EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen

Miraglia, Fabiana; Moraes, Zenaide Maria; Melville, Priscilla Anne; Dias, Ricardo Augusto; Vasconcellos, Sílvio Arruda

doi:10.1590/s1517-83822009000100033

Cited by 8 publications

(3 citation statements)

References 13 publications

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“…It is important to highlight that the sample (bovine semen) was experimentally spiked, what may have influenced the results (Miraglia et al . ). Regarding 5‐FU associated with serial dilution, a similar rate to that observed in the present study (3·8%) was reported on urine of naturally infected goats (Lilenbaum et al .…”

Section: Resultsmentioning

confidence: 97%

Usage of a selective media (EMJH-STAFF) in primary culturing of pathogenic leptospires from bovine clinical samples

Loureiro

Martins

Pinto

et al. 2015

Lett Appl Microbiol

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The slow growth of leptospires and overgrowth of co-existing micro-organisms from environmental and microbiota are the major difficult to recovery Leptospira from animal clinical samples. Implementing an efficient control programme is essential to determine circulating leptospires in the region and their reservoirs. This study evaluated the relationship of a selective media (EMJH-STAFF) on the recovery of pathogenic leptospires (Leptospira noguchii, Leptospira santarosai and Leptospira interrogans), from bovine clinical samples (urine and vaginal fluid). EMJH-STAFF seems to be an important tool in obtaining local strains for epidemiological and control purposes.

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Section: Resultsmentioning

confidence: 97%

Usage of a selective media (EMJH-STAFF) in primary culturing of pathogenic leptospires from bovine clinical samples

Loureiro

Martins

Pinto

et al. 2015

Lett Appl Microbiol

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“…The inoculum was prepared by diluting a seven-day old Leptospira culture with fresh Ellinghausen-McCullough-Johnson-Harris (EMJH) broth to reach a bacterial density of 2 × 10 6 CFU/mL by using a spectrophotometer at the 420 nm wavelength. The optical density (OD) of 0.32 at 420 nm approximately corresponded to 1 × 10 8 CFU/mL (16). The OD of 0.064 was approximately equal to 2 × 10 7 CFU/mL, and a 10-fold dilution was performed to reach a leptospiral density of 2 × 10 6 CFU/mL.…”

Section: Methodsmentioning

confidence: 99%

Anti-microbial Activity of Aqueous Quercus infectoria Gall Extract against Pathogenic Leptospira

Mustafa¹,

İsmail²,

Wahab³

2018

MJMS

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BackgroundQuercus infectoria gall extract is known to have broad spectrum anti-microbial activity in vitro. This study was conducted to determine the anti-microbial activity of Q. infectoria gall extract against pathogenic Leptospira and to evaluate the morphological changes of extract-treated cells using a scanning electron microscope (SEM).MethodsA two-fold serial microdilution broth assay was used to determine the minimum inhibitory concentration (MIC) of aqueous Q. infectoria gall extract against the L. interrogans serovar Javanica and the L. interrogans serovar Icterohaemorrhagiae, at concentrations ranging from 4.00 mg/mL to 0.0078 mg/mL. The minimum bactericidal concentration (MBC) was determined by sub-culturing the broth from the microtiter plate wells that showed no apparent growth or turbidity to the freshly prepared Ellinghausen-McCullough-Johnson-Harris (EMJH) broth media, and it was subsequently observed under a dark field microscope following three weeks of incubation for purposes of growth detection. The cell morphology of both extract-treated and untreated L. interrogans serovar Icterohaemorhagiae was analysed using the SEM.ResultsThe results of the broth microdilution assay demonstrate that the aqueous Q. infectoria gall extract possesses anti-microbial activity against both of the L. interrogans serovars with MIC values of 0.125 mg/mL. The MBC values for the L. interrogans serovar Javanica and the L. interrogans serovar Icterohaemorhagiae are 0.125 mg/mL and 0.250 mg/mL, respectively. The SEM micrograph shows changes in shape and size of the extract-treated cells (at 8× MIC) in comparison to the untreated cells.ConclusionThe Q. infectoria gall extract displays anti-microbial inhibition and killing activity against the pathogenic Leptospira isolates, and thus has the potential for further exploration of its efficacy and use in the treatment of leptospirosis.

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“…Throughout the experiment clinical monitoring of animals was done. ), termed 'selective medium' (Miraglia et al, 2009), using the serial dilution technique, in order to evaluate the percentage recovery of the added micro-organism. These cultures were incubated at an average temperature of 29 uC.…”

Section: Methodsmentioning

confidence: 99%

Cholinesterases as markers of the inflammatory process in rats infected with Leptospira interrogans serovar Icterohaemorrhagiae

Silva

Wolkmer

Silva³

et al. 2012

Journal of Medical Microbiology

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The aim of this study was evaluate changes in the cholinesterase activity in blood, lymphocytes and serum of rats infected with Leptospira interrogans serovar Icterohaemorrhagiae ('L. icterohaemorrhagiae'). Sixty adult Wistar rats were divided into six groups of 10 animals: three control groups and three test groups. The animals from the test groups were intraperitoneally inoculated with 1 ml medium containing 1¾10 8 leptospires. The activity of acetylcholinesterase in blood and butyrylcholinesterase in serum increased on days 5 (P,0.05) and 30 (P,0.021) post-infection, respectively. A decrease in lymphocyte count was observed on days 15 (P,0.01) and 30 post-infection (P,0.05). On day 15 post-infection, acetylcholinesterase activity (P,0.001) in lymphocytes decreased in infected rats. However, on day 30 post-infection there was an increase in acetylcholinesterase activity in lymphocytes. In conclusion, our results showed that the activity of enzymes of the cholinergic system in the total blood, lymphocytes and serum is altered as a result of inflammation caused by infection with L. icterohaemorrhagiae. The possible causes of these alterations will be discussed in this paper.

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EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen

Cited by 8 publications

References 13 publications

Usage of a selective media (EMJH-STAFF) in primary culturing of pathogenic leptospires from bovine clinical samples

Usage of a selective media (EMJH-STAFF) in primary culturing of pathogenic leptospires from bovine clinical samples

Anti-microbial Activity of Aqueous Quercus infectoria Gall Extract against Pathogenic Leptospira

Cholinesterases as markers of the inflammatory process in rats infected with Leptospira interrogans serovar Icterohaemorrhagiae

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