Prevalence of intestinal parasites in small ruminants and their senstivity to treatments with ethnobotanical remedies in Cholistan, Pakistan

Livestock production contributes substantially to the livelihoods of poor rural farmers in Pakistan; strengthening pastoral communities plays an imperative role in the country’s thrive for poverty alleviation. Intestinal helminths constitute a major threat for pastoral livestock keepers in the whole country because chronic infestation leads to distinct losses in livestock productivity, particularly the growth of young animals. Synthetic anthelmintics have long been considered the only effective way of controlling this problem but high prices, side effects and chemical residues/toxicity problems, or development of resistance, lead to their very limited use in many pastoral systems. Additionally, poor pastoralists in remote areas of Pakistan hardly have access to appropriate anthelmintic drugs, which are also relatively expensive due to the long routes of transportation. The search for new and more sustainable ways of supporting livestock keepers in remote areas has given rise to studies of ethno-botanicals or traditional plant-based remedies to be used in livestock health care. Plant-based remedies are cheap or free of cost, environmentally safe and generally create no problem of drug resistance; they thus might substitute allopathic drugs. Furthermore, these remedies are easily available in remote areas and simple to prepare and/or administer. Cholistan desert is a quite poor region of Pakistan and the majority of its inhabitants are practicing a nomadic life. The region’s total livestock population (1.29 million heads) is almost twice that of the human population. Livestock husbandry is the primordial occupation of the communities and traditionally wealth assessment was based on the number of animals, especially goats and sheep, owned by an individual. Fortunately, about 60% of this desert region is richly endowed with highly adapted grasses, shrubs and trees. This natural flora has a rich heritage of scientifically unexplored botanical pharmacopoeia. Against this background, the present research project that was conducted under the umbrella of the International Center for Development and Decent Work at Kassel University, focused on a development aspect: in the Cholistan desert region it was firstly examined how pastoralists manage their livestock, which major health problems they face for the different animal species, and which of the naturally occurring plants they use for the treatment of animal diseases (Chapter 2). For this purpose, a baseline survey was carried out across five locations in Cholistan, using a structured questionnaire to collect data from 100 livestock farmers (LF) and 20 local healers (LH). Most of LF and LH were illiterate (66%; 70%). On average, LH had larger herds (109 animals) than LF (85 animals) and were more experienced in livestock husbandry and management. On average LF spent about 163 Euro per year on the treatment of their livestock, with a huge variability in expenditures. Eighty-six traditional remedies based on 64 plants belonging to 43 families were used. Capparaceae was the botanical family with the largest number of species used (4), followed by Chenopodiaceae, Poaceae, Solanaceae and Zygophyllaceae (3). The plants Capparis decidua (n=55 mentions), Salsola foetida (n=52), Suaeda fruticosa (n=46), Haloxylon salicornicum (n=42) and Haloxylon recurvum (n=39) were said to be most effective against the infestations with gastrointestinal parasites. Aerial parts (43%), leaves (26%), fruits (9%), seeds and seed oils (9%) were the plant parts frequently used for preparation of remedies, while flowers, roots, bulbs and pods were less frequently used (<5%). Common preparations were decoction, jaggery and ball drench; oral drug administration was very common. There was some variation in the doses used for different animal species depending on age, size and physical condition of the animal and severity of the disease. In a second step the regionally most prevalent gastrointestinal parasites of sheep and goats were determined (Chapter 3) in 500 animals per species randomly chosen from pastoral herds across the previously studied five localities. Standard parasitological techniques were applied to identify the parasites in faecal samples manually collected at the rectum. Overall helminth prevalence was 78.1% across the 1000 animals; pure nematode infestations were most prevalent (37.5%), followed by pure trematode (7.9%), pure cestode (2.6%) and pure protozoa infestations (0.8%). Mixed infestations with nematodes and trematodes occurred in 6.4% of all animals, mixed nematode-cestode infestations in 3.8%, and all three groups were found in 19.1% of the sheep and goats. In goats more males (81.1%) than females (77.0%) were infested, the opposite was found in sheep (73.6% males, 79.5% females). Parasites were especially prevalent in suckling goats (85.2%) and sheep (88.5%) and to a lesser extent in young (goats 80.6%, sheep 79.3%) and adult animals (goats 72.8%, sheep 73.8%). Haemonchus contortus, Trichuris ovis and Paramphistomum cervi were the most prevalent helminths. In a third step the in vitro anthelmintic activity of C. decidua, S. foetida, S. fruticosa, H. salicornicum and H. recurvum (Chapter 2) was investigated against adult worms of H. contortus, T. ovis and P. cervi (Chapter 3) via adult motility assay (Chapter 4). Various concentrations ranging from 7.8 to 500 mg dry matter/ml of three types of extracts of each plant, i.e. aqueous, methanol, and aqueous-methanol (30:70), were used at different time intervals to access their anthelmintic activity. Levamisol (0.55 mg/ml) and oxyclozanide (30 mg/ml) served as positive and phosphate-buffered saline as negative control. All extracts exhibited minimum and maximum activity at 2 h and 12 h after parasite exposure; the 500 mg/ml extract concentrations were most effective. Plant species (P<0.05), extract type (P<0.01), parasite species (P<0.01), extract concentration (P<0.01), time of exposure (P<0.01) and their interactions (P<0.01) had significant effects on the number of immobile/dead helminths. From the comparison of LC50 values it appeared that the aqueous extract of C. decidua was more potent against H. contortus and T. ovis, while the aqueous extract of S. foetida was effective against P. cervi. The methanol extracts of H. recurvum were most potent against all three types of parasites, and its aqueous-methanol extract was also very effective against T. ovis and P. cervi. Based on these result it is concluded that the aqueous extract of C. decidua, as well as the methanol and aqueous-methanol extract of H. recurvum have the potential to be developed into plant-based drugs for treatment against H. contortus, T. ovis and P. cervi infestations. Further studies are now needed to investigate the in vivo anthelmintic activity of these plants and plant extracts, respectively, in order to develop effective, cheap and locally available anthelmintics for pastoralists in Cholistan and neighboring desert regions. This will allow developing tangible recommendations for plant-based anthelminthic treatment of sheep and goat herds, and by this enable pastoralists to maintain healthy and productive flocks at low costs and probably even manufacture herbal drugs for marketing on a regional scale.

Frequency of Blastocystis hominis and other intestinal parasites in stool samples examined at the Parasitology Laboratory of the School of Pharmaceutical Sciences at the São Paulo State University, Araraquara

Blastocystis hominis é um protozoário, causador de infecção intestinal denominada blastocistose humana, cujo diagnóstico é realizado pelo exame coproparasitológico e por meio de técnicas de coloração permanente. Este estudo foi desenvolvido para avaliar a freqüência da infecção por Blastocystis hominis em habitantes da região de Araraquara/SP, bem como comparar diferentes métodos para a pesquisa desse protozoário em amostras de fezes. Foram estudadas 503 amostras de fezes submetidas ao exame direto a fresco, às técnicas de Faust e cols, Lutz e de Rugai e cols, além das colorações pela hematoxilina férrica, tricrômio e de Kinyoun modificada. Entre as 503 amostras examinadas, 174 (34,6%) apresentaram-se positivas para a presença de parasitas intestinais. O protozoário e o helminto mais freqüentes foram Entamoeba coli (14,6%) e Strongyloides stercoralis (6,7%), respectivamente. Blastocystis hominis foi observado em 23 (4,6%) amostras fecais com consistência predominantemente pastosa, não caracterizando quadro diarréico. Apesar da baixa freqüência de Blastocystis hominis encontrada na região de Araraquara, comparativamente a outras regiões brasileiras, é importante a realização do diagnóstico laboratorial desse protozoário. O encontro de Blastocystis hominis em material fecal é indicativo de contaminação de alimentos e água de consumo, desde que se admita a rota de transmissão oral-fecal desse parasita, o que implica na orientação da população sobre as medidas de saneamento básico e higiene como meio para se controlar problemas de saúde ocasionados pelos enteroparasitas.

Prevalence of intestinal parasites in dogs from São Paulo State, Brazil

The prevalence of gastrointestinal parasites in stray dogs, and dogs with owners was investigated by fecal examinations from 271 dogs employing sedimentation, simple flotation and centrifugation-flotation methods. The centrifugation-flotation method, when compared to simple flotation or sedimentation methods was generally more accurate in the diagnosis of all intestinal parasites, but statistical differences were detected only in relation to Giardia spp. and Cystoisospora spp. (synonym Isospora spp.). The following parasites, with their respective prevalence, were diagnosed in the fecal samples: Ancylostoma spp. (23.6%); Toxocara canis (5.5%); Trichuris vulpis (4.8%); Spirocerca lupi (1.9%); Dipylidium caninum (0.7%); Giardia spp. (12.2%); Hammondia heydorni (2.6%); Cystoisospora spp. (8.5%); and Sarcocystis spp. (2.2%). The prevalence of most parasites was similar for dogs of mixed-breed and for dogs of a defined-breed, except for Cystoisospora spp. and T canis which showed a significantly higher prevalence in mixed-breed dogs. The prevalence of Ancylostoma spp. (17.1%) was significantly lower in stray dogs than in those with an owner (31.9%) and the prevalence of Giardia spp. and Cystoisospora spp. was higher in stray dogs (P < 0.05). No effect of season on the occurrence of the different parasite genera could be observed, except for Ancylostoma spp., for which an increase in the percentage of dogs shedding eggs was observed at the beginning of Summer with a peak occurrence during April and May (Autumn). The prevalence of Ancylostoma spp., T canis, T vulpis, Giardia spp. and Cystoisospora spp. was higher in adult males than in adult females, but significant differences between the two groups occurred only with Giardia spp. Young animals were found to more frequently shed Nematode eggs in feces than adult animals. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Comparison of three concentration methods for the recovery of canine intestinal parasites from stool samples

The aim of present study was to compare the efficiency of a commercial assay and two conventional methods for fecal concentration in detecting canine gastrointestinal parasites. Fecal samples from 254 dogs were processed by centrifugation-sedimentation (CS), centrifugation-flotation (CF) and a commercial assay for fecal concentration (TF-test (R)). The following parasites were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Isospora (3.5%), and Sarcocystis (2.7%). The calculated analytical sensitivity indicated that CF was more accurate (P < 0.01) in detecting Ancylostoma, T. canis, T. vulpis and Giardia infections. However, CF showed significantly higher sensitivity only for Ancylostoma, compared to the other two methods. The kappa index value of diagnostic agreement between TF-test and CF was high for T. canis (83%) and moderate for Giardia (72%) and Ancylostoma (63%). The advantages and limitations of each method were assessed for individual diagnosis and epidemiological investigation. (C) 2010 Elsevier B.V. All rights reserved.

Prevalence of dog intestinal parasites and risk perception of zoonotic infection by dog owners in São Paulo State, Brazil

Coprological examination was used to estimate the prevalence of gastrointestinal parasites in stray and domiciled dogs from Botucatu, São Paulo State, Brazil. Risk factors for dog infection were assessed in relation to demographic, husbandry and management data. The dog owners completed a questionnaire survey on some aspects of dog parasitism such as parasite species, mechanisms of infection, awareness of zoonotic diseases and history of anthelmintic usage. Parasites were found in the faeces of 138 dogs, with an overall prevalence of 54.3%. Dogs harbouring one parasite were more common (31.4%) than those harbouring two (18.5%), three (3.2%) or four (1.2%). The following parasites and their respective frequencies were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Dipylidium caninum (2.4%), Isospora (3.5%), Cryptosporidium (3.1%) and Sarcocystis (2.7%). Stray dogs were found more likely to be poliparasitized (P < 0.01) and presented higher prevalence of Ancylostoma, T. canis and Giardia (P < 0.01) than domiciled ones. Toxocara canis was detected more frequently in dogs with < 6 months of age (P < 0.05) and no effect of sex or breed could be observed (P > 0.05). Except for Ancylostoma, that showed a significantly higher prevalence in dogs living in a multi-dog household (P < 0.01), parasite prevalences were similar in single- and multi-dog household. The answers of dog owners to the questionnaire showed that the majority does not know the species of dog intestinal parasites, the mechanisms of transmission, the risk factors for zoonotic infections, and specific prophylactic measures. The predominance of zoonotic species in dogs in the studied region, associated with the elevated degree of misinformation of the owners, indicates that the risk of zoonotic infection by canine intestinal parasite may be high, even in one of the most developed regions of Brazil.

Automated diagnosis of human intestinal parasites using optical microscopy images

Intestinal parasitosis constitutes a serious health problem in most tropical countries. The diagnosis of enteroparasites in laboratory routine relies on the examination of stool samples using optical microscopy and the error rates usually range from moderate to high. Approaches based on automatic image analysis have been proposed, but the methods are usually specific for some species, some of them are computationally expensive, and image acquisition and focus are manual. We present a solution to automate the diagnosis of the 15 most common species of enteroparasites in Brazil, using a sensitive parasitological technique, a motorized microscope with digital camera for automatic image acquisition and focus, and fast image analysis methods. The results indicate that our solution is effective and suitable for laboratory routine, in which the exam must be concluded in a few minutes. © 2013 IEEE.

Automatic segmentation and classification of human intestinal parasites from microscopy images

Human intestinal parasites constitute a problem in most tropical countries, causing death or physical and mental disorders. Their diagnosis usually relies on the visual analysis of microscopy images, with error rates that may range from moderate to high. The problem has been addressed via computational image analysis, but only for a few species and images free of fecal impurities. In routine, fecal impurities are a real challenge for automatic image analysis. We have circumvented this problem by a method that can segment and classify, from bright field microscopy images with fecal impurities, the 15 most common species of protozoan cysts, helminth eggs, and larvae in Brazil. Our approach exploits ellipse matching and image foresting transform for image segmentation, multiple object descriptors and their optimum combination by genetic programming for object representation, and the optimum-path forest classifier for object recognition. The results indicate that our method is a promising approach toward the fully automation of the enteroparasitosis diagnosis. © 2012 IEEE.

A survey on intestinal parasites of livestock guardian dogs and herding dogs in Switzerland

The present study describes the occurrence of intestinal parasite infections in livestock guardian dogs and herding dogs. A total of 71 guardian dogs (more than half of the total number of guardian dogs in Switzerland) and 21 herding dogs were coprologically examined, using a combined sedimentation-flotation method. In 21 (23 %) of the dogs intestinal parasites were detected, and 8 (8.7 %) of these dogs shed either sporocysts of Sarcocystis sp. (n = 6) or taeniid eggs (n = 2). The evaluation of questionnaires providing data on age, origin and deworming schemes of the dogs completed the study.

New evidence for the antiquity of the intestinal parasite Trichuris (whipworm) in Europe

The whipworm, Trichuris trichiura L., is one of the most common human intestinal parasites worldwide, yet little is known of its origin and global spread. Archaeological records for this nematode have all been of Neolithic or later date, suggesting a possible association between the spread of pastoral farming and human acquisition of whipworm. This paper reports the discovery of eggs of the genus Trichuris in late Mesolithic deposits from south Wales, indicating that whipworm was present in Europe before the arrival of agriculture. This raises the possibility that human infection by Trichuris arose through contact with wild animals in parts of the landscape frequented by both human and animal groups.

Intestinal parasitic infections in HIV/AIDS patients: epidemiological, nutritional and immunological aspects

This study applied a socioeconomic questionnaire designed to evaluate the frequency of intestinal parasites and characterize epidemiological, nutritional, and immunological variables in 105 HIV/AIDS patients - with and without parasitic infections, attending the Day Hospital in Botucatu, UNESP, from 2007 to 2008. Body mass index was calculated and the following tests performed: parasitological stool examinations; eosinophil, IgE, CD4(+) T and CD8(+) T lymphocyte cell counts; albumin test; viral load measure; and TNF-alpha, IFN-gamma, IL-2, IL-5 and IL-10 cytokine levels. Results were positive for parasitic intestinal infections in 12.4% of individuals. Most patients had good socioeconomic conditions with basic sanitation, urban dwellings, treated water supply and sewage, good nutritional and immunological status and were undergoing HAART. Parasites were found at the following frequencies: Entamoeba - five patients (38.5%), Giardia lamblia-four (30.7%), Blastocystis hominis-three (23.0%), Endolimax nana-two (15.4%), and Ascaris lumbricoides - one (7.7%). There were no significant differences between the two groups for eosinophils, albumin, IgE, CD4(+) T and CD8(+) T lymphocytes, INF-gamma, IL-2, or IL-10. Most patients also showed undetectable viral load levels. Significant differences were found for TNF-alpha and IL-5. These results show the importance of new studies on immunodeficient individuals to increase understanding of such variables.

INFLUENCE OF ENTERAL PARASITES ON THE BLOOD VITAMIN-A LEVELS IN PRESCHOOL-CHILDREN ORALLY SUPPLEMENTED WITH RETINOL AND OR ZINC

A sample of 471 pre-school children who frequented schools and creches in a poor district of Manaus (Amazonas), Brazil, were randomly submitted to faecal parasitological tests. Two-hundred-and-forty children from both sexes between the ages of 3 and 7 years with Ascaris lumbricoides and/or Giardia lamblia were selected. The objective of the study was to determine the possible influence of these two intestinal parasites and vitamin A and/or zinc supplementation on the serum retinol levels of primary school children. The children were submitted to clinical and anthropometric examinations, dietary interviews and biochemical examinations of retinol and carotene in the serum and of zinc in the hair. The parasitic incidence was 85.0% and about 54% of the children were polyparasitic. During the pretreatment phase, the retinol and carotene serum levels were 36% and 57%, respectively, below the normal levels. Using the Waterlow classification, the anthropometric analyses revealed that 88% of the children showed normal growth. A significant effect was observed of the anti-parasitic medicine on the serum retinol levels.

Parasitismo intestinal em uma aldeia indígena Parakanã, sudeste do Estado do Pará, Brasil

O trabalho visa estudar a ocorrência e os aspectos epidemiológicos do parasitismo intestinal na aldeia Paranatinga da tribo indígena Parakanã, Amazônia Oriental Brasileira. Foram realizados dois inquéritos coproparasitológicos, em abril de 1992 e em fevereiro de 1995. Os métodos utilizados na identificação dos agentes parasitários foram os de Hoffman e exame direto, dois métodos simples, facilmente exeqüíveis em aldeias indígenas. Da amostra de 126 índios em abril de 1992 (população de 215 índios), 101 (80,2%) encontravam-se parasitados por pelo menos um enteroparasita. Ancilostomídeos foram encontrados em 33,3%, A. lumbricoides, em 42,8%, T. trichiura, em 0,8%, e S. stercoralis, em 5,6%. Em relação aos protozoários, E. histolytica foi encontrada em 65,0% e G. lamblia, em 46,8%. No inquérito de fevereiro de 1995, apesar do aumento da prevalência total em comparação com o de abril de 1992 (p = 0,04), houve diminuição das prevalências de ancilostomídeos, E. histolytica, G. lamblia, e ausência de S. stercoralis (p<0,05). A despeito das ações de saúde e saneamento empreendidas nas aldeias Parakanã, a prevalência de enteroparasitas ainda se encontra elevada na aldeia Paranatinga, sugerindo que as medidas de atenção primária e secundária devem ser imediatamente incrementadas.

Molecular characterization of intestinal protozoa in two poor communities in the State of São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estimating the burden of disease attributable to unsafe water and lack of sanitation and hygiene in South Africa in 2000

Objectives To estimate the burden of disease attributable to unsafe water, sanitation and hygiene (WSH) by age group for South Africa in 2000. Design World Health Organization comparative risk assessment methodology was used to estimate the disease burden attributable to an exposure by comparing the observed risk factor distribution with a theoretical lowest possible population distribution. A scenario-based approach was applied for estimating diarrhoeal disease burden from unsafe WSH. Six exposure scenarios were defined based on the type of water and sanitation infrastructure and environmental faecal-oral pathogen load. For ‘intestinal parasites’ and schistosomiasis, the burden was assumed to be 100% attributable to exposure to unsafe WSH. Setting South Africa. Outcome measures Disease burden from diarrhoeal diseases, intestinal parasites and schistosomiasis, measured by deaths and disability-adjusted life years (DALYs). Results 13 434 deaths were attributable to unsafe WSH accounting for 2.6% (95% uncertainty interval 2.4 - 2.7%) of all deaths in South Africa in 2000. The burden was especially high in children under 5 years, accounting for 9.3% of total deaths in this age group and 7.4% of burden of disease. Overall, the burden due to unsafe WSH was equivalent to 2.6% (95% uncertainty interval 2.5 - 2.7%) of the total disease burden for South Africa, ranking this risk factor seventh for the country. Conclusions Unsafe WSH remains an important risk factor for disease in South Africa, especially in children under 5. High priority needs to be given to the provision of safe and sustainable sanitation and water facilities and to promoting safe hygiene behaviours, particularly among children.

Encroachment of Echinococcus granulosus into urban areas in eastern Queensland, Australia

Objective: To investigate the prevalence of Echinococcus granulosus in wild dogs (dingos and dingo-domestic dog hybrids) living in and around human habitation on Fraser Island and in townships of the Maroochy Shire, on Queensland's Sunshine Coast, Australia. Design: Wild dogs were humanely killed on Fraser Island and in the Maroochy Shire because they were deemed a potential danger to the public. Their intestines were collected and the contents examined for intestinal parasites. Procedure: Intestines were removed as soon after death as possible, packed in plastic bags and kept frozen until examination. The intestinal contents were washed, sieved and examined microscopically for the presence of helminths, which were identified and counted. Results: Intestines from 108 wild dogs, 7 foxes and 18 Fraser Island dingoes were examined. Echinococcus granulosus was only present in the wild dogs from Maroochy Shire (46.3%) with worm burdens of between 30 and 104,000. Other helminths included Spirometra erinacei, Dipylidium caninum, Taenia spp., Ancylostoma caninum and Toxocara canis. Two specimens of a trematode (Haplorchinae sp.) usually found infecting fish and seabirds were recovered from a Fraser Island dingo. Conclusion: Dingoes on Fraser Island are not infected with E. granulosus and do not pose a hydatid disease public health risk to residents or visitors. However, wild dogs examined from the Maroochy Shire do present a potential hydatid disease public health risk.