Proteolytic activities of suparen and rennilase on buffalo, cow, and goat whole casein and beta-casein.

The proteolytic specificity and activity of Mucor miehei protease (Rennilase) and Endothia parasitica protease (Suparen) on buffalo, cow, and goat whole casein and beta-casein (CN) were studied by analyzing the degradation products. The results suggest that Rennilase hydrolyzes casein of the three species in a manner similar to that of chymosin, resulting in the formation of alpha(s1)-I and beta-I, -II, -III as initial degradation fragments of alpha(s1)- and beta-CN. alpha(s1)-I was also the initial breakdown product of alpha(s1)-CN by Suparen. Contrary to Rennilase, Suparen showed a higher affinity toward beta-CN and hydrolyzes beta-CN, giving rise to degradation products characterized by mobility lower than that of beta-CN. Increasing NaCl concentration (>3%) reduced the proteolysis of beta-CN of the three species by Rennilase but not by Suparen. The hydrolysis of alpha(s1)-CN and alpha(s1)-I by the two enzymes was enhanced in the presence of NaCl.     

蜉金龟堆肥和自然堆肥类胡敏酸的表征及特性研究

分别用焦磷酸钠和氢氧化钠浸提新鲜牛粪、蜉金龟堆肥(APCM)和自然堆肥(腐熟牛粪)产物中的类胡敏酸,采用元素组成、红外光谱及差热分析研究了其结构特征。元素分析结果表明,蜉金龟堆肥焦磷酸钠浸提物——类胡敏酸(NaHLA)的芳香性低于新鲜牛粪和自然堆肥,而碳水化合物及氮元素含量增加。蜉金龟堆肥氢氧化钠浸提物——类胡敏酸(PAHLA)的碳水化合物和氮元素含量也高于新鲜牛粪和自然堆肥,而自然堆肥PAHLA的芳香性略低于新鲜牛粪和蜉金龟堆肥。红外光谱分析结果表明,与自然堆肥NaHLA相比,蜉金龟堆肥NaHLA中羧酸脂类化合物、酮类化合物、脂肪族化合物及碳水化合物有所增加。与新鲜牛粪PAHLA相比,蜉金龟堆肥和自然堆肥PAHLA的碳水化合物、脂肪族化合物、酚类化合物及纤维素酯类减少,同时蜉金龟堆肥PAHLA的酰胺化合物在蜉金龟消化过程中被其吸收。差热分析结果表明,自然堆肥NaHLA的热稳定性高于新鲜牛粪和蜉金龟堆肥,而蜉金龟堆肥NaHLA的脂族化合物、外围官能团和分子内部芳香结构含量较高,分子结构较复杂。蜉金龟堆肥PAHLA存在两个放热峰,而新鲜牛粪和自然堆肥的中温放热峰消失。综上,蜉金龟堆肥和自然堆肥采用两种提取液浸提出类胡敏酸的结构特征存在差异性。     

牛粪好氧和蚯蚓堆肥腐熟料成型基质块制备及育苗试验

为优化蔬菜育苗成型基质的配方,提高成型基质蔬菜育苗效果,分别以牛粪好氧堆肥腐熟料和牛粪蚯蚓堆肥腐熟料为主料,草炭为辅料,吸水树脂为膨胀剂,木醋液为调节剂,黄瓜为指示植物,研究不同配方对成型基质块成型及育苗效果的影响,并对各目标指标进行综合评价,以确定较佳的成型育苗基质配方。试验结果表明:牛粪好氧堆肥腐熟料制成的成型基质块在膨胀性能、抗跌碎性及育苗期间破损情况优于牛粪蚯蚓堆肥腐熟料制成的成型基质块,但其p H值、EC值较高,使其存活率和茎粗株高等育苗特性与牛粪蚯蚓堆肥腐熟料相比较差。2种腐熟料基质配方可采取不同的调节方法改善其特性,蚯蚓堆肥腐熟料中添加适量秸秆类纤维状物质可以减小其成型基质块的跌碎率和破损率,牛粪好氧堆肥腐熟料中添加硫磺粉可以调节p H值。从基质块质量综合指数来看,既适合成型又有利于育苗效果的配方为:腐熟料和辅料的体积比为3:2~4:3;成型基质块中膨胀剂的含量以该试验中最小添加量27 m L左右最为合适,与总物料(腐熟料和辅料混合后的物料)的比值为5.5 m L/L;木醋液在2种腐熟料中的添加量不同,在牛粪好氧腐熟料中,木醋液添加量约为8.5 m L/L,而牛粪蚯蚓腐熟料中则为18 m L/L,此时两种腐熟料成型基质块的跌碎率均小于5%,破损率均小于20%和小于40%,幼苗存活率分别大于40%和大于70%,全株干质量全部大于100 mg。该研究结果可为蔬菜有机栽培成型基质的开发及其品质改善提供理论依据,具有重要意义。     

水牛胎牛、青年牛和成年牛腔前卵泡的分离及培养

本研究采用梳刮的机械法分离水牛胎牛、青年牛和成年牛腔前卵泡,并分别进行培养。每个胎牛、青年牛和成年牛的卵巢分别获得943.43±143.13、249.50±40.11和128.50±19.99个腔前卵泡,从胎牛卵巢获得的腔前卵泡,其数量明显多于青年牛和成年牛(P<0.01)。培养72h后青年牛和成年牛的腔前卵泡存活率分别为52.67%、52.03%,而胎牛仅有34.69%的腔前卵泡能够存活,其数量明显低于青年牛和成年牛(P<0.05)。结果表明,青年牛卵巢分离得到的腔前卵泡数量较多,且培养成活率高,是进行水牛腔前卵泡分离培养等研究的理想试验材料。     

Influence of cow manure vermicompost on the growth, metabolite contents, and antioxidant activities of Chinese cabbage (Brassica campestris ssp. chinensis)

The effects of cow manure vermicompost on plant growth, metabolite contents, and antioxidant activities of Chinese cabbage were investigated in pot cultures. Five treatments were designed by mixing vermicompost and soil at ratio of 0:7, 1:7, 2:7, 4:7, 7:0 (w/w). Marketable weight of Chinese cabbage was significantly (p < 0.05) higher in the 2:1 treatment than in the other treatments, while plants grown in the full soil treatment (0:7) showed the lowest marketable weight (Fig. 1a). Vermicompost application significantly increased the nutrient content of Chinese cabbage leaves (p < 0.05), especially in the 4:7 treatment, with increases in the contents of soluble sugar (Fig. 2a), soluble protein (Fig. 2b), vitamin C (Fig. 3a), total phenols (Fig. 3b), and total flavonoids (Fig. 3c) by 62%, 18%, 200%, 25%, and 17% compared to the full soil treatment, respectively. The antioxidant activities expressed by 2, 2-Dipenyl-1-picrylhydrazyl-scavenging activity (Fig. 4a), hydroxyl (OH)-scavenging activity (Fig. 4b), and iron (Fe²⁺)-chelating activity (Fig. 4c) were higher by 92%, 40%, and 36% in the 4:7 than 0:7 the treatment, respectively. Vermicompost application significantly increased (p < 0.05) the plant contents of 16 essential amino acids (Table 1); the total amino acid content showed the greatest increase in the 4:7 treatment, 90% compared to the full soil treatment.     

Effect of cow urine upon denitrification

The influence of urine application upon the zone and rate of denitrification of a pastoral soil was studied using a short term incubation technique. Under the condition of full enzyme induction, no increase in denitrification activity was measured with the addition of cow urine. The zone of maximum denitrification activity was within the top 30 mm and did not shift with increasing C and N substrate at the 60–90 mm depth.     

Vermicomposting of biosolids with cow manure and oat straw

Biosolids, mainly from textile industries and the rest from households, were vermicomposted with Eisenia fetida, cow manure and oat straw for 2 months at three different moisture contents (60%, 70% and 80% dry weight base) in triplicate to reduce pathogens and toxic organic compounds, and to find the best medium for growth of E. fetida. The vermicompost with the best stability and maturity and a weight loss of 18% was obtained with 1,800 g biosolid, no straw and 800 g manure at 70% water content. This vermicompost had the following properties: pH 7.9; organic C content of 163 g kg^–1; an electrolytic conductivity of 11 mS cm^–1; a humic-to-fulvic acid ratio of 0.5 (HA/FA); total N content of 9 g kg^–1; water soluble C (C_w) less than 0.5%; cation exchange capacity of 41 cmol_c kg^–1; a respiration rate of 188 mg CO₂-C kg^–1 compost-C day^–1; a NO₃^–/CO₂ ratio greater than 8; and a NH₄⁺/NO₃^– ratio lower than 0.16. The vermicompost gave a germination index for cress (Lepidium sativum) of 80% after 2 months while the earthworm production increased 1.2-fold and volatile solids decreased five times. In addition, the vermicompost contained less than 3 CFU g^–1Salmonella spp., no fecal coliforms and Shigella spp. and no eggs of helminths. Concentration of sodium was 152 mg kg^–1 dry compost, while concentrations of chromium, copper, zinc and lead were below the limits established by the USEPA.     

牛粪和生物炭对苹果根系生长、土壤特性和氮素利用的影响

以两年生红富士/平邑甜茶苹果为试材,采用15N同位素示踪技术,研究牛粪与生物炭不同配比对苹果根系生长、土壤特性和氮素吸收利用的影响,为苹果生产中合理施肥及可持续发展提供依据。试验共设6个处理:对照(CK)、100%牛粪(T1)、75%牛粪+25%生物炭(T2)、50%牛粪+50%生物炭(T3)、25%牛粪+75%生物炭(T4)和100%生物炭(T5)。结果表明,牛粪和生物炭混施可显著改善土壤理化性状,增加土壤有机质、碱解氮、速效磷和速效钾含量,降低土壤容重。苹果根际土壤中的微生物均以细菌占绝对优势,放线菌次之,真菌含量最少;添加牛粪和生物炭均显著提高了根际土壤的细菌、放线菌和真菌数量,其中T2处理细菌、放线菌和真菌数量均达到最高。牛粪和生物炭混施还可促进苹果根系生长,根尖数、根表面积和根系活力等指标均在T2处理达到最高,分别较对照提高47.90%、33.47%、44.67%。表明T2处理可显著促进苹果根系的生长和根系活力的提高。与对照相比,牛粪和生物炭混合处理显著提高了苹果各器官的Ndff值、全氮量和15N吸收量,提高了15N利用率和15N残留率,降低了15N损失率。各处理15N利用率和15N残留率趋势为牛粪和生物炭混合处理最高,单施牛粪或生物炭处理次之,对照最低; 15N损失率呈相反的变化趋势。其中以T2处理效果最好,15N利用率和15N残留率最高, 15N损失率最低, 15N利用率较对照提高5.51%, 15N损失率较对照降低14.52%。综合分析认为, 75%牛粪+25%生物炭处理(T2)对苹果根系生长、土壤特性和氮素吸收利用的效果最好。     

蔬菜秧与牛粪好氧堆肥试验研究

以蔬菜秧、牛粪和玉米秸秆为原料,在北京延庆县旧县镇延庆有机农业示范园内进行了为期60 d的高温好氧堆肥试验。初始堆肥混合物料的含水量为61.5%,C/N为21.3,采用翻堆机进行机械翻堆供氧。结果表明,该区蔬菜秧和牛粪的养分含量较高,重金属含量较低,但水分含量很高,C/N都很低,直接用采收后的蔬菜秧和牛粪混合堆肥,达不到堆肥的适宜条件,需添加C/N较高且水分含量较低的玉米秸秆,以实现堆肥的适宜条件。试验表明蔬菜秧和牛粪堆肥是可行的,能有效解决当地的农业废弃物处理问题和有机农业生产中的有机肥短缺问题。     

The veterinary antibiotic journey: comparing the behaviour of sulfadiazine,sulfamethazine, sulfamethoxazole and tetracycline in cow excrement and two soils

Purpose

To conceptualize the journey that veterinary antibiotics (VAs) follow between the animal stall and the field, two experiments were conducted. (1) The VAs sulfadiazine, sulfamethazine, sulfamethoxazole and tetracycline were mixed into cow excrement; prepared with three dry solid content variations. (2) Cow excrement containing the same VAs was mixed into sandy and clayey saturated soils. The aim was to quantify the solid-liquid partitioning and time-dependent behaviour of VAs at each stage of the journey, to enable mathematical replication of the process in the future.

Materials and methods

In each case, the mixtures were partitioned into their solid and liquid phases and the VA concentration in each was determined using liquid chromatography tandem mass spectrometry. Sorption isotherms (K _d values) and elimination constants (k _s: solid form, k _l: liquid form) were calculated after various incubation periods.

Results and discussion

Sulfamethoxazole exhibits fast elimination in excrement; environmental contamination is unlikely. Sulfadiazine and sulfamethazine behave similarly in excrement and soils; tetracycline is more sorptive. Small percentages of the sulfadiazine, sulfamethazine and tetracycline masses initially found in excrement may subsequently be transported to environmental compartments in liquid form. However, the majority of these VAs are speculated to accumulate in soil or be transported to surface water systems via soil erosion.

Conclusions

The VA journey has been mathematically conceptualized for the first time and is supported by sorption isotherms and eliminations constants for four commonly detected VAs. Critical findings for sulfadiazine, sulfamethazine and tetracycline are (1) the majority of an initial VA mass resides in excrement liquid; (2) following incorporation into soil, the majority of the same initial VA mass resides in soil solid; (3) VAs found in soil liquid are assumed to be eliminated within a few months; (4) VAs found in soil solid are assumed to persist and accumulate; and (5) VAs are most likely to be transported to surface water systems in solid form (via soil erosion). Due to its rapid elimination in excrement, sulfamethoxazole that stems from veterinary medicine is not assumed to be a major environmental contaminant.

     

Modeling interval-censored, clustered cow udder quarter infection times through the shared gamma frailty model

Time to infection data are often simultaneously clustered and interval-censored. The time to infection is not known exactly; it is only known to have occurred within a certain interval. Moreover, observations often occur in clusters. Consequently, the independence assumption does not hold. Here we propose an extension of the shared gamma frailty model to handle the interval censoring and clustering simultaneously. We show that the frailties can be integrated out analytically, allowing maximization of the marginal likelihood to obtain parameter estimates. We apply our method to a longitudinal study with periodic follow-up on dairy cows to investigate the effect of parameters at the cow level (e.g., parity) and also parameters that can change within the cow (e.g., front or rear udder quarter) on time to infection. Dairy cows were assessed approximately monthly for the presence of a bacterial infection at the udder-quarter level, thus generating interval-censored data. Obviously, the four udder quarters are clustered within the cow. Based on simulations, we find that ignoring the interval-censored nature of the data can lead to biased parameter estimates.     

牛粪及其蚓粪磷素生物化学特性分析

评估牛粪及其蚓粪磷素生物化学特性,为合理利用牛粪及其蚓粪,提高畜禽粪便磷利用效率提供理论依据。利用扩增子测序技术,研究腐熟牛粪及其蚓粪中磷含量、磷酸酶活性和解磷微生物的差异。与牛粪相比,蚓粪pH、电导率、有效磷含量、全碳含量、全氮含量、碳磷比和氮磷比均显著降低,其中pH降低0.46个单位,电导率、有效磷含量、全碳含量、全氮含量分别降低63.86%、20.99%、28.51%、45.57%,牛粪的碳磷比和氮磷比分别为蚓粪的4.20和5.52倍;但蚓粪全磷含量和碳氮比显著提高,全磷含量提高196.40%,碳氮比为牛粪的1.31倍。蚓粪中碱性磷酸酶活性较牛粪低,而酸性磷酸酶二者间无显著差异。蚓粪的细菌和真菌α-多样性均显著高于牛粪。蚓粪中解磷细菌节细菌属(Arthrobacter)和芽孢杆菌属(Bacillus)的相对丰度较牛粪显著降低;相反,其他解磷细菌和大多数解磷真菌丰度较牛粪均呈升高趋势。蚯蚓堆肥对有效磷含量的衰减作用主要取决于牛粪的化学性质和解磷微生物的丰度。