Cyanide is a strong toxin in many tropical forage plants that can negatively affect ruminants. The aim of this study is to determine the cyanide removal efficiency, silage quality, and in vitro rumen fermentation of fresh cassava roots ensiled without an additive (control) and with Acremonium cellulase (AC), two cyanide-utilizing bacterial inoculants (Enterococcus feacium KKU-BF7 (BF7) and E. gallinarum KKU-BC10 (BC10)), and their combinations (BF7 + BC10, AC + BF7, AC + BC10 and AC + BF7 + BC10). A completely randomized design was used with eight treatments × four small-scale silo replicates. Additionally, extra silage samples (seven silos/treatment for individually opening after 0, 1, 3, 5, 7, 15, and 30 days of ensiling) were added to observe the changes in the total cyanide concentration and pH value. The fresh cassava root contained an optimal number of lactic acid bacteria (10⁵ colony forming units/g fresh matter), and the contents of dry matter (DM) and total cyanides were 30.1% and 1304 mg/kg DM, respectively. After 30 days of ensiling, all silages demonstrated a low pH (<3.95; p < 0.01). Cyanide content ranged from 638 to 790 mg/kg DM and was highest in the control (p < 0.01). The addition of BF7 + BC10 increased the crude protein (CP) content (p < 0.01). The addition of AC decreased the fibrous contents (p < 0.01). The control had less acetic acid and propionic acid contents (p < 0.01) and a greater butyric acid content (p < 0.01). However, the degrees of in vitro DM digestibility (IVDMD) and gas production were similar among treatments. Methane production ranged between 29.2 and 33.3 L/kg IVDMD (p < 0.05), which were observed in the AC + BC10 and BF7 + BC10 treatments, respectively. Overall, our results suggested that the cyanide removal efficiency after 30 days of ensiling with good-quality cassava-root silage was approximately 39% of the initial value. The enterococci inoculants and/or AC could improve the ensiling process and cyanide removal efficiency (increasing it to between 47 and 51% of the initial value). The novel enterococci inoculants (BF7 + BC10) were associated with a decreased cyanide content and an increased CP content. They appeared to promote the methanogenesis potential of the cassava root silage. More research is required to validate the use of cyanide-utilizing bacterial inoculants in cyanogenetic plants, bioenergy fermentation, and livestock.

In this study, for the first time, a systematic review and meta-analysis were conducted to understand the prevalence and genetic diversity of haemosporidian parasites—namely, Plasmodium, Haemoproteus and Leucocytozoon—in avian species in Southeast Asia. Following the PRISMA guidelines, 14,211 studies were retrieved from PubMed, ScienceDirect and Scopus, which contain data relevant to ‘Plasmodium’ or ‘Haemoproteus’ or ‘Leucocytozoon’ and ‘birds’ or ‘chickens’. Of these, 15 articles reporting the prevalence of Plasmodium, Haemoproteus or Leucocytozoon in Southeast Asia were selected for meta-analysis. The pooled prevalence estimates with 95% confidence intervals (CIs) of Plasmodium, Haemoproteus and Leucocytozoon were analyzed using a meta-analysis of their proportions, implemented in R programming. The publication bias was checked using a funnel plot and Egger’s test. Consequently, the pooled prevalences of Plasmodium, Haemoproteus and Leucocytozoon in Southeast Asia were found to be 21% (95% CI: 18–25%), 18% (95% CI: 15–22%) and 34% (95% CI: 30–37%), respectively. The prevalence of Plasmodium in domestic poultry (37.94%) was found significantly higher than in wild birds (6.46%). There was substantial heterogeneity among studies related to Plasmodium (χ² = 171.50, p < 0.0001, I² = 94.84%), Haemoproteus (χ² = 52.20, p < 0.0001, I² = 90.4%) and Leucocytozoon (χ² = 433.90, p < 0.0001, I² = 98.80%). Additionally, this review revealed 23 lineages of Plasmodium, 35 lineages of Haemoproteus and 21 lineages of Leucocytozoon reported from both domestic poultry and wild birds in Southeast Asia. In conclusion, this systematic review suggested that the prevalence of avian haemosporidian parasites in Southeast Asia is high. Particularly, domestic poultry has a high prevalence of Plasmodium, suggesting that monitoring of this parasite should be implemented in the poultry production system. Furthermore, several parasites found in wild birds are undescribed species. Further experimental studies using combined microscopic and molecular techniques might reveal the characteristics of overlooked parasites.

Soft rot disease affects a range of crops in the field and also during transit and storage, resulting in significant yield losses and negative economic impacts. This study evaluated the in vitro antibacterial activities and mode of action of Piper betle extracts against the soft rot disease-causing bacteria, Erwinia caratovora subsp. caratovora (ECC). Dried leaves of P. betle were extracted with water, ethanol, and hexane solvents and evaluated for their antibacterial activity. The results showed the highest antibacterial activity against ECC in the ethanol extract, followed by hexane and water extracts with minimum inhibitory concentration (MIC) 1.562, 6.25, and more than 12.50mg/mL, respectively. The time-kill assay indicated a bactericidal mode of action. ECC growth was destroyed within 6 and 8hours after treatment with the ethanol extract at 4-fold MIC and 2-fold MIC, respectively. The ethanol extract of P. betle showed promising activity against ECC, with the potential for further development as a novel alternative treatment to control phytobacteria.

Leucocytozoon species are common in countries with warm climates but are an often neglected blood parasite in poultry. Although Leucocytozoon macleani is less virulent than Leucocytozoon caulleryi, it can still negatively impact production performance. In Thailand, the available reports indicate a high prevalence of Leucocytozoon spp., but detailed morphological characteristics of the parasites remain insufficiently known. In this study, Giemsa-stained blood smears and extracted genomic (g) DNA were obtained from 60 domestic chickens (Gallus gallus domesticus). Blood smears were examined for the presence of Leucocytozoon species and their morphological characteristics were examined. A total of 60 gDNA samples were used for nested-PCR amplification of the cytochrome b gene of Leucocytozoon species, followed by sequencing and phylogenetic analysis. The microscopic and molecular examinations revealed prevalence of leucocytozoonosis in chickens of 85% and 90%, respectively. Sequence analysis indicated that several infected chickens harboured multiple Leucocytozoon lineages. Leucocytozoon macleani was morphologically identified in nine samples and could be linked to the lineages GALLUS17, GALLUS34, and the new lineages GALLUS63. The found gametocytes of L. macleani morphologically resembled those reported previously, but exhibited some distinct characteristics. Phylogenetically, the lineages of L. macleani isolated in this study grouped separately from some other L. macleani lineages deposited in GenBank. In conclusion, the prevalence of Leucocytozoon infection in chickens from Northeastern Thailand was high, with frequent co-infections by multiple lineages. Leucocytozoon macleani may exhibit cryptic specification. This study is the first report of L. macleani lineages described using MalAvi database nomenclature, alongside their morphological characteristics.

In ruminant nutrition, the lignocellulosic complex is a primary constraint limiting the utilization of dietary fiber. The objective of this study was to evaluate the effects of inoculating lignin-degrading bacteria (LDB) isolated from the ruminant rectum on in vitro rumen fermentation characteristics. Rectal fecal samples were collected from healthy beef cattle, dairy cattle, buffaloes, and goats (n = 4 per species) using the grab sampling technique. Twenty-eight bacterial colonies were isolated through enrichment and screening on media containing sodium lignosulfonate. Lignin degradation efficiency was assessed spectrophotometrically, while laccase activity was determined using a 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) oxidation assay. Seven isolates exhibiting ligninolytic activity (1.4–5.6% degradation efficiency) were selected to evaluate their effects on in vitro rumen fermentation using a completely randomized design with four replicates. LDB treatments were standardized to a concentration of 2.4 × 10⁵ colony-forming units/mL of rumen fluid medium, while the control received an equal volume of a 0.85% sterile NaCl solution. A rice straw-based total mixed ration served as the substrate, with rumen fluid collected from beef cattle. All treatments were incubated for 48 h. Notably, isolate BC3 consistently enhanced in vitro dry matter digestibility (23.1%), total gas production (18.6%), and total volatile fatty acid concentrations (13.2%) relative to the control and other LDB isolates (p < 0.01). All seven LDB isolates were identified as Gram-negative, rod-shaped, facultative anaerobic bacteria that exhibit catalase activity and tolerate moderately acidic conditions. Phylogenetic tree analysis based on 16S rRNA gene sequencing identified isolate BC3 as being closely related to Escherichia coli strains. These findings demonstrate that the ruminant hindgut is a promising source of LDB with the functional potential to enhance feed digestibility and fermentation end-products in the rumen. Future research should prioritize in vivo trials to evaluate the safety and efficacy of LDB as a direct-fed microbial, specifically focusing on its impact on animal performance and health.

Background and Aim: Spotted scat (Scatophagus argus) is a euryhaline teleost of increasing importance in aquaculture, ornamental fisheries, and environmental monitoring across the Indo-Pacific region. Hematological evaluation is a valuable tool for assessing fish health, physiological status, and environmental quality; however, species-specific reference data for spotted scat remain scarce. This study aimed to characterize blood cell morphology and morphometry and establish preliminary hematological reference values for wild-caught spotted scat from Southern Thailand to support veterinary diagnostics, future research, and biomonitoring applications. Materials and Methods: Twenty-six wild-caught spotted scats were collected from Tha Sung Canal, Nakhon Si Thammarat Province, Thailand, between June and August 2025. Fish were anesthetized with clove oil before blood collection from the caudal vein into ethylenediaminetetraacetic acid tubes. Wright-stained blood smears were prepared for microscopic evaluation of blood cell morphology and morphometry. Hemoglobin (Hb) concentration was measured using an automated hematology analyzer, whereas packed cell volume (PCV), erythrocyte and leukocyte counts, differential leukocyte counts, thrombocyte counts, and erythrocyte indices were determined using standard manual hematological methods. Descriptive statistics were generated, and leukocyte morphometry was compared using the Kruskal-Wallis test followed by Dunn’s multiple-comparison test. Results: Four leukocyte types were identified: neutrophils, eosinophils, monocytes, and lymphocytes. Neutrophils and monocytes were the largest leukocytes and exhibited similar diameters (p > 0.05), although monocytes possessed a characteristically deeper blue cytoplasm that facilitated differentiation. Eosinophils occurred as small and large forms, whereas lymphocytes represented the predominant circulating leukocyte population. Juvenile male fish had a mean PCV of 0.24 ± 0.07 L/L, Hb concentration of 74.41 ± 20.94 g/L, erythrocyte count of 1.80 ± 0.68 × 10¹²/L, and leukocyte count of 32.47 ± 14.81 × 10⁹/L. Lymphocytes (16.14 ± 7.27 × 10⁹/L) were the predominant leukocyte subtype, followed by monocytes, neutrophils, and eosinophils. Conclusion: This study provides the first comprehensive description of blood cell morphology together with preliminary hematological reference values for wild-caught spotted scat from Southern Thailand. These baseline data enhance diagnostic capacity for piscine hematology, support health assessment of ornamental and cultured fish, and provide a valuable foundation for future studies investigating environmental stress, disease, and the use of spotted scat as a bioindicator of freshwater, brackish, and mangrove ecosystems within the One Health framework. Keywords: aquaculture, bioindicator, complete blood count, euryhaline fish, hematology, One Health, Scatophagus argus, Southern Thailand.