Fusarium oxysporum f. sp. niveum (Fon) is the causative agent of Fusarium wilt disease of watermelon; it is the most serious soil-borne pathogen around the globe. The yield loss is around 30–80% or even more, and is presently a major hindrance to watermelon cultivation worldwide. Initially, the infected watermelon plant shows symptoms like loss of turgor pressure of the leaves and vines that can be recovered at night. The progress of the disease in contaminated transplants turns into dull green to yellow and finally necrotic. When the fungus continues to colonize the xylem vessel, it usually forms more tyloses, finally limiting water movement and causing wilt. The correct identification of the pathogen is necessary for proper disease control. As such, the selection of a molecular marker could serve as an effective means of screening the pathogen. Additionally, different methods have also been reported for the identification of Fon. Therefore, this review focused on the comprehensive description of the biology, diversity, detection, aggressiveness, mycotoxin production, and eco-friendly management strategies of the Fusarium wilt disease of watermelon.

Methicillin-resistant Staphylococcus aureus (MRSA) biofilm producers represent an important etiological agent of many chronic human infections. Antibiotics and host immune responses are largely ineffective against bacteria within biofilms. Alternative actions and novel antimicrobials should be considered. In this context, the use of phages to destroy MRSA biofilms presents an innovative alternative mechanism. Twenty-five MRSA biofilm producers were used as substrates to isolate MRSA-specific phages. Despite the difficulties in obtaining an isolate of this phage, two phages (UPMK_1 and UPMK_2) were isolated. Both phages varied in their ability to produce halos around their plaques, host infectivity, one-step growth curves, and electron microscopy features. Furthermore, both phages demonstrated antagonistic infectivity on planktonic cultures. This was validated in an in vitro static biofilm assay (in microtiter-plates), followed by the visualization of the biofilm architecture in situ via confocal laser scanning microscopy before and after phage infection, and further supported by phages genome analysis. The UPMK_1 genome comprised 152,788 bp coding for 155 putative open reading frames (ORFs), and its genome characteristics were between the Myoviridae and Siphoviridae family, though the morphological features confined it more to the Siphoviridae family. The UPMK_2 has 40,955 bp with 62 putative ORFs; morphologically, it presented the features of the Podoviridae though its genome did not show similarity with any of the S. aureus in the Podoviridae family. Both phages possess lytic enzymes that were associated with a high ability to degrade biofilms as shown in the microtiter plate and CLSM analyses. The present work addressed the possibility of using phages as potential biocontrol agents for biofilm-producing MRSA.

Phages, which are often used therapeutically, have begun to receive interest as alternatives to antibiotic growth promoters (AGPs) for enhancing chicken growth. Another option that has been extensively studied as a growth promoter in chickens is probiotics. To the best of our knowledge, there is currently no study available on the use of phages and probiotics in combination as potential feed additives for broiler chickens. Therefore, this study demonstrated the effects of a phage cocktail, probiotics, and their combination on the growth performance and gut microbiota of broiler chickens. A total of 288 one-day-old male Cobb 500 broilers were randomly allotted to one of six treatments in a completely randomised design. The treatments were (i) C (basal diet (BD) only), (ii) 1ϕ (BD + 0.1% phage cocktail), (iii) 2ϕ (BD + 0.2% phage cocktail), (iv) P (BD + 0.1% probiotic), (v) 1ϕP (BD + 0.1% phage cocktail + 0.1% probiotic), and (vi) 2ϕP (BD + 0.2% phage cocktail + 0.1% probiotic). The 1ϕP treatment had significantly (p < 0.05) better BW (35 days), BWG (22–35 days, 1–35 days), and FCR (1–21 days, 22–35 days, 1–35 days) compared to C. Unique gut microbiota diversity was also found between the ϕP (1ϕP and 2ϕP) and non-ϕP groups (C, 1ϕ, 2ϕ, and P) in ilea, particularly in the 35-day-old chickens. Microorganisms associated with short-chain fatty acid (SCFA) producers were significantly (p < 0.05) more present in the ϕP group than in the non-ϕP group. The predicted genes related to carbohydrate and amino acid metabolism were significantly upregulated in ϕP groups compared to non-ϕP groups. These genes were involved in the digestion and absorption of nutrients, as well as the production of energy. Our findings showed that the 1ϕP treatment could be a potential alternative to AGPs for poultry, as growth performance was enhanced, and gut microbiota was positively modulated.

Fusarium wilt disease incited by Fusarium oxysporum f. sp. niveum (FON) is the utmost devastating soil-inhabiting fungal pathogen limiting watermelon (Citrullus lanatus) production in Malaysia and globally. The field disease survey of fusarium wilt was carried out during December 2019 and November 2020, in three major production areas (3 farmer fields per location) in Peninsular Malaysia namely, Mersing, Serdang and Kuantan and disease incidence of 30 and 45%, was recorded for each year, respectively. Infected watermelon plants showed symptoms such as vascular discoloration, brown necrotic lesions to the soil line or the crown, one-sided wilt of a plant, or a runner or the whole plant. Infected root and stem tissues, 1-2 cm pieces were surface sterilized with 0.6% NaOCl for 1 minute followed by double washing with sterile water. The disinfected tissues were air-dried and transferred onto semi-selective Komada’s medium (Komada 1975) and incubated for 5 days. The fungal colonies produced were placed on potato dextrose agar (PDA) to attain a pure culture and incubated at 25±2℃ for 15 days. The pure fungal colony was flat, round and light purple in color. Macroconidia were straight to slightly curved, 18.56-42.22 µm in length, 2.69-4.08 µm width, predominantly 3 septate and formed in sporodochia. Microconidia measured 6.16-10.86 µm in length and 2.49-3.83 µm in width, kidney-shaped, aseptate and were formed on short monophialides in false-heads. Chlamydospores were single or in pairs with smooth or rough walls, found both terminally or intercalary. To confirm their pathogenicity, two-week-old watermelon seedlings (cv. NEW BEAUTY) were dipped into spore suspension (1 ˟ 106 spores/ml) of representative isolates of JO20 (Mersing), UPM4 (Serdang) and KU41 (Kuantan) for 30 second and then moved into 10 cm diameter plastic pots containing 300 g sterilized soil mix. Disease symptoms were assessed weekly for one month. Control seedlings were immersed in sterile distilled water before transplanting. The inoculated seedlings showed typical Fusarium wilt symptoms like yellowing, stunted growth, and wilting, which is similar to the farmer field infected plants. However, the seedlings inoculated by sterile distilled water remained asymptomatic. The pathogen was successfully re-isolated from the infected seedlings onto Komada’s medium, fulfilling the Koch’s postulate. For the PCR amplification, primers EF-1 and EF-2 were used to amplify the tef1-α region. A Blastn analysis of the tef1-α sequences of the isolates JO20 (accession nos. MW315902), UPM4 (MW839560) and KU41 (MW839562) showed 100% similarity; with e-value of zero, to the reference sequences of F. oxysporum isolate FJAT-31690 (MN507110) and F. oxysporum f. sp. niveum isolate FON2 790-2 (MN057702). In Fusarium MLST database, isolates JO20, UPM4 and KU41 revealed 100% identity with the reference isolate of NRRL 22518 (accession no. FJ985265). Though isolate FJ985265 belongs to the f. sp. melonis, earlier findings had revealed Fusarium oxysporum f. sp. are naturally polyphyletic and making clusters with diverse groups of the Fusarium oxysporum species complex (O’Donnell et al. 2015). The isolates JO20, UPM4 and KU41 were identified as F. oxysporum f. sp. niveum based on the aligned sequences of tef1-α and molecular phylogenetic exploration by the maximum likelihood method. To the best of our knowledge, this is the first report of F. oxysporum f. sp. niveum as a causative pathogen of Fusarium wilt disease of watermelon in Malaysia. Malaysia enables to export watermelon all-year-round in different countries like Singapore, Hong-Kong, The United Arab Emirates (UAE), and Netherlands. The outburst of this destructive soil-borne fungal pathogen could cause hindrance to watermelon cultivation in Malaysia. Thus, growers need to choice multiple management tactics such as resistant varieties, cultural practices (soil amendments and solarization), grafting, cover crops and fungicide application to control this new pathogen.

Klebsiella pneumoniae are opportunistic bacteria found in the gut. In recent years they have been associated with nosocomial infections. The increased incidence of multiple drug-resistant K. pneumoniae makes it necessary to find new alternatives to treat the disease. In this study, phage UPM2146 was isolated from a polluted lake which can lyse its host K. pneumoniae ATCC BAA-2146. Observation from TEM shows that UPM2146 belongs to Caudoviriales (Order) based on morphological appearance. Whole genome analysis of UPM2146 showed that its genome comprises 160,795 bp encoding for 214 putative open reading frames (ORFs). Phylogenetic analysis revealed that the phage belongs to Ackermannviridae (Family) under the Caudoviriales. UPM2146 produces clear plaques with high titers of 10¹⁰ PFU/ml. The phage has an adsorption period of 4 min, latent period of 20 min, rise period of 5 min, and releases approximately 20 PFU/ bacteria at Multiplicity of Infection (MOI) of 0.001. UPM2146 has a narrow host-range and can lyse 5 out of 22 K. pneumoniae isolates (22.72%) based on spot test and efficiency of plating (EOP). The zebrafish larvae model was used to test the efficacy of UPM2146 in lysing its host. Based on colony forming unit counts, UPM2146 was able to completely lyse its host at 10 hours onwards. Moreover, we show that the phage is safe to be used in the treatment against K. pneumoniae infections in the zebrafish model.

Co-composting is an effective approach to biowaste management. The co-composting potential of banana pseudostem (BPS) and mushroom media waste (MMW) with chicken manure (CM) has not been explored, let alone their suitable ratios of co-composting being determined. Meanwhile, the imbalance ratios of the feedstocks used in the process severely restrict the physicochemical properties and quality of the finished product. For this reason, six different ratios of BPS, MMW, and CM, viz. 1:1:1, 1:2:1, 1:3:1, 2:1:1, 2:2:1, and 2:3:1, respectively (T₁–T₆), were composted together in aerobic conditions to identify the suitable ratio by evaluating the changes in the physicochemical properties in the composting process. According to the ratio of treatments, the feedstocks were mixed on fresh weight basis. The turning process of co-composting piles was repeated at seven-day intervals to maintain the uniform aeration throughout the composting period. The piles having BPS, MMW, and CM at ratios of 1:2:1, 1:3:1, and 2:3:1, respectively, demonstrated a longer thermophilic phase, indicating more complete decomposition and earlier maturity compared to piles with higher amount of BPS. Of the ratios, BPS:MMW:CM at 1:2:1 ratio (T₂) resulted in the highest total nitrogen (1.53%), lowest C:N ratio (12.4), organic matter loss (54.5%), and increased CEC (41.3 cmol/kg). The highest germination index (129%) was also recorded in the T₂ compost, indicating that it was toxic-free and safe for seed germination. The nutrient-rich compost with high alkaline pH (≥10) can effectively ameliorate soils of an acidic nature, for example, the acidity of Ultisols and Oxisols.