Airborne gram-negative bacterial flora in animal houses

The concentration and the species composition of airborne gram-negative bacteria were studied in four cattle houses, one pig house and one poultry barn. On average only between 0.02 and 5.2% of the total number of culturable aerobic bacteria were identified as gram-negative bacteria. Obligate anaerobic gram-negative bacteria were not isolated at all. In the airborne gram-negative bacterial flora the following bacterial families dominated: the Enterobacteriaceae, the Pseudomonadaceae and the Neisseriaceae. Within the family of the Enterobacteriaceae the species Escherichia coli and Enterobacter agglomerans were predominant. In animal houses using straw as bedding material Ent. agglomerans was most frequent, whereas in animal houses without litter E. coli was mainly found. Airborne Neisseriaceae were isolated very frequently in cow barns with Acinetobacter lowffii as the primary species. Airborne Pseudomonadaceae were found in high concentrations during periods of high air humidity. The results presented may also give some indications on the origin and sources of airborne endotoxins in animal housing.     

Air microorganisms in animal housing--4. Airborne gram-negative bacteria and airborne endotoxin in pig houses

The species composition of airborne gram-negative bacteria and the relationship between inhalable endotoxin, inhalable dust and airborne bacteria were studied in 4 pig houses. The airborne gram-negative bacterial flora was dominated by Enterobacteriaceae. Within the Enterobacteriaceae the species E. coli and Enterobacter agglomerans were predominant. Significant correlation were found between the concentration of inhalable dust and inhalable endotoxin as well as between the concentration of airborne gram-negative bacteria and inhalable endotoxin. However these correlation were not very strong. With respect to the characterization of the potential hazard of organic dust exposure, measurements of the concentration of airborne dust or airborne bacteria should not be used for the estimation of the concentration of airborne endotoxin.     

禽舍微生物气溶胶含量及其空气动力学研究

采用Andersen-微生物空气样品收集器，选用普通营养琼脂和金黄色葡萄球菌选择培养基对一个种鸡场舍环境空气进行监测。其需氧菌含量从3.12×104到9.01×105，金黄色葡萄球菌含量波动于2.0×103～3.3×104CFU/m3之间。根据微生物气溶胶颗粒在Andersen-收集器不同层级上的分离情况得知，22.5%的需氧菌、1.8%的金黄色葡萄球菌气溶胶颗粒的空气动力学直径(d50)为Φ0.65～2.1μm，它们能进入人、畜的肺泡，对人畜呼吸道构成感染威胁。     

规模化驴场空气中细菌数量与种类的调查与分析

[目的]分析规模化驴场空气环境中细菌数量与种类,指导驴场防御和控制细菌性疾病的方向。[方法]从不同开放模式的驴舍用自然沉降法收集其内部空气中细菌样本及场区周边不同距离的空气细菌样本,使用平皿法对采集的细菌进行培养、计数、鉴别。[结果]驴场内空气中细菌总数的范围8.39×10~3~3.43×10~4CFU/m~3,保育驴舍（封闭式驴舍）的细菌总数高于分娩驴舍（半开放式驴舍）及种公驴舍（开放式驴舍）,其中种公驴舍细菌总数最低。驴场对周边空气环境的污染范围为200 m。主要优势菌落是葡萄球菌、链球菌、大肠杆菌、绿脓杆菌等。[结论]规模化驴场空气环境中的细菌种类繁多,驴舍内细菌数量与其开放方式有关,且细菌数量随距驴场距离的增加而减少。     

Airborne Gram‐Negative Bacterial Flora in Animal Houses

The concentration and the species composition of airborne gram‐negative bacteria were studied in four cattle houses, one pig house and one poultry barn. On average only between 0.02 and 5.2 % of the total number of culturable aerobic bacteria were identified as gram‐negative bacteria. Obligate anaerobic gram‐negative bacteria were not isolated at all. In the airborne gram‐negative bacterial flora the following bacterial families dominated: the Enterobacteriaceae, the Pseudomonadaceae and the Neisseriaceae. Within the family of the Enterobacteriaceae the species Escherichia coli and Enterobacter agglomerans were predominant. In animal houses using straw as bedding material Ent. agglomerans was most frequent, whereas in animal houses without litter E. coli was mainly found. Airborne Neisseriaceae were isolated very frequently in cow barns with Acinetobacter lowffii as the primary species. Airborne Pseudomonadaceae were found in high concentrations during periods of high air humidity. The results presented may also give some indications on the origin and sources of airborne endotoxins in animal housing.     

Effects of chemical treatments on pH and bacterial population in poultry litter: a laboratory experiment

1. The goal of this study was to evaluate the effects of three chemical treatments on pH and bacterial populations (total aerobic bacteria and gram-negative bacteria) in poultry litter under laboratory conditions. 2. Litter obtained from poultry houses was treated with three chemical treatments (alum, AlCl(3) and FeSO(4)) at the same concentration (8 g/100 g litter), while untreated litter served as a control. The study was conducted for 3 weeks. 3. All of the chemical treatments reduced total aerobic bacteria (22 to 87% of the untreated control) and gram-negative bacteria (63 to 99% of the untreated controls) populations and lowered litter pH values (5.95 to 6.64). However, a significant difference in gram-negative bacteria did not exist among chemical treatments at 0, 1 and 2 weeks. 4. These results suggest that the reduction in total aerobic bacteria and gram-negative bacteria populations is highly related to a decrease in litter pH, and acidifying treatment (alum, AlCl(3) and FeSO(4)) of poultry litter may serve as a means to help the reduction in pathogen populations and to improve economical benefits under commercial production conditions.     

Bacterial shell contamination in the egg collection chains of different housing systems for laying hens

The bacterial eggshell contamination of eating eggs in different commercial housing systems; two conventional cages, one organic aviary system and one barn production, were compared. The total counts of aerobic bacteria and the total counts of Gram-negative bacteria on the shell were used to detect key points where contamination occurred and to study the progress of contamination in the egg collection and transportation chains. The key points in the chain were those where eggs accumulated on a short conveyor belt, initial shell contamination in the alternative housing systems and extra nest-boxes placed on the ground. The high bacterial load of floor eggs (>6.3 log CFU total aerobic flora/eggshell) explains why they cannot be used for eating. On average higher initial shell contamination with total counts of aerobic bacteria was found for eggs from the alternative housing systems compared to the conventional systems; respectively 5.46 compared to 5.08 log CFU/eggshell. However, initial contamination with total counts of Gram-negative bacteria on the shells was less in the alternative systems: 3.31 compared to 3.85 log CFU/shell. Initial bacterial shell contamination tended to correlate positively with the concentration of bacteria in the air of the poultry houses. Storing shell eggs, whether temporarily refrigerated or not, for 9 d or more, resulted in a decrease in bacterial eggshell contamination for both bacterial variables.     

不同乳牛舍环境空气中细菌含量的比较研究

采用自然沉降法对南京3个规模化奶牛场和3个奶牛小区泌乳奶牛舍内细菌含量进行监测和比较。结果显示,牛舍环境空气中的细菌菌落总数,不同的牛舍不同采样点、不同的培养基其结果均不相同。运动场细菌菌落总数少于牛舍内。NA培养基菌落数最高达33.81×103 CFU/m3,MAC和S.S培养基在多个分布点无细菌生长。6个牧场空气中的细菌菌落总数为F场＞E场＞B场＞C场＞A场＞D场。牛舍的空气细菌总数与牛舍规模化程度无关,而与各牧场牛舍卫生管理有很大的相关性。从270个样本中共分离到338株细菌,A、B牧场主要是葡萄球菌,分别占分离菌的75.34%（55/73）和60.61%（40/66）;C、E、F牧场主要是肠杆菌,分别占分离菌的51.22%（42/82）、62.16%（23/37）、61.11%（33/54）;D牧场葡萄球菌占分离菌的46.15%（12/26）,肠杆菌和链球菌分别占26.92%（7/26）。结果表明,规模化奶牛场和奶牛小区空气细菌总数无明显差异。     

Relationship between the concentration of different bioaerosol components and the general hygienic condition in two pig fattening houses

In the study presented here the relationship between the hygienic condition and the concentration of different air contaminants in two pig fattening houses was investigated. Deficiencies in routine cleaning, which were characterized by an increased soiling of animals and stable surfaces with faeces, led to increased concentrations of airborne dust, airborne endotoxins as well as increased numbers of total airborne bacteria and airborne gram-negative bacteria. However these deficiencies were not reflected by the concentration of airborne Clostridium perfringens which were earlier postulated as an indicator of faecal pollution in the air of cattle barns. Furthermore the results of this study were used to discuss advantages and disadvantages of impingement and impaction for the sampling of airborne microorganisms in animal houses.     

畜禽舍微生物气溶胶向环境扩散的研究

应用一个扩散模式估算来自畜禽舍的微生物气溶胶向环境的扩散。在此考虑到了较大悬载尘埃颗粒的沉落和气悬状态下细菌生物学活性对空气微生物传播的影响。菌源高度 (栏舍废气排放孔 )认作 3m和 1 0m。当扩散级为IV和菌源强度为 1 0 6CFU/s时 ,在距离畜禽舍 1 0 0 ,2 0 0 ,30 0 ,40 0 ,5 0 0m处细菌含量分别为 5 0 ,1 0 0 ,2 0 0 ,5 0 0和 1 0 0 0CFU/m3。研究证明 ,畜禽舍之间保持一定距离将很大程度地减少气源性感染的威胁 ;畜牧场应建设在距居民区最少 5 0 0m的地方。     

市售5种消毒剂对舍饲鸭场的杀菌效果评价

为了解贵州省某舍饲鸭场环境中菌落分布情况以及市售五种常用消毒剂(苯扎溴铵、月苄三甲氯铵、复方戊二醛、戊二醛癸甲溴铵、聚维酮碘)的灭菌效果。通过采集鸭场空气、粪便、垫料、饮水样本进行菌落总数测定;分别配制最小杀菌浓度的苯扎溴铵、月卞三甲氯铵、复方戊二醛及戊二醛癸甲溴铵4种消毒剂,采用雾线、喷洒消毒方式对舍饲鸭场空气、漏缝地板、料槽、蛋框及运输车车轮进行现场消毒;配制最小杀菌浓度的聚维酮碘,采用涂抹方式对鸭蹼部皮肤进行涂抹消毒;最后分别测定鸭场空气、漏缝地板、料槽、蛋框、运输车车轮及鸭蹼样本消毒前后的菌落总数并计算5种消毒剂对细菌的杀菌率。结果显示：舍饲鸭场空气菌落总数为3.79×10⁴ CFU/m³,超出NY/T 388-1999畜禽场环境质量标准规定数值(25000 CFU/m³);粪便菌落总数为2.67×10⁷ CFU/g,超出GB18596-2001畜禽养殖业污染物排放标准规定数值(1.03×10⁶CFU/g);垫料菌落总数为3.7×10⁶CFU/g,超出NY/T1167-2006畜禽场环境质量及卫生控制规范规定的清洁土壤数值(5×10⁴CFU/g);饮水菌落总数为12 CFU/mL,符合GB 5749-2006生活饮用水卫生标准(100CFU/mL)。5种消毒剂对鸭场进行现场消毒试验,复方戊二醛对漏缝地板、蛋框、运输车车轮的细菌杀菌率最高,分别为96.78%、81.31%、87.5%;戊二醛癸甲溴铵对鸭舍空气、料槽的细菌杀菌率最高,分别为34.85%、74.35%;聚维酮碘对鸭蹼部皮肤细菌杀菌率为72.45%;4种消毒剂对漏缝地板、料槽、蛋框、运输车车轮消毒效果良好,对空气的消毒效果不理想;聚维酮碘对鸭蹼部皮肤消毒效果不理想。结果表明：该舍饲鸭场存在大量超过国标菌落总数的安全隐患,通过对该鸭场微生物菌落计数及现场消杀试验,在一定程度将菌落总数控制在了合理范围。研究结果旨在为掌握舍饲鸭场微生物菌落分布情况,制定舍饲鸭场消毒程序以及科学防控养鸭场细菌性疾病提供参考依据。     

Effect of an electrostatic space charge system on airborne dust and subsequent potential transmission of microorganisms to broiler breeder pullets by airborne dust

High levels of dust and microorganisms are known to be associated with animal confinement rearing facilities. Many of the microorganisms are carried by dust particles, thus providing an excellent vector for horizontal disease transmission between birds. Two environmentally controlled rooms containing female broiler breeder pullets (n = 300) were used to evaluate the effectiveness of an electrostatic space charge system (ESCS) in reducing airborne dust and gram-negative bacteria levels over an 8-wk period (starting when the birds were 10 wk old). The ESCS was used to evaluate the effectiveness of reducing airborne microorganism levels by charging airborne dust particles and causing the particles to be attracted to grounded surfaces (i.e., walls, floor, equipment). The use of the ESCS resulted in a 64% mean reduction in gram-negative bacteria. Airborne dust levels were reduced an average of 37% over a 1-wk period in the experimental room compared with the control room on the basis of samples taken every 10 min. The reductions of airborne dust and bacteria in this study are comparable with earlier results obtained with the ESCS in commercial hatching cabinets and experimental caged layer rooms, suggesting the system could also be applied to other types of enclosed animal housing.     

Prevalent Serotypes of Pasteurella multocida Isolated from Different Animal and Avian Species in India

Identification and estimation of the prevalence of Pasteurella multocida organisms in different animal and avian species in India during November 2000 to July 2003 was carried out. Out of 418 samples collected from different outbreaks suspected to be caused by P. multocida, a total of 206 bacterial cultures were identified as P. multocida on the basis of cultural, morphological and biochemical characteristics. All the 206 cultures were isolated from different domestic animal species (cattle, buffalo, sheep, goat, pig and rabbit), avian species (chicken, duck, quail, turkey, goose) and wild animals such as leopard and deer. Serotyping of P. multocida cultures revealed the presence of various serotypes (A:1, A:3, A:1,3, A:4, B:2, D:1 and -:1) among the livestock population. P. multocida polymerase chain reaction (PCR) assay applied on different forms of bacterial cultures (bacterial culture lysate, direct bacterial colony and mixed bacterial culture lysate) yielded an amplified product of approximately 460 bp specific for P. multocida. The results of PCR assay correlated well with conventional methods of identification. The present investigation revealed the presence of varied serotypes among livestock and PCR assay was found to be useful for rapid, sensitive and specific diagnosis of pasteurellosis in animals and avian species.     

Bacterial eggshell contamination in conventional cages, furnished cages and aviary housing systems for laying hens

The influence of housing system on the initial bacterial contamination of the eggshell was studied. Two long-term experiments were performed. Bacterial eggshell contamination, as expressed by total count of aerobic and Gram-negative bacteria, was periodically analysed for eggs from a conventional cage, a furnished cage with nest boxes containing artificial turf or grids as nest-floor material and an aviary housing system. Results were log-transformed prior to statistical analyses. For both experiments no systematic differences were found between the conventional cage and furnished cage. The type of nest-floor material in the nest boxes of the furnished cages also did not systematically influence the bacterial contamination. A possible seasonal influence on contamination with a decrease in the winter period (up to > 0.5 log cfu/eggshell) of total count of aerobic and Gram-negative bacteria was observed in the first experiment. The contamination with total aerobic flora was higher (more than 1.0 log) on eggs from the aviary housing system compared to the conventional and the furnished cage systems. For Gram-negative bacteria this was not the case. During the entire period of both experiments, independent of housing system, shell contamination was not influenced by age of hens or period since placing the birds in the houses. For the total count of aerobic bacteria a restricted positive correlation (r2 = 0.66) was found between the concentration of total bacteria in the air of the poultry houses and initial shell contamination.     

Long-term study of aerobic bacteria of the genital tract in stud dogs.

The aerobic bacterial flora of the genital tract was characterized in 15 stud dogs in an 18-month study. The dogs represented 4 breeds and were from 3 kennels. Bacterial samples from the prepuce and semen were collected every month, except in connection with matings, when they were collected weekly (464 samples). The dogs that were included all mated at least once during the study. The mean pregnancy rate, litter size, and pup mortality for the bitches with which they had mated were all within normal limits. The most frequent bacteria isolated from the prepuce and semen were Pasteurella multocida, beta-hemolytic streptococci, and Escherichia coli. There was a tendency for breeds to differ in frequency of the most common bacterial species. Bacterial culture yielded no aerobic growth in 14.2% of the preputial samples and 69.8% of the semen samples. Bacteria were transferred between dog and bitch at mating. In this study of healthy breeding dogs, neither the fertility of the dog nor that of the bitch was affected by the bacteria transferred.     

Effect of electrostatic space charge on reduction of airborne transmission of Salmonella and other bacteria in broiler breeders in production and their progeny

Salmonella in birds is a concern because of the human foodborne illness associated with the consumption of poultry meat and eggs. One of the methods of transmission of Salmonella within a flock can be by the air. Therefore, we used reduction of transmission of Salmonella to monitor the effectiveness of the electrostatic space charge system (ESCS). During the average broiler breeder laying cycle of 40 wk, a large amount of dust becomes airborne and accumulates on walls, ceiling, and equipment. Many microorganisms adhere to these dust particles, making dust an excellent vector for horizontal disease transmission between birds.We used two environmentally controlled rooms containing commercial broiler breeders to evaluate the effectiveness of an ESCS that produced a strong negative electrostatic charge to reduce airborne dust and, subsequently, microorganism levels. The ESCS caused the dust to become negatively charged, therefore moving to the grounded floor in the treatment room. The use of the ESCS resulted in a significant reduction (P < 0.0001, 61% reduction) in airborne dust concentration levels, which resulted in a significant reduction (P < 0.0001, 76% reduction) in total airborne bacteria and gram-negative bacteria (48% reduction) in the treatment room. Significant reductions (P < 0.05) of gram-negative bacteria (63% reduction) on the egg collection belts were also recorded in the treatment room, which resulted in a significant reduction (P < 0.0001) of gram-negative bacteria (28% reduction) on the eggshell surface. The ESCS treatment resulted in fewer Salmonella enteritidis-positive hens and their progeny from the treatment room due to reductions of dust and airborne bacteria. In addition, this significant reduction in bacteria on the eggshell surface should result in less bacteria in the day-old chicks, therefore better early chick livability. There was no significant difference (P > 0.05) in egg production, male or female body weights, mortality, or reproductive performance in the ESCS room compared with the control room.     

Effects of oral administration of metronidazole on small intestinal bacteria and nutrients of cats

OBJECTIVE: To determine effects of oral administration of metronidazole on the number and species of duodenal bacteria and selective nutrients of cats. ANIMALS: 6 healthy domestic shorthair cats. PROCEDURE: Undiluted duodenal fluid was obtained for quantitative and qualitative bacterial culture to determine species and number of bacteria in healthy cats. Blood samples were assayed for taurine, total protein, albumin, cobalamin, and folate concentrations. Cats then were given metronidazole (20 mg/kg of body weight, PO, q 12 h) for 1 month, after which bacterial cultures and serum assays of nutrients were repeated. Nine months after cessation of antibiotic treatment, duodenal bacteria were re-evaluated and serum was assayed for total protein, albumin, cobalamin, and folate concentrations. RESULTS: Oral administration of metronidazole caused a significant decrease in aerobic and anaerobic bacterial counts in the duodenum of healthy cats, accompanied by emergence of Streptococcus spp and Corynebacterium spp. Serum concentrations of cobalamin and albumin increased when duodenal bacterial counts were decreased, although changes in folate or taurine concentrations were not detected. Measured variables did not differ, when comparing results obtained before and 9 months after cessation of metronidazole. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of metronidazole decreased the number of aerobic bacteria and altered indigenous flora in the small bowel of cats. Normal duodenal flora appeared to be stable, because species of bacteria were re-established by 9 months after cessation of metronidazole. Bacterial flora appeared to have an impact on nutrients, because albumin and cobalamin increased during antibiotic administration and returned to preadministration concentrations after cessation of the antimicrobial.     

应用巢氏PCR方法快速检测猪鼻拭子样品中产毒素多杀性巴氏杆菌

根据产毒素多杀性巴氏杆菌的toxA基因序列,设计了2对特异性引物,扩增的片段大小分别为864和447 bp,从而建立了一种能直接从猪鼻拭子中快速检测产毒素多杀性巴氏杆菌的巢氏PCR方法。该方法能检出26CFU菌量以上的模板DNA,且不能从猪的其它7种常见病原菌扩增到特异性条带。通过对5个不同地区阳性猪群中采集的146份临床鼻拭子样品进行巢氏PCR检测和细菌分离鉴定,结果2种方法同时为阳性的样品有44份,同时为阴性的样品有97份,另有5份样品只有巢氏PCR检测为阳性,巢氏PCR检测与细菌分离鉴定的符合率为96.58%。试验表明:巢氏PCR方法能快速、灵敏地从猪鼻拭子样品中检出产毒素多杀性巴氏杆菌,适合临床进行大规模病原学检测,具有良好的应用前景。     

CORNEO-CONJUNCTIVAL MICROFLORA OF CLINICALLY NORMAL SYRIAN HAMSTERS (MESOCRICETUS AURATUS)

This study was performed to determine the normal aerobic bacterial flora of the cornea and conjunctiva in Syrian hamsters. Eleven healthy adult Syrian hamsters were used. Collection of specimens was performed using sterile micro-swab applicators. Immediately after sample collection, microbiologic aerobic culture was initiated. Fourteen eyes (63%) showed bacterial growth and a total number of 19 different species were isolated which belonged to 7 bacterial genera. Gram-positive bacteria were the most prominent with 83.3% (20/24) of isolates. Results of this study could help veterinarians in the diagnosis and therapeutic monitoring of surface ocular disease in this species.     

Intracellular locations of dermonecrotic toxins in Pasteurella multocida and in Bordetella bronchiseptica

Location of dermonecrotic toxin (DNT) in the cells of Pasteurella multocida or Bordetella bronchiseptica was investigated. After cell lysis by various procedures, various fractions prepared from bacterial cells grown in liquid culture media were assayed for dermonecrotic activity by skin testing of guinea pigs. During the death phase of the growth tested for the 2 bacterial species, little cell-free DNT was detected in the culture supernatants. Throughout the log and stationary phases of the growth, DNT activity was cell associated, but was not seen in the culture supernatants, which indicated that DNT was not secreted by actively growing P multocida or B bronchiseptica cells. Little DNT was released by subjecting whole cells to osmotic shock, a common procedure that releases proteins from the periplasmic space of many gram-negative bacteria. After sonication and centrifugation of whole cells, a substantial amount of DNT was released; results were similar when spheroplasts were used instead of whole cells. Treatment of whole cells with trypsin did not decrease the DNT activity, but trypsin treatment of sonicated cells resulted in a significant decrease in the DNT activity (P less than 0.01). The results indicated an intracellular location of the DNT of P multocida or B bronchiseptica. The DNT of P multocida or of B bronchiseptica is probably located in the cytoplasmic space.