Microscopic evaluation and physiochemical analysis of Dillenia indica leaf

Objective

To study detail microscopic evaluation and physiochemical analysis of Dillenia indica (D. indica) leaf.

Methods

Fresh leaf sample and dried power of the leaf were studied macroscopically and microscopically. Preliminary phytochemical investigation of plant material was done. Other WHO recommended parameters for standardizations were also performed.

Results

The detail microscopy revealed the presence of anomocytic stomata, unicellular trichome, xylem fibres, calcium oxalate crystals, vascular bundles, etc. Leaf constants such as stomatal number, stomatal index, vein-islet number and veinlet termination numbers were also measured. Physiochemical parameters such as ash values, loss on drying, extractive values, percentage of foreign matters, swelling index, etc. were also determined. Preliminary phytochemical screening showed the presence of steroids, terpenoids, glycosides, fatty acids, flavonoids, phenolic compounds and carbohydrates.

Conclusions

The microscopic and physiochemical analysis of the D. indica leaf is useful in standardization for quality, purity and sample identification.     

New scope on the relationship between rotifers and Biomphalaria alexandrina snails

Objective

To investigate the effect of rotifer internalization into snail tissue on the development of schistosomes.

Methods

Susceptible laboratory-bred Biomphalaria alexandrina (B. alexandrina) snails were exposed to lab-maintained rotifers; Philodina spp., two weeks before and after being infected with Schistosoma mansoni (S. mansoni) miracidia. The consequent histopathological impact on snail tissues and cercarial biology were investigated before and after emergence from snails.

Results

Contamination of B. alexandrina snails with philodina, two weeks before miracidial exposure, was found to hinder the preliminary development of S. mansoni cercariae inside the snail tissues. Furthermore, when snails were contaminated with rotifers two weeks post miracidial exposure; growth of already established cercariae was found to be retarded. The consequent influence of internalized rotifers within the snail tissue was clearly reflected on cercarial emergence, activity and infectivity along the four weeks duration of shedding. In the present study, comparison of snail histopathological findings and altered cercarial biology observed between the experimental and control groups indicated that the rotifers may have affected the levels of snail''s energy reservoirs, which eventually was found to have had an adverse impact on reproduction, growth and survival of the parasite within the snail host, coupled with its performance outside the snail.

Conclusions

In future biological control strategies of schistosomiasis, ritifers should be considered as a parasitic scourge of humanity.     

Occurrence of Clinostomum complanatum in Aphanius dispar (Actinoptrygii: Cyprinodontidae) collected from Mehran River,Hormuzgan Province,South of Iran

Objective

To investigate the possibility of Aphanius dispar (A.dispar) acting as intermediate host for Clinostomum complanatum (C. complanatum), from Mehran River, Hormuzgan Province of Iran.

Methods

During a biological study of A. dispar in Mehran River, Hormuzgan Province, South of Iran, a total of 97 fish specimens were collected in 24 January 2006.

Results

4 specimens (4.12%) including 1 male and 3 female were found infected with C. complanatum metacercaria. These metacercaria were coiled in the epiderm on the body surface of infected fishes. The infection is known as yellow spot disease. The parasite abundance, intensity and prevalence were 0.05%, 1.25% and 4.12%, respectively. The infection was higher in females than males.

Conclusions

To the best of our knowledge, this is the first report on occurrence of C. complanatum metacercaria in A. dispar in Iran.     

In vitro antiplasmodial effect of ethanolic extracts of coastal medicinal plants along Palk Strait against Plasmodium falciparum

Objective

To identify the possible antiplasmodial compounds from Achyranthes aspera (A. aspera), Acalypha indica (A. indica), Jatropha glandulifera (J. glandulifera) and Phyllanthus amarus (P. amarus).

Methods

The A. aspera, A. indica, J. glandulifera and P. amarus were collected along Palk Strait and the extraction was carried out in ethanol. The filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 µg/mL) of leaf, stem, root and flower extracts of A. aspera, A. indica, J. glandulifera and P. amarus were tested for antiplasmodial activity against Plasmodium falciparum. The potential extracts were also tested for their phytochemical constituents.

Results

Of the selected plants species parts, the stem extract of A. indica showed excellent antiplasmodial activity (IC₅₀= 43.81µg/mL) followed by stem extract of J. glandulifera (IC₅₀= 49.14µg/mL). The stem extract of A. aspera, leaf and root extracts of A. indica, leaf, root and seed extracts of J. glandulifera and leaf and stem extracts of P. amarus showed IC₅₀ values between 50 and 100 µg/mL. Statistical analysis revealed that, significant antiplasmodial activity (P<0.01) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it showed that there were no morphological changes in erythrocytes by the ethanolic extract of all the tested plant extracts. The in vitro antiplasmodial activity might be due to the presence of alkaloids, glycosides, flavonoids, phenols, saponins, triterpenoids, proteins, and tannins in the ethanolic extracts of tested plants.

Conclusions

The ethanolic stem extracts of P. amarus and J. glandulifera possess lead compounds for the development of antiplasmodial drugs.     

Synthesis of silver nanoparticles using leaves of Catharanthus roseus Linn. G. Don and their antiplasmodial activities

Objective

To develop a novel approach for the green synthesis of silver nanoparticles using aqueous leaves extracts of Catharanthus roseus (C. roseus) Linn. G. Don which has been proven active against malaria parasite Plasmodium falciparum (P. falciparum).

Methods

Characterizations were determined by using ultraviolet-visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), energy dispersive X-ray and X-ray diffraction.

Results

SEM showed the formation of silver nanoparticles with an average size of 35–55 nm. X-ray diffraction analysis showed that the particles were crystalline in nature with face centred cubic structure of the bulk silver with the broad peaks at 32.4, 46.4 and 28.0.

Conclusions

It can be concluded that the leaves of C. roseus can be good source for synthesis of silver nanoparticle which shows antiplasmodial activity against P. falciparum. The important outcome of the study will be the development of value added products from medicinal plants C. roseus for biomedical and nanotechnology based industries.     

Toxicity profile of ethanolic extract of Azadirachta indica stem bark in male Wistar rats

Objective

To investigate the toxic implications of ethanolic stem bark extract of Azadirachta indica (A. indica) at 50, 100, 200 and 300 mg/kg body weight in Wistar rats.

Methods

Fifty male rats of Wistar strains were randomly grouped into five (A-E) of ten animals each. Animals in Group A (control) were orally administered 1 mL of distilled water on daily basis for 21 days while those in Groups B-E received same volume of the extract corresponding to 50, 100, 200 and 300 mg/kg body weight.

Results

The extract did not significantly (P>0.05) alter the levels of albumin, total protein, red blood cells and factors relating to it whereas the white blood cell, platelets, serum triacylglycerol and high-density lipoprotein cholesterol decreased significantly (P<0.05). In contrast, the final body weights, absolute weights of the liver, kidney, lungs and heart as well as their organ-body weight ratios, serum globulins, total and conjugated bilirubin, serum cholesterol, low-density lipoprotein cholesterol and computed atherogenic index increased significantly. The spleen-body weight ratio, alkaline phosphatase, alanine and aspartate transaminases, sodium, potassium, calcium, feed and water intake were altered at specific doses.

Conclusions

Overall, the alterations in the biochemical parameters of toxicity have consequential effects on the normal functioning of the organs of the animals. Therefore, the ethanolic extract of A. indica stem bark at the doses of 50, 100, 200 and 300 mg/kg body weight may not be completely safe as an oral remedy and should be taken with caution if absolutely necessary.     

Mass rearing of Lucilia sericata Meigen (Diptera: Calliphoridae)

Objective

To carry out an experimental study with the main objective of mass rearing of sheep flies (Lucilia sericata).

Methods

Hand collection and beef- or cattle liver-baited net traps were used for field fly sampling from April, 2010 to November, 2010. The samples collected from different places were placed in properly labeled tubes and sent to the Entomology Laboratory. Since maggot identification is important in inducing mortality, they were kept under insectary condition to develop to adult stage and identified using systematic keys.

Results

A total of 218 flies were collected in three rounds of sampling from the field of Tehran and Karaj Counties. In the first generation, 433 flies including 135 (31.17%) male, and 298 (68.82%) female were yielded. The female/male of parent ratio was calculated as 1.72 in Tehran and in Karaj areas, whereas it was 2.20% and 1.81%, respectively in F1 and F2 generations, respectively.

Conclusions

During this study, the mass rearing of sheep blow fly has been established at the School of Public Health, Tehran University of Medical Sciences and can be used for producing flies for maggot therapy.     

Acaricidal activity of Cymbopogon citratus and Azadirachta indica against house dust mites

Objective

To examine the acaricidal effects of the essential oil of Cymbopogon citratus leaf extract (lemongrass) and ethanolic Azadirachta indica leaf extract (neem) against house dust mites Dermatophagoides farinae (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus).

Methods

Twenty-five adults mites were placed onto treated filter paper that is soaked with plant extract and been tested at different concentrations (50.00%, 25.00%, 12.50%, 6.25% and 3.13%) and exposure times (24hrs, 48hrs, 72hrs and 96 hrs). All treatments were replicated 7 times, and the experiment repeated once. The topical and contact activities of the two herbs were investigated.

Results

Mortalities from lemongrass extract were higher than neem for both topical and contact activities. At 50 % concentration, both 24 hrs topical and contact exposures to lemongrass resulted in more than 91% mortalities for both species of mites. At the same concentration and exposure time, neem resulted in topical mortalities of 40.3% and 15.7% against D. pteronyssinus and D. farinae respectively; contact mortalities were 8.0% and 8.9% against the 2 mites, respectively. There was no difference in topical mortalities of D. pteronyssinus from exposure to concentrations of lemongrass and neem up to 12.50%; lemongrass was more effective than neem at the higher concentrations.

Conclusions

Generally, topical mortalities of D. farinae due to lemongrass are higher than that due to neem. Contact mortalities of lemongrass are always higher that neem against both species of mites.     

Use of buffy coat thick films in detecting malaria parasites in patients with negative conventional thick films

Objective

To determine the frequency of malaria parasite detection from the buffy coat blood films by using capillary tube in falciparum malaria patients with negative conventional thick films.

Methods

Thirty six uncomplicated falciparum malaria patients confirmed by conventional thick and thin films were included in the study. The patients were treated with artemisinin combination therapy at Hospital for Tropical Diseases, Bangkok, Thailand for 28 day. Fingerpricks for conventional blood films were conducted every 6 hours until negative parasitemia, then daily fingerpricks for parasite checks were conducted until the patients were discharged from hospital. Blood samples were also concurrently collected in 3 heparinized capillary tubes at the same time of fingerpricks for conventional blood films when the prior parasitemia was negative on thin films and parasitemia was lower than 50 parasites/200 white blood cells by thick film. The first negative conventional thick films were compared with buffy coat thick films for parasite identification.

Results

Out of 36 patients with thick films showing negative for asexual forms of parasites, buffy coat films could detect remaining 10 patients (27.8%) with asexual forms of Plasmodium falciparum.

Conclusions

The study shows that buffy coat thick films are useful and can detect malarial parasites in 27.8% of patients whose conventional thick films show negative parasitemia.     

Prevalence of malaria infection in Sarbaz,Sistan and Bluchistan province

Objective

To survey malaria prevalence in Sarbaz from April 2009 to October 2010.

Methods

Epidemiological data of 1 464 confirmed malarial patients were analyzed according to demographic status, sex, age, nationality, isolated species and residence place.

Results

The majority of patients were male 950 (64.8%) but 514 (35.2%) were female. 82.5% of patients were Iranian, 14% Pakistani immigrants, and 3.5% Afghan immigrants. Data collected showed that 90% of isolated species were Plasmodium vivax, 7.8% Plasmodium falciparum, and 2.2% Plasmodium malariae and mixed species.

Conclusions

Therefore, it is crystal clear that refugees should be prohibited by government and controlled by experts in health centers in order to campaign effectively with this life threating disease.     

Antiarthritic effects of Ajuga bracteosa Wall ex Benth. in acute and chronic models of arthritis in albino rats

Objective

To evaluate the antiarthritic activity of Ajuga bracteosa using albino rats.

Methods

The antiarthritic activity of 70% ethanolic extract of Ajuga bracteosa (EEAB) was evaluated against turpentine oil- and formaldehyde- induced acute non immunological and complete freund''s adjuvant (CFA)-induced chronic immunological arthritis in albino rats.

Results

EEAB showed a significant (P<0.05) and dose dependent inhibitory effect against acute and chronic models of arthritis. EEAB exhibited better antiarthritic activity than the standard aspirin.

Conclusions

EEAB exhibits a significant and promising antiarthritic activity against acute and chronic arthritis and supports the traditional use of Ajuga bracteosa for rheumatism and other inflammatory diseases.     

Incidence and histopathology of encysted progenetic metacercaria of Clinostomum complanatum (Digenea: Clinostomidae) in Channa punctatus and its development in experimental host

Objective

To study the incidence of encysted progenetic metacercariae of Clinostomum complanatum (C. complanatum) in Channa punctatus (C. punctatus), associated histopathology and the experimental infection to laboratory chicken to obtain ovigerous adult worms.

Methods

Live C. punctatus were brought from local fish market of Aligarh, India, dissected and examined on a monthly basis for the presence of C. complanatum cysts. For histochemistry, infected tissue sections with attached cysts were processed for haematoxylene and eosin staining. Cysts were aseptically fed to 4 day old leghorn chicken to obtain adult worms. Mechanically excysted metacercaria and the ovigerous adult worms were stained in carmine to prepare permanent slides.

Results

One year survey for the infection of encysted progenetic metacercaria of C. complanatum in C. punctatus revealed the prevalence, intensity and abundance of 24.7%, 2.27 and 0.608, respectively. Histopathology showed heavy infiltration of immune cells at the site of cyst attachment and some tissue damage was also evident. Following feeding to experimental chicken, about 41.07% of the encysted metacercariae were able to excyst and migrate back to bucco-pharyngeal region where they tenaciously attached and fed on blood, and transformed into ovigerous adult worms from 62 hours onwards of post infection.

Conclusions

The parasite is potentially pathogenic to the host, and the availability of a suitable intermediate host can be a contributing factor for the occurrence of C. complanatum metacercaria either in the excysted or encysted form, indicating loose host specificity and zoonotic potential.     

In vivo toxicity study of Lantana camara

Objective

To investigate the toxicity of methanol extract of various parts (Root, Stem, Leaf, Flower and Fruit) of Lantana camara (L. Camara) in Artemia salina.

Methods

The methanol extracts of L. camara were tested for in vivo brine shrimp lethality assay.

Results

All the tested extract exhibited very low toxicity on brine shrimp larva. The results showed that the root extract was the most toxic part of L. camara and may have potential as anticancer agent.

Conclusions

Methanolic extract of L. camara is relatively safe on short-term exposure.     

Hibiscus sabdariffa extract inhibits in vitro biofilm formation capacity of Candida albicans isolated from recurrent urinary tract infections

Objective

To explore the prevention of recurrent candiduria using natural based approaches and to study the antimicrobial effect of Hibiscus sabdariffa (H. sabdariffa) extract and the biofilm forming capacity of Candida albicans strains in the present of the H. sabdariffa extract.

Methods

In this particular study, six strains of fluconazole resistant Candida albicans isolated from recurrent candiduria were used. The susceptibility of fungal isolates, time-kill curves and biofilm forming capacity in the present of the H. sabdariffa extract were determined.

Results

Various levels minimum inhibitory concentration of the extract were observed against all the isolates. Minimum inhibitory concentration values ranged from 0.5 to 2.0 mg/mL. Time-kill experiment demonstrated that the effect was fungistatic. The biofilm inhibition assay results showed that H. sabdariffa extract inhibited biofilm production of all the isolates.

Conclusions

The results of the study support the potential effect of H. sabdariffa extract for preventing recurrent candiduria and emphasize the significance of the plant extract approach as a potential antifungal agent.     

In vitro treatments of Echinococcus granulosus with fungal chitosan,as a novel biomolecule

Objective

To determined the antiparasitic activity of the isolated chitosan from Penicillium viridicatum, Penicillium aurantiogriseum and commercial chitosan against protoscolicidal of hydatid cysts were determined.

Methods

After isolating chitosan from fungal cell walls, four concentrations (50, 100, 200, 400 µg/mL) of each type of prepared chitosan and commercial chitosan were used for 10, 30, 60, and 180 min, respectively.

Results

Among different type of chitosan, commercial chitosan with the highest degree of deacetylation showed high scolicidal activity in vitro. Fungal chitosan could be recommended, as good as commercial chitosan, for hydatic cysts control.

Conclusions

It seems to be a good alternative to synthetic and chemical scolicidal.     

Polymorphic patterns of pfcrt and pfmdr1 in Plasmodium falciparum isolates along the Thai-Myanmar border

Objective

To investigate the distribution and patterns of pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum (P. falciparum) isolates collected from the malaria endemic area of Thailand along Thai-Myanmar border.

Methods

Dried blood spot samples were collected from 172 falciparum malaria patients prior received treatment. The samples were extracted using chelex to obtain parasite DNA. PCR-RFLP was employed to detect pfcrt mutation at codons 76, 220, 271, 326, 356 and 371, and the pfmdr1 mutation at codon 86. Pfmdr1 gene copy number was determined by SYBR Green I real-time PCR.

Results

Mutant alleles of pfcrt and wild type allele of pfmdr1 were found in almost all samples. Pfmdr1 gene copy number in isolates collected from all areas ranged from 1.0 to 5.0 copies and proportion of isolates carrying>1 gene copies was 38.1%. The distribution and patterns of pfcrt and pfmdr1 mutations were similar in P. falciparum isolates from all areas. However, significant differences in both number of pfmdr1 copies and prevalence of isolates carrying>1 gene copies were observed among isolates collected from different areas. The median pfmdr1 copy number in P. falciparum collected from Kanchanaburi and Mae Hongson were 2.5 and 2.0, respectively and more than half of the isolates carried>1 gene copies.

Conclusions

The observation of pfmdr1 wild type and increasing of gene copy number may suggest declining of artesunate-mefloquine treatment efficacy in P. falciparum isolates in this border area.     

Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab

Objective

To determine the presence of Babesia bovis (B. bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals.

Methods

Blood samples were collected from 144 large ruminants, including 105 cattle and 39 buffaloes, from six districts in southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar and Vehari. Data on the characteristics of animals and herds were collected through questionnaires. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B. bovis on the blood and serological profile of infected animals.

Results

27 out of 144 animals, from 5 out of 6 sampling districts, produced the 541-bp fragment specific for B. bovis. Age of animals (P=0.02), presence of ticks on animals (P=0.04) and presence of ticks on dogs associated with herds (P=0.5) were among the major risk factors involved in the spread of bovine babesiosis in the study area. ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle.

Conclusions

: This study has reported for the first time the presence of B. bovis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output.     

The prevalence of Linguatula serrata nymphs in camels slaughtered in Mashhad slaughterhouse,Northeast, Iran

Objective

To evaluate the prevalence of nymphal stages of L. serrata in mesenteric lymph nodes of camels slaughtered in Mashhad slaughterhouse, Northeast of Iran.

Methods

For this purpose, mesenteric lymph nodes of 400 camels of different sex and age were examined. The lymph nodes were examined macroscopically and a digestion method was also applied for investigation of samples which was negative macroscopically.

Results

The mesenteric lymph nodes of 73 camels out of 400 (18.25%) were infected by L. serrata nymphs.

Conclusions

Prevalence of L. serrata nymphs in males and females and different age was not significantly different (P>0.05), but difference was observed between the prevalence in different seasons (P< 0.05). The potential importance of these findings to human health is discussed. This is the first report of infection with L. serrate of camels in camels slaughtered at northeast of Iran.     

Unusual developing sites of dengue vectors and potential epidemiological implications

Objective

To identify the unusual breeding sites of two dengue vectors, i.e. Aedes albopictus (Ae. albopictus) and Aedes aegypti (Ae. aegypti).

Methods

During the second half of 2010, we performed an occasional survey in rural (Teluk Tempoyak) and urban (Gelugor) areas of Penang Island, Malaysia, to identify cryptic breeding sites.

Results

In the rural area, we found heterogeneous immature stages of Ae. albopictus in the water bowl of an encaged bird. We also observed Ae. aegypti eggs deposited in the flush tank of a toilet in the urban area.

Conclusions

It can be concluded that both breeding patterns can increase contact with hosts (humans and birds) and presumably population densities of Ae. albopictus and Ae. aegypti, thereby potentially boosting the risks for spread and transmission of arboviral diseases.     

Cytotoxicity of methanol extracts of Elaeis guineensis on MCF-7 and Vero cell lines

Objective

To investigate the cytotoxic effect of Elaeis guineensis methanol extract on MCF-7 and Vero cell.

Methods

In vitro cytotoxicity was evaluated in by MTT assay. Cell morphological changes were observed by using light microscope.

Results

The MTT assay indicated that methanol extract of the plant exhibited significant cytotoxic effects on MCF-7. Morphological alteration of the cell lines after exposure with Elaeis guineensis extract were observed under phase contrast microscope in the dose dependent manner.

Conclusions

The results suggest the probable use of the Elaeis guineensis methanol extract in preparing recipes for cancer-related ailments. Further studies on isolation of metabolites and their in vivo cytotoxicity are under investigation.