Volume 10, Issue 4: 158-166; July 27, 2020  
TREATMENT TRIALS OF EPIZOOTIC LYMPHANGITIS WITH  
LOCAL MEDICINAL PLANTS: A REVIEW  
Mersha ASFAW and Tewodros FENTAHUN  
Unit of Biomedical Sciences, College of Veterinary Medicine and Animal Sciences, University of Gondar, P.O. Box 196, Gondar, Ethiopia.  
Supporting Information  
ABSTRACT: The aim of this paper was to review the use of local herbal medicines to treat Epizootic lymphangitis  
(EZL) and challenges related with safety, efficacy and quality control of herbal medicines. EZL has deleterious  
effect on both welfare and health of the horses and mules. In addition it has a serious negative impact on mainly  
the livelihoods of cart-horse owners/drivers. Basically, antifungal drugs for the treatment of EZL are costly and  
mostly unavailable in such areas especially in developing countries like Ethiopia. Medicinal herbs have a hopeful  
future since there are about half a million plants around the world, most of them have not yet been studied in  
medical practice, and current and future studies on medical activities can be effective in treating this disease.  
Furthermore, there is no gainsaying the fact that the requirements as well as the research protocols, standards  
and methods needed for the evaluation of the safety and efficacy of herbal medicines are much more complex  
than those required for conventional pharmaceuticals. These days, there are several trials on local plants like  
Xanthium strumarium (X. strumarium), Combretum molle (C. molle) seed and Phytolacca dodecandra (P.  
dodecandra) extracts inhibited the growth of Histoplasma capsulatum var farciminosum (H. capsulatum var  
farciminosum). Among these, the aqueous and n-butanol extracts of P. dodecandra with minimum inhibitory  
concentration (MIC) of (0.078%-0.156%) and (0.039%0.078%) respectively have been inhibiting the growth of  
H. capsulatum var. farciminosum. In vivo, over 58.3% horses with the disease responded to treatment then the  
other two plant extracts. In conclusion, P. dodecandra extracts showed a significant effect to inhibit the growth  
of H. capsulatum var farciminosum in vitro and EZL in vivo.  
Keywords: Combretum molle, Epizootic lymphangitis, Xanthium stramorium, Phytolacca dodecandra, Medicinal  
herbs.  
INTRODUCTION  
Epizootic lymphangitis (EZL) is a contagious, chronic disease which mainly affects horses, mules, and camels (Biyashev et  
al., 2019; Adedokun et al., 2020). It is caused by Histoplasma capsulatum var. farciminosum (H. capsulatum var.  
farciminosum). The disease is characterized clinically by a suppurative, ulcerating, and spreading pyogranulomatous,  
multifocal dermatitis and lymphangitis. It is seen most commonly in the extremities, chest wall and the neck, but it can  
also be manifested as an ulcerating conjunctivitis of the palpebral conjunctiva, or rarely as a multifocal pneumonia. The  
organism may also invade open lesions including ruptured strangles abscesses and castration wounds (OIE, 2009).  
The source of the H.capsulatum var. farciminosum can be the skin lesions, nasal and ocular exudates of infected  
animals or the soil. This organism can also spread on fomites (common utensil) such as grooming or harnessing  
equipment. Biting flies in the genera Musca and Stomoxys are thought to spread the conjunctival form. The pulmonary  
form probably develops when the animal inhales the organism (Public Health Agency of Canada, 2001).  
Epizootic lymphangitis is more common in tropical and subtropical regions than in temperate zones (Alsaad et al.,  
2016). H.capsulatum var. farciminosumis endemic in some countries in the Mediterranean region, and in parts of Africa  
and Asia including India, Pakistan and Japan (OIE, 2009). Many treatment types have been tried, largely without success.  
Parenteral iodides and amphotericin B have been reported as effective. However, although the disease is highly  
prevalentand economically important in Ethiopia (Ameni, 2006), the treatment options mentioned have not been  
employed because of the cost of the drugs and their absence in Ethiopia. This warrants for the need for other approaches  
including the use of traditional remedies.  
Traditional medicine is used throughout the world as it is heavily dependent on locally available plant species and  
plant-based products and capitalizes on traditional wisdom-repository of knowledge (Awas and Demissew, 2009). The  
wide spread use of traditional medicine could be attributed to cultural acceptability, economic affordability and efficacy  
against certain type of diseases as compared to modern medicines. Knowledge of medicinal plants of Ethiopia and of  
their uses provides vital contribution to human and livestock health care needs throughout the country (Belayneh et al.,  
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Citation: Asfaw M and Fentahun T (2020). Treatment trials of epizootic lymphangitis with local medicinal plants: a review. Online J. Anim. Feed Res., 10(4): 158-161.  
2012). The plant- based human and livestock health care persists and remains as the main alternative treatment for  
different ailments in Ethiopia, largely due to shortage of pharmaceutical products, prohibitive distance of the health  
service stations, unaffordable prices by small holder farmers and pastoralists for conventional drugs, emergence and re-  
emergence of certain diseases and appearance of drug resistant microbes and/or helminthes (Bekele et al., 2012).  
Whole plant of Xanthium strumanrium (X. strumanrium) as well as all parts separately is used in medicine  
(Bhogaonkar and Ahmad, 2012; Fan et al., 2019). The genus xanthium also possess antibacterial, antiviral, antimalarial,  
fungicidal, insecticidal and cytotoxic activities against cancer cell lines (Sravani et al., 2010; Passos et al., 2019).  
Phytolacca dodecandra (P. dodecandra) is one of the many plants claimed to have antifungal secondary metabolites. The  
antifungal effect of the crude aqueous extract of P. dodecandra was demonstrated in- vitro against different genera of  
dermatophytes of human pathogen and four clinical isolates of Candida albicans (Woldeamanuel et al., 2005; Tura et al.,  
2017).  
Combretum molle was used as a medicinal plant since ancient times (Grønhaug et al., 2008). The test of C. molle  
seed extract as antifungal property has been demonstrated in various studies (Masoko et al., 2007; Anato and Ketema,  
2018). Medicinal herbs have a hopeful future since there are about half a million plants around the world, most of them  
have not yet been studied in medical practice, and current and future studies on medical activities can be effective in  
treating diseases (Singh, 2015).  
In terms of population exposure alone, it is essential to identify the risks associated with the use of herbal  
medicines, and in this regard, the safety of these products has become an issue of great public health importance (WHO,  
2004; WHO, 2005). There is no gainsaying the fact that the requirements as well as the research protocols, standards and  
methods needed for the evaluation of the safety and efficacy of herbal medicines are much more complex than those  
required for conventional or orthodox pharmaceuticals (WHO,2005; Zhou et al., 2013). Thus, the general requirements  
and methods for quality control of finished herbal products remain far more complex than for other pharmaceuticals  
(WHO, 2003; WHO, 2004; WHO, 2005). Therefore the main objectives of this paper is to review the commonly used local  
herbal medicine to treat EZL and challenges related with safety, efficacy and quality control of local herbal medicines.  
TREATMENT TRIALS FOR EZL USING HERBAL MEDICINE  
The Xanthium strumarium leaf extract  
The X. strumarium, a rough cocklebur is broad leaved, tap rooted herbaceous annual plant. This is in a family of  
asteraceae, sub family asteroideae, tribe heliantheae, and genus Xanthium and species X. strumarium. It grows as weed  
throughout on waste lands. Cockleburs are short day plants and they can also flower in the tropics where the day length is  
constant. The herb is reputed as medicine in Europe, China, Indo-china, Malaysia and America also (Bhogaonkar and  
Ahmad, 2012). Stem is erect, ridged, rough and hairy and frequently branched which results somewhat bushy plants from  
30-120 cm tall. It has small greed unisexual flower occurring in separate cluster at the end of the brunches and main  
stems. The fruit is brown, hard, woody, bur from 0.4-0.8 inch long and coved with stout, hooked bristle. Its seed are  
produced in hard, spiny, globes or oval double chambered single seeded bur (Agharkar, 1991).  
Beside its medicinal values if a small quantity of parts of the mature plants is consumed, the seeds and seedlings  
will cause intoxication because extremely toxic chemical carboxyatratyloside is contained in them (Madalln and Sing,  
2001). Whole plant of X. strumanrium as well as all parts separately is used in medicine (Bhogaonkar and Ahmad, 2012).  
The genus xanthium also possess antibacterial, antiviral, antimalarial, fungicidal, insecticidal and cytotoxic activities  
against cancer cell lines (Sravani et al., 2010).  
Antifungal activity can be determined by the agar diffusion method. Test samples are diluted in Sabouraud dextrose  
agar followed by solidification in slanting positions. Test fungal cultures are inoculated on the slant and are incubated at  
29°C for 3-7 days (Paxton, 1991; Nisaret al., 2010). The principal compounds isolated from X. strumanriumleaves are  
found to contain, isoxanthanol, hydroquinone, caffeyolquinic acids, xanthanol, anthraquinone, cardenolide,  
leucoanthocyanin, simple phenolic striterpenoids and thiazinedione (Bhogaonkar and Ahmad, 2012). X. strumanrium  
produces secondary metabolites such as alkaloids, tannins, terpenoids, flavonoids, chloroform and n-hexane fractions  
whose activity has been demonstrated to be antifungal (Gujar and Talwankar, 2012). Antifungal activity of these  
molecules from X. strumarium exhibited 60% and 50% inhibition activity against the major dermatophyte fungi,  
Microsporum canis (Bharathi et al., 2010).  
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Citation: Asfaw M and Fentahun T (2020). Treatment trials of epizootic lymphangitis with local medicinal plants: a review. Online J. Anim. Feed Res., 10(4): 158-161.  
Figure 1 - Lesions of Epizootic lymphangitis (Wondmnew and Teshome, 2016).  
The Phytolacca dodecandra extracts  
Antifungal effect is one of the effects of secondary metabolites produced by plants. It is one of the many plants  
claimed to have antifungal secondary metabolites. Many studies indicated that, saponins are responsible for its antifungal  
effect. The antifungal effect of the crude aqueous extract of P. dodecandrais demonstrated in vitro against different  
genera of dermatophytes of human pathogen and four clinical isolates of Candida albicans (Woldeamanuel et al., 2005).  
The crude aqueous extract is also found to have effect against H. capsulatum var farciminosum both in- vitro and in- vivo  
(Ameni and Tilahun, 2003; Hadush et al., 2008). The n-butanol and aqueous extracts of P. dodecandra are evaluated for  
their effects on the isolates of H. capsulatum var. farciminosum and for the treatment of cases of epizootic lymphangitis.  
The phytochemical analysis of P. dodecandra shows the presence of saponins, alkaloids, and phenolic compounds in the  
berries of P. dodecandra. Thus, the secondary metabolites identified in the berries are all active antifungal compounds  
(Arif et al., 2009), which could imply that these secondary metabolites could be responsible for the antifungal activity of  
the berries observe in the n-butanol extract of the berries.  
The antifungal effect of n-butanol extract is observed to be much greater than that of the aqueous extract. The  
minimum inhibitory concentration (MIC) of n-butanol extracts range from (0.039%0.078%); whereas that of the aqueous  
extract is in the range of (0.625%1.250%). Similar finding for the aqueous extract is reported in which the MIC of P.  
dodecandra against the yeast forms of different Candida species is higher than 0.5% (Woldeamanuel et al., 2005).  
Another study shows that the MIC of the aqueous extract of P.dodecandra is 1% (Ameni and Tilahun, 2003). The MIC for  
novel pharmacological compounds should be <0.1% (Kuete, 2010).  
The minimum fungicidal concentration (MFCs) of aqueous and n-butanol extracts of P. dodecandra ranges from  
(1.250%2.500%) and (0.078%0.156%), respectively. The prepare ointment is topically applied and the result shows  
that, 58.3% are completely healed, while 41.7% did not cure. Comparable results are reported with the aqueous extracts  
(Ameni and Tilahun, 2003; Hadush et al., 2008). The n-butanol extract of P. dodecandrais effective against H. capsulatum  
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Citation: Asfaw M and Fentahun T (2020). Treatment trials of epizootic lymphangitis with local medicinal plants: a review. Online J. Anim. Feed Res., 10(4): 158-161.  
var. farciminosum. Previous toxicity studies on P. dodecandra indicated that human and guinea pigs can tolerate skin  
irritation of P. dodecandra. Moreover, oral LD50 are found to be 2.6 and 2.2 g/kg in mice and rats, respectively (Ameni  
Combretum molle extracts  
Combretum molle (C. molle) is used as a medicinal plant since ancient times (Grønhaug et al., 2008). The test of  
C.molle seed extract as antifungal property has been demonstrated in various studies (Masoko et al., 2007).  
Phytochemical studies carried out in the genus Combretum have shown the occurrence of many classes of constituents,  
including triterpenes, flvonlds, lignans, non-protein amino acid and tannins from different parts of the plant (Pietrovsk et  
al., 2006). C. molle has been widely used as a medicinal plant to treat various diseases such as parasitic, protozoan, and  
fungal infectious diseases in East and West Africa (Grønhaug et al., 2008). Antifungal activity is reported in numerous  
fungal models that used Candida albicans, Candida neoformans, Epidermophyton flccosum, Microsporum gypseum,  
Trichophyton mentagrophytes, Aspergillus fumigatus, Sporothrix schenckii and Microsporum canis (Masoko et al., 2007).  
The minimum inhibitory concentration (MIC) of C.molle seed extracts obtain in the study is 0.0156 %. However, the  
difference between the MIC (0.0156%) and the maximum none inhibitory concentration (0.0078%) suggests that the MIC  
could be lower. As Kuete (2010) stated, the MIC for novel pharmacological compounds should be <0.1%. So, the MIC  
(0.0156 %) is below 0.1% and hence this extract can be considered active. The positive control Ketaconazole is found to  
be more potent than C.molle seed extracts in inhibiting the growth of the mycelia form of H. capsulatum var.  
farciminosum (Asres et al., 2006). The C.molle seed extracts has inhibitory effect on H. capsulatum var. farciminosum.  
The Minimum inhibitory effect of C.molle seed extracts obtained from the study by Wondmnew and Teshome (2016) is  
also in harmony with previous studies made on fresh garlic extract (0.5 mg/ml, 0.05%) by Mesfi (2012), P. dodecandra  
(0.03%) by Mekonnen et al. (2012). According to a study by Mekonnen et al. (2012), the MICs of n-butanol and aqueous  
extracts of P. dodecandra are (0.039%-0.078%) and (0.625%-1.250%), respectively. The difference in MIC of the two  
extracts of P. dodecandracan be ascribing by the difference in the polarity of the solvent used in the extraction process.  
This is supported by Masoko et al. (2007); Eloff et al. (2005); and Cowan, (1999), as the polarity of the solvent has great  
effect on the quantity and types of bio-molecules extract.  
The main antifungal molecule in C.molle seed extract is tannin (Mishra et al., 2009). The presence of phenolic  
hydroxyl groups on the surface of tannin molecules participate strongly in the biological activities of tannins. It combines  
with protein and other polymers to form stable complexes through nonspecific forces such as hydrogen bonding,  
hydrophobic effects and covalent binding (Stern et al., 1996). This is done by hydrolysis of ester linkage between gallic  
acid which eventually affects the biosynthesis of cell wall and cell membrane. Impairment of biosynthesis of cell wall and  
cell membrane cause to increase the permeability of cell membrane and alterations of cell wall .This leads to decrease  
cell volume and disjunction of cell membrane from the cell wall (Suraya and Darah, 2002). Moreover, this leads to  
leakage of internal contents and no more exchange of molecule between cell wall and cell membrane.  
As Haslam (1996) tannins have two forms, and these are hydrolysable and condensed tannins which affect fungal  
growth. In the same study make by Ndip et al. (2007) both hydrolysable and condensed tannins have been found to  
possess antifungal effect. However, the hydrolysable tannins are found to be more effective against fungi. This is because  
hydrolysable tannins (gallic acid and ellagic acid) are linked to esters of core molecules which will be hydrolyzed easily  
while condensed tannins are not susceptible to hydrolysis (Haslam, 1996). In other research done by indicated that the  
fungicidal effect of the extract is due to the presence of high amount of hydrolysable tannins. In addition to its fungicidal  
effect, when C.molle seed extract is used topically, it will promote tissue healing, stop bleeding, stop further infection and  
heal the wound internally. As mentioned by Stephane et al. (2004), the ability of tannins to form a protective layer over  
the exposed tissue keeps the wound from being infected even more.  
Challenges associated with monitoring safety of local Herbal Medicine for EZL  
In terms of equine exposure alone, it is essential to identify the risks associated with the use of local herbal  
medicines, and in this regard, the safety of these products has become an issue of great animal health importance (WHO,  
2004; WHO, 2005). There is no doubt that the increasing cases of poisoning associated with the use of local herbal  
medicines in many parts of the world in recent times, is necessitating the need to ensure thorough toxicity assessment  
alongside active pharmacovigilance on these products in order to promote their safe use and protect animal health (Zhou  
Challenges related to the assessment of safety and efficacy of local Herbal Medicine  
There is no gainsaying the fact that the requirements as well as the research protocols, standards and methods  
needed for the evaluation of the safety and efficacy of local herbal medicines are much more complex than those  
required for conventional or orthodox pharmaceuticals (WHO, 2005; Zhou et al., 2013). A single local herbal medicine or  
medicinal plant may contain hundreds of natural constituents, and a mixed local herbal medicinal product may contain  
several times that number. Suppose every active ingredient is to be isolated from individual herb from which the local  
herbal medicine is formulated or produced, the time and resources required would be tremendous. Such an analysis may  
practically be impossible especially where local herbal product is a mixture of two or more herbs (WHO, 2005).  
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Medicinal herbs in the future perspectives  
Medicinal herbs have a hopeful future since there are about half a million plants around the world, most of them  
have not yet been studied in medical practice, and current and future studies on medical activities can be effective in  
treating such diseases (Singh, 2015). The use of medicinal plants has a long history; however, the use of the whole plant  
or raw materials for treatment or experimentation has many drawbacks, including changes in the plant’s compounds in  
different climates, simultaneous development of synergistic compounds that lead to adverse effects of antagonists, or  
other unexpected changes in bioactivity, and changes or loss of bioactivity due to the variability and accumulation,  
storage and preparation of raw materials; therefore, advancing towards the isolation of compounds and the use of pure  
substances with bioactivity, instead of the plant benefits, has certain benefits including convenient examination of  
therapeutic effects and determination of toxic doses to control the quality of the therapeutic formulation (Zhang, 2011).  
The beginning of the development of herbal medicines is concurrent with the development of chemistry and isolation,  
purification, and determination of plant compounds (Shakya et al., 2012).  
In the past, the drug discovery of the biological compounds from plant materials and the process of identifying the  
structures of active compounds from the extracts are problematic depending on the complexity of the compounds and  
might take weeks, months or even years. Nowadays, the rate of bioassay-guided fractionation has been significantly  
enhanced by the development of precision instruments such as high-performance liquid chromatography (HPLC), liquid  
chromatography mass spectrometry (LCMS), magnetic field and nuclear magnetic resonance (NMR) is a recent major  
breakthrough for the categorization (NMR) is a recent major breakthrough for the categorization of compounds that are  
extremely limited in quantity in their organisms of origin (Schroeder and Gronquist, 2006). Despite the success of  
research to produce medicinal plants over the past few decades, future efforts face many challenges. The quality of the  
herbal product has been studied. Standardization of raw materials is an important issue for the plant industry (Yadav et  
al., 2014).  
Herbaceous plants can be easily infected during growth, processing and collection. Contamination and pollution with  
heavy metals are two main problems with herbal drugs. It is therefore necessary to improve the quality and quantity of  
bioactive compounds for the production of herbal drugs while making effort to discover more new herbal drugs (Clark,  
1996). Due to expanding the use of natural substances around the world, the quality and safety ofplant-derived medicines  
should be comprehensively and accurately studied issues and the traditional and the millennial beliefs about these issues  
cannot be surely trusted; therefore, scientific and enlightening studies are essential to obtain reliable information for the  
use of medicinal plants in health care (Firenzuoli and Gori, 2007).  
Table 1 - Results of the in vitro evaluation of methanol extracts of P. Dodecandra, C. longa, and D. stramonium on H.  
capsulatum var. farciminosum  
Source: Hawi (2019)  
Table 2 - Growth of HCF in different concentrations of C.molle seed extract and ketoconazole  
Source: Wondmnew and Teshome (2016)  
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Citation: Asfaw M and Fentahun T (2020). Treatment trials of epizootic lymphangitis with local medicinal plants: a review. Online J. Anim. Feed Res., 10(4): 158-161.  
Figure 2 - The MICs of aqueous and n-butanol extracts of P. dodecandra against H. capsulatum var. farciminosum. A: MIC  
of aqueous extract: growth was observed starting at 0.625%; B: MIC of n-butanol extract: growth was observed starting  
from 0.039%; C: MIC of ketoconazole (standard): growth was observed at a concentration of 1.2×10-5%; D: Saline diluted  
Sabourauds dextrose agar (negative control): growth was observed in all agar plates (Mekonen et al., 2012).  
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Citation: Asfaw M and Fentahun T (2020). Treatment trials of epizootic lymphangitis with local medicinal plants: a review. Online J. Anim. Feed Res., 10(4): 158-161.  
CONCLUSION and RECOMMENDATION  
The X. strumarium leaf extract has strong inhibitory effect on the growth of the mycelial form of H. capsulatum var.  
farciminosum. The X. strumarium leaf extract can be included in the treatment of epizootic lymphangitis provided that  
convenient methods of preparation, dose and route of administration should be established through rigorous in -vitro and  
in -vivo trials. The n-butanol extract of P. dodecandra demonstrated minimum inhibitory concentration (MIC) and  
minimum fungicidal concentration (MFC) values that are considered to have antifungal properties. Therefore, since  
antifungals are not available for veterinary use in Ethiopia and also as they are expensive; searching for available and  
affordable antifungals such as n-butanol extract of P. dodecandra is recommended for the treatment of epizootic  
lymphangitis. The investigation of chemical compounds from natural products is fundamentally important for the  
development of new drugs. Ethanol macerated C. molle seed extract have a promising anti-fungal effect on mycelial form  
of H. capsulatum var. farciminosum. The main anti-fungal molecule in C. molle seed extract is hydrolysable tannins. The  
main action of tannin on H. capsulatum var. farciminosum is inhibition of cell wall and cell membrane biosynthesis. In  
topical application tannin has haemostatic and wound closure effect. The C. molle seed can be used for the treatment of  
epizootic lymphangitis if convenient methods of preparation, dose, and route of administration are established through  
meticulous in vitro and in-vivo trials. Medicinal herbs have a hopeful future since there are about half a million plants  
around the world, most of them have not yet been studied in medical practice, and current and future studies on medical  
activities can be effective in treating diseases. Those all parts of the plant have the chemicals upon extraction, the seed of  
the plants primarily and leaves secondly are preferable than the other parts of the particular plant because the seeds  
have higher concentration of the ingredients or chemicals required to use. The combination of the results got from both in  
vivo and in vitro trials are mandatory to witness the effectiveness of the particular plant’s medicinal value.  
Based on the above conclusions the following recommendations are forwarded: The appropriate treatment decision  
should be achieved to avoid suffering of equine from EZL. Challenges related with safety, efficacy and quality control of  
local herbal medicines for EL should be avoided. Any other local herbal medicines should be tried on EZL like ‘’embuay  
and ambacho’’. In-vivo studies must be conducted so that the safety margin, toxicity and cure rates will be known in order  
to use them commercially. Study on the mechanism action of the local herbal medicine extracts and their toxic effect on  
lab animals should be reported .Antifungal drug should be produced by using herbal medicines that serve to cure the  
disease and shall be scale up later at industry level.  
DECLARATIONS  
Corresponding author  
Authors’ contribution  
All the three authors reviewed the paper and contributed in developing the content.  
Conflict of interest  
The authors declare they have no competing of interests.  
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