Distribution of microbial biomass and phospholipid fatty acids in Podzol profiles under coniferous forest

Microbial‐derived phospholipid fatty acids (PLFAs) can be used to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. They have been used to assess microbial communities of humus layers under coniferous forest, but nothing is known of their distribution in the deeper soil. To investigate the vertical distribution we sampled nine Podzol profiles on a 100‐m‐long transect in a coniferous forest and analysed for their microbial biomass and PLFA pattern to a depth of 0.4 m. The transect covered a fertility gradient from Vaccinium vitis‐idaea forest site type to Vaccinium myrtillus forest site type. The cores were divided into humus (O) and eluvial (E) layers and below that into 10‐cm sections and designated as either illuvial (B) or parent material (C), or as a combination (BC). Two measures of microbial biomass analyses were applied: substrate‐induced respiration (SIR) to determine microbial biomass C (C_mic), and the sum of the extracted microbial‐derived phospholipid fatty acids (totPLFA). The soil fertility had no effect on the results. The C_mic correlated well with totPLFA (r= 0.86). The microbial biomass decreased with increasing depth. In addition the PLFA pattern changed with increased depth as assessed with principal component analysis, indicating a change in the microbial community structure. The composition of the PLFAs in the O layer differed from that in the E layer and both differed from the upper part of the B layer and from the rest of the BC layers. The deeper parts of the B layer (BC₁, BC₂ and BC₃) were similar to one other. The O layer had more 18:2ω6, a PLFA indicator of fungi, whereas the E layer contained relatively more of the PLFAs 16:1ω9, 18:1ω7 and cy19:0 common in gram‐negative bacteria. With increased depth the relative amount of 10Me18:0, the PLFA indicator for actinomycetes, increased. We conclude that the PLFA method is a promising discriminator between the microbial community structures of the horizons in Podzols.     

Application of an enzyme-immunoassay (EIA) for rapid screening of 5 alpha-pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus.

Populations of African (Loxodonta Africana) and Asian Elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, the development of easy and practical methods for the analysis of the relevant reproductive hormones is essential to support e.g. assisted reproduction. For the measurement of 5 alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 microliters) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, the mean recovery was 101% and the mean coefficients of variation were 7.3% (intra-assay) and 9.9% (inter-assay). In Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and up to 21 ng/ml during pregnancy. Estrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean 15.4 +/- 2.3). In two Asian elephant cows a calf was still-borne. Thereafter, the animals reassumed ovarian activity after approximately 8 and 13 weeks, respectively. In one animal estradiol implants for hormonal contraception caused a down regulation of the ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. We propose the direct DHP-EIA as a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories.     

A critical evaluation of serum methylmalonic acid and vitamin B12 for the assessment of cobalt deficiency of growing lambs in New Zealand

AIM: To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B12-responsiveness in lambs and critique existing serum vitamin B12 reference ranges. METHODS: Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B12 and MMA, and liveweight gain. RESULTS: Serum MMA concentrations were rarely elevated above the norm of <2 micromol/L when serum vitamin B12 concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 micromol/L) unless serum vitamin B12 concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 micromol/L). CONCLUSIONS: The current reference ranges used in New Zealand for serum vitamin B12 are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B12 concentrations. Serum MMA concentrations >13 micromol/L indicate responsiveness to supplementation whilst concentrations <7 micromol/L indicate unresponsiveness. In the range 7-13 micromol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable. CLINICAL RELEVANCE: The current reference ranges for vitamin B12 responsiveness are conservatively high and lead to over-diagnosis of vitamin B12 deficiency in ill-thriftiness of sheep.     

Clinical evaluation of sodium bicarbonate, sodium L-lactate, and sodium acetate for the treatment of acidosis in diarrheic calves

Thirty-six dehydrated diarrheic neonatal calves were used to study the effects of various alkalinizing compounds on acid-base status, the changes in central venous pressure (CVP) in response to rapid IV infusion of large volumes of fluid, and the correlation of acid-base (base deficit) status, using a depression scoring system with physical determinants related to cardiovascular and neurologic function. Calves were allotted randomly to 4 groups (9 calves/group). Over a 4-hour period, each calf was given two 3.6-L volumes (the first 3.6 L given in the first hour) of a polyionic fluid alone (control group) or were given the polyionic fluid with sodium bicarbonate, sodium L-lactate, or sodium acetate added (50 mmol/L). Acid-base status, hematologic examination, and biochemical evaluations were made immediately before infusion of each fluid (at entry) and after 3.6, 4.8, and 7.2 L of fluid had been given. Compared with control values, bicarbonate, lactate, and acetate had significantly greater alkalinizing effects on pH (P less than 0.01) and base deficit (P less than 0.01) after 3.6, 4.8, and 7.2 L of fluid were given. Bicarbonate had the most rapid alkalinizing effect and induced greater changes in base deficit (P less than 0.01) than did acetate or lactate at each of the 3 administered fluid volumes evaluated. Acetate and lactate had similar alkalinizing effects on blood. Rehydration alone did not improve acid-base status. The CVP was elevated in 10 (28%) of the 36 calves after 1 hour of fluid (3.6 L) administration, but significant differences in body weight, PCV, and clinical condition or depression score at entry were not found between calves with elevated CVP and those with normal CVP.(ABSTRACT TRUNCATED AT 250 WORDS)     

棉酚对奶牛胎儿发育毒性的研究

用棉酚含量４．０ ｇ／ｋｇ 的棉籽饼以每头０．５ ｋｇ／ｄ 饲喂怀孕奶牛３５ ｄ,研究棉酚对奶牛胎儿发育的影响。结果表明,在开始饲喂后１４～４６ ｄ 的流产率为９．１％ （１７／１８６）;中毒的妊娠奶牛平均怀孕期为（２７５．７５±５．２１） ｄ （ｎ＝ １４０）,其所产的犊牛平均初生质量为（３５．２７±５．２３） ｋｇ （ｎ＝ １４０）,与１９９１ 和１９９３ 年同期相比较,平均怀孕期缩短２～３ ｄ,初生犊牛平均质量降低３．７０ ｋｇ;活产母犊到６ 月龄和２４ 月龄时,淘汰率分别为３９．７％ （２７／６８）和８５． ３％ （５８／６８）,６ 月龄时平均质量为（１４６．６±３３．８） ｋｇ （ｎ＝ ４５）,平均日增质量为（０．６２±０．２０） ｋｇ （ｎ＝ ４５）,分别比１９９１ 和１９９３ 年的平均值降低３４．５ 和０．１７ ｋｇ     

Effects of gossypol from cottonseed meal and dietary vitamin E on the reproductive characteristics of superovulated beef heifers

Superovulated Hereford-Angus crossbred heifers (average 397 kg BW) were used to test the effect of feeding cottonseed meal (gossypol) and vitamin E on embryo quality and ovarian characteristics. Twenty-four heifers were assigned randomly to four treatments with six heifers per treatment. Treatments were the following dietary supplements: 1) SBM (soybean meal + 30 IU vitamin E/kg of diet DM); 2) SBM+E (soybean meal + 4,000 IU vitamin E x animal(-1) x d(-1)); 3) CSM (cottonseed meal + 30 IU vitamin E/kg of diet DM); and 4) CSM+E (cottonseed meal + 4,000 IU vitamin E x animal(-1) x d(-1)). Supplements based on cottonseed meal provided 43.5 g of total gossypol/d (37% negative isomer (-) and 63% positive isomer (+)). Blood samples were collected at the start of the experiment and every 3 wk thereafter up to 12 wk. Plasma a-tocopherol (alpha-T) concentration was affected by treatments (P < 0.05). Heifers supplemented with cottonseed meal had greater (P < 0.05) alpha-T concentration in plasma than heifers supplemented with soybean meal at each concentration of vitamin E. Supplementation at 4,000 IU vitamin E x animal(-1) d(-1) increased (P < 0.05) the concentration of a-T in plasma. Weight gain, hemoglobin and hematocrit were not affected by treatment. Erythrocyte osmotic fragility (EOF) increased (P < 0.05) in cottonseed meal-fed animals; however, EOF was lowered (P < 0.05) with vitamin E supplementation. Heifers fed CSM and CSM+E supplements had greater (P < 0.01) concentrations of (-)-, (+)-, and total-gossypol in plasma, corpora lutea (CL), liver, and endometrium than heifers fed SBM and SBM+E supplements. Tissue alpha-T concentration increased with increased dietary supplemental vitamin E, particularly in great amounts in the CL. Because there was no adverse effect of gossypol on superovulation response or embryo development despite concentrations of gossypol in endometrium that are toxic to embryos, it is likely that systems exist in the reproductive tract to limit gossypol toxicity.     

Microbial community structure in forest soils treated with a fire retardant

The influence of a fire retardant (Firesorb, an acrylic–acrylamide copolymer) on the microbial community structure determined by phospholipid fatty acid (PLFA) analysis was examined under laboratory conditions using two different textured soils under pine forest. Firesorb was added to unheated and heated soil samples (350°C for 10 min followed by reinoculation, to mimic a forest fire) at three levels of application (none, usual and three times the usual levels), and measurements were made after 12 weeks of incubation. The relative importance of the three factors considered on the PLFA profiles was as follows: soil heating ≫ soil texture ≈ Firesorb treatment. In the unheated soils, Firesorb had a larger effect than soil texture, while the opposite was found in the heated soils. Soil heating reduced the total PLFAs, while Firesorb tended to increase them in both the unheated and heated soils. Soil heating decreased the PLFAs indicative of gram-positive (G⁺) bacteria and tended to increase the fatty acids associated with gram-negative (G⁻) bacteria and, to a lesser extent, the PLFA 18:2ω6, considered to be predominantly of fungal origin. Firesorb treatment decreased the G⁻/G⁺ bacteria ratio in the heated soils but tended to increase it in the unheated soils, the effect being dose dependent.     

Callus induction and plant regeneration from cotyledonary explants of ash gourd (Benincasa hispida L.)

A protocol for the production of complete plantlets through multiple shoots from the cotyledon-derived calli of ash gourd (Benincasa hispida L.) is described. The embryos were excised from mature seeds and cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurin (BAP, 1–5 μM). After 10 days the well-developed green cotyledons from the growing embryos were isolated and cultured on MS medium fortified with 2,4-D (1–6 μM). The cultured cotyledons gave rise to luxuriantly growing calli after 6 weeks. These calli were subcultured on MS medium supplemented with various concentrations of BAP (1–6 μM) alone or in combination with naphthalene acetic acid (NAA, 0.2 and 0.5 μM) for regeneration. The regenerated shoots were multiplied and rooted on quarter strength MS medium supplemented with indole-3-butyric acid or NAA (1–5 μM). The rooted shoots were transplanted to soil with 90% success.     

Evaluation of stocking density and subtherapeutic chlortetracycline on Salmonella enterica subsp. enterica shedding in growing swine

The objective of this research was to determine the effect of stocking density and inclusion of subtherapeutic chlortetracycline in the diet on Salmonella fecal prevalence and antimicrobial resistance in growing swine. A 2 x 2 factorial design was employed on a privately owned commercial swine farm. Four finisher rooms were included in the study. Two of the rooms received 50 g/tonnes of chlortetracycline in the ration, two rooms received no antimicrobials in the feed. In each room, alternate pens were assigned to either high stocking density (0.60 m2/pig) or low stocking density (0.74 m2/pig). Pigs were placed in the finisher rooms at 10 weeks of age and followed for 6 weeks. Individual fecal samples were collected from the floors of each pen and cultured once weekly. Antimicrobial resistance phenotypes were determined. Data were analyzed using multilevel, multivariable logistic regression. Pigs fed chlortetracycline were at increased odds (OR 6.88, 95% CI 2.77-17.12) to shed Salmonellae. No other associations between treatments (CTC and stocking density) and Salmonella prevalence or reduced susceptibility to antimicrobials were identified. Variance in the odds of a fecal sample to be positive was distributed mostly at the lowest level, the individual fecal sample. The increased risk of shedding associated with inclusion of subtherapeutic chlortetracycline in swine diets is discordant with previous results by our group, suggesting farm or strain specific factors may impact this association. Understanding this risk may provide a potential intervention for controlling Salmonella pre-harvest.