Biological eciency and nutritional composition of Pleurotus djamor cultivated on bagasse of Agave salmiana

The objective of the present study was to use molecular techniques to identify a wild mushroom isolated from A. salmiana, and then evaluate its biological eciency, production rate, and nutritional and morphological characteristics when grown on A. salmiana bagasse with various concentrations of urea as a source of nitrogen. Two types of inoculum were employed: in grain (WG) and pellet (WP) form. The substrate was supplemented with total nitrogen concentrations (TN) of 0.77, 0.95, 1.14, 1.32, and 1.5% to evaluate its effect on the biological eciency (BE), production rate (PR), and morphological and nutritional characteristics of the fruiting bodies. The molecular analysis of the ITS region permitted the amplication of a product of 750 pb. The mushroom was identied as Pleurotus djamor. After supplementing the substrate with urea, a BE of 70% was obtained in the sample inoculated with WG at 1.32% TN. Observations found that the TN concentration of 1.5% produced malformations in the fruiting bodies. The analysis of the sporocarps indicated a raw protein content (RP) of 15–26% and that the mushroom’s nutritional composition changed according to the inoculum utilized and the percentage of nitrogen in the substrate. This is the rst report on the isolation of P. djamor on A. salmiana as an atypical substrate, and so represents an opportunity for further study and commercialization. To its chemical composition and high availability, A. salmiana bagasse is a suitable alternative substrate for cultivating edible mushrooms, specically P. djamor. mycelium WP 160 until 0.77% TN, T2: 0.95% TN, T3: 1.14% TN, T4: 1.32% TN, T5: 1.5% TN) and 3 repeats in which the minimum, maximum, and central values for total N in the A. salmiana bagasse were 0.95, 1.5, and 1.32% TN, respectively. Nitrogen was calculated in percentage amounts equivalent to the weight in grams of the urea. The solutions were prepared with distilled water and sterilized with a 0.2-µm cellulose membrane. A potentiometer was used to measure pH. Supplementation was performed 3 days after inoculation using a sterile syringe to cover all the bags. The amount of solution administered was calculated on the basis of the moisture of the inoculated bags, adjusted to 80%. Once all the bags were supplemented, they were kept under darkness until primordia appeared, protocol modied by Monterroso (2009).


Introduction
Cultivation of edible mushrooms has increased worldwide in recent years due to their culinary characteristics, health bene ts, biological activity (antioxidant, antihypertensive, anticancerogenic), and multiple biotechnological and environmental applications, among other reasons.
The genus Pleurotus is the second-most widely cultivated mushroom on the planet with an estimated production of 6.46 x 10 9 kg (Arango and Nieto 2013;Sardar et al. 2017;Royse et al. 2017). Some 50 species of this genus have been reported. Its cultivation requirements and nutraceutical and biodegradative properties have attracted interest, and it is now a promising agroindustrial product with expanding cultivation in Latin America and other areas of the world (Royse and Sánchez 2001;Barba et al. 2016;Salmones 2017).
In Mexico, the species with the highest production volumes are Pleurotus ostreatus, P. djamor, and P. pulmonaris (Salmones 2017). They are cultivated on a broad range of agroforest by-products: rice chaff, oats, wheat, cotton, palm oil, scrub, sawdust, pulps, fruit peels, jute waste, bagasse, and other kinds of chaff, among others (Bellettini et al. 2019), all of which are rich in the carbon, nitrogen, sulfur, and phosphorus necessary for the development of mushroom biomass (Gregori et al. 2007). Agave bagasse has these same properties, and estimates indicate that this material is produced at a rate of 4'709,000 tons per day. Accumulation of this bagasse causes environmental problems and contributes to the proliferation of rodents, insects, bacteria, and fungi (Gallardo 2018). It has been proposed as an alternative substrate for growing mushrooms (Heredia-Solís et al. 2014;Heredia-Solís et al. 2017), but more studies are required to improve cultivation and production conditions. Due to the genus Pleurotus' ability to grow in diverse substrates, it has been found on agave plants where it develops mainly on shoots near the ground around the base of plant, and in the cavity of exhausted magueys. It has been identi ed as P. opuntiae. This genus, however, is subject to controversy because not all the mushrooms that grow on these plants correspond to this species. In these circumstances, molecular studies are highly useful (Granados 1993; Barrales and Mata 2016;Heredía-Solís et al. 2017;Zervakis et al. 2019).
The objective of the present study was to use molecular techniques to identify a wild mushroom isolated from A. salmiana, and then evaluate its biological e ciency, production rate, and nutritional and morphological characteristics when grown on A. salmiana bagasse with various concentrations of urea as a source of nitrogen.

Isolation and cultivation
The wild mushroom was isolated from a fruiting body that developed on A. salmiana in the municipality of Chilcuautla, Hidalgo, Mexico (20°20′00″N 99°14′00″Or). The hymenia were cut in 1-cm 2 fragments, subjected to treatment (5 min in sterile distilled water, 5 min in a hypochlorite solution at 5%, and washing with sterile water), and then seeded on plates in potato dextrose agar (PDA) medium for 7 days at 28ºC until the mycelium characteristic of the mushroom was obtained. Re-seedings were done periodically to achieve puri cation.

DNA extraction
The mushroom's genomic DNA was obtained from 0.750-g pellets recovered by ltration, frozen, and then lyophilized from a liquid culture in Kirk medium for 7 days, utilizing the protocol proposed and modi ed by Huanca-Mamani et al. (2014). The concentration and purity of the DNA were measured in a NANODROP 2000, and integrity was veri ed by electrophoresis in agarose gel at 1% (30 min, 80V, electrophoresis chamber).

Ampli cation and sequencing
The ITS1-5.8-ITS2 region of the DNA of the mushroom was ampli ed by the polymerase chain reaction (PCR) using the universal oligonucleotides ITS1 and ITS4, following Díaz et al. (2014) modi ed protocol. The product of PCR was analyzed by electrophoresis in agarose gel at 1%. The base pairs were determined by comparison to the 1 Kb marker. The Wizard® S.V. Gel and PCR Clean Up System kit was utilized to purify the PCR products. Sequencing was performed at the Unidad de Síntesis y Secuenciación de ADN at the Instituto de Biotecnología of the National Autonomous University of Mexico (UNAM).

Preparation of the primary inoculum
The primary inoculum was obtained in two forms: wheat grains (WG) and pellets (WP). The former was prepared with wheat seeds hydrated for 12 h. Excess water was drained and 250 g of the wheat were placed in polypaper bags. These were sterilized for 15 min and then inoculated with 15, 1-cm 2 squares of the mushroom's mycelium. Incubation was performed at 28ºC under total darkness until the mycelium completely invaded the seeds. The WP were prepared in 250-mL Erlenmeyer asks containing 100 mL of potato dextrose broth. They were sterilized and inoculated with 10 mL of the mushroom's mycelium suspended in sterile water, and incubated at 28ºC under stirring at 160 rpm until uniform pellets formed.
Selection and characterization of the substrate The A. salmiana bagasse employed consisted of residues from the pulque, inulin, and honey industries. It was provided by the Coorporativo Magueyero San Isidro, S.A. de C.V. in the municipality of Nanacamilpa, Tlaxcala, Mexico (19°29′00″N 98°32′00″O). Selecting the residue required physicochemical analysis of the bagasse from the shoots, the pineapple, and a 50:50 mixture of shoots:pineapple.  (Gouleke 1977), and the C/N ratio. pH was measured in extracts of distilled water in a weight/volume ratio of 1:5 (Crespo et al. 2018).

Inoculation of the substrate
Bagasse from the shoots of A. salmiana was washed in distilled water and pasteurized at 65ºC for 60 min. Excess water was drained and the material was left to cool at ambient temperature. Once cooled, 27 bags, each containing 150 g (dry weight) of A. salmiana bagasse, were inoculated for each experiment (WG, WP) and distributed in 5 treatments for later supplementation with urea. The procedure followed the methodology proposed by López-Rodríguez et al. (2008) for inoculation with WG, while for inoculation with WP, we took the total content of the pellets from each ask, ltered the culture broth and deposited the pellets in the center of the polypaper bags that contained the previously sterilized substrate, according to Abdullah et al.'s modi ed protocol (2013).

Supplementation with urea
During supplementation of the substrate, the total nitrogen content (TN) of the bagasse of the A. salmiana shoots was taken as the initial parameter. The experimental design for WG and WP consisted in 9 treatments (T1: 0.77% TN, T2: 0.95% TN, T3: 1.14% TN, T4: 1.32% TN, T5: 1.5% TN) and 3 repeats in which the minimum, maximum, and central values for total N in the A. salmiana bagasse were 0.95, 1.5, and 1.32% TN, respectively. Nitrogen was calculated in percentage amounts equivalent to the weight in grams of the urea. The solutions were prepared with distilled water and sterilized with a 0.2-µm cellulose membrane. A potentiometer was used to measure pH.
Supplementation was performed 3 days after inoculation using a sterile syringe to cover all the bags. The amount of solution administered was calculated on the basis of the moisture of the inoculated bags, adjusted to 80%. Once all the bags were supplemented, they were kept under darkness until primordia appeared, protocol modi ed by Monterroso (2009).

Fructi cation and harvesting
After the appearance of the primordia, the bagasse was exposed to light at a temperature of 18-20ºC and a relative humidity of 80-90% until mature fruiting bodies were obtained (López-Rodríguez et al. 2008). The mushrooms were gathered manually with a sterile scalpel and measured. This procedure was repeated for each harvest (Fernández 2004).

Evaluation of productive parameters
The following parameters were recorded: running time of the mycelium on the substrate, precociousness (time of appearance of primordia), and fructi cation. The productive parameters determined were biological e ciency (BE) and production rate (

Statistical analyses
Data analyses were performed using the Minitab 19 statistical program. A one-way ANOVA was applied with a Tukey test for multiple comparisons. The con dence level was 95%.

Molecular identi cation
The molecular study of the fungus isolated from A. salmiana allowed us to obtain one ampli cation product with a size of 750 pb. It was identi ed as Pleurotus djamor. The sequence was deposited in GenBank under access number MW581271, and a phylogenetic tree was elaborated ( Fig. 1 Accession URL:http://purl.org/phylo/treebase/phylows/study/TB2:S28312) using the sequences of Pleurotus available at GenBank.
Culture and productive parameters of P. djamor Table 2 shows the results of the characterization of various sections of the bagasse of A. salmiana. The residue of the pineapples is rich in crude protein (CP) at 11.51 %, compared to just 4.8% in bagasse from the shoots. During cultivation, we analyzed two types of primary inoculum: a traditional one that utilizes wheat grains as support, and a second made from pellets of the mushroom itself. Observations showed that the type of primary inoculum used did not affect the development of P. djamor, and that the use of the inoculum had no impact on the mycelial growth of the mushroom on the substrate. The inoculum in pellet form (WP) colonized the A. salmiana bagasse in 13 days, while the inoculum in grain form (WG) required 25 days. The seeds that resulted from the WG treatment were homogeneous, especially in the wheat grain, while the WP pellets varied in size, were semi-uniform, and beige in color. Neither product showed signs of contamination. Table 3 shows the time of appearance of the primordia (precociousness), and the rst and second harvests of the experiments inoculated with WG and WP. Table 4 displays the different concentrations of urea and BE and PR. Because the bags inoculated with WG increased the concentration of nitrogen, precociousness decreased. In treatment T1 (substrate not supplemented with nitrogen), 14 days were required for the rst primordia to appear, while in T2 and T3 the time of appearance was 11 days, and for treatment T5 it was 10. In contrast, in the bags with WP precociousness increased with higher concentrations of nitrogen, reaching 14 days in T1 and T2, and 18 days in T3, T4, and T5.  the time required was 41-45 days. The treatments inoculated with WP only produced one wave of mushrooms at 27 ± 1 days after inoculation. This represents an economic disadvantage for production. It is likely that the WG treatments included nutrients that promoted the growth of the mushroom, while the WP inoculum lacked these compounds that strongly impact the number of harvests achieved. The BE and PR of the treatments inoculated with WG increased with respect to the non-supplemented treatment (T1) and as the concentration of TN in the bagasse increased. The maximum BE achieved was 70%, while the maximum PR was 1.18, both in T5. The BE and PR of the treatments inoculated with WP were lower, with T3 showing the highest values for these two parameters (44% and 0.73). The evidence from this experiment allowed us to determine that the primary inoculum based on pellets (WP) did not generate higher yields in the cultivation of P. djamor, as was expected. In contrast, the BE and PR of the bags inoculated with WG and supplemented with urea as a source of nitrogen improved.
The fruiting bodies of the WG and WP experiments are shown in Fig. 2. In both cases, they were of beige color with regular growth, a cottony texture, and smooth decurrent laminae. However, the form of the pileus of the mushrooms inoculated with WG was abelliform, while that of the mushrooms inoculated with WP was infundibuliform. In the WG treatments, 31% of the fruiting bodies from T1 reached a size of 5-8 cm, 39% measured 8-13 cm, but only 4% achieved a size of 13-16 cm. In T2 with WG, 54% of the mushrooms measured 8-13 cm. For T3 the gures were 62 for 8-13 cm and 24% between 5 and 8 cm. For T4, 39% of the carpophores reached a size of 8-13 cm. For T5, 48% of the fruiting bodies measured 8-13 cm. Although some mushrooms from T4 and T5 achieved average size, some carpophores showed malformations in the clusters and caps. In addition, almost 20% of the fruiting bodies in T4 detained their development after day 14 (Fig. 3).
Regarding the WP experiment, 45.5% of the fruiting bodies in T1 measured 5-8 cm, but only 6% reached 10-13 cm. In T2, 35% of the carpophores were classi ed in a size of 5-8 cm. In contrast, observations of T3 showed that 16% of the mushrooms achieved a size ≥ 16 cm with a predominance of carpophores between 10 and 13 cm. The average size in T4 was 10-13 cm (37%), while in T5, 21% of the carpophores detained their growth.

Nutritional composition of the fruiting bodies
The effect of the type of inoculum and the substrate supplemented with urea on the morphology and nutritional composition of the fruiting bodies are shown in Tables 5 and 6, respectively.  Means not labeled with the letter "a" are signi cantly different from the mean of the control level in the Tukey Multiple Comparison Test with a signi cance level (P ≤ 0.05). ±

Standard deviation
The moisture determined for the fruiting bodies in this study ranged from 85.22-89.83% in the two experiments and showed statisticallysigni cant differences with respect to controls (T1) (p ≤ 0.05). The highest CP content (26.27%) occurred in T1 of the mushrooms inoculated with WG without added nitrogen. We expected that in addition to improving BE and PR, supplementing the substrate with a source of N (urea) would increase CP content, but as the concentration of urea in the substrate increased in the WG treatments, the CP of the mushrooms decreased signi cantly. Although the concentration of CP in the mushrooms inoculated with WP achieved a value considered typical for this species, it was below the level of the mushrooms inoculated with WG. Treatment T4 (WP) had the highest protein content. On the other hand, the mushrooms in experiments WG and WP the fat content, ranged from 7.42-8.36%. Regarding ash content, the sporomes obtained from WG and WP in the present study contained 7.3-8.3%.

Discussion
The mushrooms that grow on plants of the genera Opuntia, Yucca, Agave, and Phytolacca, among others, are described as P. opuntiae, but because worldwide the genus Pleurotus includes several taxa and species, controversy exists due to the absence of valid sequencing data and/or phylogenetic reconstructions. This has generated signi cant ambiguities regarding the exact identity and distribution of P. opuntiae (Zervakis et al. 2019). The mushroom P. djamor has a cosmopolitan distribution that includes species identi ed as P. ostreatoroseus, P. parsonsiae, and P. salmoneostramineus. P. djamor is a pantropical species found in wild form in several countries in the America, including Mexico, though its distribution is probably much broader (Zervakis et al. 2019). Molecular studies have of 600-800 pb have been reported for P. abalonus, P. cystidiosus, P. cystidiosus var. Formosensis, P. fuscoquamulosus, P. smithii, and P. australis (Zervakis et al. 2004), P. djamor, P. cornucopiae, P. cystidiosus, and P. pulmonaris (Imtiaj et al. 2011), but recently fragments of 700 pb were found for P. djamor and P. ostreatus (Doroteo et al. 2018).
Mushrooms of the genus Pleurotus have the ability to grow on materials with low nitrogen concentrations. The results of our nitrogen analysis indicate that the bagasse of A. salmiana is a good candidate for use as an alternative substrate for cultivating edible mushrooms. Nitrogen is essential for protein synthesis, purines, pyridine, and chitin in mushrooms, but high levels can shrink yields (Chang and Miles 2004). Thus, it is extremely important to ensure that the substrates utilized to cultivate edible mushrooms contain an adequate nitrogen content that will allow correct growth. According to Heredia-Solís et al. (2014), the content of CP in A. salmiana and Agave weberi bagasse does not exceed 4%. In this study the nitrogen content in the bagasse, pineapples, and the mixture (50:50) was measured at 0.77, 1.84, and 1.40%, respectively; gures that concur with those reported for bagasse, which range from 0.1-1% of N (Obodai et al. 2003;Koutrotsios et al. 2014).
During the cultivation of fungi, the type of inoculum WP represents a competitive advantage, against other microorganisms that could compete for the substrate and contaminate the culture. Due to its origin, the inoculum in pellet does not harbor spores of other fungi or microorganisms because it exerts greater control (Rosado et al. 2002;Abdullah et al. 2013). According to Khanna et al. (1992), the precociousness in Pleurotus spp. ranges from 24-30 days, though Kalmis and Sargin (2004) and Naraian et al. (2009) reported the appearance of primordia at 22 and 20 days, respectively. For P. opuntiae, the appearance of primordia occurred at 30 days (Barrales and Mata 2016). In the present study, the time of appearance of the primordia of P. djamor was lower and similar to observations by Salmones et al. (2005); that is, greater precociousness of P. djamor after 11 and 13 days on wheat chaff and coffee pulp, respectively. Supplementing substrates with nitrogenated sources improves the production of mushrooms of the genus Pleurotus spp. (Naim et al. 2020). But other observations have shown de ciencies in mycelial growth and yields when nitrogen sources such as ammonium chloride were applied to P. ostreatus and P. cystidiosus at concentrations above 0.09 and 0.05% (Hoa and Wang, 2015), and with ammonium sulfate and urea at concentrations of 1% and 1.5%, respectively (Naraian et al. 2009).
The growth, development, quality, and quantitative aspects (biological e ciency, productivity) are closely-related to the type of nutrients present in the substrate, an equilibrium in the C/N ratio, and other conditions of cultivation (Mukhopadhyay et al. 2002;Curvetto et al. 2002;Bellettini et al. 2019). In this study the absence of a second harvest could be due to the inoculum in pellet form (WP) since it did not contain the characteristic nutrients of wheat grain that provide energy for mycelial growth and development (Royse et al. 2004). Other possibilities include the nutritional exhaustion of the substrate during mushroom growth, and/or the accumulation of toxic substances that impeded fructi cation (Upadhyay et al. 2002). The C/N ratio is a determining factor for mushroom production because excess nitrogen can affect the degradation of lignin, thus impeding the development of the mycelium and the formation of fruiting bodies. In the phase of development of the fruiting bodies, a low C/N ratio is favorable. Bellettini et al. (2015) recommend an C/N ratio of 28-30 of carbon and 1% nitrogen for growing mushrooms. Heredia-Solís et al. (2014) reported the same BE with P. ostreatus using a substrate made of bagasse of A. salmiana and 40% of BE with A. weberi. In other results, a BE of 60.2% was obtained with Agave tequilana (Soto-Velazco 1989), and of 33.24% using a substrate of bagasse of Agave angustifolia mixed with 30% nogal shavings and 5% wheat bran (Heredia-Solís et al. 2016). Deformations of the fruiting bodies -especially elongated stipes and reduced coloration-have been reported due to the effects of cultivating conditions (high luminosity, temperature during fructi cation, CO2 levels) (Urben 2004). However, considering that the environmental conditions in the present study were the same for all levels of nitrogen in the substrate, we deduced that higher N concentrations inhibited the growth of the fruiting bodies (Kanhar et al. 2007;Monterroso et al. 2009;Hoa and Wang 2015), and induced deformations in the mushrooms. High N content also produced slow growth of the stems of the fruiting bodies and delays their formation. It is possible, therefore, that the malformations found in this study are related to an imbalance of the C/N ratio.
The nutritional quality and composition of the fruiting bodies depends on both the cultivation conditions (temperature, moisture, pH, etc.) and the substrate, since this is the source of the nutrients and lignocellulosic material that supports growth, development, and fructi cation. The use of a waste lignocellulosic material as substrate is key to maximizing yields and enhancing product disposition (Chang and Miles 2004;Sardar et al. 2017). For this reason, we opted to cultivate P. djamor on the same source from which it was isolated (i.e., A. salmiana bagasse). One of the most important parameters for this type of food is the amount of protein content, since it is well-known that species of Pleurotus spp. grow on substrates with low nitrogen content yet the fruiting bodies they produce have higher concentrations of N. Protein content, however, is in uenced by factors that include the nature and nutritional components of the substrate, the strain, the stage of development, and the timing of post-harvest analysis (Cowling and Merrill 1966;Sardar et al. 2017). The genus Pleurotus sp. is characterized by low fat content, high protein content, and a low concentration of lipids in the fruiting bodies (Bellettini et al. 2019).
In this study the CP content of the fruiting bodies in our WP and WG treatments was found to be within the ranges established for the genus Pleurotus; that is, 17-25% (Lelley 2017), with 20.3% of CP for P. ostreatus , and 20.7-28% for P. djamor (Salmones 2017).
In studies of Pleurotus spp. cultivated in supplemented bagasse, Ortega et al. (1992) described an increase of nitrogen in the mushrooms that was related to the amount of N present in the initial substrate and to the inoculum. Sturion and Oetterer (1995) suggested that mushrooms might be xing this element. Wheat seeds can provide nutrients to the inoculum that enrich the substrate and de ne the development of the mycelium in early stages; however, because the inoculum with WP did not contain nutrients from wheat, it may have impeded optimal growth. Therefore, it is probable that the type of substrate and inoculum in uence the nutritional composition of the fruiting bodies (Bellettini et al. 2019). Also the percentage of moisture depends on the strain, substrate, and growing environment (temperature, relative humidity) (Guillamón et al. 2010). The moisture percentages for this species indicate 85-90.9% (Sardar et al. 2017).
The results obtained from the chemical composition analysis in this study suggest that P. djamor is a promising food for several types of diet due to its nutritional quality (Salmones 2017). Up to now, this mushroom has not been cultivated on A. salmiana bagasse, but its high adaptability to cultivation conditions could well make it attractive in international markets as an exotic mushroom that can be cultivated on maguey bagasse.
The suitability of a substrate for cultivating mushrooms depends on its chemical composition, availability, and cost. In this regard, residual A. salmiana bagasse from pulque, inulin, and agave honey production is a promising option that can compete on a small scale with other substrates, since its generation is growing daily, with reports of the generation of 0.5 tons of bagasse from inulin production, and 0.16 tons from producing just 0.4 tons of agave honey (Hoz-Zavala et al. 2017). In addition, its availability, though not speci c, contains the elements necessary for the developmental and reproductive functions of edible mushrooms (Salmones et al. 2005;Heredia-Solís et al. 2017;Gallardo 2018). Moreover, in comparison to similar substrates, when supplemented with urea it reaches a BE greater than that of bagasse of A. tequilana, A. augustifolia, and A. weberi. In general, then, exploiting A. salmiana bagasse represents a sustainable option in ecological, social, and commercial terms (Heredia-Solís et al. 2014;Heredia-Solís et al. 2017).
This is the rst report to address the isolation of P. djamor of Agave salmiana and grown on bagasse of Agave salmiana as an atypical substrate. During cultivation on A. salmiana bagasse, the growth of the fruiting bodies depended on the type of inoculum and the concentration of urea. Supplementing the substrate with urea (1.32%) to provide nitrogen improved both BE and PR by as much as 70% and 1.17, respectively, in the cultivation of P. djamor. At higher concentrations (1.5%), however, this inhibited the growth of the fruiting bodies and produced malformations. A. salmiana bagasse is a suitable alternative substrate due to its chemical composition and high availability because it permits the growth of the mushroom without altering the optimal nutritional composition of this genus, thus providing economic and environmental advantages. Results represent an invitation for further study and commercialization.

Consent for publication
Not applicable

Data availability
The datasets generated during and/or analysed during the current study are available in the GenBank repository, under access number MW581271, the phylogenetic tree was deposited in TreeBASE (Reviewer access URL: http://purl.org/phylo/treebase/phylows/study/TB2:S28312?x-access-code=288be19013aeff456bd2b892c2905a0e&format=html),; and this are available from the corresponding author on reasonable request.

Figure 1
Phylogenetic tree generated from the sequences of the ITS region of strains of Pleurotus.

Figure 2
Fruiting bodies from the different treatments of the experiments with WG and WP. The bodies are of beige color, with regular growth, cottony texture, and decurrent laminae.