Biocontrol effect of entomopathogenic fungi Metarhizium anisopliae ethyl acetate-derived chemical molecules: An eco-friendly anti-malarial drug and insecticide.

Insect pests represent a major threat to human health and agricultural production. With a current over-dependence on chemical insecticides in the control of insect pests, leading to increased chemical resistance in target organisms, as well as side effects on nontarget organisms, the wider environment, and human health, finding alternative solutions is paramount. The employment of entomopathogenic fungi is one such potential avenue in the pursuit of greener, more target-specific methods of insect pest control. To this end, the present study tested the chemical constituents of Metarhizium anisopliae fungi against the unicellular protozoan malaria parasite Plasmodium falciparum, the insect pests Anopheles stephensi Listen, Spodoptera litura Fabricius, and Tenebrio molitor Linnaeus, as well as the nontarget bioindicator species, Eudrilus eugeniae Kinberg. Fungal crude chemical molecules caused a noticeable anti-plasmodial effect against P. falciparum, with IC50 and IC90 values of 11.53 and 7.65 µg/mL, respectively. The crude chemical molecules caused significant larvicidal activity against insect pests, with LC50 and LC90 values of 49.228-71.846 µg/mL in A. stephensi, 32.542-76.510 µg/mL in S. litura, and 38.503-88.826 µg/mL in T. molitor at 24 h posttreatment. Based on the results of the nontarget bioassay, it was revealed that the fungal-derived crude extract exhibited no histopathological sublethal effects on the earthworm E. eugeniae. LC-MS analysis of M. anisopliae-derived crude metabolites revealed the presence of 10 chemical constituents. Of these chemicals, three major chemical constituents, namely, camphor (15.91%), caprolactam (13.27%), and monobutyl phthalate (19.65%), were highlighted for potential insecticidal and anti-malarial activity. The entomopathogenic fungal-derived crude extracts thus represent promising tools in the control of insect pests and malarial parasites.

Production of single-chain fragment variable (scFv) antibodies specific to plasma membrane epitopes on bull Y-bearing sperm.

Distinguishing between bull Y- and X-bearing sperm populations is advantageous for techniques using sexed bull semen. The aim of this study was to produce a single-chain fragment variable (scFv) antibody against plasma membrane epitopes on bull Y-bearing sperm. Variable heavy (VH)- and variable light (VL)-region genes generated from a hybridoma cell secreting a specific Y-bearing sperm monoclonal antibody (mAb-1F9) were cloned and expressed. The expected sizes of the DNA bands were ∼350 bp for the VH gene and ∼318 bp for the VL gene. The VH and VL genes were generated and used to construct an scFv gene (∼650 bp), which was expressed in E.coli TG1 cells and produced the corresponding soluble scFv antibody. Compared with the parent mAb-1F9, the scFv antibodies presented a high affinity for Y-bearing sperm and low cross-reactivity with X-bearing sperm. An immunofluorescence analysis confirmed that the scFv antibodies and mAb-1F9 recognize epitopes on the Y-bearing sperm surface. The fluorescence signal was strong on the plasma membrane of Y-bearing sperm but very weak for X-bearing sperm. This study aids the application and production of engineered scFv antibodies specific to Y-bearing sperm to distinguish between Y- and X-bearing sperm populations for techniques involving sexed bull semen.

In vitro screening antiviral activity of Thai medicinal plants against porcine reproductive and respiratory syndrome virus.

BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) results in economic losses in the swine industry globally. Several studies have investigated the use of plant extracts in the prevention and control of PRRS outbreaks. Thai medicinal plants may be useful for treating PRRSV infection in pigs. Therefore, we investigated the in vitro anti-PRRSV and antioxidant properties of seven Thai medicinal plants: Caesalpinia sappan Linn., Garcinia mangostana Linn., Houttuynia cordata, Perilla frutescens, Clinacanthus nutans, Phyllanthus emblica, and Tiliacora triandra. RESULTS: Using antiviral screening, we observed that T. triandra extract strongly inhibited PRRSV infectivity in MARC-145 cells [virus titer 3.5 median tissue culture infective dose (TCID50)/ml (log10)] at 24 h post-infection, whereas C. sappan extract strongly inhibited PRRSV replication [virus titer 2.5 TCID50/ml (log10)] at 72 h post-infection. C. sappan extract had the highest total phenolic content [220.52 mM gallic acid equivalent/g] and lowest half-maximal inhibitory concentration [1.17 mg/ml in 2,2-diphenyl-1-picrylhydrazyl and 2.58 mg/ml in 2,2-azino-bis (3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt]. CONCLUSION: T. triandra extract could inhibit PRRSV infectivity, whereas C. sappan extract was the most effective in inhibiting PRRSV replication in MARC-145 cells. This study elucidates the antiviral activities of Thai medicinal plant extracts in vivo. The results promise that Thai medicinal plant extracts, particularly T. triandra and C. sappan extracts, can be developed into pharmaceutical drugs for the prevention of PRRS in pigs.

Molecular characterization of pathogenesis involving the GAS 1 gene from Entomopathogenic fungus Lecanicillium lecanii and its virulence against the insect host Diaphorina citri.

The entmopathogenic fungus Lecaniicillium lecanii is a naturally available biological control and it is considered to be one of the best mycoinsecticide agents against the destructive insect pest Diaphorina citri Kuwayama. The present study aimed to extract and characterize the toxic insecticidal protein from L. lecanii and to assess the toxicity level against the Asian citrus psyllid the vector of Huanglongbing disease (HLB), also called citrus greening. Extracts of a toxic substance from submerged batch culture examined by sodium dodecyl sulfate-poly-acrylamide (SDS-PAGE), had a molecular weight of 45 kDa. The most abundant toxic metabolite was subjected to HPLC to purify and identified it by mass spectrometry. Subsequently, metabolite toxicity was tested against D. citri at three different concentrations (1%, 2%, and 3%). The results showed that the highest concentration had a significant maximum mortality at 120 h post application. Furthermore, we investigated the expression of the GAS1 gene which was previously identified to have a role in pathogenicity in in vivo studies in adult insect psyllids. Results of this study indicated that expression of the virulence factor gene was present at three concentrations of the fungal suspension post inoculation. This is the first study to provide this novel approach for the characterization of fungal mediated synthesis of a cuticle degrading soluble protein against the insect D. citri. The present results provide strong information on the in vivo expression of the GAS1 gene involved in fungal virulence pertaining to penetration of the insect cuticle, but not to inhibiting the growth of the host.

Effects of Cordyceps militaris spent mushroom substrate on mucosal and serum immune parameters, disease resistance and growth performance of Nile tilapia, (Oreochromis niloticus).

The aim of present study was determination effects of dietary administration of C. militaris spent mushroom substrate (SMS) on mucosal and serum immune parameters, disease resistance, and growth performance of Nile tilapia (Oreochromis niloticus). Two hundred twenty five fish of similar weight (37.28 ± 0.10 g) were assigned to the following diets [0 (T1- Control), 5 (T2), 10 (T3), 20 (T4) and 40 g kg-1 (T5) SMS]. After 60 days of feeding trial, growth performance, skin mucus lysozyme and peroxidase activities as well as serum innate immune were measured. In addition, survival rate and innate immune responses were calculated after challenge test (15 days) against Streptococcus agalactiae. The results revealed that regardless of inclusion levels, feeding Nile tilapia with SMS supplemented diets significantly increased skin mucus lysozyme and peroxidase activities as well as serum immune parameters (SL, ACH50, PI, RB, and RB) compared control group (P < 0.05). The highest increment of immune parameters was observed in fish fed 10 g kg-1 SMS which was significantly higher than other treatments (P < 0.05). Also, the relative percent survival (RSP) in T2, T3, T4, and T5 was 61.11%, 88.89%, 66.67, and 55.56%, respectively. Among the supplemented groups, fish fed 10 g kg-1 SMS showed significant higher RPS and resistance to S. agalactiae than other groups (P < 0.05). Regarding the growth performance, SGR, WG, FW, and FCR were remarkably improved (P < 0.05) in SMS groups; the highest improvement observed in 10 g kg-1 SMS treatment. According to these finding, administration of 10 g kg-1 SMS is suggested in tilapia to improve growth performance and health status.

Identification and characterization of novel single nucleotide polymorphism markers for fat deposition in muscle tissue of pigs using amplified fragment length polymorphism.

OBJECTIVE: This study was conducted to identify and evaluate the effective single nucleotide polymorphism (SNP) markers for fat deposition in the longissimus dorsi muscles of pigs using the amplified fragment length polymorphism (AFLP) approach. METHODS: Sixty-four selective primer combinations were used to identify the AFLP markers in the 20 highest- and 20 lowest-intramuscular fat (IMF) content phenotypes. Five AFLP fragments were converted into simple codominant SNP markers. These SNP markers were tested in terms of their association with IMF content and fatty acid (FA) composition traits in 620 commercially crossbred pigs. RESULTS: The SSC7 g.4937240C>G marker showed an association with IMF content (p<0.05). The SSC9 g.5496647_5496662insdel marker showed a significant association with IMF content and arachidonic levels (p<0.05). The SSC10 g.71225134G>A marker revealed an association with palmitoleic and ω9 FA levels (p<0.05), while the SSC17 g.61976696G>T marker showed a significant association with IMF content and FA levels of palmitoleic, eicosenoic, arachidonic, monounsaturated fatty acids, and ω9 FA levels. However, no significant association of SSC8 g.47338181G>A was observed with any IMF and FA levels in this study. CONCLUSION: Four SNP markers (SSC7 g.4937240C>G, SSC9 g.5496647_5496662insdel, SSC10 g.71225134G>A, and SSC17 g.61976696G>T) were found to be associated with IMF and/or FA content traits in commercially crossbred pigs. These findings provide evidence of the novel SNP markers as being potentially useful for selecting pigs with the desirable IMF content and FA composition.

Population structure of four Thai indigenous chicken breeds.

BACKGROUND: In recent years, Thai indigenous chickens have increasingly been bred as an alternative in Thailand poultry market. Due to their popularity, there is a clear need to improve the underlying quality and productivity of these chickens. Studying chicken genetic variation can improve the chicken meat quality as well as conserving rare chicken species. To begin with, a minimal set of molecular markers that can characterize the Thai indigenous chicken breeds is required. RESULTS: Using AFLP-PCR, 30 single nucleotide polymorphisms (SNPs) from Thai indigenous chickens were obtained by DNA sequencing. From these SNPs, we genotyped 465 chickens from 7 chicken breeds, comprising four Thai indigenous chicken breeds--Pradhuhangdum (PD), Luenghangkhao (LK), Dang (DA) and Chee (CH), one wild chicken--the red jungle fowls (RJF), and two commercial chicken breeds--the brown egg layer (BL) and commercial broiler (CB). The chicken genotypes reveal unique genetic structures of the four Thai indigenous chicken breeds. The average expected heterozygosities of PD=0.341, LK=0.357, DA=0.349 and CH=0.373, while the references RJF= 0.327, CB=0.324 and BL= 0.285. The F(ST) values among Thai indigenous chicken breeds vary from 0.051 to 0.096. The F(ST) values between the pairs of Thai indigenous chickens and RJF vary from 0.083 to 0.105 and the FST values between the Thai indigenous chickens and the two commercial chicken breeds vary from 0.116 to 0.221. A neighbour-joining tree of all individual chickens showed that the Thai indigenous chickens were clustered into four groups which were closely related to the wild RJF but far from the commercial breeds. Such commercial breeds were split into two closely groups. Using genetic admixture analysis, we observed that the Thai indigenous chicken breeds are likely to share common ancestors with the RJF, while both commercial chicken breeds share the same admixture pattern. CONCLUSION: These results indicated that the Thai indigenous chicken breeds may descend from the same ancestors. These indigenous chicken breeds were more closely related to red jungle fowls than those of the commercial breeds. These findings showed that the proposed SNP panel can effectively be used to characterize the four Thai indigenous chickens.

Bio-efficacy of insecticidal molecule emodin against dengue, filariasis, and malaria vectors.

Emodin, a compound isolated from Aspergillus terreus, was studied using chromatographic and spectroscopic methods and compound purity (96%) was assessed by TLC. Furthermore, high larvicidal activity against Aedes aegypti-AeA (LC50 6.156 and LC90 12.450 mg/L), Culex quinquefasciatus-CuQ (8.216 and 14.816 mg/L), and Anopheles stephensi-AnS larvae (6.895 and 15.24 mg/L) was recorded. The first isolated fraction (emodin) showed higher pupicidal activity against AeA (15.449 and 20.752 mg/L). Most emodin-treated larvae (ETL) showed variations in acetylcholine esterase, α and ß-carboxylesterases, and phosphatase activities in the 4th instar, indicating the intrinsic differences in their biochemical changes. ETL had numerous altered tissues, including muscle, gastric caeca, hindgut, midgut, nerve ganglia, and midgut epithelium. Acute toxicity of emodin on brine shrimp Artemia nauplii (54.0 and 84.5 mg/L) and the zebrafish Danio rerio (less toxicity observed) was recorded. In docking studies, Emodin interacted well with odorant-binding-proteins of AeA, AnS, and CuQ with docking scores of - 8.89, - 6.53, and - 8.09 kcal mol-1, respectively. Therefore, A. terreus is likely to be effective against mosquito larvicides.

Insecticidal efficacy of nanoparticles against Spodoptera frugiperda (J.E. Smith) larvae and their impact in the soil.

The present study aims to evaluate the different nanoparticles (Cu NPs, KI NPs, Ag NPs, Bd NPs, and Gv NPs) against 4th instar Spodoptera frugiperda larvae as well as the microbial toxicity, phytotoxicity, and soil pH. Nanoparticles were tested at three concentrations (1000, 10000, and 100000 ppm) using two methods (food dip and larvae dip) against S. frugiperda larvae. Results (from the larval dip method) showed that among the nanoparticles, the KI NPs caused 63%, 98%, and 98% mortality within 5 days in the treatment of 1000, 10000, and 100000 ppm, respectively. After 24 h post treatment, a 1000 ppm concentration showed 95%, 54%, and 94% germination rates in Metarhizium anisopliae, Beauveria bassiana, and Trichoderma harzianum, respectively. The phytotoxicity evaluation clearly showed that NPs did not affect the morphology of the corn plants after the treatment. The soil nutrient analysis results showed that no effect was observed in soil pH or soil nutrients compared to control treatments. The study clearly showed that nanoparticles are caused toxic effect against S. frugiperda larvae.

The inhibitory action of plant extracts on the mycelial growth of Ascosphaera apis, the causative agent of chalkbrood disease in Honey bee.

Ascosphaera apis is a fungal pathogen, which causes chalkbrood disease in bees and is threatening beekeeping worldwide. The demand for organic honey for export has lately heightened hence the biological control is the option. This study aimed at the in vitro evaluation of the potency of plant extracts against chalkbrood disease for the possibility of being employed as a biological control strategy. The results showed that the combination of plant extracts from cinnamon with spearmint, cinnamon with lemongrass, cinnamon with geranium, and cinnamon with palmarosa at a concentration of 25% and 12.5% inhibited mycelial growth of A. apis by 100%. This demonstrated the potentiality of combining different plant extracts in controlling this disease. In addition, oregano caused inhibition of up to 100% singly. Conclusively, cinnamon in combination with several extracts has a great potential in curbing this disease while oregano offers an amazing remedy and hence the best formulations should be generated for the beekeeper to utilize.

Larvicidal and Antifeedant Effects of Copper Nano-Pesticides against Spodoptera&nbsp;frugiperda (J.E. Smith) and Its Immunological Response.

This study aimed to synthesize and evaluate the efficacy of CuO NPs (copper oxide nanoparticles) with varying test concentrations (10−500 ppm) against larvicidal, antifeedant, immunological, and enzymatic activities against larvae of S. frugiperda at 24 h of treatment. Copper nanoparticles were characterized by using a scanning electron microscope (SEM) and energy dispersive X-ray (EDaX) analysis. The EDaX analysis results clearly show that the synthesized copper nanoparticles contain copper as the main element, and the SEM analysis results show nanoparticle sizes ranging from 29 to 45 nm. The CuO NPs showed remarkable larvicidal activity (97%, 94%, and 81% were observed on the 3rd, 4th, and 5th instar larvae, respectively). The CuO NPs produced high antifeedant activity (98.25%, 98.01%, and 98.42%), which was observed on the 3rd, 4th, and 5th instar larvae, respectively. CuO NPs treatment significantly reduced larval hemocyte levels 24 h after treatment; hemocyte counts and sizes changed in the CuO NPs treatment compared to the control. After 24 h of treatment with CuO NPs, the larval acetylcholinesterase enzyme levels decreased with dose-dependent activity. The present findings conclude that CuO NPs cause remarkable larvicidal antifeedant activity and that CuO NPs are effective, pollution-free green nano-insecticides against S. frugiperda.

High-Efficiency Bovine Sperm Sexing Used Magnetic-Activated Cell Sorting by Coupling scFv Antibodies Specific to Y-Chromosome-Bearing Sperm on Magnetic Microbeads.

Sperm sexing technique is favored in the dairy industry. This research focuses on the efficiency of bovine sperm sexing using magnetic-activated cell sorting (MACS) by scFv antibody against Y-chromosome-bearing sperm (Y-scFv) coupled to magnetic microbeads and its effects on kinematic variables, sperm quality, and X/Y-sperm ratio. In this study, the optimal concentration of Y-scFv antibody coupling to the surface of magnetic microbeads was 2-4 mg/mL. PY-microbeads revealed significantly enriched Y-chromosome-bearing sperm (Y-sperm) in the eluted fraction (78.01-81.43%) and X-chromosome-bearing sperm (X-sperm) in the supernatant fraction (79.04-82.65%). The quality of frozen-thawed sexed sperm was analyzed by CASA and imaging flow cytometer, which showed that PY-microbeads did not have a negative effect on X-sperm motility, viability, or acrosome integrity. However, sexed Y-sperm had significantly decreased motility and viability. The X/Y-sperm ratio was determined using an imaging flow cytometer and real-time PCR. PY-microbeads produced sperm with up to 82.65% X-sperm in the X-enriched fraction and up to 81.43% Y-sperm in the Y-enriched fraction. Bovine sperm sexing by PY-microbeads showed high efficiency in separating Y-sperm from X-sperm and acceptable sperm quality. This initial technique is feasible for bovine sperm sexing, which increases the number of heifers in dairy herds while lowering production expenses.

Association of IFNA16 and TNFRSF19 Polymorphisms with Intramuscular Fat Content and Fatty Acid Composition in Pigs.

Interferon-alpha-16 (IFNA16) and tumor necrosis factor receptor superfamily member 19 (TNFRSF19) are cytokines that may play a role in adipogenesis and fatness. Single nucleotide polymorphisms (SNPs) of the porcine IFNA16 and TNFRSF19 genes were verified and their association with intramuscular fat (IMF) content and fatty acid (FA) composition were evaluated in commercial crossbred pigs. Two non-synonymous SNPs of the porcine IFNA16 c.413G > A and TNFRSF19 c.860G > C loci were detected in commercial crossbred pigs. The porcine IFNA16 c.413G >A polymorphism was significantly associated with stearic acid, total saturated FAs (SFAs), and the ratio of monounsaturated FAs (MUFAs) to SFAs (p < 0.05). Furthermore, the porcine TNFRSF19 c.860G > C polymorphism was found to be significantly associated with IMF content and arachidic acid levels (p < 0.05). The results revealed that porcine IFNA16 and TNFRSF19 polymorphisms are related to IMF content and/or FA composition and affirmed the importance of these cytokine genes as potential candidate genes for lipid deposition and FA composition in the muscle tissue of pigs.

Microsatellite Polymorphism and the Population Structure of Dugongs (Dugong dugon) in Thailand.

The dugong (Dugong dugon) is an endangered species of marine mammals, so knowledge of genetic diversity of these populations is important for conservation planning within different habitats. In this study, six microsatellite markers were used to assess the genetic diversity and population structure of 77 dugongs from skin samples of stranded animals collected from 1994-2019 (69 from Andaman Sea and 8 from the Gulf of Thailand). Our results found that dugongs in the Andaman Sea had higher genetic variation than those in the Gulf of Thailand. Populations in Trang, Satun, and some areas of Krabi had highest diversity compared to other regions of Thailand. Bayesian genetic clustering analysis revealed that dugongs in Thailand consist of five genetic groups. Moreover, dugongs in the middle and lower Andaman Sea presented the greatest gene flow compared to other regions. However, based on calculation of inbreeding coefficients (Fis value = 0.239), dugong populations in the Sea of Thailand are experiencing some levels of inbreeding, and so may warrant special protections. These results provide important information for understanding the genetic status of dugongs that can lead to improved management and conservation of this endangered species.

Toxicity of Metarhizium flavoviride conidia virulence against Spodoptera litura (Lepidoptera: Noctuidae) and its impact on physiological and biochemical activities.

Insect pests of agricultural crops have establish immunological tolerance against fungal infection caused by pathogens via different humoral and cellular processes. Fungal infection can be prevented by insect antioxidant and detoxifying enzymes, but there is no clear understanding of how they physiologically and biochemically interact. Our study aims to examine the antioxidant and detoxifying enzyme defense systems of the pest insect Spodoptera litura in response to infection by Metarhizium flavoviride. At 48 h following exposure to M. flavoviride, antioxidant enzyme levels were modified, and phenoloxidase and total hemocyte count were decreased significantly. The amount of detoxifying enzymes increased significantly. M. flavoviride appears to directly affect the S. litura immune system and results in decreased immunity. In a bioassay, M. flavoviride was found to be harmful to S. litura larvae in their third and fourth instar stage. M. flavoviride may be an effective tool in the control of S. litura larvae. Such entomopathogenic fungi represent cheaper, pollution free, target specific, promising alternatives to synthetic chemical tools in the for control insect pests.

Improvement of Intramuscular Fat in longissimus Muscle of Finishing Thai Crossbred Black Pigs by Perilla Cake Supplementation in a Low-Lysine Diet.

This study was conducted to find out the effects of perilla cake (PC) supplementation in a low-lysine diet on Thai crossbred finishing pigs' productivity, carcass and meat quality, and fatty acid composition. For six weeks, a total of 21 barrows of finishing pigs were fed with three dietary treatments (T1: basal diet, T2: 2.5 percent PC supplementation in a low-lysine diet, and T3: 4.5 percent PC supplementation in a low-lysine diet). The results show that the intramuscular fat and marbling score was significantly increased by T2 and T3. On the other hand, it was found that the boiling loss and shear force value were significantly decreased by T2 and T3 (p < 0.05). In a low-lysine diet, dietary PC supplementation caused a significant increase in malondialdehyde levels in meat (p < 0.05) compared with the basal diet. It was also shown that alpha-linolenic acid level in backfat and the longissimus thoracis et lumborum muscle was increased considerably by T2 and T3. Therefore, supplementing PC in a low-lysine diet may be an alternative strategy for improving the meat quality of late-phase pigs.

Insecticidal Effect of Zinc Oxide Nanoparticles against Spodoptera frugiperda under Laboratory Conditions.

Fall armyworm Spodoptera frugiperda is a major pest of corn, rice, and sorghum among other crops usually controlled using synthetic or biological insecticides. Currently, the new invention of nanotechnology is taking root in the agricultural industry as an alternative source of pest management that is target-specific, safe, and efficient. This study sought to determine the efficacy of commercial Zinc Oxide (ZnO) nanoparticles (NPs) towards S. frugiperda under laboratory conditions. ZnO NPs were diluted into different concentrations (100-500 ppm), where the baby corn used to feed the S. frugiperda larvae was dipped. The development of the insect feeding on food dipped in ZnO solution was significantly (p < 0.05) affected, and the number of days that the insect took to complete its life cycle had a significant difference compared to the control. There was a significant difference in the adults' emergence in all the concentrations of ZnO NPs compared to the control, with over 90% of the eggs successfully going through the life cycle until adult emergence. Additionally, several body malformations were observed throughout the lifecycle of the insect. Also, the fecundity of the females was greatly affected. The findings of this study suggest the possibility of exploitation of ZnO nanoparticles not only to manage S. frugiperda but to significantly reduce their population in the ecosystem through body deformations, reduced fecundity, reduced oviposition, and hatchability of eggs. It will be a valuable tool in integrated pest management regimens.

Spermatological parameters of immunologically sexed bull semen assessed by imaging flow cytometry, and dairy farm trial.

This study compared the quality parameters of bull semen sexed using an immunological method with those of conventional semen by imaging flow cytometry and applied this semen in dairy farm trials. Semen samples were collected from ten ejaculates from five bulls. Each sample was divided into two treatments: conventional semen (CON) and semen sexed using monoclonal male-specific antibodies combined with the complement system for cytotoxicity reaction (IC-sexed). After obtaining frozen-thawed semen, we used imaging flow cytometry to assess acrosome integrity, sperm sex ratio and viability. Sperm morphology was evaluated using eosin-nigrosin staining. The percentage acrosome integrity did not differ between IC-sexed and CON semen (P = 0.313). The sperm sex ratio showed that the percentage of live X-chromosome-bearing sperm was higher than that of live Y-chromosome-bearing sperm in IC-sexed semen (P = 0.001). IC-sexed semen showed a higher percentage of head and tail defects than did CON semen (P = 0.019). In field trials, 585 cows were subjected randomly to AI with CON or IC-sexed semen. The pregnancy rate of the IC-sexed group did not differ from that of the CON group (P = 0.535). However, IC-sexed semen produced a significantly higher percentage of female calves than did CON semen (P = 0.031). Thus, immunological sexing did not adversely affect the acrosome integrity of sperm. Furthermore, a female calf birth rate of over 74 % can potentially be achieved using IC-sexed semen. These findings could help farmers to replace heifers in their herds.

Association of adipocytokine IL-1A and IL-6 genes with intramuscular fat content and fatty acid composition in pigs.

Several adipocytokines are involved in inflammatory and immune responses as well as regulated fat deposition and lipid metabolism in mammals. This study aimed to verify the polymorphisms of the porcine interleukin 1A (IL-1A) and interleukin 6 (IL-6) genes and to assess their association with intramuscular fat (IMF) content and fatty acid (FA) composition in commercial crossbred pigs. Two single nucleotide polymorphisms (SNPs) of the porcine IL-1A g.43722547A>G and IL-6 g.91508173C>T loci were found to be segregating in these crossbred pigs. Furthermore, the porcine IL-1A g.43722547A>G polymorphism was found to be significantly associated with myristic, palmitic, palmitoleic, and eicosadienoic acid levels. Moreover, the porcine IL-6 g.91508173C>T polymorphism was significantly associated with IMF content and homolinolenic acid levels. These results suggest that the polymorphisms of the porcine IL-1A and IL-6 genes correlated with lipid content and FA composition and confirmed the importance of the adipocytokine IL-1A and IL-6 genes as candidate genes for fatty acid composition in the muscles of pigs.

Evaluation of Native Entomopathogenic Fungi for the Control of Fall Armyworm (Spodoptera frugiperda) in Thailand: A Sustainable Way for Eco-Friendly Agriculture.

Fall armyworm, Spodoptera frugiperda, entered Thailand in late 2018 and has now spread in several regions, with devastating effects in maize and rice production, which are some of the most important cereals in the world. Since then, farmers have utilized the available chemical insecticides to try to control it, but their efforts have been futile. Instead, they have ended up using extraordinary dosages, hence threatening non-target species and other fauna and flora, as well as being costly. In this regard, research has been ongoing, aiming to come up with eco-friendly solutions for this insect. We surveyed and collected various isolates of native entomopathogenic fungi intending to test their efficacy against fall armyworm. Six isolates of entomopathogenic fungi were obtained and identified to Beauveria bassiana based on morphological characteristics and multi-gene phylogenetic analyses. Thereafter, the six isolates of B. bassiana were used to perform efficacy experiments against fall armyworm. Additionally, the glycosyl transferase-like protein 1 (GAS1) gene was analyzed. Consequently, all the isolates showed efficacy against S. frugiperda, with isolate BCMU6 causing up to 91.67% mortality. Further, molecular analysis revealed that all the isolates possess the GAS1 gene, which contributed to their virulence against the insect. This is the first report of utilizing native entomopathogenic B. bassiana to manage S. frugiperda in Thailand, with the revelation of GAS1 as a factor in inducing virulence and cuticle penetration. This study has provided valuable information on the potential development of Beauveria bassiana as an eco-friendly bioinsecticide for the management of fall armyworm in Thailand.