BACKGROUND: This study evaluated in vitro antigiardial activity in four Indonesian plants (Archidendron fagifolium, Diospyros sumatrana, Piper betle and Shorea sumatrana) extracted in methanol, methanol-tetrahydrofuran, and water. These plants exhibiting promising anti-parasitic activity were selected on the basis of collected behavioral data and their ability to decrease parasite load in Sumatran orangutans. Extracts of Arabidopsis thaliana, a plant routinely used as a laboratory model in research, were used as a negative control. METHODS: Plant extracts of different concentrations (400, 100, 25 and 6.25 µg/ml) and metronidazole (100 µg/ml), a standard giardicidal drug, were incubated with 25,000 trophozoites per milliliter of growth medium in 12-well tissue culture plates under anaerobic conditions for 72 h. Cultures were counted in a hemocytometer using a light microscope and then statistically evaluated. The cytotoxicity of the extracts was determined by the MTT Cell Viability Assay. RESULTS: We recorded a statistically-significant decrease in giardia trophozoites in two extracts: the methanolic extract of A. fagifolium (Kruskal-Wallis: p = 0.013) and the aqueous extract of P. betle (Kruskal-Wallis: p = 0.002). Effective concentrations of 400 µg/ml and 100 µg/ml, respectively, were revealed. Cytotoxicity was not demonstrated. CONCLUSIONS: The results of our study show that extracts of A. fagifolium and P. betle have potential as an alternative treatment of G. intestinalis infection. This is the first scientific proof of the biological activity of A. fagifolium, confirming the assumption that orangutans use this plant (as well as P. betle) for self-medication.
- Klíčová slova
- Giardia intestinalis, Antigiardial activity, Drug of choice, Natural antiparasitic, Parasite,
- MeSH
- antiprotozoální látky farmakologie MeSH
- Giardia lamblia * účinky léků MeSH
- rostlinné extrakty * farmakologie MeSH
- trofozoiti účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indonésie MeSH
- Názvy látek
- antiprotozoální látky MeSH
- rostlinné extrakty * MeSH
There is little evidence that the already described and accepted taxa of ascarids (Ascaris lumbricoides, A. suum, and A. ovis) infecting individuals of taxonomically distant groups (hominids, pigs, sheep, goats, and dogs) can be genetically or morphologically distinguished. However, despite described morphological differences, e.g., due to intraspecific variation, these are insufficient for species determination and may indicate differences amongst ascarids because of cross infections, hybrid production, and specific adaptations to hosts. Herein, the results of a molecular and morphological analysis of ascarids parasitising Sumatran orangutans (Pongo abelii Lesson, 1827) in native populations are presented. The research took place in the Bukit Lawang area, Indonesia, in 2009. Throughout the year, fresh faecal samples were collected regularly from 24 orangutans, and all were examined for the presence of nematode adults. Only five adult worms from two orangutan females were found during regular collection. Using the integrative taxonomic approach, the nematodes found were identified as A. lumbricoides. The significance of the find and its rarity is documented by the fact that this is the first confirmed finding of adult ascarids from an original orangutan site (not from a zoo) in more than 130 years (including the long-term study spanning the last 20 years focusing on orangutan parasites and natural antiparasitic drugs). More accurate morphometric parameters and genetic differences for the identification of ascarids were established. These parameters will be helpful for other findings in great apes and will also be suitable for further and precise determination of this parasite. The details distinguishing between male and female specimens are also stated and well defined. A comprehensive evaluation of the situation of Ascaris species parasitising orangutans, including a comparison with previously described orangutan parasite (i.e., A. satyri-species inquirenda), is discussed.
Parasitic nematodes of Oesophagostomum spp., commonly known, as 'nodular worms' are emerging as the most widely distributed and prevalent zoonotic nematodes. Oesophagostomum infections are well documented in African non-human primates; however, the taxonomy, distribution and transmission of Oesophagostomum in Asian non-human primates are not adequately studied. To better understand which Oesophagostomum species infect Asian non-human primates and determine their phylogeny we analysed 55 faecal samples from 50 orangutan and 5 gibbon individuals from Borneo and Sumatra. Both microscopy and molecular results revealed that semi-wild animals had higher Oesophagostomum infection prevalence than free ranging animals. Based on sequence genotyping analysis targeting the Internal transcribed spacer 2 of rDNA, we report for the first time the presence of O. aculeatum in Sumatran apes. Population genetic analysis shows that there is significant genetic differentiation between Bornean and Sumatran O. aculeatum populations. Our results clearly reveal that O. aculeatum in free-ranging animals have a higher genetic variation than those in semi-wild animals, demonstrating that O. aculeatum is circulating naturally in wildlife and zoonotic transmission is possible. Further studies should be conducted to better understand the epidemiology and dynamics of Oesophagostomum transmission between humans, non-human primates and other wild species and livestock in Southeast Asia.
- MeSH
- DNA helmintů genetika MeSH
- ezofágostomiáza * epidemiologie genetika veterinární MeSH
- feces parazitologie MeSH
- Hylobates parazitologie MeSH
- nemoci lidoopů * epidemiologie genetika parazitologie MeSH
- Oesophagostomum genetika MeSH
- Pongo pygmaeus parazitologie MeSH
- prevalence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Indonésie epidemiologie MeSH
- Názvy látek
- DNA helmintů MeSH
Key to the success of orangutan conservation management practices is the prevention of the introduction of infectious diseases to the remaining populations. Previous reports of Entamoeba spp. positive orangutans are of concern as Entamoeba spp. infection has been linked to morbidity and mortality in primates. It remains to be determined if the Entamoeba species infecting orangutans is the pathogenic Entamoeba histolytica. Orangutan fecal samples have been collected from orangutans from sites in Sumatra (Bukit Lawang, Ketambe, and Suaq, 241 samples from 64 individuals), and two sites in Kalimantan (Sebangau and Tuanan, 129 samples from 39 individuals). All samples were from wild orangutans except for a proportion from Sumatra which were from semi-wild (108 samples, 10 individuals). E. histolytica-specific nested PCR assays were carried out on the fecal samples. A total of 36 samples from 17 individuals tested positive for E. histolytica. When compared with published sequences using NCBI BLAST the E. histolytica positive samples showed a 98-99% concordance. The majority (76%, n = 36) of the positive isolates came from semi-wild orangutans in Bukit Lawang. This study supports the growing body of evidence that contact with humans is an important risk factor for infection of wild primates with E. histolytica.
- Klíčová slova
- Pongo, Protozoa, amoebiasis, parasite, zoonoses,
- MeSH
- entamébóza epidemiologie MeSH
- Entamoeba genetika izolace a purifikace MeSH
- feces parazitologie MeSH
- nemoci lidoopů epidemiologie parazitologie MeSH
- polymerázová řetězová reakce MeSH
- Pongo parazitologie MeSH
- zoonózy epidemiologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Borneo epidemiologie MeSH
- Indonésie epidemiologie MeSH
The emergence of cryptosporidiosis, a zoonotic disease of the gastrointestinal and respiratory tract caused by Cryptosporidium Tyzzer, 1907, triggered numerous screening studies of various compounds for potential anti-cryptosporidial activity, the majority of which proved ineffective. Extracts of Indonesian plants, Piper betle and Diospyros sumatrana, were tested for potential anti-cryptosporidial activity using Mastomys coucha (Smith), experimentally inoculated with Cryptosporidium proliferans Kváč, Havrdová, Hlásková, Daňková, Kanděra, Ježková, Vítovec, Sak, Ortega, Xiao, Modrý, Chelladurai, Prantlová et McEvoy, 2016. None of the plant extracts tested showed significant activity against cryptosporidia; however, the results indicate that the following issues should be addressed in similar experimental studies. The monitoring of oocyst shedding during the entire experimental trial, supplemented with histological examination of affected gastric tissue at the time of treatment termination, revealed that similar studies are generally unreliable if evaluations of drug efficacy are based exclusively on oocyst shedding. Moreover, the reduction of oocyst shedding did not guarantee the eradication of cryptosporidia in treated individuals. For treatment trials performed on experimentally inoculated laboratory rodents, only animals in the advanced phase of cryptosporidiosis should be used for the correct interpretation of pathological alterations observed in affected tissue. All the solvents used (methanol, methanol-tetrahydrofuran and dimethylsulfoxid) were shown to be suitable for these studies, i.e. they did not exhibit negative effects on the subjects. The halofuginone lactate, routinely administered in intestinal cryptosporidiosis in calves, was shown to be ineffective against gastric cryptosporidiosis in mice caused by C. proliferans. In contrast, the control application of extract Arabidopsis thaliana, from which we had expected a neutral effect, turned out to have some positive impact on affected gastric tissue.
- Klíčová slova
- Cryptosporidium, gastric, oocyst, pathology, treatment,
- MeSH
- Cryptosporidium účinky léků MeSH
- Diospyros chemie MeSH
- kokcidiostatika farmakologie MeSH
- kryptosporidióza prevence a kontrola MeSH
- Murinae * MeSH
- Piper betle chemie MeSH
- preklinické hodnocení léčiv veterinární MeSH
- rostlinné extrakty farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kokcidiostatika MeSH
- rostlinné extrakty MeSH
Fifteen Mongolian gerbils were inoculated with 10 × 106 viable trophozoites of Giardia intestinalis. Their faeces were examined daily by flotation method and the number of shed cysts was counted. Two animals (male and female) were euthanised at 4- to 5-day intervals (9, 14, 18 days post-infection (DPI)). The remaining nine gerbils were sacrificed and dissected at the end of the experiment (23 DPI). Their small intestinal tissues were processed for examination using histological sectioning and scanning electron microscopy and their complete blood count (CBC) was examined. The highest number of trophozoites at the total was observed in the duodenum in gerbils sacrificed on 14 DPI. Number of shed cysts was positively correlated with number of trophozoites rinsed from the intestine. Infected gerbils had lower body weight gain in comparison with control group and in three male gerbils; diarrhoea occurred during infection. Cyst shedding was negatively correlated with values of mean corpuscular haemoglobin concentration. Females showed another pattern in cyst shedding than males. This information needs to be taken into account while planning the experiments.
- Klíčová slova
- Gerbil model, Giardia intestinalis, Giardial intestinal distribution, Giardiasis,
- MeSH
- duodenum parazitologie patologie MeSH
- feces parazitologie MeSH
- Gerbillinae * parazitologie MeSH
- Giardia lamblia růst a vývoj fyziologie MeSH
- giardiáza parazitologie patologie MeSH
- lidé MeSH
- mikroskopie elektronová rastrovací MeSH
- modely nemocí na zvířatech MeSH
- trofozoiti růst a vývoj fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Piper betle has been used as a medicinal plant in traditional medical systems throughout South and South East Asia. Experimental studies have revealed its wide and diverse biological and pharmacological effects. In this study, antigiardial activity of Piper betle was tested using experimental infections of Giardia intestinalis, the most common cause of protozoal diarrhoea worldwide, in Mongolian gerbils. Plants were extracted in water, methanol and methanol:tetrahydrofuran. Gerbils were treated for ten days intragastrically twice a day, with the dose of 40 mg of the extract per 100 g of body weight. Drug metronidazole was used as a negative control. Gerbils' faeces were taken every day and examined by flotation method, the number of shed cysts were counted using a haemocytometer. After gerbils' sacrifice and dissection, their duodena were then processed for examination using histological sectioning and scanning electron microscopy. The antigiardial activity was evaluated by the course of cyst shedding throughout the entire experiment. A significant decline in cyst shedding, evaluated by linear regression was found in gerbils treated with the aqueous extract. Our results indicate that the aqueous extract of P. betle shows giardicidal effects.
- Klíčová slova
- Antigiardial activity, Drug of choice, Giardia intestinalis, Natural antiparasitics, Parasites, Piper betle,
- MeSH
- antiprotozoální látky farmakologie terapeutické užití MeSH
- feces parazitologie MeSH
- Gerbillinae MeSH
- Giardia lamblia účinky léků ultrastruktura MeSH
- giardiáza farmakoterapie MeSH
- lineární modely MeSH
- listy rostlin chemie MeSH
- lyofilizace MeSH
- metronidazol farmakologie terapeutické užití MeSH
- mikroskopie elektronová rastrovací MeSH
- Piper betle chemie MeSH
- průjem farmakoterapie parazitologie MeSH
- rostlinné extrakty farmakologie terapeutické užití MeSH
- tenké střevo parazitologie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indonésie MeSH
- Názvy látek
- antiprotozoální látky MeSH
- metronidazol MeSH
- rostlinné extrakty MeSH
The present study was conducted to evaluate the methanolic extracts from several plant leaves widely used in traditional medicine to cure digestive tract disorders and in the self-medication of wild animals such as non-human primates, namely Archidendron fagifolium, Diospyros sumatrana, Shorea sumatrana, and Piper betle leaves, with regard to their antimicrosporidial activity against Encephalitozoon cuniculi in immunocompetent BALB/c mice determined using molecular detection of microsporidial DNA (qPCR) in various tissues and body fluids of infected, treated mice. Of the plant extracts tested, Diospyros sumatrana provided the most promising results, reducing spore shedding by 88% compared to untreated controls. Moreover, total burden per 1 g of tissue in the D. sumatrana extract-treated group reached 87% reduction compared to untreated controls, which was comparable to the effect of the standard drug, Albendazole. This data represents the baseline necessary for further research focused on determining the structure, activity and modes of action of the active compounds, mainly of D. sumatrana, enabling subsequent development of antimicrosporidial remedies.
- Klíčová slova
- Antifungal, Encephalitozoon cuniculi, In vivo, Molecular detection, Plant extract,
- MeSH
- albendazol farmakologie terapeutické užití MeSH
- antifungální látky farmakologie terapeutické užití MeSH
- Cercopithecus aethiops MeSH
- dimethylsulfoxid farmakologie terapeutické užití MeSH
- Diospyros chemie MeSH
- Dipterocarpaceae chemie MeSH
- DNA fungální izolace a purifikace MeSH
- Encephalitozoon cuniculi účinky léků MeSH
- encephalitozoonóza farmakoterapie MeSH
- Fabaceae chemie MeSH
- feces parazitologie MeSH
- gastrointestinální trakt mikrobiologie MeSH
- imunokompetence MeSH
- listy rostlin chemie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- Piper betle chemie MeSH
- rostlinné extrakty farmakologie terapeutické užití MeSH
- spory hub účinky léků MeSH
- Vero buňky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indonésie MeSH
- Názvy látek
- albendazol MeSH
- antifungální látky MeSH
- dimethylsulfoxid MeSH
- DNA fungální MeSH
- rostlinné extrakty MeSH
Wild orangutan populations exist in an increasingly fragile state. As numbers continue to decline and populations became fragmented, the overall health of remaining individuals becomes increasingly at risk. Parasitic infections can have a serious impact on the health of wild orangutans, and can be fatal. It has been reported that rehabilitated individuals demonstrate a higher prevalence of parasitic diseases, and it is possible that they may spread these infections to wild orangutans upon reintroduction. In order to ensure the success of reintroduction and conservation efforts, it is crucial to understand the potential risks by fully understanding what parasites they have been reported to be infected with. Using this knowledge, future conservation strategies can be adapted to minimize the risk and prevalence of parasite transmission in the remaining orangutan populations. There is still limited information available on orangutan parasites, with several still not identified to the species level. Based on comprehensive literature review, we found 51 parasite taxa known to infect wild, semi-wild, and captive orangutans, including newly reported species. Here, we summarize methods used to identify parasites and draw conclusions relative to their reported prevalence. We also recommend fecal sample preservation and analytical methods to obtain best result in the future.
- Klíčová slova
- anthroponotic, disease, orangutan, parasitic infections, zoonotic,
- MeSH
- paraziti patogenita MeSH
- Pongo parazitologie MeSH
- prevalence MeSH
- primáti MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Excystation of sporozoites of Cryptosporidium parvum from oocysts is essential for successful in vitro assays. It has also been traditionally used as a measure for oocyst viability and infectivity. Laboratories use various excystation protocols so there is a need to clarify which method is the best. In this study, six different protocols for in vitro excystation of C. parvum oocysts were compared to find the most efficient excystation method (expressed as percentage excystation). Tested protocols differed in chemical pre-incubation steps, excystation media or time of incubation. There were significant differences in percentage of excysted oocysts among groups excysted by different methods. There were also significant differences in percentage of excysted oocysts between methods using pre-incubation with sodium hypochlorite and those without. The other variables examined; the presence of trypsin, kind of excystation medium and the incubation time, did not show statistical differences in percentage excystation among groups. Pre-incubation steps which included sodium hypochlorite, enhancing the permeability of the oocysts were found to increase the excystation ratio and methods using this step were the most effective.
- Klíčová slova
- Cryptosporidium parvum, Excystation methods, In vitro cultivation, Sodium hypochlorite, Trypsin,
- MeSH
- chlornan sodný farmakologie MeSH
- Cryptosporidium parvum účinky léků fyziologie MeSH
- kultivační média farmakologie MeSH
- oocysty účinky léků MeSH
- parazitologie metody MeSH
- permeabilita účinky léků MeSH
- trypsin farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chlornan sodný MeSH
- kultivační média MeSH
- trypsin MeSH
