Diversity and frequency of Nocardia spp. in the soil of Isfahan province,Iran

Objective

To isolate and identify Nocardia spp. from soil in different regions of Isfahan province in the center of Iran.

Methods

This study was conducted in 32 districts (16 cities and 16 villages) in Isfahan province during two years. A total of 800 soil samples from these regions were studied by using kanamycin. The isolated Nocardia species were examined by gram and acid-fast staining and were identified biochemically and morphologically. The frequency and distribution of Nocardia spp. were determined in relation to different factors such as soil pH and temperate climate.

Results

From 153 (19.1%) Nocardia isolates identified, Nocardia asteroids (N. asteroids) complex (45.5%) and Nocardia brasiliensis (N. brasiliensis) (24.7%) were the most frequently isolated species, followed by Nocardia otitidiscaviarum (2.2%), Nocardiopsis dassonvillei, Actinomadura actinomadura (each 1.7%) and Nocardia transvalensis (1.1%) and also unknown spp. (23.0%). In this study, most species (54.4%) of Nocardia, especially N. asteroides complex were isolated from soils with pH: 7.01-8, whereas in pH: 8.01-9 more N. brasiliensis was isolated. The most Nocardia spp. was detected from regions with semi-nomadic and temperate climate (41.1%).

Conclusions

N. asteroids complex is more prevalent in Isfahan province and soil can be a potential source of nocardiosis infections. It is to be considering that climate and soil pH are involved in the frequency and diversity of aerobic Actinomycetes.     

Bioactivity of seagrass against the dengue fever mosquito Aedes aegypti larvae

Objective

To identify the larvicidal activity of the seagrass extracts.

Methods

Seagrass extracts, Syringodium isoetifolium (S. isoetifolium), Cymodocea serrulata and Halophila beccarii, were dissolved in DMSO to prepare a graded series of concentration. Batches of 25 early 4th instars larvae of Aedes aegypti (Ae. aegypti) were transferred to 250 mL enamel bowl containing 199 mL of distilled water and 1 mL of plant extracts (0.01 mg – 0.1 mg). After 24 h the mortality rate was identified with the formulae [(% of test mortality – % of control mortality)/(100 – % of control mortality)] × 100. Each experiment was conducted with three replicates and a concurrent control group. A control group consisted of 1 mL of DMSO and 199 mL of distilled water only.

Results

: The root extract of S. isoetifolium showed maximum larvicidal activity with minimum concentration of extract of LC₅₀= 0.0 604 ± 0.0 040)µg/mL with lower confidence limit (LCL) – upper confidence limit (UCL) = (0.051–0.071) and LC₉₀=0.0 972µg/mL followed by leaf extract of S. isoetifolium showed LC₅₀= (0.062 ± 0.005)µg/mL. The regression equation of root and leaf extract of S. isoetifolium for 4th instar larvae were Y= 4.909 + 1.32x (R²= 0.909) and Y= 2.066 + 1.21x (R² =0.897) respectively. The results of the preliminary phytochemical constituents shows the presence of saponin, steroids, terpenoid, phenols, protein and sugars.

Conclusions

From the present study the ethanolic extracts of seagrass of S. isoetifolium possesses lead compound for development of larvicidal activity.     

Nematode parasite,Auchenacantha spp of flying lemur,Cynocephalus variegatus (Audebert, 1799) from Indonesia: Morphological study with SEM

Objective

To observe morphology of Auchenacantha galeopteri (A. galeopteri), Auchenacantha spinosa (A. spinosa) and Auchenacantha parva (A. parva) from Sukabumi and Ujung Kulon, Indonesia using scanning electron microscopy (SEM).

Methods

Specimens for light microscopy examination were fixed with warm 70% alcohol, cleared and mounted in lactophenol for wet mounting. Drawings were made with the aid of a drawing tube attached to a Nikon compound microscope. Specimens for SEM examination were processed according to Bozzola. Measurements were given in micrometers (µm) as the average of findings, followed by the range in parentheses, unless otherwise stated.

Results

The measurements of Auchenacantha spp are same with those of previously described. The striations of male and female A. galeopteri are widen towards outside and wavy. The mouth of female with 6 lips, each of them is wider at base than distal end in A. galeopteri and rectangular in A. spinosa. Both species have dentiform protrusion from inner surface of lips, such structure absent in A. parva, but the lips with transverse festoon like pattern at anterior end of each lip.

Conclusions

Using SEM, the lip and the striation pattern of three species of Auchenacantha can be clearly distinguished. Sukabumi and Ujung Kulon are new locality of A. galeopteri and A. spinosa, and A. parva is the new record in Indonesia.     

Microbial quality and associated health risks of raw milk marketed in the Tanga region of Tanzania

Objective

To evaluate microbial quality and associated health risks of raw milk marketed in the Tanga region of Tanzania.

Methods

A microbial quality assessment of marketed raw milk was undertaken by evaluating 59 samples of milk from selling points (collecting centres =15), bicycle boys (12) and kiosks/restaurants (32) in Tanga city during April-May 2005. Quality and milk-borne hazards were assessed using a combination of tests in order to quantify the occurrence of Brucellosis (milk ring test), Escherichia coli (E. coli) O157:H7 (culture), the coliform bacteria as well as standard plate count (SPC). Specific gravity (SG) determination was used as an indicator of adulteration.

Results

The mean coliform plate count (c.f.u/mL) of milk handled by bicycle boys (4.2×10⁶) was significantly higher than that handled by collecting centres (3.0×10⁶) and kiosk/ restaurants (1.4×10⁶), respectively (P < 0.05). Of the 59 milk samples collected, 33 (56%) were Brucella milk ring test (MRT)-positive and 78% and 17% of the samples graded satisfactorily based on SG and coliform plate counts as prescribed by East African Community standards for raw milk. There was no verocytotoxigenic E. coli (VTEC) O157: H7 in any of the milk samples collected and analysed during the present study.

Conclusions

It can be concluded that raw market milk in the study area is of poor bacteriological quality and hazardous for human consumption. This highlights the need to implement good hygiene practices and effective monitoring from production through the delivery chain to the consumer. Further studies are needed for detection of toxins that are produced by E. coli, other pathogenic spore forming bacteria (Bacillus spp. and Clostridium spp.) and other harmful microorganisms.     

Evaluation of aqueous and ethanol extract of bioactive medicinal plant,Cassia didymobotrya (Fresenius) Irwin & Barneby against immature stages of filarial vector,Culex quinquefasciatus Say (Diptera: Culicidae)

Objective

To evaluate aqueous and ethanol extract of Cassia didymobotrya leaves against immature stages of Culex quinquefasciatus.

Methods

The mortality rate of immature mosquitoes was tested in wide and narrow range concentration of the plant extract based on WHO standard protocol. The wide range concentration tested in the present study was 10 000, 1 000, 100, 10 and 1 mg/L and narrow range concentration was 50, 100, 150, 200 and 250 mg/L.

Results

2nd instar larvae exposed to 100 mg/L and above concentration of ethanol extract showed 100% mortality. Remaining stages such as 3rd, 4th and pupa, 100% mortality was observed at 1 000 mg/L and above concentration after 24 h exposure period. In aqueous extract all the stages 100% mortality was recorded at 1 000 mg/L and above concentration. In narrow range concentration 2nd instar larvae 100% mortality was observed at 150 mg/L and above concentration of ethanol extract. The remaining stages 100% mortality was recorded at 250 mg/L. In aqueous extract all the tested immature stages 100% mortality was observed at 250 mg/L concentration after 24 h exposure period. The results clearly indicate that the rate of mortality was based dose of the plant extract and stage of the mosquitoes.

Conclusions

From this study it is confirmed and concluded that Cassia didymobotrya is having active principle which is responsible for controlling Culex quinquefasciatus. The isolation of bioactive molecules and development of simple formulation technique is important for large scale implementation.     

Organophosphorous residue in Liza aurata and Cyprinus carpio

Objective

To determine the amount of azinphos methyl and diazinon residues in two river fishes, Liza aurata and Cyprinus carpio, in the north of Iran.

Methods

This study was done during 2006-2007. In this survey, 152 water and fish samples from Gorgan and Qarasu rivers, north of Iran, were investigated. Sampling was done in three predetermined stations along each river. Organophosphorus compounds (OPs) were extracted from the fishes and the water of rivers. After extraction, purification and concentration processes, the amount and type of insecticides in water and fish samples were determined by high performance thin layer chromatography (HPTLC).

Results

There was a significant difference in the residue of the insecticides in the water and fish samples between summer and other seasons in the two rivers. The highest amount of insecticides residue was seen during summer. In both rivers, the amount of diazinon and azinphos methyl residues in the two fishes was more than 2 000 mg/L in summer. There was no significant difference in insecticides residue between the fishes in two rivers. The diazinon residue was higher than the standard limits in both rivers during the spring and the summer, but the residual amount of azinphos methyl was higher than the standard limits only during the summer and only in Qarasu River.

Conclusions

It can be concluded that the amount of OPs in the water and the two fishes, Liza aurata and Cyprinus carpio, is higher than the permitted levels.     

Larvicidal and irritant activities of hexane leaf extracts of Citrus sinensis against dengue vector Aedes aegypti L.

Objective

To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis (C. sinensis) against the early fourth instars and female adults of Aedes aegypti (Ae. aegypti).

Methods

The larvicidal potential of the prepared leaf extract was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 h and LC₅₀ and LC₉₀ values were calculated. The efficacy of extract as mosquito irritant was assessed by contact irritancy assays. Extract-impregnated paper was placed on a glass plate over which a perspex funnel with a hole on the top was kept inverted. Single female adult, 3-day old unfed/blood-fed, was released inside the funnel. After 3 min of acclimatization time, the time taken for the first take-off and total number of flights undertaken during 15 min were scored.

Results

The citrus leaf extracts from hexane possessed moderate larvicidal efficiency against dengue vector. The bioassays resulted in an LC₅₀ and LC₉₀ value of 446.84 and 1 370.96 ppm, respectively after 24 h of exposure. However, the extracts were proved to be remarkable irritant against adults Ae. aegypti, more pronounced effects being observed on blood-fed females than unfed females. The extract-impregnated paper was thus proved to be 7–11 times more irritable as compared with the control paper.

Conclusions

The hexane extracts from C. sinensis leaves are proved to be reasonably larvicidal but remarkably irritant against dengue vector. Further studies are needed to identify the possible role of extract as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the extract could help in formulating strategies for mosquito control.     

Larval stages of digenetic trematodes in Melanopsis praemorsa snails from freshwater bodies in Palestine

Objective

To detect the species of larval trematodes (cercariae) in Melanopsis praemorsa snails from 5 different fresh water bodies in Palestine.

Methods

A total of 1 880 Melanopsis praemorsa snails were collected from different fresh water bodies in Palestine from October, 2008 to November, 2010. Cercariae in Melanopsis praemorsa snails were obtained by lighting and crushing methods. The behavior of cercariae was observed using a dissecting microscope.

Results

Three different species of larval trematodes were identified from Melanopsis praemorsa snails collected only from Al-Bathan fresh water body, while snails from other water bodies were not infected. These species were microcercous cercaria, xiphidiocercaria and brevifurcate lophocercous cercaria. These cercariae called Cercaria melanopsi palestinia I, Cercaria melanopsi palestinia II and Cercaria melanopsi palestinia III have not been described before from this snail in Palestine. The infection rate of Melanopsis praemorsa collected from Al-Bathan fresh water body was 5.7%, while the overall infection rate of snails collected from all fresh water bodies was 4.3%. Details are presented on the morphology and behavior of the cercariae as well as their development within the snail.

Conclusions

These results have been recorded for the first time and these cercariae may be of medical and veterinary importance.     

The zoonotic implications of pentastomiasis in the royal python (python regius)

Pentastomes are worm-like endoparasites of the phylum Pentastomida found principally in the respiratory tract of reptiles, birds, and mammals. They cause a zoonotic disease known as pentastomiasis in humans and other mammals. The autopsy of a Nigerian royal python (Python regius) revealed two yellowish-white parasites in the lungs, tissue necrosis and inflammatory lesions. The parasite was confirmed to be Armillifer spp (Pentastomid); this is the first recorded case of pentastomiasis in the royal python (Python regius) in Nigeria. This report may be an alert of the possibility of on-going zoonotic transmission of pentastomiasis from snake to man, especially in the sub-urban/rural areas of Nigeria and other West African countries where people consume snake meat.     

Cryptosporidiosis as threatening health problem: A review

The protozoa under the genus Cryptosporidium is a zoonotic apicomplexan obligate intracellular parasite. Cryptosporidiosis, the term used to designate infection caused by Cryptosporidium sp., is considered as one of the most common food and waterborne diseases with worldwide spread, acting as a common cause of diarrhoea in animals and man. In immunocompetent individuals, Cryptosporidium typically induces self-limiting diarrhoea, which may resolve on its own after 2-3 d. However, cryptosporidiosis may turn life-threatening and subsequently lead to death in small children, the elderly and immunocompromised person, especially in AIDS patient. The diagnosis for Cryptosporidium infection is usually carried out through examination of stool for the presence of oocysts which measured 4-6 µm with spherical appearance. Morphometric identification is often difficult because of the diminutive size and obscure internal structure of the protozoa. Often, the identification of Cryptosporidium is realised through the combination of methods incorporating data from morphometrics, molecular techniques, and host specificity. However, limitations to some of these techniques still exist whether because of cost, duration, expertise, or reliability. Drugs combination is implemented in treatment of cryptosporidiosis. The efficiency of paromomycin, an aminocyclitol antibiotic isolated from Streptomyces, can be effective when combined use with protease inhibitors or recombinant IL-12. Since there is no drug that achieves the complete removal of Cryptosporidium from the host, supportive therapy was preferred in both human and domestic animals.     

Comparative study on production,purification of penicillin by Penicillium chrysogenum isolated from soil and citrus samples

Objective

To explore various unexplored locations where Penicillium spp. would be available and study the production of penicillin from the isolated Penicillium spp. in different media with altered carbohydrate source.

Methods

The collected soil samples were screened for the isolation of Penicillium chrysogenum (P. chrysogenum) by soil dilution plate. The isolated Penicillium species were further grown in different production media with changes in the carbohydrate source. The extracted penicillin from various isolates was analyzed by HPLC for the efficacy of the product. Further the products were screened with various bacterial species including methicillin resistant Staphylococcus aureus (MRSA). And the work was extended to find the possible action on MRSA, along with characterization using other pathogens.

Results

From the various soil and citrus samples used for analysis, only the soil sample from Government General Hospital of Bangalore, India, and Sanjay Gandhi Hospital, Bangalore, India, showed some potential growth of the desired fungi P. chrysogenum. Different production media showed varied range of growth of Penicillium. Optimum production of penicillin was obtained in maltose which proved maximum zone of inhibition during assay. Characterization of penicillin on pathogens, like wild Escherichia coli strain, Klebsiella spp., and MRSA, gave quite interesting results such as no activity on the later strain as it is resistant. HPLC data provided the analytical and confirmation details of the penicillin produced. Accordingly, the penicillin produced from the soil sample of Government General Hospital had the high milli absorbance unit of 441.5 mAu compared with that of the penicillin produced from Sanjay Gandhi Hospital sample, 85.52 mAu. Therefore, there was a considerable change in quantity of the penicillin produced from both the samples.

Conclusions

The Penicillium spp. could be possibly rich in hospital contaminants and its environments. This research focuses on various unexplored sources of medical ailments, and also shows that the growth of penicillin is high in maltose rich media that could possibly enhance the growth.     

In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

Objective

To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants.

Methods

Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl₂ solutions for 1 min. After rinsing with sterile distilled water for 3–4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.

Results

Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of plants were established in the field.

Conclusions

The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.     

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.     

Saponins-rich fraction of Calotropis procera leaves elicit no antitrypanosomal activity in a rat model

Objective

To examine the in vitro and in vivo anti-Trypanosoma evansi (T. evansi ) activity of saponins-rich fraction of Calotropis procera (cpsf) leaves as well as the effect of the fraction on the parasite-induced anemia.

Methods

A 60-minutes time course experiment was conducted with various concentrations of the fraction using a 96-well microtiter plate technique, and subsequently used to treat experimentally T. evansi infected rats at 100 and 200 mg/kg body weight. Index of anemia was analyzed in all animals during the experiment.

Results

The cpsf did not demonstrate an in vitro antitrypanosomal activity. Further, the cpsf treatments did not significantly (P>0.05) keep the parasites lower than the infected untreated groups. At the end of the experiment, all T. evansi infected rats developed anemia whose severity was not significantly (P>0.05) ameliorated by the cpsf treatment.

Conclusions

It was concluded that saponins derived from Calotropis procera leaves could not elicit in vitro and in vivo activities against T. evansi.     

Syzygium cumini (L.) Skeels: A review of its phytochemical constituents and traditional uses

Syzygium cumini (S. cumini) (L.) Skeels (jambolan) is one of the widely used medicinal plants in the treatment of various diseases in particular diabetes. The present review has been primed to describe the existing data on the information on botany, phytochemical constituents, traditional uses and pharmacological actions of S. cumini (L.) Skeels (jambolan). Electronic database search was conducted with the search terms of Eugenia jambolana, S. cumini, jambolan, common plum and java plum. The plant has been viewed as an antidiabetic plant since it became commercially available several decades ago. During last four decades, numerous folk medicine and scientific reports on the antidiabetic effects of this plant have been cited in the literature. The plant is rich in compounds containing anthocyanins, glucoside, ellagic acid, isoquercetin, kaemferol and myrecetin. The seeds are claimed to contain alkaloid, jambosine, and glycoside jambolin or antimellin, which halts the diastatic conversion of starch into sugar. The vast number of literatures found in the database revealed that the extracts of different parts of jambolan showed significant pharmacological actions. We suggest that there is a need for further investigation to isolate active principles which confer the pharmacological action. Hence identification of such active compounds is useful for producing safer drugs in the treatment of various ailments including diabetes.     

TB or not TB? Difficulties in the diagnosis of tuberculosis in HIV-negative immigrants to Germany

Background

Immigrants to Germany and their children are at particular risk for tuberculosis (TB).

Methods

35 Patients (10 male/25 female aged 2 - 59 years (median 33 years) originating mostly from high incidence countries in Asia (19 [54.3%]) in Africa (14 [40.0%] and East Europe (2 [5.7%]), attended at the Tropical Medicine unit were analysed.

Results

Primary clinical presentation was most frequently lymphadenitis (13 [37.1%]). other organs involved included bones (7 [20.0%]), central nervous system (5 [14.3%]), urogenital organs (3 [8.6%]), lung (3 [8.6%]), mediastinum, (2 [5.7%]) and abdomen (2 [5.7%]). ESR was abnormal in 21/28 (75.0%), CRP in 20/35 (57.1%), and protein electrophoresis in 22/26 (84.6%) cases. The tuberculin skin test was strongly positive in all 15 cases where the test had been performed. Tuberculosis interferon gamma release assay (TB-IGRA) was positive in all 35 cases (100%). PCR for nucleic acids of Mycobacterium (M.) tuberculosis complex was positive in only 7/20 (35.0%) cases. M. tuberculosis was identified in 32/35 (91.4%), M. bovis in 2 (5.7%) cases. 1 case was diagnosed clinically. All patients were negative for HIV. Typical histopathology was seen in the 29 cases, where biopsies had been taken. Chest-X-ray did not reveal specific pulmonary lesions in the majority of cases (22/35 [62.9%]). Diagnosis of TB was mostly delayed (4 to 299 weeks, [median 8]). The most frequent primary suspicion was a malignancy (17/35 [48.6%]) while TB was initially suspected in 5 cases only. Diagnosis of TB is impeded by its multifaceted presentation especially in immigrants.     

Antiplasmodial and antiulcer activities of Melanthera scadens

Objective

To evaluate the antimalarial and antiulcerogenic activities of leaf extract and fractions of Melanthera scandens (M. scandens).

Methods

The crude leaf extract (37–111 mg/kg) and fractions (chloroform, ethylacetate and methanol; 78 mg/kg) of M. scadens were investigated for antiplasmodial activity against chloroquine-sensitive Plasmodium berghei infections in mice and for antiulcer activity against experimentally-induced ulcers. The antimalarial activity during early and established infections as well as prophylactic was investigated. Artesunate (5 mg/kg) and pyrimethamine (1.2 mg/kg) were used as positive controls. Thin films made from tail blood of each mouse were used to assess the level of parasitaemia of the mice. Antiulcer activity of the crude extract was also evaluated against indomethacin, ethanol and histamine induced ulcers.

Results

The extract and its fractions dose-dependently reduced parasitaemia induced by chloroquine-sensitive Plasmodium berghei infection in prophylactic, suppressive and curative models in mice. These reductions were statistically significant (P<0.001). They also improved the mean survival time (MST) from 9.28 to 17.73 days as compared with the control (P<0.01–0.001). The activities of extract/fractions were incomparable to that of the standard drugs i.e. artesunate and pyrimethamine. On experimentally-induced ulcers, the extract inhibited indomethacin, ethanol and histamine induced ulcers. These inhibitions were statistically significant (P<0.001) and in a dose-dependent fashion.

Conclusions

The antiplasmodial and antiulcerogenic effects of this plant may in part be mediated through the chemical constituents of the plant.