Microbiological Quality of Chicken Sold in Accra and Determination of D₁₀-Value of <i>E. coli</i>

Chicken is an excellent source of good quality protein, but it is highly susceptible to microbial contamination and often implicated in food borne disease. The microbiological quality of chicken at different retail outlets (supermarkets, local markets and farms) in Accra was investigated, and D10-values of E. coli in refrigerated and frozen retailed chicken was determined. The microbiological quality of chicken was studied by analyzing 27 chicken thigh samples collected from the retail outlets. D10-value of Escherichia coli was determined by using a linear regression model after gamma irradiation of inoculated chicken samples with doses of 0, 150, 300, 450, 600, 750 and 900 Gy. Mean total viable counts for the supermarkets, local markets and farms were 6.46, 6.91 and 6.57 log10 cfu/g respectively. Mean total coliform counts for the supermarkets, local markets and farms were 3.80, 3.46 and 3.14 log10 cfu/g respectively and the mean S. aureus counts were also 2.32, 2.28 and 2.70 log10 cfu/g respectively. There were no significant differences (p > 0.05) between the mean total viable count, total coliform counts and S. aureus count for the supermarkets, local markets and the farms. Mean counts of E. coli detected at the supermarket, local markets and farms were 1.27, 2.59 and 2.74 log10 cfu/g respectively. Salmonella spp. was detected in 2 out of the 27 samples. Fifty-two percent and 70% of samples respec-tively had total viable counts and total coliform counts within the microbial safety standards. Mean D10E. coli were 0.22 and 0.32 kGy in refrigerated and frozen chicken respectively. Presence of pathogenic bacteria in fresh chicken sold in some retail outlets in Accra was confirmed. Low D10-values of E. coli especially under refrigerated conditions suggest susceptibility to low dose irradiation and possibility of controlling spoilage and pathogenic microflora of fresh poultry.

Multiple Antimicrobial Resistance of Extended Spectrum Beta-Lactamase-Producing Escherichia coli from Small-Scaled Poultry Farms and Retail Chicken

Antibiotics used for agricultural purpose has contributed to the increased prevalence of antibiotic-resistant bacteria. The goal of this study was to investigate the prevalence and antimicrobial resistance of ESBL-producing E. coli in small-scaled poultry farms and retail chicken. The cultured E. coli isolates were subjected to phenotypic tests, susceptibility tests, and the polymerase chain reaction for detection of blacTX-M, blasHv, and blaTEM genes. From 120 samples each of chicken feces, retail chicken, soil and chicken feed, ESBL-producing E. coli isolates were detected in 75.9%, 63.6%, 39.2%, and 13.3% of the samples, respectively. Minimum inhibitory concentration （MICs） values indicated that ESBL-producing E. coli were resistance to ampicillin （MIC 〉 32 μg/mL）, gentamicin （M1C ≥ 16 μg/mL）, cefotaxime （MIC 〉 4 μg/mL） and cefhiaxone （MIC 〉 4 gg/mL）, respectively. The total resistance for imipenem was also observed at 1.0% （MIC ≥ 4 gg/mL） and none of the isolates were resistant to ceftazidime （MIC 〉 16 μg/mL）. ESBL-producing E. coli from chicken feces and retail chicken carried blasHv gene at a rate of 6.8% and 5.7%, respectively and blaCTX-M gene was also revealed at 2.9% in retail chicken. Moreover, ESBL-producing E. coli isolated from soil harbored blasnv and blaCTX-M genes at 5%. None of the feed samples yielded ESBLs genes. Twenty three resistance patterns were observed for multi-resistant ESBL-producing E. coli. This study highlights the prevalence of multi-antimicrobial resistant ESBL-producing E. coli in small-scaledpoultry farms and retail chicken, hence the need to review poultry management practices to minimize the occurrence.

单味中药水提物对鸡源耐药性E.coli地方株的体外抑菌活性研究

为了研究单味中药水提物对河北地区鸡源耐药性大肠埃希氏菌(Escherichia coli,E.coli)地方流行株QH15(O_2)、QH16(O_(38))、QH17(O_(89))的体外抑菌活性。利用平板琼脂打孔法和改良微量二倍稀释法分别测定抑菌圈直径和最小抑菌浓度(MIC)。结果表明:诃子、五味子、五倍子、黄连、黄芩、苦参6种中药均对3株鸡耐药性E.coli地方株高度敏感,抑菌圈直径在15~24 mm之间,MIC在7.825 mg/mL^31.25 mg/mL之间,其他的药物存在不同程度的敏感性。

Bacteriological and pathological studies of egg peritonitis in Commercial layer chicken in Namakkal area

Objective:To detect the various bacteriological agents and pathological changes in commercial layer chicken affected with egg yolk peritonitis in Namakkal region of India.Methods:A total of 6572 layer chicken from 85 commercial farms were subjected for the study,out of which 1715 showed various types of oviduct almoimalities.Among the 1715,264 birds from six farms were identified as egg peritonitis on the basis of postmortem examination.Trachea,lung,heart blood,liver,peritoneal exudate,oviduct(infundibulum,magnum,uterus)and cloacal swabs were collected from the 264 birds with egg peritonitis lesion for screening of bacterial agents.Signalment,clinical signs and pathological changes were recorded in the affected flocks.Result:The results of the present investigation indicated that the E.coli associated egg peritonitis was responsible for 15.39%of the reproductive tract abnormalities in commercial layers between 21 and 80 week of age.In the affected flocks egg production drop and mortality varied from 3%to 20%and 0.5%to 7.0%respectively.It was noticed during peak egg production(21 to 60week)and southwest monsoon season(58%).Statistical analysis of age,season and egg production by Chi square test of independence revealed highly significant difference.E.coli was isolated as a pure culture and concurrent with other bacterial agents in 226 and 38 birds respectively.Among the fifteen E.coli serotypes identified serotype O_(166),O_(?)and O_(111)were predominant.Necropsy examination of affected birds revealed the presence of amorphous or insipissiated yolk material in the abdominal cavity with inflammatory changes in the ovary,oviduct and intestine.Microscopically the oviduct surface epithelium showed degeneration and desquamation,moderate to marked infiltration of inflammatory cells especially heterophils and lymphocytes in various regions and lumen contained serofibrinous exudate,inflammatory and desquamated epithelial cells with bacterial microcolonies.Ovarian follicles revealed hyperemia,degeneration of granulosa cells and infiltration of inflammatory cells.Intestine showed degenerative,necrotic and inflammatory lesion.Conclusion:The findings of this study showed that the egg peritonitis might be caused by either the translocation of intestinal E.coli into the peritoneal cavity or by the movement of cloacal E.coli into the oviduct followed by ascension of these bacteria up the oviduct,through the infundibulum,and into the peritoneal cavity.To control the egg peritonitis faecal contamination with E.coli should be minimized.

Transmission identification of Escherichia coli aerosol in chicken houses to their environments using ERIC-PCR

In order to study E. coli aerosol spreading from chicken houses to their surrounding air, air samples, including indoor and outdoor air (upwind 10 and 50 m as well as downwind 10, 50, 100, 200 and 400 m away) of 5 chicken houses were collected using six-stage Andersen microbial samplers and Reuter-Centrifugal samplers (RCS). E. coli concentrations (CFU/m3 air) collected from different sampling sites were calculated. E. coli strains from chicken feces samples were also isolated. Furthermore, the enterobacterial repetitive intergenic consensus (ERIC)-PCR method was applied to amplify the isolated E. coli strain DNA samples. Through the genetic similarity analyses of the E. coli obtained from different sampling sites, the spreading of bioaerosol from animal houses to the ambient air was characterized. The results showed that the isolated E. coli concentrations in indoor air (9―63 CFU/m3) in 5 chicken houses were higher than those in upwind and downwind air, but there were no significant differences between the indoor and downwind sites 10 m away from all the 5 houses (P>0.05). The phylogenetic tree indicated that a part of the E. coli (34.1%) isolated from indoor air had 100% similarity with those isolated from feces, and that most of E. coli isolated (54.5%) from downwind at 10, 50, 100 or even 200 m had 100% similarity with those isolated from indoor air or feces too. But those isolated from upwind air had a lower similarity (73%―92%) with corresponding strains isolated from indoor air or feces. Our results suggested that some strains isolated from downwind air and indoor air originated in the chicken feces, but most of isolates obtained from upwind air samples did not come from the chicken feces or indoor air. Effective hygienic measures should be taken in animal farms to prevent or minimize downwind spreading of microorganism aerosol.

不同地区101个禽源性大肠杆菌分离株的致病性试验

从国内１８个省、市、自治区大肠杆菌病疑似病禽中分离并鉴定出大肠杆菌５９５株，鉴定出其中４４０个分离株的血清型。采用Ｒｏｓｅｎｂｅｒｇ氏报道的方法，测定了来自不同地区的６０个优势血清型分离株对ＳＰＦ鸡、３０个其他常见血清型及１１个国内少见血清型分离株对普通易感鸡的致病性。结果表明：所测定的１０１个分离株均为致病株，且９０％以上属高、中度致病株。

氧氟沙星对霉形体与大肠杆菌合并感染鸡的药效学研究

以试管两倍稀释法测得氧氟沙星及其对照药物恩诺沙星、洛美沙星和强力霉素对鸡败血霉形体的最小抑菌浓度分别为０．００６２５、０．０２５、０．４和０．４ｍｇ／Ｌ。５０、１００、２００ｍｇ／Ｌ氧氟沙星、５０ｍｇ／Ｌ恩诺沙星、５０ｍｇ／Ｌ洛美沙星和１００ｍｇ／Ｌ强力霉素连续５ｄ饮水给药，对人工合并感染败血霉形体和大肠杆菌病鸡的治愈率分别是９３．３％、９６．７％、９６．７％、１００％、９６．７％和８３．３％，感染对照组的死亡率为５３．３％。氧氟沙星及其它对照药的增重效果显著高于感染对照组（Ｐ＜０．０１），并能显著地减少气囊损伤率、抗体反应阳性率和病原再分离率。

鸡大肠杆菌病地方性致病菌株的分离鉴定及免疫研究

在吉林省３个地区患大肠杆菌病的鸡肝脏、心血中分离出１１５株致病性大肠杆菌，并进行了生理、生化、血清学、免疫学及抗药性鉴定。吉林省地方株菌体抗原为Ｏ８８、Ｏ９、Ｏ２４、Ｏ６８、Ｏ５的菌株占５３％，能抵抗多种抗生素，以菌体抗原Ｏ２４、Ｏ６８、Ｏ８８和１种未能鉴别出血清型的菌株为代表菌株制成灭活水苗和油佐剂苗，用于免疫２５万只鸡，保护期６个月，保护率达９８％以上。

禽病原性大肠杆菌1型菌毛的分离与鉴定

以旋涡混合法使禽病原性大肠杆菌分离株５６６ 、１７９４ 和ＴＫ３ 菌毛脱落，经硫酸铵沉淀、透析后进行蔗糖密度梯度离心，收集密度为１１０ 至１１５ｇ／ｃｍ３ 的蛋白带，经ＳＤＳＰＡＧＥ测定，３ 株菌菌毛蛋白的分子量分别在１７５ 、１７０ 和１７０ｋＤ；提纯菌毛保留了甘露糖敏感性凝集豚鼠红细胞的能力，证明它们为１ 型菌毛；从１７９４ 株提取的１ 型菌毛免疫ＢＡＬＢ／Ｃ 小鼠产生的高免血清在Ｗｅｓｔｅｒｎ ｂｌｏｔ 中与３ 个菌株的相应菌毛蛋白均呈阳性反应。上述结果表明，受检的３ 株禽病原性大肠杆菌均表达了１ 型菌毛，其分子量在１７５ ～１７０ｋＤ 之间，３ 个菌株的１

屠宰前鸡、猪源大肠杆菌耐药性调查

本试验旨在了解屠宰前鸡、猪源食品动物体内大肠杆菌耐药情况,分析潜在的食品安全问题。从广州市畜禽交易市场随机采集待屠宰鸡和猪的粪便样品,分离鉴定大肠杆菌,并采用琼脂稀释法检测大肠杆菌对15种抗菌药物的敏感性。结果显示,从658份猪源样品和133份鸡源样品中共分离鉴定出731株大肠杆菌,其中猪源606株,鸡源125株。药敏试验结果显示,731株大肠杆菌均表现出不同程度的耐药,耐药谱广且多重耐药现象严重。对复方新诺明和四环素的耐药率为90.0%以上,仅对头孢西丁、黏菌素和阿米卡星较敏感(耐药率均低于3%)。鸡源大肠杆菌对头孢噻肟、头孢曲松、新霉素、阿米卡星、萘啶酸和环丙沙星的耐药率显著高于猪源大肠杆菌(P<0.05)。鸡源大肠杆菌中3耐及3耐以上的菌株占97.60%,猪源大肠杆菌占94.72%。结果表明,屠宰前畜禽体内大肠杆菌对临床常用抗菌药物的耐药性非常严重,以多重耐药为主,且耐药谱丰富多样。提示屠宰前畜禽携带的耐药菌对食品安全和人类健康存在较大的安全隐患。

规模化蛋鸡养殖场大肠杆菌耐药性调查

从南宁周边某蛋鸡养殖场病死的雏鸡、5～12周龄青年鸡和蛋鸡中采集病料20份进行病原菌的分离、鉴定、动物致病性试验和血清型测定。分离出18株致病性大肠杆菌,主要有O50、O133、O88、O8、O785个血清型。用25种抗菌药物进行药物敏感性试验,结果表明,80%的分离菌株对美罗培南、亚胺培南、头孢噻肟高敏;80%的分离菌株对氨苄西林、苯唑西林、大观霉素、四环素和氯霉素产生不同程度的耐药性;100%的分离菌株对多粘菌素B、复方新诺明和链霉素耐药,说明该养殖场大肠杆菌耐药性非常严重。

大肠杆菌DNA作佐剂的鸡新城疫灭活疫苗的免疫效果

用大肠杆菌DNA作为鸡新城疫灭活苗的佐剂接种 15日龄雏鸡 ,以鸡新城疫油乳剂灭活苗为对照 ,用血凝抑制实验检测病毒特异性血凝抑制抗体。结果表明 ,大肠杆菌DNA及油乳剂两种佐剂均能增强特异性血凝抑制抗体的效价 ,二者组间抗体滴度差异不显著 ,但大肠杆菌DNA佐剂组雏鸡的体重明显高于油乳佐剂组 (P <0 0 1)。

河南省鸡致病性大肠杆菌血清型、耐药性的研究

对河南省3个不同地区的38株鸡致病性大肠杆菌进行了O血清型的鉴定、15种抗生素药物的耐药性试验。结果表明：河南省鸡致病性大肠杆菌的血清型种类众多,以O78,O1,O2为优势血清型,分别占定型菌株的35.29%,23.53%,17.65%;耐药图谱复杂,可以分为21种耐药类型,菌株耐药种类4-14种;不同来源的菌株其血清型、耐药性一般不同;相同来源的菌株,其耐药图谱相同或相似,但血清型有多种。

人工诱发鸡大肠杆菌病的接种规律

采用鸡大肠杆菌 O2标准强毒株 ,对 4 80羽宝万斯 -高兰蛋雏鸡颈部皮下注射进行人工发病 ,研究了细菌培养时间、接种量与致病力之间的关系和规律。结果表明 ,在同一毒株条件下 ,细菌致病力强弱 ,主要取决于接种感染菌数 ,只要接种菌数适宜 ,4～ 2 4 h细菌培养液均可成功发病 ,不必受普遍采用 1 8h细菌培养液接种的时间约束。这一试验结果可为灵活。

鸡舍内外环境中气载大肠杆菌同源性的分子鉴定

采用ANDERSEN-6级空气微生物样品收集器和RCS离心式采样器在5个鸡场舍内空气、舍外上风向和下风向不同距离收集气载大肠杆菌;并收集鸡的粪便,分离大肠杆菌。利用大肠杆菌基因间重复一致序列为引物的聚合酶链式反应（ERIC-PCR）分型技术,扩增不同测量点收集的大肠杆菌的DNA图谱。通过每个采样点的大肠杆菌的浓度变化以及大肠杆菌遗传相似性分析确认动物舍微生物气溶胶向舍外环境的传播。结果显示：5个鸡舍内空气中大肠杆菌的浓度远远高于舍外上风和下风向的大肠杆菌浓度（P〈0.05或P〈0.01）,但是舍外不同距离间的大肠杆菌浓度差异并不显著（P〉0.05）。同样,ERIC-PCR结果表明,从鸡的粪便中分离的大肠杆菌与从舍内空气中分离的部分大肠杆菌（34.1%）,以及从鸡场舍外下风方向分离到的多数大肠杆菌（54.5%）与从舍内空气或粪便中分离的大肠杆菌相似性可达100%。而从鸡舍上风向分离到的大肠杆菌与从舍内空气或粪便中分离的大肠杆菌相似性为73%-92%。从而说明来自动物体的大肠杆菌既能污染舍内空气,又能对其周围的环境构成污染。本研究揭示了微生物气溶胶的传播规律,具有公共卫生及流行病学意义。

乌梅中化合物V对鸡大肠杆菌的体内和体外杀灭作用研究

通过现代色谱方法应用硅胶、SephadexLH-20凝胶等材料分离了乌梅活性成分;应用波谱方法解析化合物结构,以体外试管倍比稀释法测最小抑菌浓度、体内试验评价防治鸡大肠杆菌病的潜在价值,对乌梅中提取化合物V的抗鸡大肠杆菌活性进行了初步评价。结果表明,以抗大肠杆菌活性为导向,从中药乌梅中分离得到一个单体成分,3-羟基-3-羧基戊二酸二甲酯(化合物V),体外试验显示化合物V的最小抑菌浓度为3.125mg/mL;体内试验显示肉鸡人工感染大肠杆菌后,胶膏、化合物V组、硫酸新霉素组、阴性感染对照组成活率分别为40%,60%,60%和20%;剖检后无菌取肝脏组织培养,以上各测试组大肠杆菌检出率分别为100%,80%,100%和100%。由此得出结论:通过体内、体外试验证明化合物V对肉鸡大肠杆菌病具有一定的预防效果。

鸡源性大肠杆菌产超广谱β-内酰胺酶基因型分布

检测鸡大肠杆菌超广谱β-内酰胺酶(ESBLs)的基因型并了解辽宁省鸡源性大肠杆菌产ESBLs基因型分布。对大连4个地区不同鸡场临床分离32株产ESBLs鸡大肠杆菌分别用TEM、OXA、CTX-M和SHV等4种通用引物进行PCR扩增确定其基因型。结果显示,32株鸡大肠杆菌均检出TEM型基因、12株检出CTX-M型基因、2株检出OXA型、未检出SHV型;TEM+CTX-M双阳性检出11株、TEM+OXA双阳性检出2株。TEM基因亚型占检出比例最高(100%)。目前辽宁省产ESBLs鸡大肠杆菌基因亚型以TEM为主,其次为CTX-M型,未发现SHV型;以TEM+CTX-M双阳性检出率最高(34.4%)。

不同省区鸡源大肠杆菌耐药性调查与系统进化群分析

为有效控制鸡大肠杆菌病,对不同省区鸡源大肠杆菌致病菌株的分布情况及耐药特征进行了调查分析,分别采用PCR法和MIC法进行了系统进化群分群检测和药敏试验,采用皮下注射进行了动物致病性试验,并采用SPSS17.0软件进行差异显著性分析。结果显示:低致病群B1(38.13%)和非致病群A(34.02%)菌株分布相对较多,其次是高致病群D(21.23%)和B2(6.16%);鸡源大肠杆菌耐药严重,12种药物的耐药率在50%以上,多重耐药问题突出,多重耐药率高达98.86%;不同省区鸡源大肠杆菌菌株的耐药程度存在差异,个别药物(如奥格门丁、头孢噻呋等)差异显著;高致病群B2和D群菌株的耐药程度较B1和A群菌株严重,其中氧氟沙星、庆大霉素、大观霉素、头孢噻呋、磺胺异噁唑、复方新诺明、奥格门丁等7种药物差异显著(P<0.05)。试验表明,我国鸡源大肠杆菌高致病菌株检出率高,且耐药程度相对严重,在适宜条件下更易导致鸡群发病且较难控制。

氟哌酸、环丙氟哌酸、泰妙灵治疗鸡败血霉形体与大肠杆菌合并感染的疗效比较

本研究对氟哌酸、环丙氟哌酸、泰妙灵作了体外抑菌试验与人工发病治疗试验。结果表明，环丙氟哌酸对鸡败血霉形体的体外抑菌强度为氟哌酸、泰妙灵的８倍，对鸡大肠杆菌的体外抑菌强度分别为氟哌酸、泰妙灵的４倍、８倍。在人工发病治疗试验中，环丙氟哌酸（１ｍｇ／ｋｇ）对鸡败血霉形体与大肠杆菌合并感染的疗效与泰妙灵（２０ｍｇ／ｋｇ）相当，明显优于氟哌酸（１０ｍｇ／ｋｇ），显示出广阔的临床推广应用前景。

3株鸡大肠埃希菌地方分离株I型菌毛的免疫原性

选择宁夏地区的大肠埃希菌3个优势血清型O15、O78、O18菌株,大规模培养后提取I型菌毛,制备天然菌毛亚单位疫苗,免疫接种7日龄雏鸡后通过攻毒试验和抗体水平测定评价疫苗的免疫效果。结果表明,未免疫鸡的死亡率为40%～100%,而免疫鸡用同源菌株攻毒后死亡率为0～10%,用异源菌株攻毒后死亡率为20%～70%,免疫组鸡肝脏、心包和气囊的病变程度均显著轻于未免疫鸡,免疫鸡血清内出现高滴度的ELISA抗体。