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Abstract

The aim of the study was to estimate the effect otthe composting process in the container technology Kneer on E. coli inactivation. The bacteria placed in the special carriers were introduced into the composted material. The elimination rate of E. coli differed depending on both the carriers· location in the biomass and the thermal conditions. The most effective hygienization, nr the material was noticed in summer - after 48 h in the middle layer, 6 days in the top layer and 10 days in the bottom layer. In spring and autumn, the bacteria survived the longest in the bottom layer - 85 and 45 days, respectively. Apart from the high temperature, the research points out the action of other factors such as competition, antagonism and antibiosis.
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Authors and Affiliations

Beata Szala
Zbigniew Paluszak
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Abstract

Escherichia coli producing extended-spectrum beta-lactamase (ESBL) are antimicrobial- -resistant Enterobacteriaceae important in the livestock production sector, especially dairy cows because these antimicrobial-resistant bacteria can be transferred to consumers via consumption. If antimicrobials are continually used in dairy farms, this may result in antimicrobial resistance. Therefore, investigation of antimicrobial resistance and finding new alternative methods for inhibiting ESBL-producing E. coli is essential. Hence, the aim of this study was to examine the ability of selected antimicrobials to inhibit E. coli ATCC 25922, control bacteria and ESBL-producing E. coli isolated from dairy farms. We also investigated the capacity of polyphenol extract from 10 varieties of medicinal plants to inhibit ESBL-producing E. coli using a broth microdilution method. It was found that control bacteria were susceptible to all antimicrobial agents, i.e., ampicillin, cefotaxime, ciprofloxacin, chloramphenicol, gentamycin, imipenem, nalidixic acid, tetracycline, and sulfamethoxazole/ trimethoprim. However, ESBL-producing E. coli exhibited both susceptibility and resistance to selected antimicrobials. The polyphenol extracted from Psidium guajava Linn at the lowest concentration was 4.5 mg/mL, which could inhibit control bacteria, but at the same concentration could not inhibit ESBL-producing E. coli. These phenomena indicated that ESBL-producing E. coli had both susceptibility and resistance to antimicrobials. Polyphenol, which could inhibit non-resistant E. coli, could not inhibit ESBL-producing E. coli.
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Authors and Affiliations

A. Konputtar
1 2
W. Aengwanich
1 3
B. Saraphol
1
M. Yossapol
1 2

  1. Faculty of Veterinary Sciences, Mahasarakham University, 79 Nakhonsawan Road, Talad, Maung, Maha Sarakham, 44000, Thailand
  2. Bioveterinary Research Unit, Mahasarakham University, 79 Nakhonsawan Road, Talad, Maung, Maha Sarakham, 44000, Thailand
  3. Stress and Oxidative Stress in Animals Research Unit, 79 Nakhonsawan Road, Talad, Maung, Mahasarakham University, Maha Sarakham 44000, Thailand
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Abstract

Members of Enterobacteriaceae are known to produce extended-spectrum beta-lactamases (ESBL) which hydrolyze the beta-lactam group of antibiotics. The existence of ESBL-producing Salmonella enterica ( S. enterica) and Escherichia coli ( E. coli) harbored by urban avifauna was investigated in this study. Dropping samples (n= 180) were collected from six different bird species in the district Jhang, Punjab province, Pakistan. Isolation and identification of ESBL isolates were made by using cefotaxime- (4 mg/L) supplemented MacConkey agar and double disc synergy test (DDST). Polymerase chain reaction (PCR) was performed for the detection of four different ESBL genes including bla CTX-M, bla TEM, bla SHV and bla OXA. A total of 42.69% isolates were confirmed as ESBL via DDST including 30.64% S. enterica and 49.54% E. coli. The incidence of ESBL S. enterica and ESBL E. coli was found highest in egret ( Ardea alba) and pigeon ( Columba livia) as 64.28% and 78.95%, respectively. The bla CTX-M gene was detected in 57.89% and 64.81% of isolates of S. enterica and E. coli, respectively. Among other genes in S. enterica and E. coli, bla TEM (21.05%, 20.4%); bla SHV (15.78%, 9.26%), and bla OXA (5.26%, 5.56%) were detected, respectively. All of the tested isolates were found resistant to at least one of the thirteen antimicrobial agents except meropenem. To the best of our knowledge, this is the first study reporting the incidence and genetic diversity of ESBL bacteria associated with urban avifauna in Pakistan. The urban avifauna can serve as a potential subject of bio-surveillance to monitor the emergence of antimicrobial-resistant bacteria.
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Authors and Affiliations

M.A. Saeed
1
U. Waheed
1
S. Ehtisham-ul-Haque
1
A.U. Khan
1
M. Kashif
2
M.F. Qamar
1
A. Ghafoor
3
M. Saqlain
1
J. Asghar
1

  1. Department of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang, 35200, Pakistan
  2. Department of Clinical Sciences, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang, 35200, Pakistan
  3. Institute of Microbiology, University of Veterinary and Animal Sciences, Outfall Road, Lahore, 54000, Pakistan
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Abstract

Neonatal calf diarrhea (NCD) is one of the most important concerns in cattle production. Escherichia coli is the most important bacterial agent of NCD. Although vaccination and antibiotic treatment are common in NCD, the high antigenic diversity of E. coli and the increase in antibiotic resistance cause difficulties in the control. The study aimed to investigate the rate of E. coli in calf diarrhea, isolate an agent of the NCD E. coli strain, determine antimicrobial resistance, and find out about some surface antigens. Fecal samples (n=115) were analyzed to isolate pathogenic E. coli strains with nine mixed infections; sixty-one strains isolate from fifty diarrhoeic calves. Among the isolates from diseased animals, 22 K99+STa+F41, 3 K99+STa, 3 strains F41, 2 strains Stx1, one strain K99, one strain eae, and one strain Stx2+eae were detected. 27 strains of F17- associated fimbriae have been identified. 17 strains F17a, 6 strains F111, 3 strains F17c, one strain carrying the F17a and F17c gene regions, whereas subfamily typing of one strain could not be performed. Serotypes were determined by molecular and serological methods: 32/61 (52.5%) isolates were O101 and 2/61 (3.3%) isolates were O9 serotypes. But 27 strain serotypes could not be detected. The antibiotic resistance profiles of the isolates were determined by the disc diffusion method. The resistance rates to antibiotics were trimethoprim- sulphamethoxazole 91.7%, ampicillin 86.7%, enrofloxacin 86.7%, gentamicin 45%, tobramycin 41.7%, cefotaxime 3.3%, and ceftazidime 1.7%. Due to increasing antibiotic resistance, prophylaxis is gaining importance. In further research, E. coli surface antigenic structures should be examined in detail, and it should form the basis for vaccine and hyperimmunization studies to be developed.
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Authors and Affiliations

M.R. Coşkun
1
M. Şahin
2

  1. Department of Microbiology, Faculty of Veterinary Medicine, Kafkas University, 36100, Kars, Turkey
  2. Department of Microbiology, Faculty of Veterinary Medicine, Kyrgyz-Turkish Manas University, 720038, Bishkek, Kyrgyzstan
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Abstract

Montmorillonite (MMT), a natural absorbent agent, has widely been accepted for its antidiarrhea function in human and farm animals; however, its specific physicochemical property limits its biological function in practical use. In the current study, raw MMT was loaded by andrographolide, namely andrographolide loaded montmorillonite (AGP-MMT). The microstructure of AGP-MMT was observed by scanning electron microscope (SEM) and X-ray diffraction (XRD). The effect of AGP-MMT on the growth performance, intestinal barrier and inflammation was investigated in an enterotoxigenic Escherichia coli (ETEC) challenged mice model. The results show that the microstructure of MMT was obviously changed after andrographolide modification: AGP-MMT exhibited a large number of spheroid particles, and floccule aggregates, but lower interplanar spacing compared with MMT. ETEC infection induced body weight losses and intestinal barrier function injury, as indicated by a lower villus height and ratio of villus height/crypt depth, whereas the serum levels of diamine oxidase (DAO), D-xylose and ETEC shedding were higher in the ETEC group compared with the CON group. Mice pretreated with AGP-MMT showed alleviated body weight losses and the intestinal barrier function injury induced by ETEC challenge. The villus height and the ratio of villus height/crypt depth, were higher in mice pretreated with AGP-MMT than those pretreated with equal levels of MMT. Pretreatment with AGP-MMT also alleviated the increased concentration of serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), and the corresponding genes in the jejunum induced by ETEC infection in mice. The protein and mRNA levels of IL-1β were lower in mice pretreated with AGP-MMT than those with equal levels of MMT. The results indicate that AGP-MMT was more effective in alleviating intestinal barrier injury and inflammation in mice with ETEC challenge than MMT.
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Bibliography

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Authors and Affiliations

P. Wang
1
L. Li
1
L. Gan
1
Q. Chen
1
H. Qiao
1
W. Gao
1
Y. Zhang
1
J. Wang
1

  1. College of Biology Engineering, Henan University of Technology, Zhengzhou, China
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Abstract

Studies were carried out in the summer seasons of l 995-1999 (from June to September) on the quantitative and qualitative composition of Enterobacteriaceae bacteria (including Escherichia coli and Salmonella sp.), and potentially pathogenic bacteria Aeromonas hydrophila, Pseudomonas aeruginosa and Staphylococcus sp. in the water of 8 bathing sites of the Lake Wigry. Aeromonas hydrophila occurred in the all samples of studied water and was the most numerous in water sampled from sites of increased trophic levels. Irrespective of the site and time of sampling Pseudomonas aeruginosa was rarely isolated. In the total of 160 samples of water analysed Salmonella sp.. Escherichia coli and Staphyloccocus aureus were determined in 32 (22.6%), 68 (42.4%) and 90 (55%) samples, respectively. Pathogenic bacteria of the genus Salmonella sp. and potentially pathogenic Staphylococcus aureus and Aeromonas hydrophila identified in the analysed offshore waters (also when Escherichia coli were absent) suggest that the use of the Lake Wigry waters for swimming, falling into account faecal bacterial counts, may not be sufficient to fully reflect safety conditions for bathers. The results of the research suggest that the evaluation of the Lake Wigry surface waters for recreational use should include the frequency of the occurrence of Salmonella sp., Staphylococcus aureus, Aeromonas hydrophila and Pseudomonas aeruginosa. These three species, which arc not directly linked to faecal contamination, can cause various diseases of the skin, nasal and oral cavities, eyes, internal car and other problems in people swimming in contaminated water.
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Authors and Affiliations

Ewa Korzeniewska
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Abstract

Enterotoxigenic Escherichia coli (ETEC) is the causative agent of a wide range of diseases, which are the important cause of illness and mortality in piglets. ETEC strains expressing F4 fimbriae are frequently associated with post-weaning diarrhea (PWD) and lead to great economic losses in swine production industry worldwide. The aim of this study was to establish a rapid and effective isothermal amplification method for detection of F4 fimbriae. Loop-mediated isothermal amplification (LAMP), Polymerase spiral reaction (PSR) and cross-priming ampli- fication (CPA) were used to develop and optimize the detection method first time. Subsequently, the specificity and sensitivity of these methods were evaluated, and the clinical samples were detected with these methods. All the F4-positive samples could produce ladder-like amplifica- tions products and lead the chromogenic substrate SYBR Green I produce green fluorescence, while in blank control and negative samples lack of this pattern or remained orange. The sensi- tivity of LAMP and CPA were 10 times higher than PSR method. Meanwhile, these three methods were validated with clinical samples, 7 were found positive, while 125 samples were negative, the testing results were consisted with the real-time PCR method. These findings suggested that the isothermal amplification based on the F4 fimbriae is a rapid, effective and sensitive method under resource constrains.

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Authors and Affiliations

L.Y. Zhao
J.H. Niu
X.L. Gao
C.N. Liu
S.M. Liu
N. Jiang
X.P. Lv
S.M. Zheng
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Abstract

Koumiss has beneficial therapeutic effects on bacterial diseases. Four antibacterial com- pounds from yeasts ( Kluyveromyces marxianus and Saccharomyces cerevisiae) in koumiss were evaluated for their antibacterial effects against three Gram-negative bacteria, three Gram-positive bacteria and five pathogenic Escherichia coli strains. The antibacterial compounds from yeasts in koumiss were extracted, and their main components were determined. The inhibition zones were analyzed, and their minimum inhibition concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined. Aqueous phases of Kluyveromyces marxianus and Saccharomyces cerevisiae at pH 2.0 and 8.0 produced larger inhibition zones than those in other phases, and then antibacterial compounds from K. marxianus (K2, pH=2.0; K8, pH=8.0) and S. cerevisiae (S2, pH=2.0; S8, pH=8.0) were obtained. Their main components were organic acids and killer toxins. K2 had more propanoic acid and S2 had more oxalic acid than others. The inhibition zones of K2, K8, S2 and S8 against three Gram-negative bacteria and three Gram-positive bacteria were 12.03-23.30 mm, their MICs were 0.01-0.13 g/mL, and MBCs were 0.03-0.50 g/mL. Meantime, the inhibition zones of K2, K8, S2 and S8 against five pathogenic E. coli were 16.10-25.26 mm, their MICs were 0.03-0.13 g/mL, and MBCs were 0.13-1.00 g/mL. These four antibacterial compounds from yeasts in koumiss had broad antibacterial spectrum. In addition, K2 and S2 were better than K8 and S8.
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Authors and Affiliations

Y.J. Chen
1 2
C.G. Du
1 3
Y.Q. Guo
1
Y.F. Zhao
1
C. Aorigele
2
C.J. Wang
3
H. Simujide
2
W. Aqima
2
X.Y. Zhang
1

  1. Vocational and Technical College, Inner Mongolia Agricultural University, Baotou 014109, P.R. China
  2. College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, P.R. China
  3. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, P.R. China

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