规模养殖场粪污治理和综合利用

20世纪80年代前,养猪业以分散养殖为主,但随着农业机械化耕作迅速推进,耕地规模化经营加快,畜禽粪便的使用因季节性用肥和劳动力成本提高等因素受到了限制.畜禽进行标准化规模养殖后,粪便集聚度提高,对环境污染越来越严重,笔者认为,解决畜禽粪便污染,变废为宝,已成为畜牧业可持续发展的新课题.     

禁止在饲料和动物饮水中使用的物质

中华人民共和国农业部公告第1519号为加强饲料及养殖环节质量安全监管,保障饲料及畜产品质量安全,根据《饲料和饲料添加剂管理条例》有关规定,禁止在饲料和动物饮水中使用苯乙醇胺A等物质（见附件）。各级畜牧饲料管理部门要加强日常监管和监督检测,严肃查处在饲料生产、经营、使用和动物饮水中违禁添加苯乙醇胺A等物质的违法行为。     

Microbial contamination in human and animal drinking water

Water plays an important roll in the epidemiology of a lot of infectious diseases. Groundwater as well as surface water contains generally microorganisms of several species, which cannot always differentiated properly in autochthonous flora and contaminants with health significance. Sources of bacterial or viral contaminants may be feces from man and/or animals but with different counts in ground and surface water. With respect to water used for supplying farm animals it is stated that it must have initially drinking water quality. Since it generally looses this quality in this supply system the following requirements shall be met: Free of Salmonellas and/or Campylobacter in 100 ml, no E. coli in 10 ml, total bacterial count at 37 degrees C less than 1000 cfu/ml and total bacterial count at 20 degrees C less than 10,000 cfu/ml.     

几种禾本科牧草内生细菌的分布特性

不同禾本科牧草植株内生细菌含量差异显著,杂交狼尾草闽牧六号内生细菌含量最高,达26.800×10⁴CFU/g。具体到各个部位,禾本科牧草不同部位内生细菌含量差异也显著。通过微生物鉴定系统对内生菌进行鉴定的结果表明,64株内生细菌中与MIDI数据库比较相似性指数大于0.500,可确定其菌种名称的有50株,属于19个种,隶属于12个属。以分离到的19种内生菌为样本,以各菌株在各部位出现与否为指标,用类平均法对数据进行系统聚类分析,当λ=8.35时,可将19种内生细菌分为4个大类群。从聚类结果显示,内生菌与宿主植株和不同分离部位都存在相关性,与宿主植株的关系更密切。     

饮用水水质与动物生产

论述了水的感官性状指标、化学指标和细菌学指标对动物健康与生产的影响,介绍了各项指标的卫生要求与 卫生标准以及保证饮用水水质的措施     

Prevalence of shiga toxin-encoding bacteria and shiga toxin-producing Escherichia coli isolates from dairy farms and county fairs

Shiga toxin-encoding bacteria (STB) and shiga toxin-producing Escherichia coli (STEC) were detected and isolated from dairy cattle and their farm environment and from manure piles at Minnesota (MN) county fairs from 2001 to 2002. A total of 2,540 samples were collected from 28 dairy cattle farms (8 organic and 20 conventional), 17 calf pens (5 organic and 12 conventional), and 12 county fairs. STB were detected from 71 (3.2%) of 2208 fecal samples with 20 (71.4%) of 28 dairy farms having at least one positive animal sample. In samples collected from conventional farms, 41 (2.3%) of 1750 fecal samples were STB-positive and 13 (65%) of 20 farms had at least one positive animal. Thirty (6.6%) of 458 fecal samples from organic farms were STB-positive and 7 (87.5%) of 8 farms had at least one positive animal. STB was detected from 31 (17.4%) of 178 samples and 7 (58.3%) out of 12 manure piles at county fairs. A total of 43 STEC isolates were recovered and belonged to 26 different serotypes (19 O and 18 H types). Among STEC, 60.5% possessed only stx1, 30.2% stx2, and 9.3% both stx1 and stx2. The genes eae and hlyA were detected in more than 50% of the STEC isolates. STB can be found on most dairy cattle farms including organic and conventional herds and county fairs. The presence of these potentially pathogenic bacteria in county fairs may pose a risk to the public who have contact with cattle or their environment.     

Antimicrobial resistance in fecal Escherichia coli isolated from growing chickens on commercial broiler farms

To investigate the effects of rearing practices of commercial broiler chickens on the incidence of antimicrobial resistance in commensal Escherichia coli isolates, fecal E. coli isolates obtained in 4 farms were screened for anitimicrobial resistance. Ten E. coli isolates were recovered from each of the fecal samples collected from 10 birds in the farms at the ages of 2 days, 14-17 days, and 47-50 days. In 2 out of the 4 farms, no antimicrobials were used during the rearing period. In the other two farms, following collection of the fecal samples at 14 and 15 days of age, oxytetracycline (OTC), sulfadimethoxine (SDMX), and tylosin were given to birds on one farm and SDMX was used in the other. Isolates resistant to ampicillin and OTC that were obtained from an untreated flock at different sampling times were closely related to each other by pulsed-field gel electrophoresis patterns (PFGE) of XbaI-digested chromosomal DNA. PFGE analysis together with in vitro conjugation experiments suggested that diversity of resistance phenotypes within a clone may be resulted from the acquisition and loss of R-plasmids in an untreated and a treated flock. The numbers of resistance phenotypes observed among fecal isolates increased during the growth of the chickens in all the farms. The results in the present study suggest that persistence of commensal E. coli strains resistant to antimicrobials even in the absence of antimicrobial administration. It is also hypothesized that horizontal transmission of resistance determinants resulted in the emergence of different resistance phenotypes in those farms.     

Parasitic zoonotic disease agents in human and animal drinking water

Human- and veterinary important parasites of the subkingdom of protozoans and helminths infect humans and animals by ingestion of parasites in contaminated water. The parasites are excreted from the body of infected humans, livestock, zoo animals, companion animals or wild animals in the feces. Recreational waters, agricultural practices and wild animals serve as vehicles of transmission of the parasites in the water supplies. The following topics are addressed: a) the life cycles of parasitic diseases-causing agents with proven or potential transmission via water b) the development and the current research status of the analytical techniques for the detection of parasitic diseases-causing agents from water c) the occurrence of Cryptosporidium and Giardia in surface water supplies and in treated water d) the possible water sources and transmission ways of the parasites into the water supplies e) the behaviour and the possibilities for the removal or elimination of the parasites by water treatment.     

Antimicrobial resistance of Salmonella and generic Escherichia coli isolated from surface water samples used for recreation and a source of drinking water in southwestern Ontario,Canada

Antimicrobial resistance (AMR) in the aquatic environment represents an important means of introduction and dissemination of resistance genes, and presence of resistant pathogens in surface waters may pose a public health concern to recreational and drinking water users. The purpose of this study was to explore antimicrobial resistance patterns in water samples collected from the Grand River watershed (southwestern Ontario, Canada) to describe the composition, trends and potential risks of AMR in the aquatic environment. As part of FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), stream water samples were collected bi‐weekly from sampling sites within the Grand River watershed in the Waterloo, Ontario sentinel site and tested for the presence and antimicrobial susceptibility of Salmonella spp. (2005–2013) and generic Escherichia coli (2012–2013). Of all samples tested, 16% of Salmonella and 22% of E. coli isolates were resistant to at least one antimicrobial, including three Salmonella isolates and two E. coli isolates that were resistant to Category I antimicrobials, which are classified as very high importance for the treatment of serious bacterial infections in humans. The greatest proportion of resistant E. coli isolates were observed from the river site upstream of the drinking water intake, while the greatest proportion of resistant Salmonella isolates were from sites upstream in the watershed, and at one recreational water site. Salmonella resistance trends remained fairly stable between 2007 and 2013, with the exception of streptomycin and tetracycline which increased in 2010 and 2013. Continued surveillance of antimicrobial resistance patterns and exploration of risk factor data will allow for a better understanding of resistance transmission in the aquatic environment.     

尖叶胡枝子青贮微生物数量变化及发酵特性

以尖叶胡枝子（Lespedeza hedysaroides）为原料,对其表面附生的2株乳酸菌（LH29、LH33）的生化特性、生长特性、产酸速率进行了研究,经传统鉴定方法及16S rRNA分析方法鉴定,LH29为戊糖片球菌,LH33为植物乳杆菌;发酵初期戊糖片球菌产酸速率优于植物乳杆菌,发酵中后期后者产酸速率优于前者;将SNOW LACT L (SL)和纤维素酶(AC) 2种添加剂添加至尖叶胡枝子中,研究了不同发酵时间微生物数量的动态变化及发酵品质。发酵初期,各种微生物的数量较青贮原料上的数量有较大的增长,发酵第2天,添加AC处理组的乳酸菌数量最高,SL+AC添加组的乳酸菌的数量显著高于对照组（P＜0.05）,发酵第30天,乳酸菌、好氧细菌、酵母菌、霉菌及大肠杆菌的数量有所减少,大肠杆菌减少更明显;各添加组与对照组相比,氨态氮及丁酸含量显著降低（P＜0.05）,乳酸含量明显提高（P＜0.05）。通过Flieg青贮饲料评分方案得出直接青贮饲料的发酵品质较差,等级为劣,而添加AC及SL+AC处理的青贮饲料发酵品质Flieg等级为优。     

Molecular characteristics of ESBL-producing Escherichia coli isolated from chickens with colibacillosis

BackgroundAvian pathogenic Escherichia coli (APEC) causes colibacillosis, resulting in significant economic losses in the poultry industry.ObjectivesIn this study, the molecular characteristics of two extended-spectrum beta-lactamase (ESBL)-producing APEC isolates were compared with previously reported ESBL-producing E. coli isolates.MethodsThe molecular characteristics of E. coli isolates and the genetic environments of the ESBL genes were investigated using whole genome sequencing.ResultsThe two ESBL-producing APEC were classified into the phylogenetic groups C and B1 and ST410 and ST162, respectively. Moreover, the ESBL genes of the two isolates were harbored in different Inc plasmids. The EC1809182 strain, harboring the bla_CTX-M-55 gene on the plasmid, exhibited extensive homology to IncFIB (98.4%) and IncFIC(FII) (95.8%). The EC1809191 strain, harboring the bla_CTX-M-1 gene, was homologous to IncI1-I (Gamma) (99.3%). All chromosomes carried the multidrug transporter, mdf(A) gene. Mobile genetic elements, adjacent to CTX-M genes, facilitated the dissemination of genes in the two isolates, analogous to other ESBL-producing E. coli isolates.ConclusionsThis study clarifies the transmission dynamics of CTX-M genes and supports strengthened surveillance to prevent the transmission of the antimicrobial-resistant genes to humans via the food chain.     

ESBL- and plasmidic class C beta-lactamase-producing E. coli strains isolated from poultry, pig and rabbit farms

This study aims to determine the presence of extended-spectrum (ESBL) and plasmidic class C beta-lactamase-producing Enterobacteriaceae in poultry, pig and rabbit farms of Catalonia (Spain). PFGE typing showed a low clonal relationship among strains carrying these mechanisms of resistance. Ninety-three percent of them were resistant to two or more of the non-beta-lactam antimicrobials tested and harboured ESBL and plasmidic class C beta-lactamases. Greater diversity of these enzymes was found in strains from poultry farms, the CTX-M-9 family, especially CTX-M-14, with CMY-2 being the most frequent. The isolation of TEM-52 and SHV-2-producing Escherichia coli strains from these animal farms is noteworthy. In contrast, 73% of the strains from pig farms had CTX-M-1, and neither the CMY-type nor CTX-M-9 family enzyme was found. Likewise, it is the first time that CTX-M-1 and SHV-5 encoding strains have been isolated in pigs. On the other hand, in rabbit farms CTX-M-9 family was also the most frequent, being detected in three of a total of four strains. The last one showed a CMY-2, for the first time detected in these animals, too. In conclusion, commensal E. coli strains of food-producing animal farms are a reservoir of ESBL and plasmidic class C beta-lactamases.     

Antimicrobial resistance in generic Escherichia coli isolated from swine fecal samples in 90 Alberta finishing farms

The objective of this study was to determine the prevalence of antimicrobial resistance in generic Escherichia coli isolates obtained from 90 Alberta finisher swine farms. Up to 5 isolates were obtained from each of 269 pooled fecal samples and were classified as susceptible or resistant according to Clinical and Laboratory Standards Institute guidelines. Of the 1322 isolates, 166 (12.6%) were susceptible to all 15 antimicrobials. No resistance to amikacin, ceftiofur, ceftriaxone, or ciprofloxacin, antimicrobials of importance in human medicine, was observed. Relatively low frequencies of resistance were observed to gentamicin (1.1%), amoxicillin/clavulanic acid (0.7%), and cefoxitin (0.7%). Higher frequencies of resistance were observed for tetracycline (78.9%), sulfisoxazole (49.9%), streptomycin (49.6%), ampicillin (30.6%), chloramphenicol (17.6%), kanamycin (10%), and trimethoprim/ sulfamethoxazole (6.4%). Among the isolates resistant to > or = 2 antimicrobial classes, 20.8%, 20.6%, 18.2%, 7.0%, 1.8%, 0.2%, and 0.2% were resistant to 2, 3, 4, 5, 6, 7, and 8 antimicrobials, respectively. The most common multidrug-resistance patterns (resistance to > or = 2 antimicrobial classes) were streptomycin-tetracycline (9.4%), streptomycin-sulfisoxazole-tetracycline (6.2%), and ampicillin-streptomycin-sulfisoxazole-tetracycline (6.1%). More clustering (higher intra-class correlation coefficients) in antimicrobial resistance was observed for isolates at the same visit than for isolates from different visits in the same farm, indicating that sampling more farms, testing fewer isolates per visits, and taking longer periods between visits may be appropriate and more efficient for a better understanding of potential shifts in resistance over time.     

奶牛场废弃物研究态势的全球文献计量分析

为了全面了解有关奶牛粪便研究的发展状况及该研究领域的历史动态,运用CitespaceⅢ信息可视化软件对1925—2016年内Web of Science核心合集系列数据库收录的2 395条有关奶牛粪便研究领域的共引文献、研究机构、高频热点词汇进行了分析,并绘制了关于该领域的科技文献知识图谱。结果表明:国际上有关奶牛粪便研究的关注在不断提高,美国、加拿大、瑞典、荷兰等国家在该研究方向处于领先水平;同时该领域向着系统性、多学科的方向发展,说明国际间以及机构合作有利于加强并推动奶牛养殖业与奶牛粪便研究的发展。     

西藏地区藏北嵩草附着乳酸茵的理化特性

本研究以藏北嵩草（Kobresia littledalei）上附着的乳酸菌为研究对象，分离得到肠膜明串珠菌肠膜亚种两株，融合魏斯氏菌11株，食物魏斯氏菌6株，然后对这3种试验菌进行不同温度及pH条件下的生长试验、耐盐试验及糖发酵试验等生理生化特性的研究。结果表明，与常规条件下分离得到的相同菌株相比较，青藏高原极端环境中分离的魏斯氏乳酸菌和明串珠菌株都能在4~40 ℃的条件下生长，部分融合魏斯氏菌还可在50 ℃下微弱生长；耐酸碱试验表明，魏斯氏乳酸菌株都能在pH值为3.0和9.5的条件下生长；在耐盐性方面，部分魏斯氏菌可以在18%的NaCl下生长；在碳源利用方面，本研究提取的食物魏斯氏菌可利用半乳糖；融合魏斯氏菌可利用阿拉伯糖、蜜二糖、甘露醇。因此，藏北嵩草中所分离的乳酸菌比常规环境中的乳酸菌有较强的耐温性和耐酸碱性，而且能更广地利用碳源，从而为其在饲料青贮、酸奶发酵等方面的应用奠定基础。     

广西不同养殖模式猪源大肠杆菌耐药性和耐药基因检测分析

本研究目的是了解广西不同养殖模式猪源大肠杆菌的耐药情况和耐药基因流行情况,为临床合理用药治疗猪大肠杆菌病以及减缓细菌耐药性产生提供参考。采用K-B纸片法测定了180株分离自广西3种养殖模式猪源大肠杆菌的耐药情况,并通过PCR检测其携带EABLs(bla_(TEM)、bla_(CTX))、PMQR因子(qnrA、oqxA、oqxB、aac(6′)-Ib-cr)、大环内酯类耐药基因(ermB)、磺胺类耐药基因(suL)和氟苯尼考耐药基因(floR)的情况。结果显示,大规模、中小规模和散养户养猪场的大肠杆菌对头孢西丁、头孢他啶和阿米卡星的敏感率高于70%,而对青霉素G、氨苄西林、阿莫西林、利福平、林可霉素和复方新诺明的耐药率均高于90%。大规模养猪场大肠杆菌对恩诺沙星的耐药率极显著低于散养户(P<0.01),对氧氟沙星、庆大霉素和氟苯尼考的耐药率显著低于散养户(P<0.05);中小规模养猪场大肠杆菌对恩诺沙星、强力霉素和氟苯尼考的耐药性显著低于散养户(P<0.05);散养户大肠杆菌对环丙沙星、卡那霉素和壮观霉素的耐药率极显著高于大规模养猪场和中小规模养猪场(P<0.01),而对头孢噻肟的耐药率极显著低于大规模养猪场和中小规模养猪场(P<0.01)。大规模养殖场、中小规模养殖场和散养户大肠杆菌的多重耐药指数(MARI)分别为0.62、0.63和0.68,分离自散养猪场的83.3%菌株(50株)表现为对13-23种药物的多药耐药,极显著高于大规模养猪场(50%,30株,P<0.01),显著高于中小规模养猪场(61.7%,37株,P<0.05)。在3种模式养猪场的大肠杆菌中均检测到bla_(TEM)、bla_(CTX、)qnrA、oqxA、oqxB、aac(6′)-Ib-cr、ermB、suL和floR 9种耐药基因,其中bla_(TEM)和suL的阳性率高达100%,bla_(CTX)的阳性率最低,各模式猪场均低于57%。3种养殖模式之间的耐药基因阳性率相比差异显著,但与耐药表型不具有显著相关性。结果表明,3种养殖模式的猪场分离的大肠杆菌均具有严重的耐药问题,且菌株可携带多种耐药基因,表现为多重耐药,而不同养殖模式猪场分离的菌株耐药性存在显著性差异。     

福建南平地区猪场饮用水中肠球菌耐药特性研究

旨在探讨不同猪场来源的饮用水肠球菌对6种抗菌药的耐药情况,了解南平地区猪场水源与饮水器出水在肠球菌耐药表型及耐药谱方面的差异。水样中富集培养肠球菌,通过生化、PCR方法鉴定肠球菌菌株。微量肉汤稀释法开展药物敏感性试验,测定肠球菌对抗菌药的最小抑菌浓度(MIC)。结果显示:南平地区11个猪场46株饮用水分离菌对抗菌药的耐药率由高到低分别为:红霉素土霉素氯霉素环丙沙星氨苄西林万古霉素,对前4种药的耐药率分别为:84.78%、76.09%、67.39%和50%,对氨苄西林的耐药率较低,为19.57%,分离出2株耐万古霉素肠球菌(VRE),耐药率为4.35%。蓄水池分离菌总体表现为对土霉素、红霉素、氯霉素耐药,耐药率较饮水器饮用水分离菌略低,为72.73%、72.73%、63.64%,对环丙沙星、氨苄西林的耐药显著低于饮水器分离菌,分别为27.27%、9.09%。保育舍饮水器分离菌对所有药物都有更高的耐药率,其次为育肥舍、哺乳舍、怀孕舍饮水器。南平地区11个猪场一共有13种耐药谱,有19株分离菌对4~5种的抗菌药耐药,其中5株来自于保育舍饮水器。水池进水口及蓄水池分离菌的多重耐药率均低于饮水器分离菌,保育舍饮水器多重耐药率最高。研究结果提示,猪场源头饮用水与饮水器饮用水分离菌对抗菌药耐药存在差异,饮水管网内耐药菌的存在可能是主要原因。