Microencapsulation of lemongrass and mangosteen peel as phytogenic compounds to gas kinetics, fermentation, degradability, methane production, and microbial population using in vitro gas technique.

The purpose of the current study was to evaluate the impact of various doses of microencapsulated lemongrass and mangosteen peel (MELM) on gas dynamics, rumen fermentation, degradability, methane production, and microbial population in in vitro gas experiments. With five levels of microencapsulated-phytonutrient supplementation at 0, 1, 2, 3, and 4% of substrate, 0.5 g of roughage, and a concentrate ratio of 60:40, the trial was set up as a completely randomized design. Under investigation, the amount of final asymptotic gas volume was corresponding responded to completely digested substrate (b) increased cubically as a result of the addition of MELM (P < 0.01) and a cubic rise in cumulative gas output. The amount of MELM form did not change the pH and NH3-N concentration of the rumen after 12 and 24 h of incubation. However, methane production during 24 h of incubation, the levels were cubically decreased with further doses of MELM (P < 0.01) at 12 h of incubation. Increasing the dosage of MELM supplementation at 2% DM resulted in a significant increase in the digestibility of in vitro neutral detergent fiber (IVNDF) and in vitro true digestibility (IVTD) at various incubation times (P < 0.05), but decreased above 3% DM supplementations. Moreover, the concentration of propionic acid (C3) exhibited the variations across the different levels of MELM (P < 0.05), with the maximum concentration obtained at 2% DM. The populations of Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens, and Megasphaera elsdenii revealed a significant increase (P < 0.05), while the quantity of Methanobacteriales decreased linearly with increasing doses of MELM. In conclusion, the inclusion of MELM at a concentration of 2% DM in the substrate which could enhance cumulative gas production, NDF and true digestibility, C3 production, and microbial population, while reducing methane concentration and Methanobacterial abundance.

Influences of Thermal Stress During Three Weeks Before Market Age on Histology and Expression of Genes Associated With Adipose Infiltration and Inflammation in Commercial Broilers, Native Chickens, and Crossbreeds.

The objectives of this study were to examine the effects of cyclic thermal stress on histological characteristics of breast muscle and gene expression regarding adipose infiltration and inflammation in breast muscles collected from different breeds of chickens. The birds, from commercial broilers (CB, Ross 308, 3 weeks), native (NT, 100% Thai native Chee, 9 weeks), H75 (crossbred; 75% broiler and 25% NT, 5 weeks), and H50 (crossbred; 50% broiler and 50% NT, 7 weeks), were equally assigned into control or treatment groups. The control samples were reared under a constant temperature of 26 ± 1°C, while the treatment groups were exposed to 35 ± 1°C (6 h per day). After a 20-day thermal challenge, 12 male birds per treatment group were randomly collected for determination of live body weight, breast weight, numbers of growth-related myopathies, and breast meat chemical composition. Histological lesions were evaluated in the pectoralis major muscle immediately collected within 20 min postmortem based on hematoxylin and eosin staining. The results indicated that despite interaction between thermal stress and breed effects, thermal challenge significantly reduced feed intake, live body weight, and breast weight of the birds and increased moisture content in breast meat (p < 0.05). An interaction between the two main factors was found for protein content (p < 0.05) for which control CB showed less protein than the other groups. Heat stress decreased histological scores for adipose infiltration in CB (p < 0.05), but it did not significantly influence such scores in the other groups. CB received histological scores for adipose tissue at greater extent than those for the other groups. Differential absolute abundance of CD36, FABP4, LITAF, PDGFRA, PLIN1, PPARG, POSTN, SCD1, and TGFB1 in the muscle samples well-agreed with the trend of histological scores, suggesting potential involvement of dysregulated fibro-adipogenic progenitors together with imbalanced lipid storage and utilization in the breast muscle. The findings demonstrated that the cyclic thermal challenge restricted growth performance and breast mass of the birds, but such effects attenuated infiltration of adipose tissue and inflammatory cells in the CB breast muscle.

Effects of Cyclic Thermal Stress at Later Age on Production Performance and Meat Quality of Fast-Growing, Medium-Growing and Thai Native Chickens.

The present study aimed at assessing the impact of cyclic thermal stress on production performance and meat quality of commercial broilers (BRs), Thai native chickens (NT) and the hybrids between BR and NT (H75; crossbreed 25% NT). At the age of 3, 5 and 9 weeks for BR, H75 and NT, respectively, each strain was equally divided (n = 50) into control and treatment groups. The controls were raised at a constant 26 ± 1 °C, while the treatments were subjected to thermal stress (35 ± 1 °C, 6 h daily) for 3 weeks. The results indicated that final weight and average daily gain of BR and NT treated groups were significantly lower than those of their control counterparts. Reduced body weight gain of BR and H75, as well as feed intake of H75, was observed in the treatment groups (p < 0.05). The stressed BR breasts showed decreased moisture, fat and carbohydrate, accompanied by increased protein, ash, L *-value, b*-value and shear force (p < 0.05). No significant effects (p ≥ 0.05) of the thermal stress on meat quality indices were found for H75 and NT breast samples. Pectoral myopathies were observed in BR and H75 chickens, but the numbers of cases were decreased in the thermally treated groups.

Factors affecting gene expression associated with the skin color of black-bone chicken in Thailand.

The objective of this study was to investigate the effect of breed, sex, and age on the gene expression level of melanocortin 1 receptor (MC1R), DOPA chrome tautomerase (DCT), tyrosinase-related protein 1 (TYRP1), tyrosinase (TYR), and agouti signaling protein (ASIP) genes in Thai commercial chicken lines. All chicken have received Newscastle vaccination, and no antibiotics or any drugs were used in this study. Four chicken breeds including Black-Chinese, KU-Phuparn, Sri Mok, and Pradu Hang Dam were used in this study. These breeds can be classified by their skin color into 3 group including black (Black Chinese and KU-Phuparn), light black (Sri Mok), and yellowish white (Pradu Hang Dam). One hundred chickens per breed were used in this study. Breast skin tissue was randomly collected from 8 chickens (4 males, 4 females) per breed at 4, 8, 12, and 16 wk of age. The mRNA expression was analyzed using qRT-PCR and the gene expression level was calculated as 2-ΔΔCT. From the results, breed significantly (P < 0.01) affected the expression level for the 5 genes evaluated. Birds with the black skin color had greater TYRP1 and TYR gene expression when compared to chickens with light black and yellowish-white skin color, respectively. Whereas, chickens with yellowish-white skin color had greater ASIP gene expression when compared to chickens having the other skin colors. Sex significantly affected DCT, TYRP1, and TYR gene expression where the gene expression in males was greater when compared to females (P < 0.05). Age affected all gene expression levels (P < 0.01). At 4 wk of age, MC1R, DCT, TYRP1, and TYR gene expression was the highest and decreased as bird age increased (P < 0.05); however, ASIP gene expression was greatest at 8 wk of age. After 8 wk of age all gene expression for the genes evaluated in this study decreased as age increased. In addition, an interaction between breed and sex (P < 0.05) impacted DCT and ASIP gene expression. The results from this study showed that all genes evaluated can be used as candidate markers to further improve the blackness of the chicken's skin because the most desired skin color is black in the Thai black-bone chicken population.

Carnosine Content and Its Association with Carnosine-Related Gene Expression in Breast Meat of Thai Native and Black-Bone Chicken.

This study aimed to investigate the carnosine content and ATP-grasp domain-containing protein 1 (CARNS1) gene expression and their relationship with breast meat of Black Chinese (BC), KU-Phuparn (KP), Pradu Hang Dam (PD), and Black Chinese × Pradu Hang Dam (Sri Mok: SM) to aid in the selection and mating programs for developing functional meat in Thai chicken populations. The results show that the carnosine content in each breed and breed group varied from 428.08 to 553.93 mg/100 g, whereas the relative expression of CARNS1 ranged from 0.84 to 1.56. The BC and KP chicken breeds had a higher carnosine content (p < 0.01) and higher CARNS1 expression level (p < 0.05) than the SM and PD chicken breeds. The carnosine content and relative gene expression for each age ranged from 423.02 to 577.83 mg/100 g and 0.68 to 1.83, respectively. At 4 weeks of age, the carnosine content (p < 0.01) and gene expression (p < 0.05) were the highest. However, they decreased as chicken age increased further. The carnosine content and gene expression linearly decreased as chicken age increased (p < 0.01). The correlation coefficient between the level of gene expression and carnosine content was moderately positive. The results from this study showed that different breeds and ages of chickens have different amounts of carnosine, and CARNS1 could act as a biomarker to study marker-assisted selection to improve functional meat in the chicken population in Thailand.

Droplet digital polymerase chain reaction assay for identifying and quantifying pork products.

Halal products are growing in consumer markets worldwide, and pork meat is classified as non-halal. Manufacturers of processed foods and products must ensure that their products follow Islamic dietary law because pork is prohibited for Muslims. Droplet digital polymerase chain reaction (PCR) (ddPCR) is a novel method for identifying pig species and quantifying pork products. This experiment aimed to investigate pork species and establish the proportion of pork in meat products using the mitochondrial cytochrome b gene (CYTB). The study found that the correlation coefficient between the meat weight and DNA concentration of pork was 0.997, and the correlation coefficient between the DNA concentration and the target DNA copy number of pork was 0.998. The accuracy of the ddPCR assay was verified using a sample of a known proportion of pork, and it was revealed that this method is highly precise in quantifying pork products. Nine products contained an undeclared meat proportion (90%). The limit of detection for pork was 0.0001 ng. The analysis indicated that the ddPCR assay has high accuracy and sensitivity for quantifying pork products. Therefore, the predictive model can be used in routine laboratories for quality assurance of halal food products.

Increasing Fat Deposition Via Upregulates the Transcription of Peroxisome Proliferator-Activated Receptor Gamma in Native Crossbred Chickens.

This study aimed to study the role of PPARs on fat deposition in native crossbred chicken. We studied the growth, abdominal, subcutaneous, and intramuscular fat, and mRNA expression of PPARA and PPARG in adipose and muscle tissues of four chicken breeds (CH breed (100% Thai native chicken), KM1 (50% CH background), KM2 (25% CH background), and broiler (BR)). The result shows that the BR chickens had higher abdominal fat than other breeds (p < 0.05) and the KM2 had an abdominal fat percentage higher than KM1 and CH respectively (p < 0.05). The intramuscular fat of BR was greater than KM1 and CH (p < 0.05). In adipose tissue, PPARA expression was different among the chicken breeds. However, there were breed differences in PPARG expression. Study of abdominal fat PPARG expression showed the BR breed, KM1, and KM2 breed significantly greater (p < 0.05) than CH. In 8 to 12 weeks of age, the PPARG expression of the CH breed is less than (p < 0.05) KM2. Crossbreeding improved the growth of the Thai native breed, there was also a corresponding increase in carcass fatness. However, there appears to be a relationship between PPARG expression and fat deposition traits. therefore, PPARG activity hypothesized to plays a key role in lipid accumulation by up-regulation.

Gene expression of fatty acid binding protein genes and its relationship with fat deposition of Thai native crossbreed chickens.

OBJECTIVE: The objectives of this study were to investigate the relationship between the mRNA expression of adipocyte type fatty acid binding protein (A-FABP) and heart type FABP (H-FABP) in Thai native chicken crossbreeds and evaluate the level of exotic inclusion in native chicken that will improve growth while maintaining its relatively low carcass fat. METHODS: The fat deposition traits and mRNA expression of A-FABP and H-FABP were evaluated at 6, 8, 10, and 12 weeks of age in 4 chicken breeds (n = 8/breed/wk) (100% Chee breed [CH] [100% Thai native chicken background], CH male and broiler female [Kaimook e-san1; KM1] [50% CH background], broiler male and KM1 female [Kaimook e-san2; KM2] [25% CH background], and broiler [BR]) using abdominal fat (ABF) and muscular tissues. RESULTS: The BR breed was only evaluated at 6 weeks of age. At week 6, the CH breed had a significantly lower A-FABP expression in ABF and intramuscular fat (IF) compared with the other breeds. At 8 to 12 weeks, the KM2 groups showed significant upregulation (p<0.05) of A-FABP in both ABF and IF compared to the CH and KM1 groups. The expression of H-FABP did not follow any consistent pattern in both ABF and IF across the different ages. CONCLUSION: Some level of crossbreeding CH chickens can be done to improve growth rate while maintaining their low ABF and IF. The expression level of A-FABP correlate with most fat traits. There was no consistency of H-FABP expression across breed. A-FABPs is involved in fat deposition, genetic markers in these genes could be used in marker assisted studies to select against excessive fat accumulation.

Effect of Supplemental Kluyveromyces marxianus and Pichia kudriavzevii on Aflatoxin M1 Excretion in Milk of Lactating Dairy Cows.

The objective of this study was to determine the effect of supplementing Kluyveromyces marxianus CPY1, K. marxianus RSY5 and Pichia kudriavzevii YSY2 isolated from ruminal fluid of dairy cows on transfer of aflatoxin B1 (AFB1) from feed into aflatoxin M1 (AFM1) in milk, DMI, milk production and nutrient digestibility. Four multiparous Holsteins in mid-lactation were used in a 4 × 4 Latin square design trial consisting of 14 days in each experimental period for sample collection. Between each period, 14 clearance days prior to the next treatment were allowed to minimize carryover effects. In each treatment, subsequent supplementation of isolated yeast was compared, i.e., (1) control (without yeast supplementation), (2) K. marxianus CPY1 (K1Y), (3) K. marxianus RSY5 (K2Y) and (4) P. kudriavzevii YSY2 (PY). All diets contained 22.28 µg of AFB1/kg. Treatments were individually fed at the rate of 2 g/day (1 × 109 CFU/g) of yeast biomass or corn meal in the control group. Concentrations of AFM1 in milk was reduced with yeast and averaged 1.54, 0.36, 0.43 and 0.51 µg/L for control, K1Y, K2Y and PY, respectively (p < 0.01). The transfer of AFB1 from feed into AFM1 in milk was higher in control compared with K1Y, K2Y and PY (7.26% vs. 1.18%, 1.44% and 1.69% respectively, p < 0.01). Supplementation of yeast also improved DMI and milk compositions, but no differences were observed in nutrient digestibility or milk yield among treatments. Concentration and yield of milk protein, fat, lactose, solid-not-fat (SNF) and total solids were greater in cows fed yeast compared with the control (p < 0.01). These results indicate that K. marxianus CPY1, RSY5 and P. kudriavzevii YSY2 shows promise as a dietary supplementation to detoxify AFB1 and improve DMI and yield of milk components.

Different concentrations of cysteamine, ergothioneine, and serine modulate quality and fertilizing ability of cryopreserved chicken sperm.

The aim of this study was to evaluate the effects of freezing diluents supplemented in three potential amines/amino acids, namely, antioxidant cysteamine (2-aminoethanethiol [AET]), ergothioneine (ERG), and serine (SER), in optimization of chicken sperm cryopreservation. The semen of 36 Pradu Hang Dum males, selected based on their motility vigor score, was frozen by a simple freezing method using nitrogen vapors and dimethylformamide (DMF). In a first experiment, a wide range of AET, ERG, and SER doses were tested. Semen quality was evaluated after incubation at 5°C or after cryopreservation in straws in the Blumberger Hahnen Sperma Verdünner (BHSV) diluent + DMF (6% v/v) with or without AET, ERG, or SER. The best targeted doses of AET, ERG, or SER were then selected for experiment 2 that was focused on cryopreserved semen. Frozen-thawed sperm quality was evaluated by different in vitro tests and by evaluation of fertility. Objective motility parameters were evaluated by computer-assisted sperm analysis. Membrane integrity, acrosome integrity, and mitochondria function were evaluated using appropriate dyes and flow cytometry. Lipid peroxide production was assessed by the thiobarbituric acid test (malondialdehyde production). Fertility obtained with frozen-thawed semen supplemented or not in AET, ERG, or SER was evaluated after artificial insemination of laying hens. ERG and AET decreased sperm lipid peroxidation and decreased fertility, even at low doses. The presence of 4 mmol of SER significantly decreased lipid peroxidation, increased the frozen-thawed sperm quality, and increased fertility after sperm cryopreservation (90% vs. control 84%, P < 0.05). In a third experiment, the use of 1 mmol of sucrose (the best result of our previous study) added to 4 mmol of SER-supplemented extender was tested. This addition allowed to the highest levels of fertility (93%). In conclusion, the addition of 4 mmol of SER in semen cryopreservation diluents decreases peroxidation and improves the efficiency of the process.

Comparison of TNC and standard extender on post-thaw quality and in vivo fertility of Thai native chicken sperm.

Semen extender has a vital role in preservation of sperm cells properties in terms of sperm viability, motility, acrosome integrity, and mitochondrial membrane potential. The objective of the present study was to evaluate a new extender, known as Thai native chicken (TNC) extender compared to BHSV-based and modified Sasaki extenders for freezing chicken semen. Semen from Thai native roosters was collected, pooled, and randomly divided into three groups. Semen was frozen with a simple freezing method using nitrogen vapor and dimethylformamide. In the first experiment, post-thaw motion parameters, viability, acrosome integrity, mitochondrial function, and lipid peroxidation levels were analyzed using computer-assisted sperm analysis, propidium iodide, fluorescein isothiocyanate-conjugate peanut agglutinin, JC-1, and the thiobarbituric acid reaction. Results showed that the type of extender had no effect on the percentage of total motile and curvilinear velocity. The percentage of progressive motile, straight-line velocity, and average path velocity of post-thawed semen were significantly lower in TNC compared to the modified Sasaki extender. However, the percentages of post-thawed acrosome integrity and active mitochondria were significantly higher in TNC extender (P < 0.05). For the second experiment, semen was thawed by using each of extenders thereafter, was inseminated to 48-layer breeder hens to determine the fertility rate. Among the three extenders used, the highest fertility rate was found in TNC extender. In conclusion, TNC extender can be recommended as an appropriate and useful cryopreservation media for native chicken semen since it maintains the quality of rooster semen and fertility after freezing and thawing process.

Sucrose increases the quality and fertilizing ability of cryopreserved chicken sperms in contrast to raffinose.

Chicken semen conservation is an important tool for programs of genetic diversity management and of endangered breeds' conservation. However, the method still needs to be improved in order to be applied in a wide variety of environments and breeds. Our objective was to compare the effects of 2 external cryoprotectants saccharides (sucrose and raffinose) on the sperm freezability of a Thai local breed, Pradu Hang Dum, in which semen was frozen with a simple freezing method using nitrogen vapors and dimethyl formamide (DMF). Thirty-six males were selected on their motility vigor score for the experiments. In a first experiment, a large range of sucrose and raffinose doses were tested. Semen quality was evaluated after incubation at 5°C or after cryopreservation in straws in the saline Blumberger Hahnen Sperma Verdünner diluent + DMF (6% v/v) with or without sucrose/raffinose. The best targeted doses of sucrose and raffinose were then kept for experiment 2 that was focused on cryopreserved semen. In this experiment, semen quality was measured on frozen-thawed sperm: different objective motility data evaluated by computer-assisted sperm analysis (CASA), membrane integrity, acrosome integrity, mitochondria function evaluated using flow cytometry, lipid peroxide production assessed by the thiobarbituric acid test. Fertility obtained with frozen-thawed semen supplemented or not with sucrose or raffinose was also evaluated after artificial insemination of laying hens. The presence of sucrose at the osmotically inactive dose 1 mmol significantly increased the vigor motility, membrane integrity, acrosome integrity, and mitochondrial functions of frozen-thawed sperm (P < 0.05), and showed the highest levels of fertility after sperm cryopreservation (91% vs. control 86%, P < 0.001). Raffinose showed negative effects on in vitro semen quality from 1 to 100 mmol. Fertility was also negatively (P < 0.001) affected by raffinose (fertility rate 66 to 70%). We thus showed in the present study the high success of a simple chicken sperm cryopreservation method with an external cryoprotectant easily available and cheap, the sucrose, used at an osmotically inactive low concentration.

VITRIFICATION OF THAI NATIVE CATTLE OOCYTES: EFFECTS OF ETHYLENE GLYCOL CONCENTRATIONS AND EXPOSURE TIME, LINOLEIC ACID ALBUMIN AND CHOLESTEROL-LOADED METHYL-B-CYCLODEXTRIN.

The present study aimed to improve the oocyte vitrification procedure for preservation of Thai native cattle genetic resources. In Experiment I, oocytes were exposed to various doses (2%, 4% and 6%) of ethylene glycol (EG) in vitrification solution I (VS-I) for different equilibration times (10 or 20 min) before being exposed to VS-II and then subjected to vitrification. Experiment II was divided into two parts: (a) oocytes were matured in medium supplemented with linoleic acid albumin (LAA) (1% or 2%) and then vitrified; (b) matured oocytes were preincubated with cholesterol-loaded methyl-ß-cyclodextrin (CLC) (1% or 2%) and then vitrified. Equilibration of oocytes by exposure to 6% EG in VS-I for 10 min (Experiment I), and in vitro maturation of immature oocytes in medium supplementation with 2% LAA (Experiment II) were the most effective methods; vitrified/thawed oocytes showed higher rates of survival and subsequent embryonic development compared with the other experimental groups.

Genetic characterization of Bhutanese native chickens based on an analysis of Red Junglefowl (Gallus gallus gallus and Gallus gallus spadecieus), domestic Southeast Asian and commercial chicken lines (Gallus gallus domesticus).

The genetic diversity of Bhutanese chickens needs to be understood in order to develop a suitable conservation strategy for these birds in Bhutan. In this, work, we used microsatellite markers to examine the genetic diversity of Bhutanese chickens. Four Bhutanese chicken varieties (Black plumage, Frizzle, Naked neck and Red Junglefowl-like, corresponding to Yuebjha Narp, Phulom, Khuilay and Seim, respectively), two subspecies of Red Junglefowl (Gallus gallus gallus and Gallus gallus spadecieus), two varieties of Thai native chickens (Pradhu Hang Dam and Chee; Gallus gallus domesticus) representing the Southeast Asian domestic chicken, and two commercial lines (Broiler and Single Comb White Leghorn) were genotyped with 18 microsatellites that included 16 loci recommended by the FAO/ISAG for investigations of genetic variability in chickens. All loci were polymorphic, with the number of alleles ranging from six (MCW0111) to 23 (MCW0183). Substantial genetic variation was observed in all populations, with the Bhutanese native chicken Yuebjha Narp (Black plumage chicken) showing the lowest genetic variability. Despite extensive intrapopulation variation, the genetic differentiation among 10 populations was moderate. A neighbor-joining tree revealed the genetic relationships involved while principal component analysis showed that Bhutanese native chickens should be given priority in conservation efforts because of their genetic distinctiveness. Chee chickens are especially valuable as a reservoir of predomestic diversity, as indicated by their greater genetic variation and their position in the phylogenetic tree.

Genetic characterization of Thai indigenous chickens compared with commercial lines.

A total of 210 chicken samples, from seven strains, were genotyped using 20 microsatellite loci of which 16 are recommended by the Food and Agriculture Organization. The genetic variability and divergence of four Thai indigenous strains and three commercial lines were assessed to generate baseline information for conservation, promotion, and make sustainable utilization of indigenous chicken resources in Thailand. A total of 227 alleles were distributed ranging from six (MCW 111) to 16 (MCW 183 and LEI 166) alleles per locus. The highest (0.81) and lowest (0.77) average of expected heterozygosities were observed in indigenous chicken (Dang) and commercial layer (Isa Brown), respectively. All microsatellite loci were in the Hardy-Weinberg equilibrium, except for MCW111 and ADL372 in the Isa Brown line. The subpopulation division coefficient (F(ST)) was strong with the value of 0.183 indicating the genetic differentiation among the studied groups. Four genetic clusters were detected: the first group consisted of layers (Isa Brown and White Leghorn); the second group was broiler; the third group consisted of non-black feather indigenous chicken (Chee, Dang, and Leung Hang Khoa); and the fourth group was black feather indigenous chicken (Pradu Hang Dam). The results of this study also suggested that Pradu Hang Dam is suitable to be developed as a meat type chicken due to lower genetic distance between Pradu Hang Dam and broiler.