages by solubilization in acetyl bromide as a standard for lignin analyses

Lignin concentration can be measured in plants by the acetyl bromide-soluble lignin spectrophotometric method; however, as with any spectrophotometric method, a reliable standard is needed. In the present experiments, lignin was extracted from each of the forages under study with the acetyl bromide reagent. The lignin isolated with acetyl bromide (LIAB) was then used as the reference standard in the acetyl bromide-soluble lignin (ABSL) analysis, which was compared with the acid detergent lignin (ADL) and potassium permanganate lignin (PerL) lignin analyses. Two maturity stages of each of the following forages were analyzed: Medicago sativa, Cynodon dactylon var. Coastal, Panicum maximum var. Centenário and var. Coloni?o, Cynodon plectostachyus, Pennisetum purpureum, Setaria nandi, and Avena sativa. In addition, one wood sample, Eucalyptus sp., was analyzed. In general, ABSL values were highest (P < 0.001), followed by PerL and ADL, which also differed from each other (P < 0.001). Correlations with in vitro dry matter digestibility of samples were highest with the ABSL method. Absorption spectra of LIAB, either from plants of different maturity stages or from different vegetable species, suggested the presence of differences among some of the lignins.     

Lactic acid-utilizing bacteria in ruminal fluid of a steer adapted from hay feeding to a high-grain ration.

A mature, rumen-cannulated steer fed Coastal Bermuda grass hay for 8 weeks was adapted to a high-grain ration by stepwise increases in grain over a period of 4 weeks. The grain rations had concentrate-to-roughage ratios of 40:60, 70:30, and 85:15 and were fed for 10, 7, and 11 days, respectively. Numerical estimates of lactate-utilizing bacteria in ruminal fluid of the steer the last 3 days each ration was fed were made by colony counts. Lactate-utilizing bacteria were identified as Megasphaera elsdenii, Peptococcus asaccharolyticus, and Selenomonas ruminantium. Maximal numbers of lactate-utilizing bacteria were observed in the ruminal fluid of the steer during feeding the concentrate-to-roughage ration of 70:30.     

Peanut agglutinin as a surface marker for canine T lymphocytes

Peanut agglutinin (PNA) and surface immunoglobulin (SIg) were investigated as markers for T and B lymphocytes in blood and lymphoid tissues of dogs of various ages. In the blood study, 4 age groups (n = 8 dogs/group) were used. The mean (+/- SD) percentages of PNA-positive (PNA+) cells were 68.4 +/- 8.6% (group 1, less than 1 year old), 70.3 +/- 9.2% (group 2, 1 to 2 years old), 72.0 +/- 3.7% (group 3, 5 to 6 years old), and 63.8 +/- 10.1% (group 4, 10 to 11 years old). The mean percentages of SIg-positive (SIg+) cells in blood were 32.1 +/- 10.6% (group 1), 43.2 +/- 7.0% (group 2), 34.3 +/- 4.8% (group 3), and 35.0 +/- 6.8% (group 4). The mean total percentages of PNA+ and SIg+ cells were 100 +/- 6.0% (group 1), 113.5 +/- 4.9% (group 2), 106.3 +/- 5.3% (group 3), and 98.9 +/- 9.2% (group 4). The proportions of PNA+ and SIg+ cells in dogs of group 2 were significantly (P less than 0.05) different from those in dogs of the other groups. Serial changes in PNA+ and SIg+ cells were investigated in blood of 6- to 29-week-old pups (n = 8). A significant (P less than 0.05) transient decrease in PNA+ cells and a corresponding increase in SIg+ cells was observed in pups between 14 and 17 weeks old. Lymphoid tissue specimens and blood samples were obtained from 2- to 6-month-old dogs (n = 11) and from 6- to 12-month-old dogs (n = 10).(ABSTRACT TRUNCATED AT 250 WORDS)     

Desmin as a possible immunohistochemical marker for feline hypertrophic cardiomyopathy

Desmin has been suggested as a possible histopathological marker for hypertrophic cardiomyopathy (HCM) in humans. To test whether a similar pattern of desmin staining applies to HCM in cats, we conducted an immunohistochemical study on myocardial samples from 13 cats (HCM 4, other cardiomyopathies (OCM) 4, and control 5). The pattern of staining for desmin in HCM cats was not the same as that reported in humans, but was weaker than in OCM cats and controls. This suggested that desmin may be a possible histochemical marker for feline HCM, but our data was insufficient to clearly confirm this.     

Telomerase activity as a marker for malignancy in feline tissues

OBJECTIVE: To establish the diagnostic significance of the telomeric repeat amplification protocol (TRAP) assay in detecting feline malignancies. SAMPLE POPULATION: Solid tissue specimens collected from 33 client-owned cats undergoing diagnostic or therapeutic procedures at the University of Illinois Veterinary Medical Teaching Hospital between July 1997 and September 1999 and an additional 20 tissue samples were collected from 3 clinically normal control cats euthanatized at the conclusion of an unrelated study. PROCEDURE: The TRAP assay was used for detection of telomerase activity. Each result was compared to its respective histopathologic diagnosis. RESULTS: Twenty-nine of 31 malignant and 1 of 22 benign or normal tissue samples had telomerase activity, indicating 94% sensitivity and 95% specificity of the TRAP assay in our laboratory. CONCLUSIONS AND CLINICAL RELEVANCE: The diagnostic significance of telomerase activity has been demonstrated in humans and recently in dogs by our laboratory. We tested feline samples to determine whether similar patterns of telomerase activity exist. On the basis of our results, the TRAP assay may be clinically useful in providing a rapid diagnosis of malignancy in cats. The telomerase enzyme may also serve as a therapeutic target in feline tumors.     

D-Lactic acidosis in calves as a consequence of experimentally induced ruminal acidosis

In order to test the hypothesis that ruminal drinking in calves can lead to D-lactic metabolic acidosis, ruminal acidosis was induced in nine calves by intraruminal application of untreated whole milk via a stomach tube. The amount of the daily force-fed liquid was 3 x 1 l. The experimental design called for an end of intraruminal applications if two or more of the following signs were observed: severe depression, estimated degree of dehydration >10%, absence of sucking reflex, lack of appetite for two consecutive feedings, severe metabolic acidosis with calculated Actual Base Excess (ABE) <-15 mmol/l. The procedure was scheduled to be discontinued on the 17th day of experiment. The onset of ruminal acidification occurred rapidly, and mean pH value fell from 6.70 (+/-0.48) to 4.90 (+/-0.38) after the first application. The following days the pH values varied between 4 and 5. Rumen acidity was characterized biochemically by a significant increase in both isomers of lactic acid. The effects of the intraruminal administration on the calves were detrimental; eight of nine calves showed an acute disease process. According to the pre-established clinical standard, seven of nine calves were removed from the intraruminal feeding schedule. All but one of the calves developed severe systemic acidosis. The increase in anion gap demonstrated the net acid load. In all the calves D-lactate levels were found to show a significant and rapid increase. On the contrary, L-lactate never deviated from physiological levels. These observations confirm that, in young calves as in adult cattle, ruminal acidosis may lead to a clinically manifested D-lactic metabolic acidosis.     

Lipid coating as a mode of protecting free methionine from ruminal degradation

A method of protecting free methionine from partial ruminal degradation utilizing a lipid-protein matrix was developed. Eight wether lambs were fitted with abomasal cannulae and utilized in a 4 X 4 Latin square design experiment to determine the effectiveness of the protection matrix. The squares were blocked by animal and time, with each animal receiving each of the four diets. The diets were: (1) a negative control tall fescue and corn-based diet containing no added methionine; (2) a positive control diet that contained 3 g of methionine with ground corn, zein, coconut oil and methionine each added individually; (3) the control diet 1 supplemented with a methionine, ground corn, zein and coconut oil matrix that provided 3 g methionine/d; and (4) the control diet 1 containing the methionine matrix to provide 6 g methionine/d. Digestibility and balance data were obtained by collecting feces and urine over a 7-d period, followed by a day of blood sampling at 2, 4 and 6 h postfeeding. Abomasal samples were then subsequently collected over 3 d on a time schedule that represented every 2 h. Feeding protected methionine decreased (P less than .08) urinary N by .69 g/d and increased (P less than .08) N retention by 1.07 g/d. Plasma urea N was decreased (P less than .003) by 2.06 mg/100 ml and plasma free methionine increased (P less than .001) by 1.94 mumol/100 ml in lambs fed the protected methionine matrix. These data indicate that coating free methionine with the preparation described herein was partially effective in delivering methionine to the absorptive sites and subsequently to the tissues of the ruminant animal.     

Determination of bacteria constituting ruminal fibrolytic consortia developed on orchard grass hay stem

To determine the relationship between Fibrobacter succinogenes and other rumen bacteria, the bacterial community structure on fiber was analyzed by using two different materials. These were ruminally incubated orchard grass hay stems without and with preincubation with F. succinogenes (natural and artificial consortia, respectively). The natural consortium mainly consisted of Firmicutes (56.6%) and Bacteroidetes (33.1%), while the artificial consortium showed a significantly higher proportion of Firmicutes (85.5%) and a lower proportion of Bacteroidetes (4.6%). At species or genus level, Butyrivibrio fibrisolvens, the U2 group, Ruminococcus albus and Lachnospiraceae incertae sedis made up a higher proportion in the artificial consortium. The most dominant bacterial group was the Butyrivibrio‐Pseudobutyrivibrio‐Lachnospiraceae incertae sedis group, which accounted for 19.7% in the natural and 29.5% in the artificial consortium. Within the genus Butyrivibrio, the phylogenetic groups SA and VA2 and phylogeny‐undefined Butyribivrio, but not VA1, were detected at high frequency in the artificial consortium. These results suggest that ecological and possibly functional relationships exist in the rumen among F. succinogenes, a subset of B. fibrisolvens, the U2 group, R. albus and Lachnospiraceae incertae sedis.     

Feasibility of serological bulk milk testing as a method for BVD surveillance

A commercial ELISA detecting antibodies against bovine viral diarrhoea Virus (BVDV) was analysed for its applicability for bulk-milk screening. Detection limits were analysed using native and concentrated milk samples (milk treated with rennet and ammonium sulfate precipitated) from 10 cows whose sera showed different reactivity levels in the ELISA and from two cows which gave birth to persistently infected calves during the last year. Further this and a second commercial ELISA were used to screen 591 randomly selected bulk-milk samples. To clarify discrepancies thirty-nine herds were included in a follow-up study. A second bulk-milk sample and serum samples from 10 young cattle of 6 to 28 month of age per herd were analysed for antibodies against BVDV. The results of this second testing and the detection of viremic animals in 4 herds confirmed the results from initial bulk-milk testing with both tests. The analysed test is suitable for bulk-milk testing although its application is limited by vaccination.     

不同粗精比日粮对山羊瘤胃微生物氨基酸组成的影响

文中比较研究了不同粗精比日粮对山羊瘤胃微生物氨基酸组成的影响,结果表明,液相微生物和固相微生物的氨基酸组成有较大差异,前者含有更高比例的天冬氨酸、甘氨酸和丙氨酸,后者含有更高比例的亮氨酸和苯丙氨酸等必需氨基酸以及谷氨酸和酪氨酸。粗料比例增加增大了微生物赖氨酸和脯氨酸的含量,但减少了蛋氨酸和酪氨酸的百分含量,尤其是对与饲料颗粒疏松结合的细菌影响更大。     

Microsatellite marker C04107 as a diagnostic marker for copper toxicosis in the Danish population of Bedlington terriers

The linkage phase of marker C04107 was evaluated before implementation of the marker in a diagnostic test. Blood samples from 68 dogs were collected and genotyped by PCR. Two alleles were detected with sizes of 160 bp and 164 bp and allele frequencies of 0.45 and 0.55 respectively. Genotyping revealed that 35 dogs were heterozygous (51.5%), 22 dogs were homozygous for the normal allele (32.3%) and 11 dogs were homozygous for the disease allele (16.2%). Liver biopsies were taken from 14 selected dogs and the copper content was evaluated histologically. Biopsies from 8 dogs homozygous for the disease allele showed many copper granules along with single cell necrosis, haemosiderosis and cellular infiltration. In liver biopsies from 6 dogs genotyped to be heterozygous or homozygous for the normal allele, copper granules were absent or moderate in number and no lesions were present. The survey demonstrates that the linkage phase of marker C04107 in the Danish population of Bedlington terriers is similar to the linkage phase detected in other countries. Thus, the marker can be used in a diagnostic test for copper toxicosis in Denmark.     

Effects of replacing dietary starch with neutral detergent–soluble fibre on ruminal fermentation,microbial synthesis and populations of ruminal cellulolytic bacteria using the rumen simulation technique (RUSITEC)

A rumen simulation technique (RUSITEC) apparatus with eight 800 ml fermenters was used to investigate the effects of replacing dietary starch with neutral detergent–soluble fibre (NDSF) by inclusion of sugar beet pulp in diets on ruminal fermentation, microbial synthesis and populations of ruminal cellulolytic bacteria. Experimental diets contained 12.7, 16.4, 20.1 or 23.8% NDSF substituted for starch on a dry matter basis. The experiment was conducted over two independent 15‐day incubation periods with the last 8 days used for data collection. There was a tendency that 16.4% NDSF in the diet increased the apparent disappearance of organic matter (OM) and neutral detergent fibre (NDF). Increasing dietary NDSF level increased carboxymethylcellulase and xylanase activity in the solid fraction and apparent disappearance of acid detergent fibre (ADF) but reduced the 16S rDNA copy numbers of Ruminococcus albus in both liquid and solid fractions and R. flavefaciens in the solid fraction. The apparent disappearance of dietary nitrogen (N) was reduced by 29.6% with increased dietary NDSF. Substituting NDSF for starch appeared to increase the ratios of acetate/propionate and methane/volatile fatty acids (VFA) (mol/mol). Replacing dietary starch with NDSF reduced the daily production of ammonia‐N and increased the growth of the solid‐associated microbial pellets (SAM). Total microbial N flow and efficiency of microbial synthesis (EMS), expressed as g microbial N/kg OM fermented, tended to increase with increased dietary NDSF, but the numerical increase did not continue as dietary NDSF exceeded 20.1% of diet DM. Results suggested that substituting NDSF for starch up to 16.4% of diet DM increased digestion of nutrients (except for N) and microbial synthesis, and further increases (from 16.4% to 23.8%) in dietary NDSF did not repress microbial synthesis but did significantly reduce digestion of dietary N.     

Kinetics of ruminal degradation of wheat and potato starches in total mixed rations

Wheat and potato are rich in starch but their starches differ in their rate of ruminal degradation. Kinetics of in sacco disappearance and profiles of ruminal fermentation were studied for these two concentrates in total mixed rations based on grass silage or corn silage. Wheat starch was more rapidly (34%/h) degraded by rumen microorganisms than potato starch (5%/h). The differences in starch degradation in sacco were found again in the VFA concentrations, mainly in grass silage-based diets. Overall ruminal pH, total VFA concentration, and proportions of acetate, propionate, and butyrate are more variable for wheat during the kinetic (amplitude and quickness) than for potato in grass silage-based diets. In these diets, risks of acidosis were more elevated with wheat than with potato but the VFA concentrations were also higher. These differences of fermentation profile were so reduced in corn silage-based diets that, in this case, wheat can be substituted by potato without any effect on digestion and no risk of acidosis.     

Feline lymphangiosarcoma--definitive identification using a lymphatic vascular marker

Abstract   Three cases of feline exudative dermatitis associated with lymphangiosarcoma are described. The animals, an 11-year-old, neutered male and two 10-year-old, neutered female short hair European cats, presented with a 2-month history of transparent liquid oozing from the skin of the groin and caudal abdomen. On physical examination the neutered male cat and one of the females were slightly depressed and showed loss of weight. Skin lesions were similar in all cats and characterized by the presence of alopecia and moist dermatitis in the ventral abdomen, groin and inner thigh. The hair at the periphery appeared matted by the fluid. In all three cases, histopathological examination of skin biopsies from the abdomen identified poorly defined neoplasia involving dermis and subcutis, characterized by proliferation of spindle cells aligned along pre-existing collagen bundles. The dissection of collagen bundles gave rise to irregular shaped anastomosing, often blind-ending vascular channels and trabeculae. Vascular spaces were mostly optically empty. These histological features were strongly suggestive of lymphangiosarcoma. Neoplastic cells were positive for the blood vascular marker Von Willebrand factor, and a lymphatic vascular marker LYVE-1 (Lymphatic Vessel Endothelial receptor – 1), demonstrating the mixed vascular origin of the tumour. Ultrastructural findings confirmed the final diagnosis of lymphangiosarcoma.     

Relative contributions of ruminal bacteria and protozoa to the degradation of protein in vitro

Mixed ruminal microorganisms from a cow fed timothy hay and concentrate supplement (50:50) were incubated with various protein sources for 15 h (no carbohydrates or growth), and deamination was studied under enzyme-limiting substrate-excess conditions (n = 3). Addition of amphotericin (10 micrograms/ml) killed protozoa and decreased (P less than .05) ammonia production from killed bacteria but it had no effect (P greater than .05) on casein deamination. Monensin (5 micrograms/ml) also killed protozoa; however, it decreased (P less than .05) casein deamination to a much greater extent than amphotericin. Antibacterial antibiotics (penicillin G, polymixin B, cephalosporin C and streptomycin) greatly reduced (P less than .05) ammonia formation from casein. Isolated bacteria always produced more ammonia than isolated protozoa, but the difference was less with heat-treated, particulate proteins. Heated soybean protein was as soluble as heated casein but it was deaminated (P less than .05) at a faster rate by bacteria. Nonammonia-nonprotein N accumulation was greater (P less than .05) with the protozoa than bacteria. When incubations containing bacteria or protozoa were compared with combinations of protozoa and bacteria, the combinations always caused a synergistic increase in ammonia and decrease (P less than .05) in nonammonia-nonprotein N. These results suggest: soluble proteins were primarily degraded by bacteria; protozoa could contribute to the degradation of insoluble, particulate proteins; protozoa were limited in their ability to assimilate peptides (or amino acids); low molecular weight products could be fermented more readily by bacteria and monensin was toxic to protozoa, but decreases in ammonia were primarily due to action of monensin on bacteria.