scholarly journals Comparison of Methods for Processing Drinking Water Samples for the Isolation of Mycobacterium avium and Mycobacterium intracellulare

2008 ◽  
Vol 74 (10) ◽  
pp. 3094-3098 ◽  
Author(s):  
Rachel Thomson ◽  
Robyn Carter ◽  
Chris Gilpin ◽  
Chris Coulter ◽  
Megan Hargreaves
Keyword(s):  
Mycobacterium Avium ◽  
Water Samples ◽  
Culture Media ◽  
Tap Water ◽  
Cetylpyridinium Chloride ◽  
Sodium Thiosulfate ◽  
Mycobacterium Intracellulare ◽  
Solid Media ◽  
Lowenstein Jensen ◽  
Mycobacterial Growth

ABSTRACT Several protocols for isolation of mycobacteria from water exist, but there is no established standard method. This study compared methods of processing potable water samples for the isolation of Mycobacterium avium and Mycobacterium intracellulare using spiked sterilized water and tap water decontaminated using 0.005% cetylpyridinium chloride (CPC). Samples were concentrated by centrifugation or filtration and inoculated onto Middlebrook 7H10 and 7H11 plates and Lowenstein-Jensen slants and into mycobacterial growth indicator tubes with or without polymyxin, azlocillin, nalidixic acid, trimethoprim, and amphotericin B. The solid media were incubated at 32°C, at 35°C, and at 35°C with CO2 and read weekly. The results suggest that filtration of water for the isolation of mycobacteria is a more sensitive method for concentration than centrifugation. The addition of sodium thiosulfate may not be necessary and may reduce the yield. Middlebrook M7H10 and 7H11 were equally sensitive culture media. CPC decontamination, while effective for reducing growth of contaminants, also significantly reduces mycobacterial numbers. There was no difference at 3 weeks between the different incubation temperatures.

scholarly journals Multicenter Evaluation of the BACTEC MGIT 960 System for Recovery of Mycobacteria

1999 ◽  
Vol 37 (3) ◽  
pp. 748-752 ◽  
Author(s):  
Bruce A. Hanna ◽  
Adeleh Ebrahimzadeh ◽  
L. Bruce Elliott ◽  
Margie A. Morgan ◽  
Susan M. Novak ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Liquid Medium ◽  
Mycobacterium Avium ◽  
Culture Systems ◽  
Solid Media ◽  
Lowenstein Jensen ◽  
The Mean

We evaluated the BACTEC MGIT 960 system, which is a fully automated, noninvasive system for the growth and detection of mycobacteria with a capacity to incubate and continuously monitor 960 7-ml culture tubes. We studied 3,330 specimens, 2,210 respiratory and 1,120 nonrespiratory specimens, collected from 2,346 patients treated at six sites. Processed specimens were inoculated into the BACTEC MGIT 960 and BACTEC 460 TB systems, as well as onto Lowenstein-Jensen slants and Middlebrook 7H11/7H11 selective plates. From all culture systems, a total of 362 isolates of mycobacteria were recovered; these were recovered from 353 specimens collected from 247 patients. The greatest number of isolates of mycobacteria (289, or 80% of the 362 isolates) was recovered with the BACTEC MGIT 960, followed by the BACTEC 460 TB (271, or 75%) and solid media (250, or 69%). From all culture systems a total of 132 isolates of Mycobacterium tuberculosiscomplex were recovered. The greatest number of isolates of M. tuberculosis complex was recovered when liquid medium was combined with conventional solid media; the number recovered with BACTEC 460 TB plus solid media was 128 (97%), that recovered with BACTEC MGIT 960 plus solid media was 121 (92%), that recovered with BACTEC 460 TB was 119 (90%) and that recovered with all solid media combined was 105 (79%). The recovery with BACTEC MGIT 960 alone was 102 (77%). The mean times to detection (TTD) for M. tuberculosis complex were 14.4 days for BACTEC MGIT, 15.2 days for BACTEC 460 TB, and 24.1 days for solid media. The numbers of isolates of Mycobacterium avium complex (MAC) recovered were 172 (100%) for all systems, 147 (85%) for BACTEC MGIT 960, 123 (72%) for BACTEC 460 TB, and 106 (62%) for all solid media combined. The TTD for MAC in each system were 10.0 days for BACTEC MGIT 960, 10.4 days for BACTEC 460 TB, and 25.9 days for solid media. Breakthrough contamination rates (percentages of isolates) for each of the systems were 8.1% for BACTEC MGIT 960, 4.9% for BACTEC 460 TB, and 21.1% for all solid media combined.

scholarly journals Hospital water filters as a source of Mycobacterium avium complex

2010 ◽  
Vol 59 (10) ◽  
pp. 1198-1202 ◽  
Author(s):  
Joseph O. Falkinham
Keyword(s):  
Water Supply ◽  
Mycobacterium Avium ◽  
Water Samples ◽  
Repetitive Sequence ◽  
Mycobacterium Avium Complex ◽  
Mycobacterium Intracellulare ◽  
Fingerprint Pattern ◽  
Water Filters ◽  
Mycobacterium Malmoense

Bronchoscopes and the filters used for washing them were found to yield high numbers of Mycobacterium avium isolates sharing the same repetitive sequence-based PCR (rep-PCR) fingerprint pattern as M. avium isolates recovered from patient samples collected by bronchoscopy. Water and biofilm samples collected from the bronchoscopy preparation laboratory yielded M. avium, Mycobacterium intracellulare, Mycobacterium malmoense and Mycobacterium gordonae. Several M. avium and M. intracellulare isolates from water samples in the bronchoscopy laboratory had rep-PCR patterns matching those of patient bronchoscopy isolates. Five of the 22 (23 %) M. avium patient bronchoscopy isolates and 42 of the 56 (75 %) M. intracellulare patient bronchoscopy isolates could have been due to contamination from the water supply.

Culture of Helicobacter pylori from domestic water samples – the impact of strain variation on growth on solid and in liquid media

2006 ◽  
Vol 54 (3) ◽  
pp. 147-152 ◽  
Author(s):  
R.J. Owen ◽  
S.A. Chisholm ◽  
G. Brick ◽  
J.V. Lee ◽  
S. Surman-Lee ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Culture Media ◽  
Tap Water ◽  
Clinical Isolates ◽  
Strain Variation ◽  
Liquid Media ◽  
Domestic Water ◽  
Source Of Infection ◽  
Solid Media ◽  
The Impact

Helicobacter pylori is an important global human pathogen and there is growing evidence from PCR assays that contaminated drinking water might be a possible source of infection in some circumstances. There are no validated protocols for direct isolation but various culture media have been developed for possible environmental sampling. Our aim here was to investigate how inter-strain variation might affect the interpretation of results with such media. Two laboratory adapted reference strains and four recent clinical isolates were tested on four solid media and in ten liquid media. Considerable variation was found between strains in their ability to recover on the different media after stress exposure (suspension in sterile tap water). Generally, clinical isolates were less robust than the laboratory-adapted strains and, overall, the former required longer recovery times. Our findings highlighted the importance of using a range of isolates for evaluations, as examination of laboratory-adapted strains alone did not provide an accurate representation of the utility of media that may be used to recover H. pylori from water.

scholarly journals Effect of Cetylpyridinium Chloride (CPC) on Colony Formation of Common Nontuberculous Mycobacteria

Pathogens ◽  
2018 ◽  
Vol 7 (4) ◽  
pp. 79 ◽  
Author(s):  
Myra Williams ◽  
Joseph Falkinham
Keyword(s):  
Water Samples ◽  
Colony Formation ◽  
Nontuberculous Mycobacteria ◽  
Cetylpyridinium Chloride ◽  
T Test ◽  
Solid Media

Cetylpyridinium chloride (CPC) is widely used to decontaminate water samples for the cultivation of nontuberculous mycobacteria (NTM). The rationale for using CPC is that it kills more non mycobacteria than NTM and thereby prevents the outgrowth and detection of mycobacterial colonies on solid media. The few CPC-susceptibility measurements that have been published, suggest that CPC-decontamination does kill significant numbers of NTM. We confirm that observation here and further demonstrate that CPC-susceptibility varied significantly by one log between representative NTM species and between strains of the same species. CPC-susceptibility was the same for cells collected from cultures or water-acclimated (P = 0.6485, T-test) and CPC-susceptibility was relatively similar over the range of commonly employed CPC dosages. We conclude that use of CPC as decontaminating agent may lead to failure to recover an NTM isolate and considerable underestimates of NTM numbers.

scholarly journals Bacterial survival rate on toothbrushes and their decontamination with antimicrobial solutions

2004 ◽  
Vol 12 (2) ◽  
pp. 99-103 ◽  
Author(s):  
Sandra Sato ◽  
Izabel Yoko Ito ◽  
Elza Helena Guimarães Lara ◽  
Heitor Panzeri ◽  
Rubens Ferreira de Albuquerque Junior ◽  
...  
Keyword(s):  
Survival Rate ◽  
Culture Media ◽  
Tap Water ◽  
Cetylpyridinium Chloride ◽  
Anaerobic Bacteria ◽  
Colony Count ◽  
Black Pigment ◽  
Bacterial Survival ◽  
Gram Negative ◽  
Bacterial Counts

The purpose of this study was to evaluate bacterial survival rate on toothbrushes after brushing and the efficacy of their decontamination by spraying antimicrobial solutions. Thirty subjects were instructed to spray the solutions on toothbrush bristles after brushing. Each volunteer tested three sprays, one solution per week; the sprays were labeled spray 1 (cetylpyridinium chloride - CPC - and basic formulation), 2 (basic formulation only) and 3 (control - sterile tap water). At the end of each week, the brushes were collected and sonicated in Letheen Broth®; the suspensions were ten-fold diluted and the dilutions were plated on various culture media. Anaerobic bacteria, evaluated by colony count of black pigment producing organisms on Ask medium, were recovered from 83.3% of the samples, Streptococci from 80% and aerobic Gram-negative bacilli from 46.7% of them in the control tests. There was a significant decrease in toothbrush contamination with antimicrobial sprays 1 and 2, the first showing the greatest decrease on bacterial counts.

scholarly journals Comparison of Four Different Culture Media for Isolation and Growth of Type II and Type I/III Mycobacterium avium subsp. paratuberculosis Strains Isolated from Cattle and Goats

2006 ◽  
Vol 72 (9) ◽  
pp. 5927-5932 ◽  
Author(s):  
Luc�a de Juan ◽  
Julio �lvarez ◽  
Beatriz Romero ◽  
Javier Bezos ◽  
Elena Castellanos ◽  
...  
Keyword(s):  
Incubation Period ◽  
Mycobacterium Avium ◽  
Culture Media ◽  
Egg Yolk ◽  
Disease Diagnosis ◽  
Type I ◽  
Type Ii ◽  
Sodium Pyruvate ◽  
Tissue Samples ◽  
Solid Media

ABSTRACT Culture is considered the definitive technique for Johne's disease diagnosis, and it is essential for later applications of certain molecular typing techniques. In this study, we have tested four solid media (Herrold's egg yolk medium [HEYM] with sodium pyruvate and mycobactin [HEYMm-SP], HEYM with mycobactin and without sodium pyruvate [HEYMm], Middlebrook 7H11 with mycobactin [Mm], and L�wenstein-Jensen with mycobactin [LJm]) for isolation of Mycobacterium avium subsp. paratuberculosis strains in 319 tissue samples from cattle herds and goat flocks. We have shown that each of the two main groups of M. avium subsp. paratuberculosis (type II and type I/III) has different requirements for growth in the culture media studied. The recommended solid media for isolation of type I/III strains are LJm and Mm, since the combination of both media allowed the recovery of all these strains. The most widespread culture medium, HEYM, is not suitable for the isolation of this group of M. avium subsp. paratuberculosis strains. Regarding the type II strains, HEYMm-SP was the medium where more strains were isolated, but the other three media are also needed in order to recover all type II strains. The incubation period is also related to the strain type. In conclusion, because the type of strain cannot be known in advance of culture, coupled with the fact that cattle and goats can be infected with both groups of strains, we recommend the use of the four solid media and the prolongation of the incubation period to more than 6 months to detect paratuberculous herds/flocks and to determine the true prevalence of the infection.

scholarly journals Comparison of four culture media for isolation of Mycobacterium avium complex from porcine tissues

1979 ◽  
Vol 9 (2) ◽  
pp. 194-196
Author(s):  
C O Thoen ◽  
E M Himes ◽  
J L Jarnagin ◽  
R Harrington
Keyword(s):  
Mycobacterium Avium ◽  
Mycobacterium Avium Complex ◽  
Culture Media ◽  
Agar Medium ◽  
Egg Yolk ◽  
Sodium Pyruvate ◽  
Acid Fast Bacilli ◽  
Lowenstein Jensen ◽  
Time Required ◽  
Porcine Tissues

The efficiency of four culture media was compared for the isolation of Mycobacterium avium complex from 197 procine tissues. In 82 tissues with microscopic granulomas and acid-fast bacilli, a significantly greater number of isolates were obtained on Middlebrook 7H10 medium with sodium pyruvate than on Stonebrink medium, Herrold egg yolk agar medium, or Lowenstein-Jensen medium (P=0.01). In 46 tissues in which no microscopic granulomas or acid-fast bacilli were observed, a significantly greater number of isolates were made on Middlebrook 7H10 medium or Herrold egg yolk agar medium than on Stonebrink medium or on Lowenstein-Jensen medium (P=0.01). The time required to grow M. avium complex on Lowenstein-Jensen medium was significantly greater than the time required to observe growth on Stonebrink, Middlebrook 7H10, or Herrold egg yolk agar medium (p=0.001).

Acanthamoeba Species in Tap Water, Egypt

2017 ◽  
Vol 9 (1) ◽  
Author(s):  
Ahmad Z Al-Herrawy ◽  
Mohamed A Marouf ◽  
Mahmoud A. Gad
Keyword(s):  
Water Samples ◽  
Tap Water ◽  
Pcr Analysis ◽  
Pulmonary Infections ◽  
Clinical Syndromes ◽  
Acanthamoeba Species ◽  
Amebic Encephalitis ◽  
Acanthamoeba Culbertsoni ◽  
Autumn And Winter

Genus Acanthamoeba causes 3 clinical syndromes amebic keratitis, granulomatous amebic encephalitis and disseminated granulomatous amebic disease (eg, sinus, skin and pulmonary infections). A total of 144 tap water samples were collected from Giza governorate, Egypt. Samples were processed for detection of Acanthamoeba species using non-nutrient agar (NNA) and were incubated at 30oC. The isolates of Acanthamoeba were identified to species level based on the morphologic criteria. Molecular characterization of the Acanthamoeba isolates to genus level was performed by using PCR. The obtained results showed that the highest occurrence percentage of Acanthamoeba species in water samples was observed in summer season (38.9%), then it decreased to be 30.6% in spring and 25% in each of autumn and winter. PCR analysis showed that 100% of 43 Acanthamoeba morphologically positive samples were positive by genus specific primer. In the present study eight species of Acanthamoeba can be morphologically recognized namely Acanthamoeba triangularis, Acanthamoeba echinulata, Acanthamoeba astronyxis, Acanthamoeba comandoni, Acanthamoeba griffini, Acanthamoeba culbertsoni, Acanthamoeba quina and Acanthamoeba lenticulata. In conclusion, the most common Acanthamoeba species in tap water was Acanthamoeba comandoni

ECOLOGICAL AND PHYSIOLOGICAL ASSESSMENT OF CHEMICAL COMPOSITION OF DRINKING TAP WATER IN KHANTY-MANSIYSK AND SALEKHARD

2020 ◽  
pp. 159-167
Author(s):  
V.V. Lapenko ◽  
L.N. Bikbulatova ◽  
E.M. Ternikova
Keyword(s):  
Drinking Water ◽  
Chemical Composition ◽  
Water Samples ◽  
Tap Water ◽  
Iron Concentration ◽  
Absorption Spectrometry ◽  
Northern Region ◽  
Transport Processes ◽  
Magnesium Concentration ◽  
Geochemical Environment

Water is very important for humans, as it is a solvent for metabolic products. Moreover, it is necessary for metabolism, biochemical and transport processes. The elemental status in persons depends on the geochemical environment and consumption of bioelements with food and water. The aim of the paper is to conduct a comprehensive assessment of chemical composition of drinking tap water in Khanty-Mansiysk and Salekhard. Materials and Methods. The chemical composition of 100 samples of drinking tap water was analyzed by atomic absorption spectrometry, spectrophotometry and capillary electrophoresis. All in all, there were 50 samples from Khanty-Mansiysk and 50 samples from Salekhard. The results were compared with Sanitary Rules and Norms 2.1.4.1074-01. Results. Drinking tap water in Salekhard contains a significantly higher concentration of iron, which is much above the maximum allowable concentration, if compared to water samples in Khanty-Mansiysk (p=0.03). In the cities under consideration, the water undergoes high-quality reagent-free treatment. However, the deterioration of the water supply networks in Salekhard is 3 times as high as in Khanty-Mansiysk. Calcium and magnesium concentration in water samples from Khanty-Mansiysk is 5.6 and 3.9 times lower than the MAC; in water samples from Salekhard calcium concentration is 6.3 (p=0.008) and magnesium concentration 4.6 (p<0.001) times lower than the MAC. Conclusion. The consumption of ultra-fresh drinking water leads to low intake of bioelements, which are a part of enzymes contributing to the human antioxidant defense and can result in manifestation of cardiovascular diseases. This is especially true for Salekhard with very soft drinking water and high iron concentration, which excess can exhibit prooxidant properties. Keywords: tap water, bioelements, northern region, antioxidants. Вода является важнейшим соединением для человека: необходима в качестве растворителя продуктов метаболизма и протекания обменных, биохимических и транспортных процессов. Элементный статус организма человека зависит от геохимического окружения и поступления биоэлементов с пищей и водой. Цель. Провести комплексную оценку химического состава водопроводной воды городов Ханты-Мансийск и Салехард. Материалы и методы. Методами атомно-абсорбционной спектрометрии, спектрофотометрии и капиллярного электрофореза проанализирован химический состав 100 проб водопроводной воды: по 50 из Ханты-Мансийска и Салехарда. Результаты сравнивали с СанПиН 2.1.4.1074-01. Результаты. В водопроводной воде Салехарда установлена превышающая ПДК и достоверно более высокая концентрация железа сравнительно с водой Ханты-Мансийска (р=0,03). При условии качественной безреагентной водоподготовки в изучаемых городах это обусловлено изношенностью водопроводных сетей в Салехарде, более чем в 3 раза превышающей этот показатель в Ханты-Мансийске. Концентрация кальция и магния в воде Ханты-Мансийска в 5,6 и 3,9 раза ниже ПДК; в воде Салехарда – в 6,3 (р=0,008) и 4,6 (р<0,001) раза ниже ПДК соответственно. Заключение. Употребление ультрапресной питьевой воды на фоне очень малого поступления с водой биоэлементов, входящих в состав ферментов антиоксидантной защиты организма человека, может привести к манифестации кардиоваскулярных заболеваний. Это особенно актуально для г. Салехарда с очень мягкой питьевой водой с повышенным содержанием железа, избыток которого может проявлять прооксидантные свойства. Ключевые слова: водопроводная вода, биоэлементы, северный регион, антиоксиданты.

Performance of Different Solid and Liquid Culture Media for the Improvement of Tuberculin Production

2020 ◽  
Keyword(s):  
Culture Media ◽  
Synthetic Medium ◽  
Chemical Properties ◽  
Economic Loss ◽  
Delayed Type Hypersensitivity ◽  
Basic Medium ◽  
Primary Methods ◽  
Lowenstein Jensen ◽  
Time Required ◽  
Modified Media

Tuberculosis (TB) is one of the most important zoonotic bacterial diseases. A huge economic loss which could be direct or indirect are associated with the disease. Currently, the primary methods used for detection of TB in humans and cattle include the measurement of a delayed type hypersensitivity to purified protein derivative (PPD). So, the need for preparation of purified PPD with adequate properties and increasing the final PPD yield with decreasing the time of tuberculin production has stimulated the interest in the development of its preparation. Our study was performed to compare between the standard and modified media for improving tuberculin production. Middle brook 7H10 agar medium was used as a modified basic medium for mycobacterial growth, followed by cultivation of mycobacteria on Middle brook 7H9 broth medium. For the production, strains were inoculated onto the culture medium (Dorest Henly synthetic medium). Other steps for tuberculin production was done according to standard Weighbridge protocol. The results demonstrated that the using of both Middle brook 7H10 agar and Middle brook 7H9 broth instead of Lowenstein-Jensen (LJ) and glycerin broth media which used in currently produced tuberculin, have better physical and chemical properties. In addition, reducing the time required for production by accelerating the time of microbial growth. Also, it was found that the tuberculin produced using modified media was slightly more potent or the same as currently tuberculin produced. So, both Middle brook 7H10 agar and Middle brook 7H9 broth media are recommended for production of tuberculin saving time and increasing potency of the product but more investigation was recommended for estimation types of protein present in both locally prepared and modified tuberculin.

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