RECORD OF MEDICINAL JEW’S (Auricularia auricula-judae (Bull.) Quél) EAR MUSHROOM GROWING IN THE GREATER ACCRA REGION, GHANA AND ITS POSSIBLE HEALTH VALUES

Medicinal mushrooms are part of total health delivery system of a nation. A mushroom of recognised health benefits was seen for the first time on a pencil cactus (Euphorbia turicalli L.) in the Greater Accra Region (Adentan Municipal Assembly) in SeptemberOctober 2021. The fruiting body was gelatinous with the shape of a human ear. The basidiomata were either solitary, densely rosette with caestipose habit either astipitate or with vestigial stipe. The general morphological features were akin to that of Auricularia auricula-judae. Anatomical and morphometric studies showed that the fruiting body had smooth hymenium with a layer of basidia which were cylindrical or club-shaped. The dimensions of the basidia (70.86±4.27 μm long; 9.54± 0.60 μm wide) closely agreed with the range reported for A. auricula-judae. The white/greyish basidiospores were generally sausage-shaped (allantoid) ranging in length (18.47±0.73μm) and width (7.49±0.3μm) closely to reported range. The economical and medicinal values of the mushroom are described and future commercial exploitation through biotechnological technique highlighted.


Introduction
The genus Auricularia consists of mushrooms originally described in Europe and belongs to the Phylum: Basidiomycota; Family: Auriculariaceae; Order: Auriculariales (Kirk et al., 2008). It is well known in Europe, China and other Eastern Asian countries, especially Auricularia heimuer (Wu et al. 2014). This fungus, A. heimuer, is believed to be a taxon reported 2000 years ago in the Chinese Shennong's Compedium of Materia Medica.
Auricularia auricula-judae (=A. auricula) is recorded in the Chinese Icons of Medicinal Fungi (Ying et al. 1987). The sporophore is gelatinous and shallow. The shape of the basidiomata resembles the human ear. When humid, the texture is cartilaginous, or waxy. As fruiting body dries out (desiccate), the gelatinous basidiomata shrinks considerably and becomes very tough (Piepenbring, 2015). The hymenium (fertile layer) on the upper surface is smooth or slightly wrinkled varying in colour from reddish, to golden brown, to dark brown. The lower sterile surface is dark olive brown, hairs short, continuous, more or less flexuous gradually tapering towards the apex. Basidia are cylindrical to barrel-shaped with about three transverse septa, 50-62 x 3-5.5 μm wide. The spore is hyaline to white, often curved, cylindrical or oblong 9-18 μm long x 5-7.5 μm wide (Ying et al., 1987;NCBI, 2020;Mohanan, 2011).
This mushroom grows either solitary, densely in groves and dead older trees such as oak, elm, poplar, banyan, acacia or rotten wood. In Europe, China and Australia where it is found, it occurs throughout the year but most prevalent in late summer and autumn (Li et al., 2021).
The genus Auricularia recorded in Africa are summarised in Table 1. There is hardly any reference in the pertinent literature on the occurrence of A. auriculatajudae in Ghana. The only record in Ghana is by Apertorgbor et al. (2006). But the exact location where this fungus was found during the survey in Southern Ghana was not stated; neither did they mention the type of wood on which it was found. Mushrooms have become an integral part of traditional cuisine and health practices in households in East, Central and West Africa (Dijk et al. 2003;Rammeloo & Wallleyn 1993) as well as China, Europe, Americas and the South East Asian-region (Wu et al., 2015;Oli et al., 2020;Li et al., 2021).
The etymology of Auricularia in Latin meaning "ear" and "judae" means Judas-the man who was said to have betrayed Jesus in the Garden of Gethsemane as recorded in the Bible. This legend which dates back more than 400 years suggests that, the fungal ear that emerged from older wood are visible residue of Judas tormented spirit (www.first.nature. com, 2021).
The fruiting bodies of Auricularia are waxy, rubbery, cartilaginous and colour ranges widely from purplish-brown, yellow brown to black especially when dry. In Africa, just as in Europe and elsewhere, the fungus can be seen throughout the year.
A.auricula-judae was previously considered as a single species but has recently been demonstrated to be as species complex using phylogenetic analyses by ITS sequence and combined ITS, LSU and rpb2 sequences (Wu et al., 2015). Seven species were delineated by this method and it included 3 species new to science; A. angiospermarum, A. minutissima, A. tibetica, in addition to A. heimuer and A. villosula. Mushrooms have enormous biologically active secondary metabolites of high therapeutic application (Sevindik, 2018;Gebreyohannes et al., 2019a, b;González-Palma et al. 2016;Badshah et al. 2015;Gargano et al., 2017). These mushrooms also have a wide range of secondary metabolites of high therapeutic value such as antioxidants, antidiabetes, antiviral, antithrobolic, antiinflammatory and anti-tumour properties (Iftekhar et al., 2011;Alves et al., 2012;Waktola & Temesgem, 2018;Bills and Glover, 2016;Onyango et al., 2016). Recent reports indicate that over 136 mushroom extracts also have shown antimicrobial activity against gram positive and gram-negative bacteria (Oli et al., 2020;Deka et al., 2017;Islam et al., 2021).
Polysaccharides present in A. auriculajudae makes it qualified to be classified aptly as a functional food. These functional ingredients are greatly beneficial to human health (Lin et al., 2021). It also contains high levels of insoluble fibres with varied bioactivities. A. auricula-judae also contains several phenolic compounds including epicatechin and catechin and is also known to exhibit antioxidant properties to prevent free radical induced oxidative damage, balance the blood glucose and lipid levels to alleviate diabetes, hyperlipidaemia, obesity and stimulates macrophage and inflammatory cytokines to prevent inflammation, exhibit anti-cancer properties by stimulating cellular apoptosis and prevents gut microbial dysbiosis by stimulating the frominum of healthy microbiota (Islam et al., 2021). The main sugar components of basidiocarp are mannose, galactose, glucose and also contains wound healing polysaccharides (Mapoung et al. 2021).
In terms of medicinal value this mushroom is good for the treatment of piles, is a good stomach tonic which regulates the flows of vital energy (Ying et al., 1987). The sporophore enriches energy mild to sweet tart and nourishes lungs, stops haemorrhage and invigorates blood circulation; has the effect of embellishment and invigorating bowel movement. The Chinese use it for rheumatic pains in the legs and lumbago, for weakness after childbirth, cramp and numbness, for pains and injury of wound, obstruction in arteries and veins, and tetany (Ying et al., 1987). Furthermore, the Chinese and Europeans use it to treat malignant dysentery, and enteritis, for menorrhagia with leucorrhoea, for haemation and haemorrhoids and uterine bleeding, for gastric disorders causing nausea and excessive phlegm. (Ying et al., 1987).
In West Africa this mushroom A. auricula-judae is used in folk medicine for complaints of illnesses including sore throat, sore eyes, jaundice, astringent etc. (m.wikipedia, org). In Chinese restaurants, it is used to prepare dishes such as hot and sour soup which is also available in Ghana. In Ghanaian folklore medicine, it is used as a blood tonic (m. wikipedia, org; Apetorgbor et al., 2006).
There has not been any study of morphological, anatomical features and phytochemistry of the fungus which will delimit A. auricula-judae in Ghana and indeed Africa for future biotechnological cultivation and exploitation for the pharmaceutical industry and to our health benefits. This paper provides some vital morphometric and anatomical diagnostic features which could act as a springboard for future biotechnological cultivation and exploitation of its bioactive compounds for health delivery improvement in Africa.

Location of study area
This edible mushroom was spotted in a house at a residential area, Lakeside Estate, in a rock garden within the Ga, Greater Accra, Adentan Municipality with coordinates N4°33' 28° E 0° 55' 56" Latitude 4.5576511, Longitude 0.99322071.

The fruiting body/ basidiomata
When fresh, the basidiomata was gelatinous, orange brown to vinaceous to reddish brown. (Figs. la and b). The fruiting body was either solitary, gregarious, rosette or cespitose (growing in dense clusters) (Figs. 2a and b), sessile or sub stipitate, discoid or auriculate with lobbed margin projecting up to form an ear-like lobe. The fruiting body was 2-6mm thick when moist (Figs. la and b) becoming greyish upon drying smeared with white spores (Fig. 3a-c).

Anatomical studies
The anatomy of hymenium was studied by cutting a longitudinal section by hand and sliding microtome (Reichart Nr.15917, Austria) and staining with lactophenol and mounting on a glass slide with a cover slip using conventional anatomical methods. Photographs were taken under photomicroscope (Computer Model Leica 1CC50W) with a software (Leica LAS EZ Version 1.8.0). Dimensions of at least 30 basidiospores and 30 basidium were estimated by stage micrometre and eyepiece graticule measurements and then confirmed with a measuring software ImageJ Version 1.53. (NIH/LOCI, University of Wisconsin, USA).

Host plant for the edible mushroom
The host plant for this mushroom A. auriculajudae is commonly known as naked lady, aveloz, milk bush, pencil cactus or fire sticks (Anon, 2010). Its botanical name is Euphorbia tirucalli Linn. (Euphorbiaceae, Malpighiales). It is a tree which grows in semi-arid and tropical climates. It is usually used to feed cattle or as a hedge especially in the savanna zones (Haevermans, 2017). In Ghana, it is used as a hedge or decorative plant. This plant is a hydrocarbon plant which produces a poisonous latex milk can cause temporary blindness if not handled properly (Wikipedia, anon 2017). According to the ICUN it is placed in category 3.1 of least concern Conservation status (Haevermans, 2017). Euphorbia tirucalli is a plant that has been promoted as an anti-cancer agent (Aveloz, 2015). Euphorbia tirucalli has a wide distribution in Africa in black clay soils, being prominently present in Northeastern, Central and Southern Africa. It has been introduced to many other tropical regions, such as Brazil, India, Vietnam, the Philippines and Ghana. (Haevermans, 2017).

Fruiting body/ basidiomata
The initial stages of the development of the mushroom were cryptic under the hard-older bark of the host plant was not visible until its prolific appearance on 4 th -5 th October 2021 after a heavy downpour of rain in Accra. The gelatinous orange brown to vinaceous reddishbrown gelatinous fruiting body was seen on the host in the rock garden (Figs. 1a and b). There was an intervening dry spell of 10-12 days which caused the basidiomata to shrivel by losing moisture attended by discharge of copious amounts of basidiospores (Figs. 3b and c). Fig. 3a shows microecological surroundings creating a humid condition with palm trees and pencil cactus in the foreground. Figs. 2a and b show the morphology of the fruiting body as seen in front and behind with the subsessile stipe attachment.

Anatomic studies
The hymenium (fertile layer) on the upper surface of the basidiomata (fruiting body) was smooth, wrinkled varying in colour from reddish brown to golden brown to dark brown. All the basidia were seen as cylindrical or club-shaped creamy white in colour at various stages of development (Fig. 4a). The basidia were variable in size. Table 2 summarizes the dimensions of basidia (70.86±4.27 µm long; 9.54± 0.60 µm wide). The spore prints on the fruiting body were white to greyish white in colour (Figs. 3b and c); generally, sausageshaped (allantoid) to oval ranging in the length (18.47±0.73µm) and 7.49±0.3µm in width (Table 2). Figs. 4a and 4b show the morphology of the basidia and basidiospore respectively in the hymenium layer of the fruiting body. Table 1 shows the occurrence of Auricularia species in Africa. These include A. auriculajudae, A. delicata, A. fuscociccinea, A. mesenterica, A. polytricha and A. tenuis. There is a preponderance of A. auricula-judae over the other species in Africa (Table 1).

TABLE 1
Record of Auricularia species occurring in Africa.

Discussion
The fruit body of this mushroom found in Accra resembles that of Auricularia auricula-judae and is waxy and cartilaginous, resembling human ear with a wide range of colours ranging from purplish brown, yellow brown, vinaceous to sometimes black (Figs. 1-3). This is akin to what is stated in the pertinent literature as having wide occurrence in Africa (Table 1) and is the first record of its occurrence in the Adentan Municipal Assembly, in the Greater Accra Region, in a residential house within the Lakeside Estate, Ashalley Botwe. Previous record was reported by Apetorgbor et al. (2006), in an unnamed location of Southern Ghana. They also did not state the host plant. This is the first record of A. auricula-judae on dried older wood of pencil cactus, Euphorbia turicalli L. or fire sticks. This data also extends the list of host plants supporting the growth and development of the A. auricula-judae. It has been previously recorded growing solitary, or densely in groves, rosette, caestipose on the dead wood such as oak, elm, poplar, bayan, sycamore, acacia, eucalyptus etc. in Europe, South East Asia, Australia and England (Li et al., 2021). The shape of the basidiomata which resembles human ear shrunk considerably and became very tough (Fig. 3) as occurred in this study under drought conditions. This agrees with the findings of Piepenbring (2015) and Li et al. (2020) for the same fungus under drought conditions.
The longitudinal section of the hymenium (Figs. 4a and b) shows that the basidia were at various stages of development and were club shaped or cylindrical with at least three transverse septa. The basidia length was 70.86±4.27µm long and 9.54±0.60µm wide (Table 2). These values fairly agree with the reported values in the pertinent literature by Ying et al (1987), Mohanan (2011) andNCBI (2020). They stated a range of 60-72µm long and 4.0-7.5µm wide ( Table 2). The spore print of the hymenium was hyaline to white in colour (Fig. 3) and were often curved sausage shaped (or allantoid) (Fig. 4b). This observation agrees with that of other workers (Ying et al., 1987;NCBI 2021;Mohanan, 2011).
This mushroom occurs throughout the year in Africa and Ghana (Apetorgbor et al. 2006) and agree with what obtains in Europe, Asia and Australia (Oli et al. 2020;Li et al. 2021).
The perennial nature of flushing and occurrence of this mushroom should lend the fungus the opportunity for cultivation. However, before developing the local techniques for commercial cultivation in Africa and Ghana in particular, there should be some preliminary biotechnological screening of the local isolates. This group has never been studied in the detail in Africa and most of the literature data are probably based on macroscopic resemblances (Rammelo & Walleym, 1993). It will be instructive to study the local isolated using phylogenetic analyses by ITS sequence and combined ITS and 150 and rpb2 sequences as was done by Wu et al. (2015) for the Chinese and European species. This aforementioned study is more relevant as mushroom have a plethora of biologically active secondary metabolites of high therapeutic applications (Sevindik, 2018;Bahshah et al., 2015;González-Palma et al., 2016;Gargano et al., 2017;Gebreyahannes et al., 2019a, b) and may differ in concentration of active ingredients based on genetic lines. It is also well known that Auricularia also have a wide range of secondary metabolites of high therapeutic value such as antioxidants, anti-diabetic, antiviral, antithrombotic, antiinflammatory and anti-tumor properties worthy of exploitation for health improvement (Bill and Glover, 2016;Gryango et al., 2016;Iftehekhar et al., 2011;Alves et al., 2012;Walktola & Temesgen, 2018). Recent studies indicate that, over 136 mushroom species extracts have also been shown to have antimicrobial activity against gram positive and gram-negative bacteria (Islam et al., 2021;Oli et al, 2020;Deka et al., 2017). The genera Auricularia and Termitomyces are prominent among these mushrooms with biologically active secondary metabolites of high therapeutic applications (Gebeyohannes et al., 2019a, b) as well as antimicrobial activity against both gram negative and gram-positive bacteria (Islam et al., 2021;Oli et al., 2020;Deka et al., 2017).
There are other economically beneficial compounds in A. auricula-judae such as polysaccharide serving as functional food for human health (Liu et al., 2020;Miao et al., 2020), insoluble fibres with varied bioactivities not excepting sugars (mannose, galactose, glucose etc.) as well as woundhealing polysaccharides (Mapoung, 2021). In summary, this mushroom is such a potent source of pharmaceutical factory for bioactive healing compounds in the health delivery system that it cannot be discounted. This paper has provided morphological and anatomical evidence that this mushroom occurs in Ghana and future studies should aim at ascertaining its biotechnological physiology and molecular characteristics with the view to providing a spring board for its commercial cultivation, as explained, for better health.