ORIGINAL RESEARCH

International Journal of Recycling of Organic Waste in Agriculture (2021)10: 29-42Doi:10.30486/IJROWA.2020.1904599.1104

Valorization of vermicompost with bacterial fermented chicken feather hydrolysate for the yield improvement of tomato plant: A novel organic combination

Ishita Biswas1, Debasis Mitra1, Ansuman Senapati 2, Debanjan Mitra1, Sourav Chattaraj1, Murshed Ali1, Goutam Basak1,3, Periyasamy Panneerselvam2, Pradeep K. Das Mohapatra1,3*

Received: 28 July 2020 / Accepted: 01 December 2020 / Published online: 15 February 2021

AbstractPurpose Chicken feather protein hydrolysate (CFPH) has drawn a significant attention as a component/type of biofertilizer in recent years, because of the beneficial impact on the growth of the plant. The current study aims to evaluate the potential influence of the combination of CFPH with vermicompost (VC) on growth-promotion and yield improvement in tomato plants. Method Feather degrading bacteria were isolated and characterized using 16s-rDNA sequencing, and assessed for biochemical reactions, growth-promoting attributes and keratinase activity. The medium used for feather degrada-tion studies consisted of 0.75% (w/v) of raw feather, with 1% (v/v) of inoculum at 37°C, pH 7.5 and at 120 rpm. A field study was done by randomized block design (RBD) with five treatments in tomato.Results Keratinolytic and feather degrading bacteria isolated and used in this study were identified as Bacillus cereus PKID1 with accession number MT158702. The bacterium gave the highest keratinase activity of 80±0.28 U/ml. The CFPH showed the potential to promote remarkably the germination % of tomato (84.13), rice (87.24), onion (84.13), chilli (84.13), chickpea (73.24) seeds; field experiment significantly increased plant growth and yield compared with control.Conclusion The principal component analysis of the field experiment as a result of tomato plant-growth, the order of best treatment efficacy for improvement of parameter estimates was as follows: CFPH and VC > CFPH > VC > recommended dose of fertilizers (RDF) > control. Thus, the application of CFPH with VC could improve the productivity of crops and decrease the use of chemical fertilizers.

Keywords Fertilizer, Fermentation, Keratinase, 16S-rDNA

Department of Microbiology, Raiganj University, Raiganj - 733 134, Uttar Dinajpur, West Bengal, IndiaCrop Production Division, ICAR - National Rice Research Institute, Cuttack - 753 006, Odisha, IndiaProfessor A. K. Bothra Environment Conservation Centre, Raiganj University, Raiganj – 733 134, Uttar Dinajpur, West Bengal, India

1

2

3

Pradeep K. Das Mohapatra pkdmvu@gmail.com

Introduction

Application of chemical fertilizers for the production of crops creates serious environmental hazards to human health. For reducing these harmful impacts of chemical fertilizers, organic fertilizers are preferred (Sharma et al. 2019). VC is among those organic soil amendments

which have been widely used in agriculture with a high market demand globally. VC is enriched manure with essential nutrients and contains a number of agriculturally important microbes (Arancon et al. 2005), thus considered as a biofertilizer for the application in agricultural productivity (Banu et al. 2001; Jones et al. 2005). Besides that, VC improves soil health, increases plant productivity and also limits the diseases caused by soil-borne pathogenic organism in plants (Chaoui et al. 2002; Scheuerell et al. 2005; Paul et al. 2014a). Many reports have shown that soil supplemented with VC can increase the growth and yield of some crops and plants (Joshi et al. 2013; Kumar et al. 2018a; Rekha et al. 2018; ). Utilization of organic fertilizer produced from biological waste materials enriched with plant growth-promoting (PGP) supplements is the noble practice

International Journal of Recycling of Organic Waste in Agriculture (2021)10: 29-4230

for present day agriculture (Vessey 2003; Ahemad and Kibret 2014). The use of organic wastes as fertilizer provides a potential way of waste management and recycling (Chew et al. 2019; Sharma et al. 2019). Researchers have focused on the utilization of organic wastes in agronomic improvement because the compost produced from animal wastes are environment friendly and will be the supplement of commercial fertilizers that have costly and intensive production process (Joardar and Rahman 2018; Nagarajan et al. 2018; Nurdiawati et al. 2019).

Feathers are the most abundant waste generated by the poultry industry and it is gradually increasing everyday (worldwide annual production is around 8.5 billion tons) due to the large consumption of poultry products. The production rate of the chicken feathers in India is about 350 million tons (Agrahari and Wadhwa 2010; Sahoo et al. 2012). Due to its recalcitrant nature, assimilation of such a great amount of feathers leads to wastage of feather protein (Paul et al. 2014a) and environmental pollution because they are the source of various disease-causing microorganisms (Jayathilakan et al. 2012). The chief nutritional constituent of the feather is tough, insoluble protein keratin [over 90% (w/w)] along with larger amounts of phenylalanine, glycine, arginine, cysteine, and glutamate (Veerabadran et al. 2012; Paul et al. 2014a). Keratin is difficult to degrade by pepsin, trypsin and papain for its massive cross-linked polypeptide chains and the presence of several disulfide bonds that provide mechanical resistance (Gupta and Ramnani 2006).

Recently, researchers have reported that CFPH could serve as N2 fertilizer (Genç and Atici 2019) due to its high-protein content but its recalcitrant nature results in decreasing rates of its decomposition and mineralization of N in the soils (Thuriès et al. 2001). Traditional biochemical and thermal feather degradation processes involve steam pressure cooking and alkali hydrolysis that not only denature the amino acids but also utilize a great amount of energy. The microbial enzymatic catalysis converts keratin into free amino acids, low molecular weight peptides that could be easily available to animals and plants for their uptake, and the protein hydrolysates (PH) thus produced might be used for agriculture (Thuriès et al. 2001; Pillai and Archana 2008; Kumar et al. 2018a). The objectives of the present work are to: isolate and identify keratinolytic bacteria to be used for CFPH production; study the effects of soil amendment with CFPH and VC on tomato growth and

yields under controlled field conditions; and determine seed germination assays on tomato, rice, onion, chilli and chickpea plants.

Materials and methods

Collection of chicken feather and soil sample

The feathers of chicken were collected from the slaughter shops. Feathers were washed repeatedly with the help of normal tap water and then dried for a period of 2 days and further kept at room temperature for further processing. The soil sample was obtained from the dumping site of poultry feathers of Raiganj town, Uttar Dinajpur district, West Bengal in a sterile polythene bag and brought to the laboratory.

Isolation of keratinolytic bacteria from the soil sample

To isolate feather-degrading bacteria, a 10 g sample of soil was mixed with 250 ml of sterile saline water and were shaken properly by placing it on a rotary shaker for 4 h. Then 1 ml was taken from the solution and serially diluted up to 10−9. The diluted samples were individually spread on the casein agar plate of the medium composition of 0.5 g (w/v) of NaCl; 0.1 g (w/v) of MgSO4; 5.0 g (w/v) of casein; 1.5 g (w/v) of agar (pH 7.5) and allowed to incubate at the temperature of 37°C for 2 days. The plates were then examined for the appearance of colonies. Colonies with a comparatively larger size were selected for further study. The pure isolates were then grown in keratin agar medium of 0.5 g (w/v) of keratin, 0.03 g (w/v) of K2HPO4, 0.04 g (w/v) of KH2PO4, 0.05 g (w/v) of NaCl, 0.01 g (w/v) of MgCl2, 6H2O, 1.5 g of agar and incubated at 37°C (Paul et al. 2014a, b). The isolate that produces the largest colonies was selected and named PKID1 and stored in the slant of keratin agar medium for further use.

Identification of the selected bacterial strain

The isolated bacterial strain above was identified based on its molecular, biochemical, and morphological character is t ics (Mitra 2017) . Morphological characterizations involved morphology, pigmentation, and Gram staining whereas biochemical characterization was based on the IMViC tests, starch, casein, urea hydrolysis, production of indole, MR/VP test, lipid

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hydrolysis, catalase, oxidase and carbohydrate fermentation test by HiCarboTM Kit (KB009B1).

DNA extraction, gene amplification and sequencing

The genomic DNA of the isolate was extracted by the help of DNA isolation kit (Zymo™, USA). Approximately, 10 mL of 24 h grown culture (nutrient broth) was centrifuged at the rpm of 8000 for 15 min and the extraction of DNA from cell pellet was done as per the specified protocol of the kit. The purity and quantity of DNA were determined by using a UV-spectrophotometer (Nanodrop, Thermo Scientific). 27f (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492r (5′-GGTTACCTTGTTACGACTT-3′) primers were utilized for the amplification of 16S-rRNA gene (Suzuki and Giovannoni 1996) and purified samples of PCR were sequenced through Sanger sequencing. Contigs were created from the resulting forward and reverse reading sequences using the CAP3 assembly software (Huang and Madan 1999) and the existence of chimera was tested using DECIPHER v2.0 (Wright 2016).

Sequence analysis and deposition of isolates

Alignment of sequence data was done by using BLASTN 2.8.1 (https:/blast.ncbi.nlm.gov/Blast.cgi) and the sequencing was deposited in the NCBI (https:/submit.ncbi.nlm.gov/subs/genbank/) for the accession number. The study of the phylogenetic analysis was performed via. Sequence program MEGA6-ClustalW followed by evolutionary tree construction using MEGA6 software based on the Tamura-Nei model viz. 1000 bootstrap replications and a standard pattern of partly removed nucleotide modifications (Tamura and Nei 1993; Kumar et al. 2018b).

Production of CFPH through submerged fermentation

The fermentation medium was prepared with the composition (g/L) 5 g NaCl, 1 g MgSO4, 0.5 g K2HPO4, and 10 g chicken feather in 250 ml of normal water in Erlenmeyer flasks. Fermentation was carried out by addition of 1% inoculum followed by incubation at 37°C and 120 rpm for about 72 h. Inoculum medium was the same as the isolation medium except that agar was not added. After the centrifugation of fermented broth at

5,000 rpm for 15 min, the supernatant was collected and analyzed further for the keratinolytic activity.

Production of VC

Organic wastes such as kitchen waste and cow dung were utilized as substrates for VC production. Kitchen waste was collected from the hostel of Raiganj university campus and cow dung were collected from the local area of Raiganj, Uttar Dinajpur, West Bengal. Earthworms (Eisenia foetida Savigny) were collected from West Bengal CADC Keotan, Baghan, Kaliyaganj, Uttar Dinajpur, 733 129 West Bengal. Cylindrical tanks were prepared using cement rings and the size of the vermicompost unit was 0.18 m3. The base of the tanks was filled with sand to make a thick bed for proper percolation of water. After that different organic wastes (combination of cow dung, kitchen waste and wheat straw) were collected and put in different ratios above the sand bed. Heavy species from kitchen wastes were avoided. All these were sun-dried for 7-8 days. In one tank only cow dung was used, in the second tank cow dung and kitchen waste were mixed in 4:3 ratios, in third tank cow dung, kitchen waste and paddy straw were mixed in 4:3:1 ratio. All organic waste was decomposed for at least 20 days and vermicomposted for 40 days with Eisenia foetida. Table 1 represents the details of requirements, handling and harvesting for VC production.

Study of physicochemical parameters of field soil and VC

The pH, soluble salts, organic carbon, phosphate, nitrogen, potassium, sulphate sulphur (Sharma et al. 2014; Shukla et al. 2016) and micro nutritional parameters of soil and VC were analyzed (Nurdiawati et al. 2019).

Test plant, site, experimental layout and treatments

The field experiment was carried out at Professor A. K. Bothra Environment Conservation Centre, Raiganj University, Raiganj - 733 134 Uttar Dinajpur, West Bengal India (25.6329° N, 88.1319° E) from January to March 2020. The soil physio-chemical properties at 0 - 30 cm depth were measured. Soil was fine loamy texture with limited clay and humus accumulation having the

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Table 1 Requirements, handling and harvesting of VC

Description ValueUnit 0.18 m3

Species used for VC production Eisenia foetida SavignyType/ nature of worm EpigeicEarthworm inserted 200Earthworm released after composing 350Substrate used Organic wastes (combination of cow dung, kitchen waste and wheat straw)Temperature for earthworm production 28-30°CCompost period 40 daysWatering schedule for the unit 2times / dayMoisture content should be maintained 70-80%Quantity of VC recovered (kg) 60

following physio-chemical properties listed in Table 3. Tomato plants were chosen for field experiment and five plots were prepared. Initially germination of tomato var. Karan (F1 hybrid) seeds were allowed in earthen pots for a week and then they were transferred to the field. The experiment field was arranged in a randomized block design (RBD) with five treatments. There were four replicates of each treatment arranged within each block. Each block was located 75×150 cm apart, consisted of five treatments and total of three plants/block in each plot. The five treatments were as follows;

(T1) Control, (T2) CFPH, (T3) Vermicompost (VC); 5 t per ha(T4) RDF- 200:250:250 kg of NPK per ha, and (T5) 1000 gm VC + 100 ml CFPH. Before application, CFPH and VC were dispersed

in water at the rate of 1:1 and drenched basally. The plots were watered every day to keep sufficient water content of the soil. The data on agronomic parameters, i.e. plant height of shoots and root (in centimeters) by metric rule, number of leaves by counting, fruits number and crop yield by weighing scale, were determined at the regular time interval of 10 days up to harvest of the crop. The height of the plant was measured as the distance between the plant base and the first branch of the tassel (Badu-Apraku et al. 2010). Field experiment, all data were analyzed through R software. Other plant growth promoting experiment of % seeds germination of rice, tomato, onion, chilli, chickpea seeds was performed. Seeds were surface sterilized with bleach solution (0.02% Triton X-100

and 30% commercial bleach) for 15 min., washed 3 times with the help of sterile water and treated with CFPH for 30min. followed by the towel method and observed the results at regular intervals. Another, one pot (5 kg soil/pot; 4 replication) experiment was conducted on rice (surface sterilized) seeds treatment with CFPH for 20days.

Statistical analysis

Principal component analysis (PCA) was performed by using the R software (Team 2000). Experimental data were statistically analyzed using WASP 2.0, Goa online statistical analysis software. Standard errors were calculated for all mean values. Differences were considered significant at the p ≤ 0.05 level.

Results and discussion

Isolation of bacteria

In the present study, 12 bacteria were isolated from a sample of soil, of which one potential isolate (PKID1) was selected based on the biochemical characterization viz. starch, casein, lipid hydrolysis and keratinase enzyme activity (Appendix Table 1). Similarly, Nagarajan et al. (2018) documented that isolates B. subtilis strain FW12 showed the highest keratinase and plant growth promoting activity. Paul et al. (2014a) isolated a Paenibacillus woosongensis TKB2 strain on the basis of biochemical, and morphological characterizations and feather hydrolysate which were applied as a useful biofertilizer.

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Screening for biochemical, morphological and keratinase production

The identified isolate PKID1 was characterized on the basis of their morphological, physiological, biochemical, and cultural properties as presented in Table 2 and keratinolytic enzyme activity, respectively.

Table 2 Morphological, cultural and biochemical characteristics of the bacterial isolateCharacteristics ResultGram’s staining +veSize RodTemperature 28±2.00CpH 7.4±0.2Colour WhiteMotility +Spore forming + (>95%)Starch hydrolysis +Casein hydrolysis +Urea hydrolysis +Production of indole -MR/VP test -/-Lipid hydrolysis +Catalase -Oxidase -Keratinase enzyme activity (U/ml) 80±0.28Carbohydrate fermentation test: (KB009B1)Inulin -Sorbitol +Sodium gluconate +Mannitol -Salicin +Dulcitol +Glycerol +Inositol -Adonitol +Arabitol -Erythritol -alpha-Methyl-D-glucoside -

(+) positive, (-) negative

The isolate PKID1 strain was identified to be a Gram positive ‘rod’ as it showed positive outcome on starch hydrolysis, motility, casein hydrolysis, urease and lipid hydrolysis activity. Carbohydrate utilization test was performed by HiCarboTM Kit (KB009B1) and positive results showed in sorbitol, sodium gluconate, salicin, dulcitol, glycerol and adonit.

Identification of bacterial strain

The potential isolate was identified as B. cereus PKID1 with the following Genbank accession numbers MT158702. 16S rRNA gene sequence and accession number of the related strain were retrieved from nBLAST and MSA was performed using ClustalW.

Neighbors joining molecular phylogenetic tree showing the relationship between PKID1 (MT158702) with other strains, based on their 16S rRNA gene sequence (Fig. 1). Bootstrap values are denoted as randomization of 1000. The accession number of reference strain from the GenBank are indicated in Fig. 1.

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Fig. 1 Phylogenetic tree on the basis of 16S rRNA sequence of gene, showing the distance between strains PKID1 with other strains of Bacillus species

The bootstrap values are denoted at the point of branch (percent values from 1000 replicate bootstrap samplings). The bar represents the distance of evolution by 0.0005

VC and physico-chemical properties

Physico chemical properties of the experimental field soil, vermicompost that was prepared at the PAKBECC (VERMITECH), Raiganj University, Raiganj and the chicken feather protein hydolysate (CFPH) that was produced using raw feather by B. cereus PKID1 (NCBI GenBank accession No. MT158702) are represented in

Table 3 Physico-chemical properties of experimental field soil and VC

Characteristics Soil Vermicompost (compost tea)pH 6.7n 6.2sa

Soluble salts 0.65n 0.75n

Organic carbon (%) 1.07h 1.43h

Phosphate (kg/h) 276h 552h

Potassium (kg/h) 255m 405h

Sulphate sulphur (ppm) 14.2m 20.1h

Other elements (N2) 642h 858h

Zinc (ppm) 1.8s 3.05s

Manganese (ppm) 1.5s 15s

Copper (ppm) 0.68s 0.81s

Iron (ppm) 10.2s 25.5s

Boron (ppm) 0.35d 0.58s

(n) normal; (h) high; (m) medium; (l) low; (d) deficient; (s) sufficient; (sa) slightly acidic

Table 3. Parameters were studied to find out the pH, total soluble salts, organic carbon (TOC), phosphate (P), potassium (K), sulphate sulphur (S), organic nitrogen (TN), zinc (Zn), manganese (Mn), iron (Fe), copper (Cu), boron (Huang et al. 2013b; Table 2). Results have shown that vermicompost produced from organic waste has shown comparatively higher amount of organic and inorganic nutrients compared to the experimental soil (Fig. 2; Table 3). VC imparts positive effects on improving the physicochemical properties of soil and provides structural stability to soil and also prevents deterioration of soil quality (Tejada et al. 2009; Piya et al. 2018).

Field application results of CFPH and VC treatment on growth promotion of tomato plant

The PCA was capable of explaining 41.71% in overall

variations of eight parameters estimated across five different treatments (Fig. 3). The first dimension (PC1) was capable of explaining 23.24% of variation followed by the second dimension (PC2), which

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Fig. 2 (A) Earthworms (Eisenia foetida) and (B) VC

explained 18.47% of variations. With the correlation values corresponding to the individual parameters, LLt (0.745) and Lbt (0.604) were better explained by PC1 as compared to Lbt (0.63), Lll (0.592) and Lno (0.529), which were better explained by PC2. All the correlation coefficients had p < 0.001 signifying their appropriateness in quantifying variances across the treatments. The following parameters viz. Pht, Flo and

Fru had p > 0.1 suggesting their inappropriateness for use in analyzing responses of different treatments. However, with lower correlation values, Pht (0.353) was explained by PC1 and Fru (0.273) was explained by PC2. Not shown in the biplot was Flo (0.835) which was found to be better explained by the third dimension (PC2) but was not used for analyzing treatment responses as PC3 explained 15.51% of the variance, which was

Fig. 3 PCA analysis of field experiment growth parameters of tomato plant after 50 days The following treatments were applied (T1) control, (T2) CFPH, (T3) VC, (T4) RDF, and (T5) 1000 gm VC + 100 ml CFPH. Abbreviations: (Pht) plant height in cm, (Lno) leaves no, (Llt) leaf length in cm, (Lbt) leaf breadth in cm, (Lll) leaflet length in cm, (Llb) leaflet length breadth in cm, (Flo) flower number, (Fru) fruit number

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lower than PC2. In addition, the eigenvalue variance decreased with increase in principal components starting from PC1 (1.859) to PC2 (1.478) and PC3 (1.241). Furthermore, there were significant variations within treatments. Represented in large symbols are the mean of eigenvalue variances of different treatments. From the biplot representation, it was visualized; the bottom right quadrant had T1, followed by T4 in the bottom left quadrant, and the treatments T2, T3 and T5 in the top left quadrant. From this analysis, the order of treatment effectiveness on improvement in parameter estimates were as follows: T5>T2>T3>T4>T1.

The analysis confirms the parameters represented by PC2 viz. Lbt, Lll and Lno, were strongly influenced by the treatments T5, T2 and T3 as compared to T1. Comparisons of mean values response deciphered reduction in the estimates of the following parameters Lbt and Llt under the effect of treatment T4.

The agronomic parameters such as plant height, leaves number per plant, leaf length and breadth, leaflet length and breadth and flowers number and fruits per plant at different treatments are shown in Fig. 4. It can be seen from the result that at 50 days T4 treatment did not significantly increase the height of tomato plants compared to control (T1) while the greatest height (91.75 cm) was observed at T5 treatment (a combination of CFPH and VC) and then in T2 treatment (90.58 cm) (Fig. 4A). The leaves number in plants tends to raise along with the increase in the duration of treatment. The amalgam of CFPH and VC was found to increase the mean number of leaves in a promising way thus greatest leaves number per plant was determined in T5 treatment (39.25) at 50 days (Fig. 4E). Though T3 treatment was also showing a positive influence in increasing the leaf number but it was not remarkably differed in comparison with control. Again after 50 days of treatment, combined treatment T5 showed the highest increase in leaf length and breadth (30.25 cm and 26.41 cm, respectively) however, compared to that T2 treatment showed the highest increase in leaflet length and breadth (22.08 cm and 14.66 cm, respectively) (Figs. 4B, 4C, 4D and 4F). Flower and fruit number per plant was also represented in Fig. 4G and 4H. The highest number of flowers was recorded at 40 days of treatment and T2 treatment (17.83) was found to produce a significantly large number of flowers however in case of fruit, highest yield was recorded at 50 days of treatment in case of treatment T5 (31.25).

Findings of field experiment from two time harvested tomato fruit yields per plant contain the T1 (3.54 kg), T2 (4.3 kg), T3 (4.7 kg), T4 (4.23 kg), and maximum T5 (5.2 kg) (Fig. 5).

Percentage of seed germination in different crop and vegetables, i.e. rice, tomato, onion, chilli, chickpea and growth promotion on rice results showed in Table 4 and development of rice plant root through the intervention of CFPH treatment results significantly enhanced as compared to control (Fig. 6).

However, several researchers reported that a viable alternative route of feather hydrolysis is feather biodegradation by the help of microbial keratinase (Paul et al. 2014b; Nagarajan et al. 2018; Genç and Atici 2019). Keratinase is reported to be produced by several microorganisms including bacteria such as B. licheniformis (Beg et al. 2003), B. pumilus CBS (Jaouadi et al. 2008), B. subtilis (Pillai and Archana 2008), Paenibacillus woosongensis TKB2 (Paul et al. 2014b); fungi such as, Rhizomucor, Absidia, and Aspergillus (Friedrich et al. 1999); and a few actinobacteria such as Streptomyces flavis and Microbispora aerata (Gushterova et al. 2005). Similarly, another eco-friendly and cost-effective approach for the management of organic waste is vermicomposting. Though the health benefits of fruits and vegetables have enormous evidence, however, their large consumption results in the accumulation of organic wastes because 50% of it get rotten (Huang et al. 2013a). The rotten products cause pollution to the environment and also influence the climate and biological diversity. Through vermicomposting an alternative manure is produced from such organic waste through the combined action of earthworm and microorganism inhabiting the digestive tract of earthworm (Huang et al. 2013b). Vermicompost is an enriched manure with essential nutrients and contains a number of agriculturally important microbes (Edwards 1983; Arancon et al. 2005). Sobucki et al. (2019) have reported the utilization of feather hydrolysate as a source of nitrogen for greenhouse in the cultivation of lettuce. Similarly, protein hydrolysate produced by B. amyloliquefaciens 6B had a positive impact on the germination of mung bean, and enhanced agronomic parameters as compared to untreated soil (Bose et al. 2014). Feather hydrolysate produced by B. pumilus KHS-1 also showed promising results in case of carrot and chinese cabbage (Kim et al. 2005). Wang et al. (2017) reported on the role of vermicompost as

International Journal of Recycling of Organic Waste in Agriculture (2021)10: 29-42 37

Fig. 4 Growth promotion of tomato plant in different treatments

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Fig. 5 Yield of tomato (kg / plant) after 50 days by different treatments

a potential soil supplement for improving the quality and yield and of tomato plants. Nurdiawati et al. (2019) determined the role of feather protein hydrolysate in liquid form as a potential biofertilizer which raises the yield and growth of mung bean (Vigna radiata) and patchouli (Pogostemon cablin Benth). Influence of feather hydrolysate produced through the biological method on the cultivation of banana was reported by Gurav and Jadhav (2013). Other studies on Ipomoea aquatica growth also showed a positive influence of composted feather on the yield and growth of plants (Joardar and Rahman 2018). Paul et al. (2014b) reported the impact of feather hydrolysate on the development of microorganisms in the soil and growth of Cicer arietinum. Similarly, Ansari and Sukhraj (2010) have revealed a great impact of vermiwash and vermicompost on plant growth parameters and productivity of okra (Abelmoschus esculentus). Our novel organic combinations with all aspects have shown that VC combined with CFPH can raise the crop growth and yield and use as a biofertilizer in comparison with all previous studies.

Conclusion

The present study was done for the management of feather waste by PKID1 strain and can be efficiently used for plant growth promotion. The results revealed that PKID1 strain can degrade feather waste in the

form of CFPH and can improve the soil quality and production of plants in a single inoculum and with VC combined. CFPH treatment with tomato, rice, onion, chilli and chickpea seeds showed a higher percentage of germination compared to control. Therefore, CFPH and VC enrich micro and macro -nutrients in a new mixed compost that promotes multifunctional plant growth capacity for agricultural and horticultural crops.

Acknowledgement The authors are grateful to Raiganj

University, Raiganj 733 134, Uttar Dinajpur, West Bengal, India

and (DM, DM and SC) Government of West Bengal, India for

Swami Vivekananda Merit Cum Means Ph.D. Scholarship. The

authors wish to thank reviewers and editor for their valuable

suggestions to increase the scientific quality of the manuscript.

Compliance with ethical standards

Conflict of interest The authors declare that there are no

conflicts of interest associated with this study.

Open Access This article is distributed under the terms of

the Creative Commons Attribution 4.0 International License

(http://creativecommons.org/licenses/by/4.0/), which permits

unrestricted use, distribution, and reproduction in any medium,

provided you give appropriate credit to the original author(s) and

the source, provide a link to the Creative Commons license, and

indicate if changes were made.

International Journal of Recycling of Organic Waste in Agriculture (2021)10: 29-42 39

Tabl

e 4

Gro

wth

pro

mot

ion

and

% o

f see

ds g

erm

inat

ion

on d

iffer

ent s

eeds

trea

ted

by C

FPH

Fig. 6 Treatment of rice seeds with CFPH and root development after 20 days; (a) CFPH treated, (b) control

International Journal of Recycling of Organic Waste in Agriculture (2021)10: 29-4240

Tab

le 1

Mor

phol

ogic

al a

nd b

ioch

emic

al sc

reen

ing

of 1

2 is

olat

es

Appendix

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