Ethno-veterinary practices of Poaceae taxa in Punjab, Pakistan


 Background: Plant species of Poaceae family are not only used as fodder and forage but also contribute substantially in the treatment of various health disorders, particularly in livestock. Consequently, present study was aimed to document the therapeutic uses Poaceae taxa by the inhabitants of the Punjab Province to treat various veterinary health disorders. Methods: Semi structured interviews, group discussion and field walks were conducted to collected data. Furthermore, quantitative indices includingcultural significance index (CSI), relative frequency of citations (RFC), fidelity level (FL), and relative popularity level (RPL) andJaccard Index (JI) were adopted for data analysis. Results: Traditional uses of 149 plants belonging to 60 genera, 16 tribes of 5 sub families of Poaceae were recorded. Whole plant and leaves were the most consistent grazing parts with 40.94 and 29.53% contribution and decoction (35 reports) was the most preferred mode of administration. Majority of the plant species were employed to treat infectious diseases (25.93 %). and digestive disorders (14.10 %). Triticum aestivum depicted highest CSI, RFC and RPL levels at 8.00, 0.96, 1.00, respectively, followed by Oryza sativa and Poa annua . Likewise, T. aestivum and Saccharum spontaneum had 100 % FL and ROP. Jaccard index ranged from 12.25 to 0.37. Twelve plant species namely Chrysopogon zizanioides (anti-inflammatory), Pennisetum lansatum (improve bull fertility), Cymbopogon citratus (glandular secretion), Sorghum saccharatum and Themeda triandra (malaria), Aristida funiculate (anticancer) , Koeleria argentia (skin allergies), Tetrapogon villosus (antibacterial), Cynodon radiates (eyes infection), Sporobolus nervosa (Jaundice), Enneapogon persicus (antifungal), and Panicum repens (dysfunctional cattle organs) were reported for the first time with novelethnoveterinary uses. Conclusion: Inhabitants of the study area had strong association with surrounding plant biodiversity and possess significant knowledge on therapeutic uses of grasses and other members of Poaceae to treat various health disorders in animals. Plant species with maximum cultural and medicinal values could be a potential source of novel drugs to cure health disorders in animals and human as well.

especially in the dry winter season when other feed sources are not available [21]. Indigenous people with long histories of livestock rearing may have developed precious information about potential forage resources and they prefer to use grasses as fodder because they are highly plateable than other form of fodders [22]. Their platabilityalso depends on animal choice and may be linked with their seasonal availability and morphological and chemical nature of plant [23]. Animals generally prefer fresh foliage over dried because leaves of grasses are rich source of protein and cellulose and have low lignin than forbs and shrubs [24]. Grasses beside the nutritional and healthcare services also reduce the grazing pressure on other platable species and improve the productivity in livestock [25].
Though, different workers have reported the traditional uses of plant species from different areas of Punjab [26][27][28][29][30][31][32][33][34], but little is known about thetherapeutic potential of grasses and other members of family Poaceae [4], particularly for the treatment of various diseases in livestocks. Consequently, present studywas intended with the aim to document theethnomedicinal uses of plantspecies of Poaceae family from Punjab province of Pakistan, traditionally used to treat various health disorders in livestock and to explore the cultural significance of species and their popularity among different tribes on the basis of their usage in animal healthcare.

Materials And Methods Study area
Punjab is the second largest province of Pakistan after Balochistan. It encompasses of 205,344 km 2 area, located between latitudes 27.42º and 34.02º N and longitudes 69.81º and 75.23º E at the north western edge of the geological Indian plate in South Asia [35]. Punjab is comprised of 36 districts which are grouped into 5 agro-ecological zones [36] i.e. southern irrigated zone, arid desert zone, river zone, northern irrigated zone, thal sandy deserts zoneand baranizone are representative districts (Fig. 1). Northern irrigated zone and south desert zone are the two largest zone, with huge difference in cultural groups and ethnobotanical practices [4]. Majority of the area in Punjab consists of fertlile alluvial plain heavily irrigated with 5 rivers namely Jehlum, Ravi, Chenab and Sutlej. Spare deserts can be found in sourthern Punjab and Sulaiman Range. The variation in temperature and rainfall occurs all over the year. Soil is sandy, clay and loamy [37]. Most of the area experience foggy weather during winter and hot weather insummer. The average annual temperature ranges from − 2 °C to 45 °C. June is the hottestes and January is the coldest month of the year. Average annual rainfall of last five year is 479.8 mm. Nothern parts of the province receive reasonable amount of rainfall throught the year as compared to the sourthern part. Almost half of the rainfall occur during the month of July and Agust averaging about 255 mm.
Punjab is home to over half of the total population of Pakistan. The ethnic composition of the area is quite diverse comprising of different tribes and communities. Rana, Gujjar, Butt, Rayain are the major ethnic groups. Most of the people speak Punjab language followed by Saraiki and Pashto. Urdu and English languages are used in government offices.Compared to the other provinces, it has highest literacy rate. Punajb contributes major share in the economy of Pakistan in terms of GDP. The economy of the people in province is based on agriculture and wheat is the widely cultivated crop with significant production of rice, cotton, corn, sugarcane, pulses and jute. The major occupation of the rural communities is farming and they depend on agricultural means and livestock management to support livelihood.The inhabitants of the Punjab province have diverse traditional knowledge and practices because of linguistic and cultural variations. In agricultural lands like Punjab, grasses are preferred over other medicinal herbs and shrubs [4] because they are common, highly palatable and easy to process in order to cure livestock ailments [38].

Data collection
Data on ethnomedicinal application of Poaceae members to treat ethnoveterinary diseases were collected based on group discussion, semi-strcuture interviews and open and closed ended questionnaires during field visist in 2016-17, following the methods reported previously [39][40][41]. Prior to collect information, proper moral agreement was obtained from the head of local government and local informants. In total, 271 participants including both men and women, village leaders, shepherds, cattle holders who worked in local farms and some senior household animal owners were interviewed.
Demographic information about the participants was gathered by adopting a method of [42].
Questionnaires were first developed in English, afterwards translated in local languastics i.e. Punjabi and Saraiki. Before conducting interviews, prior informed consents were also obtained from all the participants after briefing the objectives of the current study and no further ethical approval was required as there is lacking of explicit rules or regulations pertain to the practices of ethnomedicinal uses of plants or animals in Pakistan. However, Participants were allowed to discontinue the interviews at any time.
Collected plant specimen were identified with the help of different flora of Pakistan, whereas botanical and family names were furteh verified from literature [43], www.efloras.org/index.aspx and Kew grass data base (https://www.kew.org/data/grasses-db/index.htm). For voucher specimen satnadrd herbarium techniques as explained before [44,45] were strictily followed. All plants were labelled and deposited in the herbarium at the Department of Botany, University of Gujarat, Punjab, Pakistan and the voucher specimens were preserved for record.

Cultural Significance Index (CSI)
The relationship between use reports of a given species and agreement among the informant knowledge was attributed through cultural significance index (CSI). It was calculated following a method of [46] using formula: Where i is the management of species having considerable impact on community (a species cultivated, managed or operated by any mean is awarded score of 2 and the value 1 is awarded if species is yet free from any kind of manipulation), e is the use preference of informant for one plant species over another species for a specific purpose (value 2 is for preferred species and value 1 is for non-preferred species), c is use frequency of a plant species (value 2 is attributed to a high potential plant species being considerably used by informants and value 1 is awarded to a rarely cited species), correction factor (CF) is level of informant consensus which comes from species citation divided by the number of citations of the most mentioned species.

Relative frequency of citation (RFC)
RFC is to set up the priority order among the listed species and its value is depended on the numbers of participantswho have mentioned a particular species as a medicinal plant or good fodder indicating its significance. The RFC was estimated with the help of following equation following [47].
Where, FC is number of participants who stated a particular plant species as an excellent medicinal plant and N is the total number of participants included in the study.
Use Value (UV) Use value (UV) was calculated by applying standard procedure reported previously [48].
Where U is total number of use reports mentioned by informants for a given plant and n is the total number of informants interviewed for a given plant species. UV close to 1 indicates many use reports for a given plant and its importance among informants.

Fidelity level (FL)
FL comes from the percentage of informant knowledge who report the uses of a plant species for an ailment and was determined using formula as reported previously by Where, Ip is the number of participants who reported the use of a grass for specific purpose; and Iu is the sum of participants who claimed the use of a grass for any purpose. High level of FL reflects the high use of plant species in specific disease in the study area.

Relative popularity level (RPL)
Plant healing potential can't be differentiated when they show same fidelity level. In order to differentiate the healing potential of species with same FL values, relative popularity level is calculated, which is the ratio between ailments cured by a specific plant and total number of informants who reporting that disease. Base of RPL, plant species are divided into popular and nonpopular groups. Popular speciesare those reported by more than half number of informants or above and rest of the species are declared as non-popular. For popular plant species RPL was arbitrarily selected equal to 1 that represents the complete popularity of a species for the cure of ailments and 0 value represents that no ailment was treated by this species [36] Rank order priority (ROP) Plant  and less educated people and rest of informations were shared by the educated informants but well educated informants were less conversant on the ethnomedicinal uses of plant species, particularly of Poacease taxa. High level of exposure to modernization and dependence of allopathis medicines could be the reason behind this, which have already been reported [49,50]. Though, previous studies were focused on a single community having one ethic group and same culture [51], but in the last few decades ethnobiologists are more interested in cross cultural variation of traditional knowledge of different communities and ethnic groups [52]. Because of this, we also collected data on ethnoveterinary uses of poaceae taxa from different ethnic groups i.e. Punjabi, Gujjar, Butt, Khawaja, Arayeen, and Rana. These groups have diverse culture and speak didderentlanguistics (Fig. 2)  were of perennial nature and rest of the 46% were annual herbs (Table 2). Plant part (s) used As depicted in Fig. 3, about 41% recepies were based on whole plant, followed by leaves, aerial parts and stem (29.53, 20.13 and 5.370%, respectively). As majority of the Poaceae taxa are small annual herbs with shallow roots, therefore they are easy to pull out as a whole plant and utilized to treat various diseases [4]. Likwiese, leaves are also easy to collect, rich in health beneficial secondary metabolites that contribute significantly in the treatment and prevention of health disorders [49,53,54].Leaves have also been reported previously as one of the most consistently used plant part for grazing and medicinal purposes [26,36,54].

Method of preparation and administration
As mentioned in Fig. 4, decoction was the common method of herbal preparation with 35 reports, followed by juice (31 reports), paste (26 reports), extract (24 reports), powder (10 reports), grains (5 reports), smoke (2 reports) and herbal tea, oil and raw material (1 report each). Crude preparation of decoction by boiling the plant parts in water for the treatment of various ailment is a common practice among the ethnic communities of in Punjab. The powder is prepeared by grinding the shade dried plant parts and paste is made from crushing the fresh or dried plant parts with water or oil [34].Mode of administration falls into two main categories viz. oral and topical. Herbal preparations used to treat internal diseases i.e. gastrointestinal disorders, fever, pain etc. are usually administrated orally, while for joint pain, skin infections topical method is common. Most of the plants were given orally as fodder (59 reports) without processing them in crude preparation.Offering plant as fodder to animals is the best way to treat a specific disease without having side effects.
Interestingly, there is a significant trend of multi-plant formulation devised by semiprofessional herbalists and traditional practitioners. In that case, powder of more than one plants/plant part is orally administered with water known as "Phakki". There are certain cases with such recipes where overdose and malpractice resulted in adverse drug reactions.

Ethno-veterinary uses of Poaceae taxa
In Pakistan, ethno-veterinary studies are an important source of indigenous information associated with animal healthcare system. The present study is a continuance of earlier explorations for the improvement of records on the ethno-veterinary medication in Pakistan. In Pakistan, many documentations of ethno-veterinary knowledge have been develop so far [9], [60],    [105], America [106], Canada [7] and Brazil [107]. Besides these studies, literature on Poaceae members used in ethno-veterinary practices is still missing in Pakistan. Therefore, this paper contains an important information of biological resources used in ethnomedicines (EM) and ethnoveterinary practices (EVPs) in Punjab, Pakistan.
Ethno-veterinary implies all traditional methods used to treat common diseases in livestocks [55].
Nutritional value and pharmaceutical properties of Poacea taxa make them an ideal candidate among indigenous communities to cure and feed the domestic cattle [4]. Inhabitants of the study area use 149 plant speices to treat various health disorders in cattle, which were grouped into 12 major disease categories (Tables 2). As shown in Fig. 5 We reported that Dactyloctenium aegyptium as detoxifier and antiallergic plant but according to [63] this plant has anti-inflammatory, anti-cancer, anti-microbial properties and is widely used to treat small pox and ulcer in children. Leucorrhoea, anemia, skin cleaner, diabetes and herpes are treated with Hordium vulgare in different parts of Punjab. Its stem is grinding and mixed with water in order to gain the weight in Hawassa Zuria District, Sidama zone, Southern Ethiopia [64].This plant is also used to treatfever in some parts of India while in Salt Range of Pakistan, flour made from this plant is used to cure jaundice [65,66]. Desmostachya bipinnata is diuretic, anti-amenorrhea and is used to treat manydigestive disorders. [67] documented that powder made from the roots of this plant is used to cure rheumatism in Soon Valley of Pakistan while root infusion of the same plant is used to treat is urinary infections and whole plant is taken orally to treat dysentery [68] Vetiveria zizanoidesis is antiseptic, demulcent and anti-inflammatory. It is rich in aroma which is used to treat stomach disorders and inflammation [69]. Paste made from the leaves of Cynodon dactylon effective against skin injuries while juice of the plant is used in healing bone fractures. According to [66], root decoction of Cynodan dactylon is given to cattle having repiratory problem.Decoction made from Sorghum bicolor is given to the children in case of typhoid and water extracted from the plant is used to bath the babies [70]. Themeda anathera is refrigerant and juice extracted from leaves is used to reduce depression. However, decoction made from the leaves is also used as blood purifier [71].
Desmostachya bipinnata is anti-amenorrhea and diuretic. Decoction made from the leaves is given to asthma patient to relieve from pain [72]. Decoction made from the whole plant of Cymbopogon jwarancusa is used in typhoid fever. Other documented ethnomedicinal uses are; abdominal pain, tumor and unconsciousness [73]. Eleusine indica is used in abdominal pain which is in line with the study of [7], who reported that grain flour made from the aerial parts of the same plant is used to treat digestive problems. Apluda mutica is used to get relieve from stomach pain while [74] reported that paste made from this plant is also used to cure fungal diseases in livestock. [38] describe that besides imporving digestion, Bothriochloa bladhii is also used as stored food and fodder for both livestock and wild ruminants. Extract made from aerial parts of Zea mays is detoxifier, nerve tonic and antiseptic. [56] reported that seedsof the same plant are mixed with oil to manage tick infection in livestock. Paste made from leaves of Imperata cylindrica is given to cattle in order to control microbial infections. [75] reported that this plant possess antioxidant, neuroprotective and anticer properties. Juice extracted from leaves and roots of Saccharum spontaneum improves appetite and gives relieve in inflammation, urinary problems and abdominal pain. According to [60], stem of this plant is chewed to relieve stromach pain. Whole plant of Dichanthium annulatum is used in indigestion but [76] also reported that whole plant is also used in dysentery and menorrhagia.

Cultural significance index (CSI)
Cultural significance index was used to calculate the importance of individual plant used by indigenous people.In CSI, the recognition or reputation of species is linked to its functions to the people and are considered auxiliary element in the cultural recognition of a plant [108,109]. It was observed that cultural importance of each species varies between local communities. This difference is influenced by level of knowledge, the particular cultural settings and the local conditionsIn the present study, CSI values varied from 0.13 to 8.00 which was markedly affected by the preference, manangement and frequency of use by the local inhibitants ( Table 2).
The highest CSI value was obtained byTriticum aestivum ( [111], who observed that plants which are not easily availability touser community for the disease managementarelesssignificant and usually havelower CSI value. However [112] proposed a weak correlation between use and availability of plants and suggested that species with greater cultural significance have a tendency tobecome vulnerable or rare locally.

Relative frequency of citations (RFC)
Relative frequency of citations reveals the importance of each grassamong indigenous communities of Province Punjabin ethno-veterinary medicines and primary health care of animals to make them healthy and productive. It is calculated from the citation frequency of informants claiming the use of a plant species divided by the total number of informant who participated in the survey to share their indigenous knowledge [5]. In our work, RFC ranges 0.96 to 0.14 ( Table 2).
Maximum The poor educational background is the major reason that directly affects the learning system in this concern. The study is in agreement with [113]. Relative frequency of citation highlighted the importance of individual species among local communities based on the number of uses [114].It has been suggested [55] that plants with high RFC values should be involved in biological, phytochemical and pharmacological studies for further investigation of drug development. Such kind of plants must be conserved on priority basis due to the threat of over exploitation and extensive use of these plants in community [19]. Values of RFC are very dynamic as it changes with area to area and depends on the folk knowledge of the native people. It is well known that species with low RFC values are not unavoidably insignificant [115]. Their Use Value (UV) UV index was used to evaluate the importance of plant species among the indigenous communities [116]. UV value ranged from0.02 to 0.93 ( Table 2) inhabitants are well aware off these plants [57], [118]. Species with low use value were, Chloris dolicostachya (0.02), Enneapogon persicus (0.04), Panicum repens (0.05), Pennisetum lansatum (0.06) and Tragus racemosus (0.06).This might be due to their less availability but they are not necessarily less effective [57]. These plants can be employed inthe development of human pharmaceuticals [119].

Correlation among used indices
The relationship among ethnobotanical indices i.e.CSI, RFC and UV. is given in Fig. 6. A strong positive correlation was present between CSI and RFC (r 2 = 0.60), followed by CSI and UV (r 2 = 0.52) while between RFC and UV a relatively weak positive correlation was recorded (r 2 = 0.25). About 60% of the values of CSI and RFC and52% of the CSI and UV values fall in the same region, representing high association among these indices whereas, only 25% values of RFC and UV showed weak correlation between these two indice.

Fidelity level
Fidelity level of 24 most important species ranged from 14.3 to 100% (Table 3). In general, the highest fidelity level of a species highlighted the existence of a particular disease in the study area and utilization of plant species by the local people in order to treat it [93,94]. and 70%, respectively. Use of plants by human beings for animal health care is an old practice. These species are not only used for feeding livestock instead these are an important and cheap source of medicine for cattle to treat multiple health issues [120]. The traditional ethno-veterinary system has played a significant role in animal production especially in the rural areas where livestock diseases are locally treated [121,122].

Relative popularity of species
One hundred and forty-ninespecieswere mentioned by 271 informants, interviewed during this study for different kind of diseases. Of these, 125 specieswere reported by fewer than 8 informants and therefore were excluded for further discussion. The rest of the 24 specieswere reported by more than 7 informants are presented in Table 3. For speciescited by 8 to 62 informants, the number of uses per species increased progressively (Fig. 7) with increase in the number of informants interviewed showing positive correlation (r, 0.14, coefficient of variation, 0.46). Conversely, speciesmentioned by more than 65 informants, the average number of uses per speciesdid not increased with increasing number of informants. About sixteen plant species mentioned by 62 informants were grouped as unpopular whereas, eight species reported by 65 or more informants were classified as popular.
Specieswith high popularity level (1.0 RPL) were; Triticum aestivum, Saccharum officinarum, Sorghum helepense; Cymbopogon jwarancusa, Saccharum bengalense, Saccharum spontaneum, Oryza sativa and Vetiveria zizanioides. The healing potential of each species may vary and is expressed by its FL value [36]. Rank order priority index can be used as correction factor to rank plants properly with different fidelity level [48]. Out of 24 species only 9 attained 50% or above ROP values. This can be attributed to the decreasing popularity of herbal medicines in the study area. Triticum aestivum and Saccharum spontaneum were reported with highest ROP value (100%), trailed by Cymbopogon jwarancusa (98%), Vetiveria zizanioides (94%), Saccharum officinarum (84%), Saccharum bengalense (76%), Oryza sativa (74%), Sorghum halepense (69%) and Arundo donax (58%). Rest of the specieswere presented by less than 50% of ROP values. The high popularity of these speciescan be attributed to their high nutritional values [4] and may be linked to the fact that local farmers are well aware of these speciesand they frequently used them for the treatment of various ailments in the livestock. This is an agreement with similar findings of previously reported studies conducted in same province (Punjab) [4,36]. Our study is also consistent with the findings of on the status of healing potential of medicinal plants in Palestinian area [48] and medicinal plants among Bedouins communities in Negev desert [46].

Jaccard Index (JI)
Novelty index was done (Table 4) in order to compare the reported taxa with the other studies conducted in different parts of same province (Punjab), other provinceof Pakistan andin neighbouring countries like India, Bangladesh and Nepal.Ethobotanical information may have cultural differences and different origin among rural populations as this knowledge greatly varied from region to region [52,54]. Therefore, a comprehensive research with highunderstandingof ethnobotanical folk knowledge is mandatory for better judgement [123], and exploration of traditional knowledge in order to find novelty in work and possible drug discovery [98,124]. Jaccard index in our study ranged from 12.2 to 0.37. Within Pakistan, highest Jaccard index (12.2) was found with previous report from Hafizabad Punjab, Pakistan [4], followed by the study conducted by [125], from district Layyah,   [41]. It has been recorded that neighbouring indigenous communities share more common traditional practice of plants as medicines in order to cure various ailments and this is because of more social trade and sharing of ethnomedicinal knowledge among native groups [132,133]. In contrast, a low similarity index indicates less sharing of medicinal knowledge and low social interaction that could have been happened in the past bringing more difference in ethnobotanical practices [50]. Geological isolation of ethnic groups and plants resulted a significant change in vegetation structure and therapeutic uses of indigenous plants and this may be a reason for loss of ethnobotanical information [44]. A low degree of similarity index of our study with other studies conducted in same province or in other parts of Pakistan indicated that either a little attention has been paid towards grasses in these studies and zero (0) percent similarity index show that grasses were totally ignored. Therefore, this is the first comprehensive report on ethno-veterinary knowledge of indigenous grasses of whole province of Punjab.
Similarities and differences in ethnobotanical use of plant species were compared with studies conducted within and outside the Pakistan. The maximum diversity in ethnobotanical uses of plants was found with the study of [4] from Central Punjab, Pakistan (13 uses) and data is presented in Table 5. It was trailed by other studies conducted in surrounding areas; [118] from Toba Tek Singh, Punjab, Pakistan (13 uses), [134] from Peshawar, Pakistan (3 uses), and [135] from Uige, Northern Angola (3 uses) and [136] from Guimaras Island, Philippines (3 uses). In rest of the studies, diversity in therapeutic uses of medicinal plants was found 1 to 2 or 0. The maximum similarity in ethnobotanical uses was found in [4] from Central Punjab, Pakistan (38 uses), followed by [125] from Layyah, Punjab Paksitan (7 uses), [126] from Gujrat, Punjab, Pakistan (7 uses) and [36] from Hafizabad, Punjab, Paksitan (6 uses).

Ethics approval and consent to participate
Before conducting interviews, prior informed consent was obtained from all participants. The study was carried out according to the set rules and recommendations of the Code of Ethics of the International Society of Ethnobiology. No further ethics approval was required.

Consent for publication
This manuscript does not contain any individual person's data and further consent for publication is not required.

Competing interest
The authors declare that they have no competing interest.

Funding
No funding is available for this work. Linguistic wise classification of informants.  Methods of herbal preparations.  Correlation between number of informants and ailments.