Low-cost liquid medium for in vitro cultivation of Leishmania parasites in low-income countries

Background: Cutaneous Leishmaniasis (CL) induced by Leishmania aethiopica has two clinical manifestations: ulcerating, self-healing CL and non-ulcerating, non-healing CL. The grossly disfiguring multiple nodules on the face and exterior surface of limbs during non-ulcerative CL are sometimes misdiagnosed as other skin infections. Thus the need for definitive and prompt laboratory diagnosis will be required. Identifying Leishmania parasite by culture method is considered as a definitive method for initiation of treatment and as an effective component of leishmaniasis control methods. Recently the involvement of Fas (CD95) and Tumor Necrosis Factor (TNF) Related Apoptosis Inducing Ligand (TRAIL) induced apoptotic pathways were proposed to be involved in tissue destruction and ulceration during L. major induced CL. Aims: 1) to develop an alternative culture media that could minimize the cost for culturing Leishmania from patient lesions. 2) to investigate if the expression of FasL and TRAIL differs in ulcerating and non- ulcerative CL. Methods: GALF-1 media was formulated in our lab and compared to RPMI 1640 medium and conventional Locke s semi solid media (LSSM) which is one of the modifications of Novy-MacNeal-Nicolle (NNN) culture media. Amastigotes transformation, cryopreservation, recovery of parasites, cost and mass cultivation were analysed. Expression of Fas ligand (FasL), TRAIL and apoptosis were assessed by immunohistology in human skin biopsies from L. aethiopica induced ulcerative or non-ulcerative CL. FasL and TRAIL blocking experiments were performed in a murine model of CL. Results and discussion: GALF-1 is cheap and its ingredients available in a low income country such as Ethiopia. GALF-1 was able to transform amastigotes from Ethiopian patients samples and could be used to cultivate promastigotes in large quantities. Cost analysis showed 80% to 95 % decreased costs as compared to conventional media. Promastigotes cultured with GALF-1 could be cryopreserved in liquid nitrogen with comparable re-culture potential to conventional media. Affordability of diagnostic assays is a key issue for resource poor countries and the possibility to cut the cost of the efficient culture method for diagnosis through the use of inexpensive local formulated reagents could improve the diagnosis of leishmaniasis in low income endemic countries. More FasL expressing cells were detected in dermis of ulcerative CL as compared to non-ulcerative CL and controls. TRAIL expression was higher in ulcerative CL as compared to non-ulcerative CL and controls in both epidermis and dermis. Increased dermal expression of FasL and TRAIL was associated with ulcer formation during CL. This correlated with an inhibition of the ulcerative process in a murine CL model during FasL and TRAIL neutralisation.The mechanisms of the involvement of FasL and TRAIL in ulceration was not elucidated and putative reason(s) for the difference in dysregulation of apoptosis are discussed

Treatment of head lice with dimeticone 4% lotion: comparison of two formulations in a randomised controlled trial in rural Turkey

<p>Abstract</p> <p>Background</p> <p>Dimeticone 4% lotion was shown to be an effective treatment for head louse infestation in two randomised controlled trials in England. It is not affected by insecticide resistance but efficacy obtained (70-75%) was lower than expected. This study was designed to evaluate efficacy of dimeticone 4% lotion in a geographically, socially, and culturally different setting, in rural Turkey and, in order to achieve blinding, it was compared with a potential alternative formulation.</p> <p>Methods</p> <p>Children from two village schools were screened for head lice by detection combing. All infested students and family members could participate, giving access to treatment for the whole community. Two investigator applied treatments were given 7 days apart. Outcome was assessed by detection combing three times between treatments and twice the week following second treatment.</p> <p>Results</p> <p>In the intention to treat group 35/36 treated using dimeticone 4% had no lice after the second treatment but there were two protocol violators giving 91.7% treatment success. The alternative product gave 30/36 (83.3%) treatment success, a difference of 8.4% (95% CI -9.8% to 26.2%). The cure rates per-protocol were 33/34 (97.1%) and 30/35 (85.7%) respectively. We were unable to find any newly emerged louse nymphs on 77.8% of dimeticone 4% treated participants or on 66.7% of those treated with the alternative formulation. No adverse events were identified.</p> <p>Conclusion</p> <p>Our results confirm the efficacy of dimeticone 4% lotion against lice and eggs and we found no detectable difference between this product and dimeticone 4% lotion with nerolidol 2% added. We believe that the high cure rate was related to the lower intensity of infestation in Turkey, together with the level of community engagement, compared with previous studies in the UK.</p> <p>Trial Registration</p> <p>Current Controlled Trials ISRCTN10431107</p

AFRICAN JOURNAL OF MICROBIOLOGY RESEARCH

Leishmaniasis is still a problem for many countries including developed ones. The subgroups of the serum complements have significant roles on the onset of the infection. The aim of this study was to determine the serum complement (C1, C3, C4) levels of cases with definite diagnosis of leishmaniasis and compare them with healthy controls. The study group included 43 visceral leishmaniasis (VL) and 13 cutaneous leishmaniasis (CL) cases. Two control groups were formed for the study. The first control group included 50 individuals of the same age group having no health complaints, admitted for routine control and found to be serologically negative. The second control group included 28 individuals, consistent with the diagnostic criteria of VL. The serum complement levels of C1, C3 and C4 were measured by the nephelometric method. The average levels of C1 were found to be high in VL cases (78.98%) and low in CL (41.69%) group, which was statistically significant (p=0.011). In addition, the average levels of C3 were high in CL group (96.08%) and low in VL group (62.00%), which was also statistically significant (p=0.010). C4 levels were found to be high in the control group, while similar in VL and CL groups. C1, C3, C4 levels were found to be lower in the first control group of healthy individuals. The levels of the subgroups of complement system show statistically significant in both VL and CL cases, which suggested that they could be useful in verifying the results of the serological examinations

PARASITOLOGY RESEARCH

Definitive diagnosis of Trichomonas vaginalis, one of the common causes of sexually transmitted diseases in Turkey, relies on the microscopic examination of both fresh preparations and culture material of genital secretions and urine. The aim of this study was to compare the diagnostic efficacies of two culture media, serum-free TB1 and TB2, including iron and vitamin B12, respectively, with the well-known medium, TYM. Growth rate reached peak levels 48 h in TYM and 72 h in both TB1 and TB2 after inoculation. The highest amount of viable trophozoites has been obtained from TB1, almost equal to TYM but significantly higher than TB2. Fresh preparations obtained from the vaginal secretions of 119 patients have been examined and vaginal samples have been inoculated in TB1, TB2, and TYM. Viable T. vaginalis trophozoites have been detected in ten (8.4%) of fresh preparations and 11 (9.9%) of each medium. It is concluded that serum-free TB1 medium could be used effectively in both the isolation and maintenance of T. vaginalis culture in vitro

INTERNATIONAL JOURNAL OF CLINICAL PHARMACOLOGY RESEARCH

Intestinal parasitic infections are major causes of childhood diseases in rural regions. The significant causes of this problem are the insufficient resources to improve the infrastructure in these regions and inadequate health services in both the diagnosis and treatment of diseases, The aim of this study was to assess the association between intestinal parasitic infections and clinical symptoms of infected children. Cellophane tape and stool samples of 100 children, aged between 7-10 years, from two villages of Manisa city, were examined for intestinal parasites and their symptoms were recorded. Although some symptoms, such as abdominal pain, anorexia and fatigue were found to be common in infected children, no significant relationship was found between symptoms and intestinal parasitic infections

JOURNAL OF CLINICAL MICROBIOLOGY

A new liquid culture medium prepared with chemicals that can be obtained economically and commercially was tested in in vitro cultivation of Leishmania promastigotes to obtain a large number of organisms to use in serological studies, The number of Leishmania infantum and Leishmania tropica promastigotes taken from Novy-MacNeal-Nicolle (NNN) medium reached 1 x 10(7)/ml at the end of the 8th day In our nem medium, though in NNN medium the number of organisms reached only 5 x 10(6)/ml. After 10 subsequent passages, the culture medium prepared was evaluated as being quite inexpensive, simple, and successful compared with other commercially available liquid culture media