Botanical ethnoveterinary therapies in three districts of the Lesser Himalayas of Pakistan

Background Ethnoveterinary knowledge is highly significant for persistence of traditional community-based approaches to veterinary care. This is of particular importance in the context of developing and emerging countries, where animal health (that of livestock, especially) is crucial to local economies and food security. The current survey documents the traditional veterinary uses of medicinal plants in the Lesser Himalayas-Pakistan. Methods Data were collected through interviews, focus groups, participant observation, and by administering questionnaires. A total of 105 informants aged between 20–75 years old who were familiar with livestock health issues (i.e. farmers, shepherds, housewives and herbalists) participated in the study. Results A total of 89 botanical taxa, belonging to 46 families, were reported to have ethnoveterinary applications. The most quoted families were Poaceae (6 taxa), Fabaceae (6), Asteraceae (5), and Polygonaceae (5). Adhatoda vasica was the most cited species (43%), followed by Trachyspermum ammi (37%), and Zanthoxylum armatum var. armatum (36%). About 126 medications were recorded against more than 50 veterinary conditions grouped into seven categories. The highest cultural index values were recorded for Trachyspermum ammi, Curcuma longa, Melia azedarach, Zanthoxylum armatum var. armatum and Adhatoda vasica. The highest informant consensus factor was found for pathologies related to respiratory and reproductive disorders. Comparison with the local plant-based remedies used in human folk medicine revealed that many of remedies were used in similar ways in local human phytotherapy. Comparison with other field surveys conducted in surrounding areas demonstrated that approximately one-half of the recorded plants uses are novel to the ethnoveterinary literature of the Himalayas. Conclusion The current survey shows a remarkable resilience of ethnoveterinary botanical knowledge in the study area. Most of the species reported for ethnoveterinary applications are wild and under threat. Thus, not only is it imperative to conserve traditional local knowledge of folk veterinary therapies for bio-cultural conservation motives, but also to assist with in-situ and ex-situ environmental conservation initiatives, which are urgently needed. Future studies that focus on the validation of efficacy of these ethnoveterinary remedies can help to substantiate emic concepts regarding the management of animal health care and for rural development programs.


Introduction
Ethnoveterinary medicine is a broad field encompassing people's beliefs, skills, knowledge and practices related to veterinary health care [1]. Medicinal plants traditionally used in the treatment of animal diseases play a crucial role in local health modalities. Specifically, phytotherapeutics often represent the primary form of therapy in rural veterinary care as allopathic modalities remain inaccessible, especially in the developing world [2]. Therefore, local knowledge of ecological resources for veterinary care is of particular importance to pastoral and agro-pastoral communities that rely heavily on livestock for their livelihood and food security. However, traditional ethnoveterinary knowledge is still mainly orally transmitted from generation to generation (i.e., in the form of traditional remedies, poems, drawings stories, folk myths, proverbs and songs). Due to the nature of oral transmission, this form of local knowledge remains fragile and threatened, and presents an urgent need for being recorded and documented.
An increasing number of studies have very recently focused on the documentation of local ethnoveterinary practices in South Asia . These studies hold potential for having a tremendous impact on the Himalayan region, in particular, where efforts for sustaining endogenous development and ultimately improving the health and well-being of both animals and humans is still largely neglected. Pakistan has a very large livestock population composed of a number of local breeds that are well adapted to local conditions. In particular, there are an estimated 27 million buffaloes, 30 million cattle, 27 million sheep, 54 million goats, one million camels, 0.3 million horses, 4 million asses, 0.2 million mules and 74 million poultry in Pakistan [25].
The objectives of this field study were multifold: 1. to record the local knowledge related to medicinal plants used for treating animal diseases in the Lesser Himalayan region in Pakistan; 2. to compare the collected data with the traditional medical knowledge devoted to humans in the same region; 3. to compare the collected data with those of other ethnoveterinary studies conducted in the Himalayan region over the last decades; 4. to assess their cultural importance and the consensus among the informants regarding cited veterinary pathologies; and 5. to examine local perceptions of factors that threaten wild medicinal plant resources.

Study site
An ethnobotanical study was conducted from March 2010 to April 2013 in different locations of the Lesser Himalayas, which is a hotspot for plant biodiversity in Pakistan. Fifty-five localities in three districts (Haripur, Abbottabad and Mansehra) within the Khyber Pakhtunkhwa (KPK) province were selected for inclusion in the study (Figure 1). The Lesser Himalayan range in Pakistan lies between 33°-44′ and 35°-35′ north latitude and between 72°-33′ and 74°-05′ east longitude, comprising an area of 23,295 km 2 . The climate of the area is subtropical in the lowland plains and foot-hills zone and subtropical-sub alpine in middle Himalayas, Siwalik, Murree and entire Hazara hills. The average rainfall varies from 70-90 mm in southern and 100-130 mm in the northern parts. The vegetation of the Lesser Himalayas falls within the subtropical, temperate, sub-alpine and alpine zones. The region is divided into six vegetation zones, namely: the subtropical sub-humid zone, the subtropical humid zone, the temperate humid zone, the sub alpine zone and the zone of the glaciers/snowfields. This area is populated by several ethnic groups (Syed, Abbasi, Karaal, Jadoon, Tanoli, Ghakar, Gujar, and Awan), all speaking the Hindko dialect of the Western Punjabi, and belonging in turn to the Indo-Aryan (Indic) language family spoken in Northern Pakistan.

Ethnobotanical data collection
Ethnobotanical surveys were conducted in all four seasons. Participatory rural appraisal (PRA) approaches were adopted during fieldwork and prior informed consent was obtained before conducting interviews. Information regarding ethnoveterinary practices was collected through semistructured interviews and guided fieldtrips with the help of traditional healers. A total of 105 informants (75 males and 30 females), ranging from 20-75 years old and including farmers, shepherds, housewives and herbalists familiar with livestock problems and use of conventional recipes, were interviewed and their responses recorded in detail.
Information regarding the vernacular plant names, part (s) used, methods of preparation, mode/route of application and treated diseases were documented during each interview. Taxonomic identification of the collected plant samples was carried out with the help of Flora of Pakistan [26], The Plant List [27] and by one of the authors (MAK, plant taxonomist). Family nomenclature follows the Angiosperm Phylogeny Group III designations [28]. Additionally, 15 key informants were selected at four locations within three study districts (Haripur, Abbottabad and Mansehra) and specific information regarding the perceived threats for the local medicinal flora was obtained. Following identification of 5 key perceived threats (agricultural land expansion, overharvesting, overgrazing, fuel, fire), we employed pair-wise ranking techniques in which respondents were presented with two threats and chose one from the two threats at a time [29]. Respondent scores were then summed up and ranks for each threat determined by region.

Data analysis
Cultural importance index (CI) values for each species and mean cultural importance values for each family (mCIf ) were calculated as described in a previous quantitative ethnobotanical work [30]. Briefly, CI values of species were calculated based on previously described methods [31] and express the sum of the proportion of informants that mention each species used. The CI values for each species were calculated using the following formula, with UR i : use reports in each use-category and N: total number of survey participants: Moreover, we calculated the mean cultural importance (mCI) index of plant species as measured in three study districts (Haripur, Abbottabad, Mansehra) within the Khyber Pkahtunkhaw Province of Pakistan, on the basis of their cultural importance index (CI) calculated for each single district. To calculate the mCIf, CI values of all reported species within a family were added. Regression analysis was performed upon comparison of mCIf with the number of species in each respective family.

Result and discussion
Taxonomic diversity of the species A total of 89 plant species belonging 81 genera and 46 families were reported by the study participants against veterinary aliments have been gathered and documented alphabetically along with their local names, parts used, preparations, applications, indications and citation numbers (Table 1). Among the most utilized botanical families, Poaceae and Fabacaeae were ranked first with highest number of species (6 taxa), followed by Asteraceae (5), Polygonaceae (5), and Apocynaceae (4) (Figure 2).

Plant parts used, their preparations and applications
Among the plant parts included in veterinary applications, leaves were most commonly used (26%), followed by seeds (13%), whole plant (13%), and fruits (11%). The methods of preparation of the therapeutic materials sometimes varied from individual to individual (e.g., the same plant material for the same ailment could be prepared in different ways, depending upon the preferences of different healers). A list of 126 ethnoveterinary remedy preparations is presented in Table 1. The large majority of recipes being were prepared from single plants (70%) rather than mixtures. In most cases, water was the solvent employed in preparation of the remedy. Besides plants and water, some other materials were also commonly incorporated in the preparations: salt, sugar, milk, oil, eggs and ghee. The most common therapeutic formulations fall into eleven main categories, the most popular of which were pastes (35%), fresh plant parts (15%), decoctions (14%), and powders (13%).

Ethnoveterinary plant uses
Study participants identified more than 50 veterinary ailments that could be grouped into seven general categories: gastrointestinal disorders (these were treated by 47 formulations); skin infections (30), parasites, fever and respiratory diseases (10), reproductive disorders (9), lactation (4), and musculoskeletal system disorders (3). More than 40 taxa were documented for their application to treat more than two veterinary conditions. By comparing the present data with all of the available ethnoveterinary literature concerning the surrounding geographic areas, it appears that nearly half of the quoted plants have never been described before as useful in folk veterinary practices. The other half has already been reported in the literature, but in some cases, for different ethnoveterinary purposes. In this section, we explore some reports on other ethnoveterinary applications of these species in the literature. This discussion is organized by plant family.

Acanthaceae
Regarding Adhatoda vasica, the leaves are used to treat stomach pains, fever, dehydration, diarrhea, dysentery, indigestion and gas troubles. The leaf paste of this plant has been reported for uses in the treatment of hoof rot in the literature [46]. Interestingly, aqueous extracts from the leaves have shown significant activity against Bacillus bacteria [47,48].

Amaranthaceae
Paste prepared from whole plant of Amaranthus viridis is used here against weakness in cattle. The leaves of the same plant were reported as emollient in amenorrhea, scorpion sting and snake bite in a study conducted in Islamabad, Pakistan [49].

Amaryllidaceae
The crushed bulbs of Allium cepa are administered to treat indigestion, stomach gripe, fever and for lactation in the study area, whereas in Italy, they are used to prevent pestilence [50]. The leaves, flowers and bulb extracts of A. cepa have demonstrated activity against pathogens such as Staphylococcus aureus, Salmonella typhi, Candida albicans, and nematodes [48,51].

Anacardiaceae
The fruit pickle of Mangifera indica is used for mouth infections. Others have reported that the leaves of same plant are fed to livestock to treat retained fetal membrane [20]. Chloroform, ethanolic, water and petroleum ether extracts of M. indica were found to have antibacterial, anti-viral anti-fungal activities, as well as antiinflammatory properties [52,53].

Apiaceae
Aerial parts of Foeniculum vulgare were used to treat indigestion and diarrhea. Flowers and fruit of the same species have been reported as galactagogues and ruminative [50]. Seeds of Trachyspermum ammi are given to cattle as appetite stimulant and to increase milk production. In the Sargodha district of Pakistan, seed powder and decoctions of the same plant were reported for treatments against genital prolapse and to treat retained fetal membrane [20]. Alcoholic and aqueous extracts of this plant species have shown antibacterial activity [54].

Apocynaceae
Leaves, stems and twigs of Calotropis procera are applied to cure mouth and eye watering, colic, indigestion, pain and inflammation. Other reports regarding use of this plant include crushed leaves for the relief of flatulence, latex to increase lactation and bark decoction for hoof rot [46]. The leaves and seeds are also reported to be useful for silent estrus and delayed puberty [20]. Alcoholic and aqueous extracts of C. procera have shown antibacterial activities [54]. Powder prepared from the roots and leaves of Carissa opaca is given to cattle to treat infected or sore throats and to heal wounds. In Uttar Pradesh, India, aerial parts of C. opaca were reported to be administered orally to kill pest in cattle [46].

Araliaceae
An aqueous extract of Hedera nepalensis is applied to remove leech in cattle. In Italy, the use of fresh leaves and plant decoctions for abortive and anti-inflammatory purposes have been reported [50].

Asteraceae
The seeds and paste made from the whole plant of Saussurea heteromalla are used to treat edema and to purify the blood. In Islamabad, the seeds were reported as carminatives and used also in tonics for horses and camels [49]. In the present study, we found that decoctions and pastes of Senecio chrysanthemoides are used for the treatment of sore joints and arthritis, whereas other work has reported the use of roots and leaves for treating blackleg disease and Evil-eye [55].

Boraginaceae
The leaf paste of Trichodesma indicum is used to treat stomach disorders and intestinal worms in cattle in the study area, whereas others have reported the use of this paste in the treatment of mastitis and for uterine prolapse [46].

Brassicaceae
Brassica campestris seed oil is used for skin, eye and stomach infections. Other studies in Pakistan and India [20,46] have reported the use of this oil in topical applications for sores and the treatment of genital prolapses. Eruca sativa seed oil is used to treat dysentery in the study area. E. sativa seed powder has been reported for diarrhea in other work [46].

Cannabaceae
Paste from the leaves, seeds and floral buds of Cannabis sativa are applied as an appetite stimulant, anti-leech, anti-lice, and for abdominal swelling and indigestion. Other studies have reported the use of decoctions and infusions for measles and East coast fever [56] and leaves for genital prolapse [20].

Convolvulaceae
Paste prepared from Cuscuta reflexa is fed to cattle for treatment of swelling (rumination problems), indigestion and short mammary glands. Other studies have documented its use as a galactagogue food (after being fried) [57].

Euphorbiaceae
Seed oil of Ricinus communis is administered to treat constipation. Other studies have documented the use of R. communis for intestinal obstruction, hoof problems, digestive problems, wounds, abscesses, to expel retained placenta and for silent estrus/delayed puberty in cattle [20,46,58]. The stem/leaf hexane extract of R. communis was suggested to be active against Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus [59].

Fabaceae
Acacia nilotica bark decoctions are used for the treatment of stomach pains in livestock. The bark of this same plant has been reported to be used in the case of hoof rot and genital prolapse in cattle [20,46]. The seeds of Trigonella foenum-graecum are reported to treat diarrhea here, whereas in other areas of Pakistan they are used for treatment of genital prolapse, silent estrus and delayed puberty [20].

Lythraceae
The fruit rind of Punica granatum is used to cure dysentery. Other work reports the use of leaf paste for enteritis, bark powder for helminthic infection, flowers as a tonic and the rind as an astringent and to treat diarrhea [60]. Antibacterial studies on the alcoholic and aqueous extracts of this plant have demonstrated activity against Bacillus subtilis, Escherichia coli, Proteus vulgaris, Salmonella typhimurium, Pseudomonas aeruginosa and Staphylococcus aureus [51].

Meliaceae
The leaves and fruit of Melia azedarach are used against foot, mouth, skin infections, gas trouble and indigestion in the study areas while according to other studies; it is used as a cooling agent and for genital prolapse [20,46].

Ptderidaceae
Adiantum incisum leaf paste is used for abdominal pain in the study area, whereas in Italy, a decoction of the plant is used to expel the placenta following delivery [50].

Poaceae
Paste prepared from the seeds of Oryza sativa is used to treat weakness and respiratory infection. It was reported [20,50] that seeds of the same plant are also used against diarrhea and to treat retained fetal membrane. Triticum aestivum seeds are used against dysentery, sore mouth and to increase milk production in livestock. Other studies have reported its use as a ruminative, laxative, for dermatitis, delayed puberty, silent estrus and to treat retained fetal membrane [20,50]. Zea mays inflorescences are given to cure urinary inflammation in cattle. Z. mays has been reported for applications in wound healing and treating genital prolapse in other studies [20,50].

Polygonaceae
Local people use the roots and leaves of Rumex nepalensis for treating diarrhea, dysentery, intestinal worms, allergies and to stop bleeding in cattle. Crushed roots of this plant have been reported for treatment of blackleg disease (an infectious disease attributed to Clostridium spp.) [55].

Rutaceae
Citrus limon juice is used in the treatment of mastitis. Others have reported the use of citrus juice for uterine prolapse in cattle [20].

Sapindaceae
The powder and juice of Aesculus indica fruit and seeds is used against cough, fever, abdominal pain and to heal wounds in animals in the study area. However, in other regions of Pakistan, the seed endocarp is given to horses to relieve stomach pain, colic and swelling [61][62][63].

Saxifragaceae
Fresh leaves and powder derived from the rhizomes of Bergenia ciliata are topically applied for use in wound healing. Dried and fresh leaves of the same plant have been used to treat diarrhea in animals [64]. Alcoholic and aqueous extracts of B. ciliata rhizome has shown antibacterial and antifungal activities [65].

Scrophulariaceae
The fresh leaf paste of Verbascum thapsus is used to treat diarrhea. Others report the use of a leaf ointment for the treatment of rectal prolapse [50].

Solanaceae
Solanum surattense is used for healing wounds, fever, indigestion, cough and as a tonic. Others have reported the use of the leaves for genital prolapse [20]. A leaf extracts of S. surattense was found to be active against Staphylococcus aureus, Salmonella typhi, Candida albicans and nematodes [48,51]. The root paste of Withania somnifera is topically applied to treat bovine mastitis in this study area, whereas the crushed roots of this same species are used against an evil spirit (Wan laffa) in animals in Ethiopia [55]. Alcoholic and aqueous extracts of W. somnifera have shown antibacterial activity against Bacillus subtilis, Escherichia coli, Proteus vulgaris, Salmonella typhimurium, Pseudomonas aeruginosa and Staphylococcus auerus, as well diuretic and anti-hypercholesterolemic activities [48,54].

Theaceae
Decoctions of Camellia sinensis leaves are used to cure fever in cattle in this region, while another study in Sargodha district (Pakistan) reported the use of this decoction for treating retained fetal membrane in cows [20]. Fermented tea has been shown to be hypolipidemic and to reduce high blood pressure [49].

Xanthorrhoeaceae
The leaf pulp of Aloe vera is administered orally as ruminative. The pulp of this same species has also been reported for similar use in the treatment of digestive problems [58]. Alcoholic and aqueous extracts of this plant have shown significant activity against Bacillus subtilis, Escherichia coli, Proteus vulgaris, Salmonella typhimurium, Pseudomonas aeruginosa and Staphylococcus aureus [54,66,67]. Leaves of Asphodelus tenuifolius were used to cure weakness in horses in our study, while others have reported that root paste of this plant is applied to wounds in cattle [46].

Zingiberaceae
Turmeric powder (from Curcuma longa rhizomes) is topically applied for wound healing in cattle in the study area, while a study on equine medicines has mentioned that roots of this plant are used for hoof problems and  sore joints [58]. Alcoholic and aqueous extracts of C. longa have shown antibacterial activity [54]. Chloroform, ethanol, water and petroleum ether extracts of C. longa rhizome were also found to be active against bacteria, viruses, and fungi, and have shown anti-inflammatory activities [52,53].
Cultural importance of the species The Cultural Importance index (CI) of species is useful for estimating the significance of certain plants to a given culture [68] and takes into account not only the spread of the use (number of informants) for each species, but also its versatility, i.e. the diversity of its uses [68]. Based on medicinal applications, Trachyspermum ammi was found to be the most cited species followed by Curcuma longa, Melia azedarach, Zanthoxylum armatum var. armatum, Adhatoda vasica, Allium cepa, Foeniculum vulgare, Prunus persica, Punica granatum, Trichodesma indicum, Berberis lycium, Triticum aestivum and Peganum harmala (Table 1). It is notable that the top ten species of medicinal plants used to treat various livestock conditions were cited in all three major study sites (Haripur, Abbottabad, and Mansehra).

Cultural importance of the families
With regards to the diversity of species used, Fabaceae and Poaceae were the most important, with 6 species cited. Like the study by Pardo-de-Santayana et al. [30], we also elected to add the sum of CI of species in each family in order to measure the mean cultural importance of the families (mCIf). Unlike the aforementioned study, however, the number of species reported here did not strongly correlate with the number of species (R 2 = 0.211). This could be explained, perhaps, by the greater diversity of families (with a more limited number of species per family, average of 1.9 species/family) quoted for ethnoveterinary applications. Of the families reported, Apiaceae had the highest mCIf value, despite having only three species in this group ( Figure 2).

Informant consensus
Perceived efficacy of medicinal plants can be assessed by ICF values, with those plants that are supposed to be effective in curing diseases having elevated ICF levels [69]. We identified seven major disease categories and the highest ICF values were recorded for respiratory disorders and fever (0.68), followed by reproductive disorders (0.63), worms and other parasitic diseases (0.63) ( Table 2).

Comparison with human medicine
A large number of the veterinary plant reports share commonalities with the folk medical practices used in traditional ethnomedicine for humans in surrounding sites (last column in Table 1). This overlap may be a reflection on how folk veterinary remedies may be the diachronic result of a deep observation of the efficacy of certain plants used in animal diseases or at least of intense transfers of local knowledge between the folk veterinary and the ethnomedical domains.

Bio-conservation concerns
Various human activities may be implicated in placing some of the local medicinal flora under a state of threat within their natural habitat. The perceptions that local people share regarding this phenomenon of threats to local ecological resourcesmedicinal plants, in particular, was examined based on interviews with 15 key respondents in study districts. We examined these perceived threats using pair-wise ranking [29] of five central factors: agricultural land expansion, over-harvesting, over-grazing, uncontrolled fire setting and fuel wood collection. It was observed that agricultural land expansion was perceived as the dominant threat to medicinal plants used in ethnoveterinary medicine, followed by over-harvesting, overgrazing, fire and fuel wood collection ( Figure 3). Current conservation efforts concerning medicinal plants in this region are very limited, and as a result, the majority of them have no protection. This a major issue to be considered in future research and in local rural development initiatives.

Conclusions
A remarkable heritage of folk veterinary knowledge has been preserved within the framework of local knowledge and practices in the Pakistani communities of the Lesser Himalayas. However, like many other studies in this discipline have found, local knowledge is fragile and susceptible to rapid erosion with the expansion of biomedical paradigms and replacement of traditional resources with modern allopathic medicines. This is increasingly the case in both human and veterinary medicine. Nevertheless, as the majority of the reported species are wild and sometimes rare or under threat, much heed must be taken not to diminish these plant populations.
It is more urgent now than ever to record this rich body of knowledge not only for the purpose of biocultural conservation, but also to provide insights to scientists engaged in the search for new herbal veterinary therapies and especially to local stakeholders, who work on fostering endogenous trajectories of communitybased rural development projects in mountainous areas. The latter perspective is of crucial importance in the possible implementation of ethnobiological studies in disadvantaged areas, such as the mountain regions of Pakistan [70][71][72][73] as it may have a tremendous impact in sustaining and/or revitalizing communal forms of natural resource management [74]. Moreover, emic visions of environmental protection and provision of health and dietary care both for humans and animals may represent the key to environmental and social sustainability of social-ecological systems [75]. The validation and eventual application of this knowledge into concrete, comprehensive and culturally appropriate participatory initiatives aimed at fostering the sustainable use of local natural resources would promote the well-being of both animals and local communities.