scholarly journals The Microscopic Detection and Identification of Acanthamoeba Spp. In Water Samples Following Enrichment by in Vitro-Cultivation Under Two Distinct Temperature Conditions.

Author(s):  
Sakhi Elmahdi ◽  
MOUMNI MOSTAFA ◽  
Radid Horia ◽  
ARAHOU MOHAMED ◽  
FEKHAOUI MOHAMED

Abstract Acanthamoeba is a genus of free-living amoeba commonly found in environmental sources such as water, soil and air, and can infect humans. There is a significant challenge in the detection and identification of members of this genus in water samples. In order attempt to overcome this challenge, we investigated using in vitro culture, under distinct temperature conditions, to grow and enrich amoeba prior to detection and identification. Aliquots of 150 water samples, collected from Rabat (30 from each river, fountain, sea, public bath and tap water), were individually inoculated into standard culture medium non-nutritive agar and incubated for two weeks at 25 °C and 30 °C under otherwise standard conditions. PCR was used to confirm the presence of Acanthamoeba DNA in positive samples. The findings showed that Acanthamoeba grew more rapidly at 30 °C than 25 °C, allowing improved microscopic detection and identification at the former temperature. This investigation shows clearly that the diagnostic sensitivity of an in vitro based culture system is temperature-dependent.

2000 ◽  
Vol 41 (7) ◽  
pp. 189-196 ◽  
Author(s):  
K. Guyot ◽  
M. F. Gireaudot-Liepmann ◽  
A. Cabon ◽  
I. Riveau-Ricard ◽  
M. Lange ◽  
...  

Viable Cryptosporidium parvum oocysts were processed by the US EPA 1622 method to determine if the procedure that requires successive filtration, elutionand centrifugation alters their integrity and viability (determined by in vitro excystation). Oocyst seeded in tap water samples were also used to evaluate recovery efficiencies and impact of the whole procedure on oocyst viability. Filtration through Envirochek Gelman cartridge was found not to damage oocysts. The use of Laureth-12 buffer during the elution step was shown to lead to greater spontaneous oocysts excystation than other phosphate buffers containing between 80 and/or SDS (like the Gelman buffer). However, this drawback was widely balanced against the best efficiency of this buffer to elute oocysts captured by the cartridge filter and therefore against its high recovery efficiency. Thus, in water samples in which the oocyst concentration is expected to be low, it is more advantageous to employ the Laureth-12 buffer for the elution through it can influence viability. Centrifugation speeds (1,000–5,000 g) did not alter oocysts.


2006 ◽  
Vol 52 (6) ◽  
pp. 584-590 ◽  
Author(s):  
Priscilla Declerck ◽  
Jonas Behets ◽  
Elke Lammertyn ◽  
Ilya Lebeau ◽  
Jozef Anné ◽  
...  

The presence of high levels of Legionella pneumophila in man-made aquatic systems correlates with the incidence of nosocomial Legionnaires' disease. This requires a rapid, reliable, and sensitive quantification of L. pneumophila concentrations in suspected water systems. In this research, a homologous competitor was developed and evaluated in a L. pneumophila competitive polymerase chain reaction (cPCR) to quantify this human pathogen in a quick, cost-effective, and reliable way. Accuracy of cPCR was evaluated by analyzing cooling tower and tap water samples spiked with known concentrations of L. pneumophila bacteria, in parallel with the standard culture method. Legionella pneumophila amounts detected and calculated from cPCR and culture correlated very well: r = 0.998, P = 0.002 for tap water and r = 0.990, P = 0.009 for cooling tower water. Nevertheless, for both kinds of water samples, mean numbers of L. pneumophila calculated from cPCR results were always higher than those obtained by culture. This study makes it clear that the rapid, sensitive, and cost-effective L. pneumophila cPCR is a promising alternative to the standard time-consuming culture method and expensive real-time PCR to enumerate L. pneumophila bacteria in environmental water samples.Key words: Legionella pneumophila, competitive PCR, cost-effective, cooling tower water, tap water, sensitive detection.


Author(s):  
Ahmad Z Al-Herrawy ◽  
Mohamed A Marouf ◽  
Mahmoud A. Gad

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


Author(s):  
V.V. Lapenko ◽  
L.N. Bikbulatova ◽  
E.M. Ternikova

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.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sujit Shah ◽  
Krishna Chand ◽  
Bhagwan Rekadwad ◽  
Yogesh S. Shouche ◽  
Jyotsna Sharma ◽  
...  

Abstract Background A plant growth-promoting endophytic bacterium PVL1 isolated from the leaf of Vanda cristata has the ability to colonize with roots of plants and protect the plant. PVL1 was isolated using laboratory synthetic media. 16S rRNA gene sequencing method has been employed for identification before and after root colonization ability. Results Original isolated and remunerated strain from colonized roots were identified as Bacillus spp. as per EzBiocloud database. The presence of bacteria in the root section of the plantlet was confirmed through Epifluorescence microscopy of colonized roots. The in-vitro plantlet colonized by PVL1 as well as DLMB attained higher growth than the control. PVL1 capable of producing plant beneficial phytohormone under in vitro cultivation. HPLC and GC-MS analysis suggest that colonized plants contain Indole Acetic Acid (IAA). The methanol extract of Bacillus spp., contains 0.015 μg in 1 μl concentration of IAA. PVL1 has the ability to produce antimicrobial compounds such as ethyl iso-allocholate, which exhibits immune restoring property. One-way ANOVA shows that results were statistically significant at P ≤ 0.05 level. Conclusions Hence, it has been concluded that Bacillus spp. PVL1 can promote plant growth through secretion of IAA during root colonization and ethyl iso-allocholate to protect plants from foreign infections. Thus, this study supports to support Koch’s postulates of bacteria establishment.


2021 ◽  
Vol 44 (1) ◽  
pp. 194-202
Author(s):  
Funda Demir ◽  
Meral Yildirim Ozen ◽  
Emek Moroydor Derun

Abstract In this study, essential (Ca, Cr, Cu, Fe, K, Mg, Na, P, Zn), and non-essential (Al, Ni, Pb) element contents of the drinking and baby water samples which are sold in the local market and tap water samples in Istanbul were examined. It was determined that elements of Cr, Cu, Fe, P, Zn, Al, and Ni were below detection limits in all water samples. Among the non-essential elements analyzed in water samples, Pb was the only detected element. At the same time, the percentages that meet the daily element requirements of infants were also calculated. As a result of the evaluations made, there is no significant difference in infant nutrition between baby waters and other drinking waters in terms of the element content.


2021 ◽  
Vol 1 (2) ◽  
Author(s):  
Diane G. Edmondson ◽  
Steven J. Norris

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