Muhammad Abdul Basit^1,2, Arifah Abdul Kadir^1*, Loh Teck Chwen³, Annas Salleh⁴, Ubedullah Kaka⁵, Sherifat Banke Idris^1,6, Abdul Asim Farooq⁷, Muhammad Arshad Javid², Saeed Murtaza⁷

¹Department of Preclinical Sciences, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

²Department of Biosciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan 60000, Punjab, Pakistan

³Department of Animal Sciences, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

⁴Department of Veterinary Laboratory Diagnostics, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

⁵Department of Companion Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universiti Putra

Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

⁶Department of Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, Usmanu Danfodiyo

University, 2346 Skoto, Nigeria

⁷Department of Clinical Sciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan 60000, Punjab, Pakistan

Abstract

Plants, their extracts, and plant essential oils are considered prominent sources of new therapeutic substances. Nowadays, medicinal plants like herbs attain the keen interest of consumers and researchers. The present study evaluated Piper betle (P. betle) and Persicaria odorata (P. odorata) leaf extracts for qualitative and quantitative phytochemical screening. The phytochemical analysis of P. betle and P. odorata leaf extracts presented the occurrence of tannins, flavonoids, saponins, phenols, glycosides, and volatile oils. The higher total phenolic content and total tannins were quantified from P. betle methanolic leaf extract. Additionally, it showed increased antioxidant activity compared to P. odorata leaf extracts. The in vitro antibacterial potential of both herbs was estimated against Salmonella enterica, Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Candida albicans, and Aspergillus brasiliensis. The methanolic leaf extract of Piper betle showed antibacterial and antifungal activity against these selected strains.

Keywords: Piper betle, Persicaria odorata, Phytochemical analysis, Antibacterial activity, Antioxidant activity

Amina Rani¹, Gulnaz Afzal¹, Yasir Mahmood¹, Sana Alam¹, Zahid Iqbal², Mahboob Akbar³, Ahsan Nadeem Malik³, Muhammad Rizwan¹, Rabia Akram⁴, Mushtaq Ahmad³, Riaz Hussain^5*

¹Department of Zoology, Islamia University of Bahawalpur-63100, Pakistan

²Department of Pharmacology and Toxicology, Faculty of Veterinary and Animal Sciences, Islamia University of Bahawalpur-63100, Pakistan

³Livestock and Dairy Development Department (Poultry Production), Punjab, Pakistan

⁴Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University, Multan, Pakistan

⁵Department of Pathology, Faculty of Veterinary and Animal Sciences, Islamia University of Bahawalpur-63100, Pakistan

Abstract

Ethoxysulfuron is frequently used as a post-emergent herbicide in different food crops to control broad leaf weeds. This study aimed to investigate the potential adverse effects of ethoxysulfuron herbicide on male Japanese quail. 48 adult male birds were divided into 4 groups and orally administered with varying doses of the herbicide for 45 days. Quail were exposed to four sub-lethal concentrations (0 mg/kg, 0.70 mg/kg/day, 0.90 mg/kg/day, and 1.25 mg/kg/day) of ethoxysulfuron starting from day 1 to 45 of trial and different samples from each quail were obtained at days 15, 30 and 45 of trial following exposure. Results exhibited that with increasing ethoxysulfuron concentrations different nuclear abnormalities in erythrocytes such as the formation of micronuclei, binucleated erythrocyte, notched nuclei, blabbed nuclei, lobbed nuclei, condensed nuclei, nuclear remnants and the erythrocytes without nuclei increased in treated birds. Results showed that exposure to herbicide led to genotoxicity, as evidenced by increased values of DNA damage in different visceral organs of the exposed quail compared to the control group. Various antioxidant enzymes and oxidative stress biomarkers were significantly reduced and escalated in the liver tissues of quail. Histological examination revealed prominent histoarchitectural changes in the liver (degeneration of hepatocytes and necrosis) and kidneys (degeneration of renal tubules, necrosis of renal tubules, and necrosis of renal tubular epithelial cells) of the exposed quail. In conclusion, this study demonstrates that exposure to ethoxysulfuron herbicide can cause significant morphological and nuclear abnormalities in erythrocytes, DNA damage, and histoarchitectural changes in different visceral organs of Japanese quail. These findings suggest that exposure to herbicides may pose a health risk to both humans and wildlife.

Keywords: Ethoxysulfuron, Japanese quail, Oxidative stress, DNA damage, Histopathology

Phan Nha Hoa^1*, Ngo Duc Hoa¹, Tran Kim Diep¹, Pham Van Huyen¹, Truong Binh Nguyen²

¹Taynguyen Institute for Scientific Research, Vietnam Academy of Science and Technology, 116 Xo Viet Nghe Tinh, Da Lat, Vietnam

²Da Lat University, 01 Phu Đong Thien Vuong, Da Lat, Vietnam

Abstract

Isaria tenuipes (Peck.) Samson is an entomopathogenic fungus that has potential in pharmacology and biocontrol. The study aimed to determine the effects of temperature, pH, and media on mycelium growth of Isaria tenuipes (Peck.) Samson (DL0099) from Lang Biang Mountain, Lam Vien Plateau, Vietnam. In this study, we used the single-factor method to culture DL0099 mycelium on both surface liquid and agar media. The results indicated that the optimal temperature for the mycelium growth was 20~25 ^oC. This mushroom mycelium can tolerate temperatures at 35 ^oC for 8 days. The initial media pH range of 6~9 was found to be the most favorable to mycelia growth. Sabouraud’s agar (SA) and Sabouraud’s dextrose agar plus yeast (SDYA) media were the optimal agar media for the mycelium growth rate. Sabouraud broth (S) medium was the most suitable medium for mycelia biomass production in surface liquid culture. Yeast malt agar (YMA), malt agar (MEA), maltose agar (MA), and malt extract yeast agar (MYA) media were suitable for fruit body formation. Our results provided optimal temperature, pH, and media conditions for Isaria tenuipes (DL0099) mycelium growth on surface liquid and agar media. They lay the basis for effective propagation, biomass production, and fruit body formation of the Vietnam native Isaria tenuipes (Peck.) Samson.

Keywords: Entomopathogenic fungi, Isaria tenuipes Peck. (DL0099), Lang Biang Mountain, Mycelium growth

Johnmel A. Fabros¹, Rich Milton R. Dulay^1,2*, Kathlene Claire O. Ganareal¹, Sofronio P. Kalaw^1,2, Mark Adrian G. del Rosario¹, Renato G. Reyes^1,2

¹Center for Tropical Mushroom Research and Development, Tuklas Lunas Development Center, Central Luzon State University, Science City of Muñoz, Nueva Ecija, Philippines

²Department of Biological Sciences, College of Science, Central Luzon State University, Science City of Muñoz, Nueva Ecija, Philippines

Abstract

Agaricomycetous mushrooms are widely utilized as a source of food and or traditional medicine in the world as they exhibit both nutritional and pharmaceutical properties. In order to establish the optimal culture conditions of the 2 Coprinopsis, 2 Leucoagaricus, and 1 Leucocoprinus mushroom, we evaluated the optimum culture medium, pH condition, aeration, illumination, and temperature requirements for the luxuriant mycelial growth of these wild mushrooms. The fruiting body and mycelial biomass production were also carried out in this study to measure mushrooms’ biological efficiency. Among commercially-available culture media, malt extract agar (MEA) was found to be the most suitable for the 4 mushrooms (except C. cinerea), followed by potato dextrose agar (PDA) for 3 mushrooms (C. cinerea, C. verticillata, L. cretaceous). Potato sucrose gulaman (local crude agar) or PSG and corn grit decoction gulaman or CGDG were also found favorable for the growth of Coprinopsis verticilatta and Leucoagaricus americanus, respectively. In terms of pH requirement, L. americanus (pH 5) and Leucoagaricus meleagris (pH 5-6) favored slightly acidic, while the other three mushrooms showed a wide range of pH requirements. C. verticillata and L. meleagris favored sealed condition, while Coprinopsis cinerea favored unsealed condition. However, L. americanus and Leucocoprinus cretaceous showed efficient mycelia growth in both sealed and unsealed conditions. Dark condition were found to be favorable for mycelial growth of both C. cinerea and L. cretaceous, whereas lighted condition was found appropriate for L. americanus mycelia. However, illumination was found to be not important factor for C. verticillata and L. meleagris. All evaluated mushrooms grew best at room temperature conditions (30-32ºC), but C. cinerea, L. meleagris, and L. cretaceous could also thrive at lower temperatures (23-25ºC). Three mushrooms namely, L. amaericanus, L. meleagris, and L. cretaceous successfully produced fruiting bodies in fruiting bags containing rice straw and sawdust at a 7:3 ratio by volume with biological efficiencies (BE) of 5.75%, 5.75%, and 2.3%, respectively. However, the mycelia of C. cinerea and C. verticillata were mass-produced in potato broth in submerged cultivation with 18.18% and 23.86% BE, respectively. Generally, the optimum culture condition for both mycelial growth and fruiting body production were found to be species-dependent.

Keywords: Biological efficiency, Coprinospsissp., Leucoagaricus, Leucocoprinus, Mycelial biomass, Optimization study

Jian Wang¹, Yong Pan¹, Liping Liu¹, Chuang Wu¹, Youzhi Shi^1*, Xiaolong Yuan^2*

¹China Tobacco Hubei Industry Co., Ltd., Wuhan 443100, China

²Tobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao 266100, China

Abstract

Cigar filler leaves are the most important component of cigar because they determine its quality. Therefore, the volatile components of eight cigar filler tobacco leaves were studied and compared using gas chromatography–ion mobility spectrometry (GC–IMS). In this study, 84 compounds with high levels of nitrogenous and ketone compounds were identified. Based on the chemometric principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), the eight cigar samples were significantly distinguished. Meanwhile, we performed a discriminant analysis of volatile organic compounds in the eight cigar samples based on the variable importance in the projection (VIP) scores of the PLS-DA model, and revealed significant differences in the volatile compounds between the different varieties. 11 volatile compounds (VIP > 1) were screened and compared, among which triamine, acetic acid, acetone, and cyclopentanone were the main differential compounds/flavor substances. This study showed that GC–IMS can rapidly identify and compare the volatile compounds of various cigars, providing a theoretical basis for studying the differences in the volatile aroma of cigars, and laying a foundation for the breeding selection of subsequent varieties.

Keywords: Cigar filler tobacco leaves, Volatile flavor compounds; Gas chromatography–ion mobility spectrometry, Principal component analysis, Partial least squares-discriminant analysis

Toan Le Thanh^1,2*, Nguyen Huy Hoang², Kanjana Thumanu³, Channon Saengchan², Jayasimha Rayalu Daddam⁴, Rungthip Sangpueak², Narendra Kumar Papathoti², Kumrai Buensanteai²

¹Crop Protection Department, College of Agriculture, Can Tho University, Can Tho city, 94000, Vietnam

²School of Crop Production Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

³Synchrotron Light Research Institute, Nakhon Ratchasima, 30000, Thailand

⁴Department of Animal Science, Agriculture Research Organization, Volcani Center, Rishon Lezion 7505101, Israel

Abstract

Bacterial leaf blight (BLB) of rice has a high epidemic potential and usually causes severe damage. This research was conducted to assess the efficacy and characterize the mechanism of the systemic resistance of rice plants induced by the Bacillus subtilis strain CaSUT007 to BLB. The results revealed 30% reduction in the severity of BLB in the treated rice plants, and real-time PCR measurements indicated a significant 1.1–1.2-fold increase in their concentrations of the defense genes of phenylalanine ammonia-lyase (PAL) and ascorbate peroxidase (APX). In addition, Fourier transformed infrared spectroscopy characterization of the biochemical changes in the rice leaves indicated alterations to the lignins, pectins, and amide I vibrations – these lead to the generation of defense barriers and the reinforcement of cell walls against Xanthomonas infection and invasion, thereby contributing to disease reduction. Phylogenetic trees of pal and apx revealed a significant number of polytomies among these two gene families. Moreover, analysis of the active sites of the protein PAL and APX showed one serine rotamer and a single mutation-sensitive glutamic acid residue in the region of the binding site/pocket. The possible interactions of PAL and APX with other proteins revealed insight into the defense mechanism: APX6 interacts directly with MDAR5, MDRA3, DHAR1, and other important defense proteins, while PAL has direct interactions with 4CL4, 4CLL9, and 4CL3, among other defense proteins. Therefore, treatment with the B. subtilis strain CaSUT007 promoted faster, stronger and more intense responses in rice plants against BLB.

Keywords: Bacillus subtilis, Defense genes, Leaf blight, Protein interaction

Amanda Putri Irawan¹, Amalia Rahmawati¹, Ulfa Abdila Fahmi¹, Arief Budiman², Khusnul Qonita Maghfiroh¹, Tia Erfianti¹, Dea Putri Andeska¹, Renata Adaranyssa Egistha Putri¹, Istini Nurafifah¹, Brilian Ryan Sadewo², Eko Agus Suyono^1*

¹Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia. Jl. Teknika Sel., Sendowo, Sinduadi, Kec. Mlati, Kabupaten Sleman, Daerah Istimewa Yogyakarta 55281, Indonesia

²Department of Chemical Engineering, Faculty of Engineering, Universitas Gadjah Mada, Indonesia. Sendowo, Sinduadi, Kec. Mlati, Kabupaten Sleman, Daerah Istimewa Yogyakarta 55284, Indonesia

Abstract

Microalgae harvesting is critical to remove water from algal growth media with solid-liquid separation. Bioflocculation has the same principle as flocculation. Using solid-liquid separation, microalgae harvesting removes moisture from the algal growth substrate. The same idea underlies flocculation and bioflocculation. Using fungal and bacterial bioflocculants requires a special medium that is different from the microalgae medium, that fungi and bacteria can contaminate microalgae, so it is not recommended to be used as a bioflocculant agent. Microalgae Anabaena sp. was chosen in this study as a bioflocculant agent since it can produce exopolysaccharides (EPSs). Dissolved proteins and carbohydrates make up EPSs. This investigation looked into employing Anabaena species to extract Chlorella species. The harvest day was used to measure the parameters. A spectrophotometer was used to measure the precipitation percentages. Bligh and Dyer’s methods were used to measure lipid contents. The phenol-sulfate was used to perform carbohydrates. Bradford method was used to quantify proteins. The ratio of 1:1.25 was determined to have the best proportion of flocculation and carbohydrate content (Chlorella sp. : Anabaena sp.). The ratio of 1:1 was determined to have the maximum cell lipid and protein content (Chlorella sp.: Anabaena sp.). The application of this study will be beneficial to design effective methods for harvesting microalgae using biological materials such as other microalgae.

Keywords: Bioflocculation, Anabaena sp., Chlorella sp., Exopolysaccharides

Leilidyn Y. Zurbano^1*, Chester C. Deocaris², Carmelita P. Mapanao³, Arnel O. Rendon³, Lourdes V. Alvarez³

¹Agriculture Department, Polytechnic University of the Philippines, Lopez, Quezon Branch, Quezon Province, Philippines

²Department of Physical Science, College of Science, Polytechnic University of the Philippines, Sta. Mesa, Manila, Philippines

³Department of Biology, College of Science, Polytechnic University of the Philippines, Sta. Mesa, Manila, Philippines

Abstract

Plant-parasitic nematodes are serious pests causing important crop losses worldwide. Hence, this study was conducted to determine the occurrence of plant-parasitic nematodes (PPNs) in healthy and unhealthy pineapple crops. To determine if the infestation is prevalent, a soil metabarcoding analysis was done. Six soil samples were obtained from the rhizospheres of Red Spanish pineapple farm in Mabitac, Laguna, Philippines. They were freeze-dried and brought to the laboratory for metagenomics analysis using Primers NemF and 18Sr2b. The results showed uncultured Eukaryotes (43.1%) nematodes (31.2%), Platyhelminthes (6.9%), Apicomplexa (6.5%), Annelida (5.0%), Rotifera (3.6%), Arthropoda (1.8%), Ascomycota (1%), unidentified (0.4%) and Basillariophyta, Cercozoa, Chytridiomycota, Mollusca and Mucoromycota with 0.1%. In total, 374,410 sequence reads were obtained and were clustered into 117 OTUs at 97% similarity. In assessing the nematode community structure, it yielded 26,565 nematodes; 5,315 nematodes were obtained from the rhizosphere of healthy samples, and 21,250 were from the unhealthy ones. The fungivore Apelenchus avenae (Bastian 1865) was the most prevalent (47.8% and 43.9%) in both locations. Other nematodes found on the plant rhizospheres were Rhabdolaimus aquaticus (de Man), Acrobeloides varius, Rotylenchulus reniformis (Linford and Oliveira, 1940), Aphelenchoidinae sp., Panagrolaimoidea sp., and two unidentified nematodes. The Basiria sp., Alaimus sp., and two other unidentified nematodes were the only ones found in the rhizosphere of unhealthy pineapple plants. Meanwhile, Mesocriconema onoense, Aphelenchoides sp., Ditylenchus gilanicus, and an unidentified nematode were found in the rhizosphere of healthy pineapple plants. Phylogenetic analyses of the nematode communities show that only Orders Rhabditida and Enoplida are associated with the crop and the Family Aphelenchoididae is distant from Rhabditida, thus, dividing the clade into three. The first clade consists of diverse nematode communities of fungivores, bacterivores, predators, and plant and insect parasites. The second clade consists of bacterivore nematodes found in moist environments, and the third clade comprises mainly Aphelenchoididae species, mostly plant parasites.

Keywords: Metabarcoding, Acrobeloides varius, Aphelenchus avenae, Ditylenchus gilanicus, Mesocriconema onoense, Rhabdolaimus aquaticus, Rotylenchulus reniformis

Otie Victoria^1,2, Udo Idorenyin³, Matsuura Asana⁴, Liu Jia², Liang Shuoshuo², Shao Yang⁵, Itam Michael Okoi⁶, An Ping^2*, Eneji Anthony Egrinya¹

1Department of Soil Science, Faculty of Agriculture, Forestry and Wildlife Resources Management, University of Calabar, P.M.B. 1115, Calabar, Nigeria

2Arid Land Research Centre, Tottori University, Hamasaka 680-0001, Tottori, Japan

3Department of Crop Science, Faculty of Agriculture, Forestry and Wildlife Resources Management, University of Calabar, P.M.B. 1115, Calabar, Nigeria

4Faculty of Agriculture, Shinshu University, 8304, Kamiina County, Nagano 399-4598, Japan

5College of Plant Science & Technology, Huazhong Agricultural University, Wuhan 430070, China

6Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing 48824, MI, USA

Abstract

Salt stress is a key ecological challenge to wheat establishment at the early stage of germination, especially in drylands. A germination experiment was conducted to determine whether an exogenous seed treatment with 24-epibrassinolide could mitigate salinity stress effects on wheat germination. Seeds of the Sudanese wheat cv. Imam were treated with 24-epibrassinolide (BR1) or without (BR0) at eleven concentrations of sodium chloride (NaCl) (0.00, 1.56, 3.13, 4.69, 6.25, 7.81, 9.38, 10.94, 12.50, 14.06 and 15.63 dSm-1), in a 2 x 11 factorial experiment arranged into a completely randomized design. Seed germination was progressively delayed with increasing salinity and the daily germination was reduced significantly. The germination average time and relative injury rate increased considerably (p≤0.05) at salt levels of 7.81 dSm-1or more. The inhibitory effects of salinity on germination were significantly (p≤0.05) reversed by seed treatment with BR1. Wheat cv. Imam tolerated salt stress up to 6.25 dSm-1 at BR0 with respect to velocity of germination, germination rate, final germination rate, germination percentage and germination index, relative to no salt (0.00 dSm-1). An early uniform establishment of wheat in saline media could be enhanced by seed treatment with BR.

Keywords: Arid region, Plant growth hormone, Triticum aestivum (L), Abiotic stress

Lily Ishaq*, Agnes V Simamora, Peters O. Bako, Yoke I Benggu, Moresi M. Airthur, Effy Roefaida, Ellias St O Nguru

Department of Agrotechnology, Faculty of Agriculture, Nusa Cendana University. Jl. Adi Sucipto Penfui Kupang, East Nusa Tenggara, Indonesia

Abstract

Citrus is an important commercial crop in Timor Tengah Selatan, East Nusa Tenggara, Indonesia, but many trees are suffering a decline in health. As citrus is heavily dependent on arbuscular mycorrhizal fungi (AMF), this study investigated the abundance of AMF in the rhizosphere of healthy and declining trees in citrus orchards at 12 geographical locations. In each orchard, 6 soil and 6 root samples representing 3 healthy and 3 declining trees were collected. The soil was analyzed for AMF spore abundance as well as physical (texture) and chemical properties (organic carbon, nitrogen, phosphorus, potassium, pH, and soil exchangeable capacity), while the fine roots of citrus were assessed for colonization. The data were analyzed using analysis of variance (ANOVA) where the health condition of the trees was under the geographical location/site factor. The results showed that the abundance of AMF spores was significantly affected by the geographical location from where the soils were collected, but the health condition of the trees had no effect. However, AMF colonization was significantly affected by both site and tree health. The number of AMF morphotypes tended to be higher under healthy trees than under declining trees. Soil analysis indicated that soil fertility (N and organic C) may be important for tree health. These results provided a new perspective on the possible involvement of AMF and soil nutrients in citrus decline. Further studies are required to define the interactions between AMF, soil fertility, and disease incidence to identify strategies for managing citrus decline in the region.

Keywords: Arbuscular mycorrhizal fungi, Citrus decline, Orchard management, Orchard location, Southeast Asia