Amphotericin B Resistant Strain Development

Can I Request a Specific Writer?

Yes, if you have worked with a writer before, you can request them again for consistency and familiarity with your needs. This ensures a smooth, tailored experience. Writer continuity helps maintain consistent style and understanding of your preferences. Browse profiles or contact support to make your choice. Essay writer relationships build over time for increasingly personalized academic support.

In vitro activity of Malia azedarach Linn. Against amphotericn B resistant Leishmania tropica strain

Abstract

Cutaneous leishmaniasis is still a big health problem around the world and drug resistance has emerged as a major problem in treatment. The purpose of the study was to generate Amphotericin B resistant strain and evaluate the crude extract of Melia azedarach against wild and lab generated resistant strain of Leishmania tropica. The result obtained revealed that the continuous increase of drug pressure for 60 days cause resistance in L. tropica by 09 fold compared to wild type. The LC₅₀ value recorded for wild and resistant type strain was 0.024 and 0.224µg/ml, respectively against Amp B drug. The antileishmanial activity of crude extracts against wild and resistant type strain was 0.57 and 123.3 µg/ml for green fruit and 1220.8 and 4010.1µg/ml for ripe fruit, respectively. A significant difference (P<0.05) in activities was observed between green and ripe fruit against wild and resistant type of strain. Based on the results obtained, it can be concluded that green fruit show promising results against resistant type of L. tropica and could be a novel candidate for anti-protozoal activity.

Keywords: Leishmania tropica, Melia azedarach, Resistance.

How Do You Ensure On-Time Delivery?

We assign writers immediately and monitor progress, with backup plans to guarantee your paper arrives on schedule. Your deadlines are our priority. Our project tracking system alerts us to potential delays for proactive intervention. Rest assured, we will deliver when you need it. My assignment help includes stringent deadline monitoring for reliable punctual delivery.

Introduction

Infections associated with protozoa are a global health problem, predominantly in the Third World countries (Kondrashin, Baranova et al. 2010, Tengku and Norhayati 2011, Waldron, Ferrari et al. 2011, Al Musayeib, Mothana et al. 2013), and roughly 14% of the world population are at risk. Protozoal infections are considered as a neglected tropical disease and therefore, immense concern has been shown by WHO (Organization 2010). These deadly tropical diseases have negative impacts on the socioeconomic status of the patients (Meshnick 1997). There is a study so far conducted on protozoan infections, including charges, malaria, sleep sickness and leishmaniasis. These diseases considered a major killing factor due to different difficulties are connected with controlling the parasite, high cost, resistant, poor safety, low efficacy and toxicity to the drug (Nwaka and Ridley 2003). Among these protozoan infections leishmaniasis got a special interest in various developing countries due to the increase of resistance to the drug (Pérez-Victoria, Pérez-Victoria et al. 2001).

Leishmania is a fatal disease caused by parasite Leishmania and transmitted through female phlebotomines sand fly bite. Leishmaniasis present in three different clinical forms, i.e. cutaneous leishmaniasis, visceral leishmaniasis and muco-cutaneous leishmaniasis. It is present in tropical and subtropical areas of 88 countries of the world and approximately 10 million population of the world is suffering from cutaneous leishmaniasis (Organization 2007).

Toxicity, inconsistent efficacy and resistant between species or strains of the drug mostly used or for some of them, the long lasting parenteral administration is the need of the day led the researcher for novel antileishmanial drug, most particularly from plant source used in traditional medicine, as a foundation of new leads with new mechanisms of actions (Hoet, Stévigny et al. 2004).

Can You Help with Reflective Essays?

Our writers craft thoughtful reflective essays, blending personal insight with academic structure to meet your goals. We capture your voice perfectly. Reflective writing requires balancing personal experience with scholarly analysis. Share your story or guidelines to get started. Research study bay professionals excel at combining personal narrative with academic rigor.

The present study focused on the in vitro antileishmanial activity of crude extract of M. azedarach against wild and lab produced resistant type L. tropica strain.

Material and methods

Collection of Plant Materials

Fresh and ripe fruit of the plant was collected from the vicinity of Quaid-i-Azam University, Islamabad. The plant materials were washed with distilled water and kept in the shade till process. The plant materials were authenticated by the Department of Plant Sciences, Quaid-i-Azam University. A voucher specimen was then submitted to the herbarium.

Extraction

What Is Your Quality Control Process?

Every paper is reviewed by editors for accuracy, style, and originality, ensuring it meets high academic standards. We use advanced tools to guarantee quality. Our multi-tiered review system includes both automated checks and expert human evaluation. Your paper will exceed expectations every time. Paper writing quality assurance involves multiple verification stages before delivery.

Fruit (1 Kg) was macerated using mortar and pestle. The powdered obtained were soaked in 5 liter plastic beaker containing distilled water for 6 days with occasional shaking. The materials were filtered through muslin cloth and dried under reduce pressure using a rotary evaporator (Heidolph Laborta4000 efficient). The paste like materials obtained were dried at 40^oC in water bath and stored at 4^OC for further use.

Production of resistant strain

Resistant strain of L. tropica was developed according to the protocol described by Seifert et al. with slight modification (Seifert, Matu et al. 2003). The wild type culture was maintained in M199 medium with 10% heat inactivated Fetal Bovine Serum. Briefly, drug pressure was generated in step wise manner for 60 days and the sensitivity of strain to Amp B was investigated. The culture showed resistance was maintained in drug pressure for further use at 24^oC.

In vitro Antileishmanial Assay

Antileishmanial assay was performed according to the procedure described by Khan et al. 2014. Promastigotes of L. tropica strain were cultured in M199 media (Invitrogen, USA) with 10% Fetal Bovine Serum (PAA, GmbH). Briefly, stock solution of the test samples and standard drug Amphotericin B drug (10,000 ppm) were prepared in distilled water. Promastigotes were harvested at 1×10⁶ cells/ml and inoculated in 96 well plate. The assay was performed at different concentration (1000-0.001 ppm). For negative control, sterilized distilled water was used. The 96 well plate was incubated for 48 h at 24^OC. The live promastigotes were counted using improved Neubauer chamber under light microscope (40x magnification). The results obtained were statistically analyzed using SPSS Version 21 software (Khan, Yasinzai1 et al. 2014).

Can You Include Specific Sources?

Yes, provide your sources or let our writers find credible ones, properly cited to support your paper. Every source is cited perfectly. We integrate your required readings seamlessly while maintaining academic citation standards. Share your materials in the order form for best results. Assignment writer expertise ensures proper source integration and citation accuracy.

Statistical Analysis

All the experiment was carried out in triplicate. For comparison Post Hoc Multiple comparison test was applied using SPSS. Ver. 21 software. All the data were given mean ± standard deviation (M±SD).

Results

Resistance in L. tropica against Amp B was developed by a step wise increase of drug pressure. The sensitivity of Amp B against wild and resistant type given in Table 1. A significant different (P<0.05) in LC₅₀ value was observed for wild and resistant type 0.024 and 0.224 µg/ml, respectively. An increase of 09 fold was observed for resistant type compared to wild type.

The antileishmanial activity of green fruit (LC₅₀ value 0.57µg/ml) significant different (P<0.05) when compared to ripe fruit (LC₅₀ value 1220.8µg/ml). The LC₅₀ values for green and ripe fruit against wild type strain are depicted in Table 2. At concentration 0.05µg/ml, the percent survival for green fruit is 95±2.22%, whereas 75.0±3.51% for ripe fruit is recorded at 500µg/ml concentration (Figure 1).

Can You Write for Professional Programs?

We create papers for MBA, law, or medical programs, with writers who understand their unique requirements. Our expertise ensures top-quality results. Professional programs demand industry-specific knowledge and specialized writing conventions. Provide your program details for a customized paper. Ace tutors with professional credentials deliver field-appropriate academic writing.

A significant difference (P<0.05) in activity was also observed between green and ripe fruit against resistant type strain. The LC₅₀ value established for green and ripe fruit is 123.3µg/ml and 4010.1µg/ml, respectively as shown in Table 2. The percent survival of the parasite against green and ripe fruit is depicted in Figure 2. The results indicated that at concentration 50µg/ml, the percent survival of the green and ripe fruit is 85.0±3.5% and 100%, respectively.

Discussion

Parasitic protozoan infection constitutes a major health problem in developing countries. Resistance shown by the parasite and limited knowledge on the mechanism(s) on which these parasites acquire resistance cause additional burden in the development of practical public health policies for parasite control (Grögl, Oduola et al. 1989). The term “resistance” describes the irresponsiveness of a microorganism to an antimicrobial drug as investigated in vitro and compared with other isolates of the same species. While, clinical failure of a drug for appropriate treatment may include antimicrobial resistance, impaired immune function, accelerated metabolism and poor availability of the given drug. Primary resistance is the resistance that occurs in an organism never before to that specific drug of choice, while secondary resistance, also known as acquired resistance occurs after exposure to the drug (Rex, Pfaller et al. 1997). The resistance of the Leishmania to the Amp B was studied. Amp B a polyene antibiotic is the second line treatment for leishmaniasis, binds specifically to ergosterol, the main sterol present of Leishmania, trypanosome cruzi and fungi (Davidson, Scott et al. 1991). Amp B resistant promastigotes lines were selected by step by step increase in drug pressure, as previously shown by other antileishmanial drugs (Arana, Pérez–Victoria et al. 1998). A resistant type of parasite was generated at 0.1 µg/ml in M199 medium as parasite found dead at 0.2 µg/ml of concentration. In this context, plant originated drug showed promising results to overcome resistance. We have previously reported the potential application of green and ripe fruit of M. azedarach against wild type L. tropica strain and the results indicated that green fruit significantly (P<0.05) reduced the parasites than ripe fruit (Khan, Yasinzai1 et al. 2014), which is parallel to our findings. The effect of green fruit on resistant type L. tropica strain was significantly higher than ripe fruit. The elevated activity of the green fruit may be attributed to the high amount of lemonoids and azadirachtin present or may be due to some novel compounds (Khan, Yasinzai1 et al. 2014).