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.     

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.     

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.     

Evaluation of berries of Phytolacca dodecandra for growth inhibition of Histoplasma capsulatum var. farciminosum and treatment of cases of epizootic lymphangitis in Ethiopia

Objective:To evaluate the berries of Phytolacca dodecundra(P.dodecandra) for its effect on Histoplasma cupsulatum var.farciminosum(HCF) and for the treatment of cases of epizootic lymphangitis(ELi.Methods:Samples were collected from un-ruptured nodules of cases of EL at Debre Zeit and Akaki(central Ethiopia).Mycologieal culture and isolation of HCF were performed at the Akliln Lemma Institute of Pathobiology.Phytochemical screening was done for n-butanol extract of P.dodecandra to delect alkaloids,saponins,phenolic compounds and flavonoids.The minimum inhibitory concentrations(MICs) and minimum fungicidal concentrations(MFCs) ol aqueous and n-butanol extracts of P.dodecandra against FICF were determined by agar dilution assay.For the in vivo trial.5%simple ointment was prepared from n-butanol extract and applied topically to 24(twelve early and twelve moderate) cases of F.L.Results:Phytochemical screening showed that n-butanol extract ol P.dodecandra was positive lor alkaloids.saponins and phenolic compounds but negative for flavonoids.The MFCs of n-butanol and aqueous extracts of P.dodecandra were(0.039%-0.078%) and(0.625%-1.250%),respectively.The MFCs of n-butanol and aqueous extracts of P.dodecandra were(0.078%t-0.156%)and(1.250%-2.500%),respectively.The MIC and MFC of ketoconazole(positive control) was(1.200×10~(-5)%-2.500×10~(-5)%) and(5.000× 10~(-5)%-1.000×10~(-4)%),respectively while growth was observed on free medium(negative control).From the total of 24 treated cases of EL,14(58.3%) responded lo treatment;however,10(41.7%) did not respond to treatment.There was no significant difference in the degree of response to treatment between early and moderate cases(χ~2=0.086:P=0.408.Conclusions:It can be concluded that n-butanol extract of P.dodecandra demonstrates antifungal effects while the aqueous extract shows no antifungal activity.     

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.     

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.     

Studies on the anti-diarrheal properties of leaf extract of Desmodium puchellum

Objective

To evaluate the pharmacological activity against diarrhea of methanol and petroleum ether extract of Desmodium puchellum (Family: Fabaceae) leaves.

Methods

The extract was evaluated for castor oil-induced diarrhea and enteropooling as well as intestinal motility in rats. Both of the extracts were given to the rats at 200 mg/kg orally. Loperamide was used as a standard drug for diarrhea.

Results

The diarrheal severity was reduced significantly (P<0.05) by methanol and petroleum ether extracts by 31.95% and 28.33%, respectively, whereas 54.75% inhibition was found for standard drug loperamide at 5 mg/kg. The two extracts also significantly (P<0.05) reduced the intestinal volume in case of castor oil induced enteropooling.

Conclusions

It is concluded that both fractions contain some biologically active ingredients that are active for anti-diarrheal actions whereas methanol fraction has better potential.     

Hepatoprotective effect of leaves of Erythroxylum monogynum Roxb. on paracetamol induced toxicity

Objective

To investigate the hepatoprotective activity of methanolic extract of leaves of Erythroxylum monogynum (E. monogynum) on paracetamol induced toxicity.

Methods

Methanolic extract of leaves of E. monogynum was given in doses of 100 mg/kg, 200 mg/kg and 400 mg/kg for 7 d and toxicity was induced by paracetamol (2 mg/kg) on Day 8. Silymarin (50 mg/kg) was used as reference standard. After 24 h of toxicity induction blood samples were collected from retro-orbital plexsus and analyzed for serum parameters like serum glutamic pyruvic transaminase, serum glutamic oxaloacetate transaminse, alkaline phosphatase and total bilirubin. Livers isolated were studied for histopathological changes.

Results

Phytochemical analysis of methanolic extract of E. monogynum leaves showed the presence of carbohydrates, flavonoids, phenols and saponins. Prior administration of this extract restored the elevated levels serum markers as compared to toxic group which is also confirmed by the histopathological changes observed.

Conclusions

The present study showed that methanolic extract of leaves of E. monogynum possess hepatoprotective action against paracetamol induced hepatotoxicity.     

First detection of Leishmania infantum DNA in wild caught Phlebotomus papatasi in endemic focus of cutaneous leishmaniasis,South of Iran

Objective

To identify the vectors and reservoirs of cutaneous leishmaniasis in the endemic focus of Farashband, Fars Province, South of Iran.

Methods

Sticky papers and Sherman trap were used for collection of sand flies and rodents, respectively. Polymerase chain reaction (PCR) of kDNA, ITS1-rDNA were used for identification of Leishmania parasite in sand flies as well as rodents.

Results

Totally 2 010 sand flies were collected and the species of Phlebotomus papatasi Scopoli was the common specimen in outdoors and indoors places. PCR technique was employed on 130 females of Phlebotomus papatasi. One of them (0.76%) was positive to parasite Leishmania major (L. major) and one specimen (0.76%) was positive to Leishmania infantum. Microscopic investigation on blood smear of the animal reservoirs for amastigote parasites revealed 16 (44%) infected Tatera indica. Infection of them to L. major was confirmed by PCR against kDNA loci of the parasite.

Conclusions

The results indicated that Phlebotomus papatasi was the dominant species circulating two species of parasites including L. major and Leishmania infantum among human and reservoirs. Furthermore, Tatera indica is the only main host reservoir for maintenance of the parasite source in the area.     

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.     

Serological investigation of vector-borne disease in dogs from rural areas of China

Objective

To evaluate the Anaplasma phagocytophilum (A. phagocytophilum), Ehrlichia canis (E. canis), Dirofilaria immitis (D. immitis) (canine heartworm), Borrelia burgdorferi (B. burgdorferi) infections in countryside dogs from Yunnan, Hainan and Anhui provinces.

Methods

Serum samples were collected from 26 dogs in Yunnan, Hainan and Anhui provinces. The samples were tested using a commercial ELISA rapid diagnostic assay kit (SNAP^® 4Dx^®; IDEXX Laboratories, Inc. U.S.A.). Meanwhile, indirect immunofluorescence assay (IFA) recommended by WHO was conducted to detect IgG to A. phagocytophilum. Two methods were analyzed and compared.

Results

The number of serologically positive dogs for IgG to A. phagocytophilum was only 2 which was from Hainan province and none of the 26 dogs responded positive for E. canis, D. immitis (canine heartworm), and B. burgdorferi by ELISA rapid diagnostic method. The number of serologically positive dogs for IgG to A. phagocytophilum was 13 (50%) by IFA method. Data of the two methods were analyzed by statistical software and the difference was statistically significant (P=0.002).

Conclusions

It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method. However, we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including A. phagocytophilum, E. canis, D. immitis (canine heartworm), and B. burgdorferi which have public health significance.     

A case of misdiagnose of malaria infection

A case of malaria infection in a 42-year-old woman in rural area of Mahmodabad, Mazandaran Province, North Iran was reported and discussed elaborately. She was complaining about recurrent fevers, sweating, headache and myalgia in back. After her first admission to hospital due to misdiagnose she did not receive proper treatment and the patient suffered from clinical manifestations again. Eventually in the second admission to another hospital, after a precise examination on her thick and thin blood smear the agent of disease was recognized appropriately as Plasmodium vivax and treated accordingly.     

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.     

Anti-inflammatory activity of the leaf extacts of Gendarussa vulgaris Nees

Objective

To evaluate the anti-inflammatory property of the leaf exacts of Gendarussa vulgaris (G. vulgaris) Nees.

Methods

G. vulgaris Nees of the family Apocynaceae is a medium sized tree grown in semishade or no shade and is common in the Ernad and Nilambur taluks of Kerala.Various parts of this plant have been used in the treatment of ulcers, sores, inflammation, dyspepsia, healing of wounds, etc. The present study aimed at the evaluation of anti-inflammatory property of the aqueous and alcoholic extracts of the leaves by both in vitro and in vivo methods. In vitro method was estimated by human red blood cell membrane stabilisation (HRBC) method and in vivo method was estimated on the carrageenan induced paw oedima.

Results

Both the methods showed significant anti-inflammatory property of the different extracts tested.

Conclusions

The alcoholic extract at a concentration of 300 mg/mL showed potent activity on comparing with the standard drug diclofenac sodium.     

Effect of ethanolic extract of Ecballium elaterium against Staphylococcus aureus and Candida albicans

Objective

To evaluate the antimicrobial activity of ethanolic extract of Ecballium elaterium (E. elaterium) fruits alone against Staphylococcus aureus (S. aureus) strains and Candida albicans (C. albicans) strains, or in combination with penicillin against Staphylococcus areus strains.

Methods

Evaluation of the antimicrobial activity or synergy interaction was carried out using microdilution method.

Results

The results showed that ethanolic extract of E. elaterium fruits has antimicrobial activity against methicillin resistant S. aureus (MRSA), methicillin sensitive S. aureus (MSSA) and C. albicans. This extract showed a significant decrease in minimum inhibitory concentrations (MIC) of penicillin against both MRSA and MSSA strains. Fractional inhibitory concentration index (FIC) between penicillin and ethanolic extract of E. elaterium fruits against these test strains was less than 0.5.

Conclusions

This study suggests that ethanolic extract of E. elaterium fruits has antimicrobial activity against S. aureus and C. albicans and there is a possibility of concurrent use of penicillin and E. elaterium extract in combination in the treatment of infections caused by MRSA and MSSA strains. A wider study is needed to identify the effective components, the mode of action and the possible toxic effect in vivo of these ingredients.