Enzymatic hydrolyzation of Cordyceps militaris mushroom extracts and its effect on spent hen chicken.

Objective: This study was aimed to investigate the effect of fresh and dried hydrolyzed Cordyceps militaris (CM) mushroom with proteolytic enzymes; bromelain (CMB), flavorzyme (CMF), and mixture of bromelain: flavorzyme (CMBF) on quality properties of spent hen chicken. Methods: Mushroom extract (CME) were combined with three proteolytic enzyme mixtures that had different peptidase activities; stem bromelain (CMB), flavorzyme (CMF), and mixture of stem bromelain:flavorzyme (CMBF) at (1 : 1). The effect of these hydrolysates was investigated on spent hen breast meat via dipping marination. Results: Hydrolyzation positively alters functional properties of CM protease. in which bromelain hydrolyzed group (CMB) displayed the highest proteolytic activity at 4.57 unit/mL. The antioxidant activity had significant increment from 5.32% in CME to 61.79% in CMB. A significantly higher emulsion stability index (ESI) and emulsification activity index (EAI) compared to CME were another result from hydrolyzation (p<0.05). Texture properties along with the shear force value and myofibrillar fragmentation index (MFI) were notably improved under CMB and CMBF in fresh condition. Marination with CM mushroom protease that previously hydrolyzed with enzymes were proven to also increase the nucleotide compounds, indicated by higher AMP and IMP in hydrolysate groups (p<0.05). The concentration of both total and insoluble collagen remained unchanged, meaning less effect from CM protease. Conclusion: This study suggested the hydrolyzation of CM protease with bromelain or a mixture of bromelain:flavourzyme to significantly improve functional properties of protease and escalate the taste-related nucleotide compounds and texture profiles from spent hen breast meat.

Quality of pork after electron-beam irradiation: A meta-analysis study.

Background and Aim: Irradiation has become a preferred method for pork preservation in recent years. Electron-beam irradiation is notably recognized for its feasibility and safety among various irradiation methods. This meta-analysis study aims to elucidate the impact of electron-beam irradiation on oxidation parameters, color, sensory attributes, and microbiological conditions in pork. Materials and Methods: A total of 79 data from 22 articles were aggregated into an extensive database. The irradiation dose ranged from 0 to 20 kGy in this current meta-analysis. The observed parameters encompassed oxidation, color, sensory attributes, and microbiological conditions. A mixed-model approach was used to perform the meta-data analysis, in which irradiation dose was treated as fixed effects and distinct studies (articles) as random effects. Results: Electron-beam irradiation resulted in an increase in thiobarbituric acid-reactive substances levels and peroxide-oxygen value of pork (p < 0.01). Conversely, total volatile-base-nitrogen values (p < 0.05) were observed. Following irradiation, the pH value, lightness (L*), redness (a*), and yellowness (b*) remained unaffected. Pork color tended to decrease after irradiation treatment (p = 0.095 and p = 0.079, respectively) at 7 and 14 days of storage. The irradiation process resulted in an increase in the values of texture and juiciness parameters (p < 0.05). However, electron-beam irradiation resulted in decreased overall acceptability (p = 0.089). In terms of microbiological status, electron-beam irradiation led to a reduction in the populations of Salmonella (p < 0.01), Escherichia coli (p < 0.01), Listeria monocytogenes (p < 0.05), and coliforms (p < 0.05) at 7 and 14 days of storage. Conclusion: Electron-beam irradiation enhances lipid peroxidation in porcine meat. The color of the meat remained unchanged after treatment. However, with regard to sensory properties, electron-beam irradiation showed a tendency to decreased overall acceptability. Most microbiological parameters decreased following electron-beam irradiation.

Validating duplex-PCR targeting ND2 for bovine and porcine detection in meat products.

Food authentication is a mandatory effort to assure the fair-trade. This study developed a duplex polymerase chain reaction (PCR) from the NADH dehydrogenase subunit 2 (ND2) gene to amplify specific segments of a cattle and porcine DNA. A universal forward primer composed of nineteen base pairs (bp) (3'-CCAAACACAACTCCGAAAA-5') and species-specific reverse primers composed of twenty (3'-CCAAACACAACTCCGAAAA-5') and twenty-one (3'-TGGCAAGAATTAGGACGGTTA-5') bp were used to limit the amplified DNA segment for porcine and cattle. The PCR reaction would generate a product with a profile of 168 and 227 bp, respectively. To investigate the accuracy and limit of detection, an in vitro experiment was conducted using simplex and duplex PCR on commercial meatballs randomly purchased from a commercial market in Surakarta, Indonesia. The findings of this study indicated that ND2 could be used as an alternative genetic marker for the identification of porcine and beef species in meat-derived products.

Artemisia vulgaris anthelmintic activities to ova and adult stages of Fasciola gigantica in vitro.

Background and Aim: Fasciolosis due to Fasciola gigantica is endemic to tropical countries and Fasciola hepatica in temperate climates, highly detrimental to livestock and known as foodborne zoonotic diseases. The strategic control of the disease is mainly the use of chemical anthelmintic. This study aimed to evaluate the anthelmintic properties of Artemisia vulgaris extract on the ova and adult stages of F. gigantica. Materials and Methods: Samples were collected from the Ampel Abbatoir, Boyolali District, Central Java, Indonesia. The ova from 20-gallbladders of cattle which were naturally infected with F. gigantica and 270 living F. gigantica worms were used in this study. The ovicidal assay was performed by incubating the ova with A. vulgaris in different concentrations, that is, 5%, 2.5%, and 1.25% for 5, 9, 11, 14, and 16 days. The efficacies were evaluated by quantification of ova degeneration during developmental stages in different time points and egg-hatch assay. The flukicidal effects were observed by mortality assay in 5, 10, 20, 40, 80, 160, 320, and 640 min incubations followed by scanning electron microscopy for surface morphology and histology of the fluke's transversal sections. Results: The concentration of 5% A. vulgaris showed the strongest ovicidal activities. The percentage of hatching ova on day 16 at concentrations of 5%, 2.5%, and 1.25% were 3.33%, 6.67%, and 16.67%. These ova hatch assay showed a significant reduction (p < 0.001) compared to untreated control. The flukicidal effect was significant (p < 0.001) at a concentration of 20%, with a mortality rate reaching 66.67% in the 40 min of incubation time. The surface properties of the adult worms, including the spine, tegument, acetabulum, intestine, and vitelline follicles, were disintegrated. Conclusion: The results showed that A. vulgaris has the potential ovicidal and flukicidal properties to F. gigantica. The active compounds remained necessary to be elucidated further and its modes of action would be interesting to be predicted by molecular docking modeling.

High prevalence of liver fluke infestation, Fasciola gigantica, among slaughtered cattle in Boyolali District, Central Java.

Background: Fasciolosis is a foodborne disease caused by Fasciola sp. infecting ruminants, especially cattle. Fasciolosis remains a significant concern for Veterinary Public Health because of its zoonosis risk and transmission mode. Aim: This study aimed to determine the prevalence and risk factors associated with Fasciola gigantica infestation in cattle at Ampel abbatoir, Central Java, Indonesia. Methods: A cross-sectional study was performed on 585 cattle from February to August 2022. Visual observation postmortem was used to assess Fasciola infection based on adult flukes in liver parenchyma and ductuli biliferi. Results: The overall prevalence of fasciolosis in Ampel abbatoir is high, reaching 25.12% (147/585). The highest prevalence was observed in the Ongole breed, 42.1% (24/57), female cattle, 38.72% (115/297), body condition score criteria of 2 50% (21/42), cattle aged >3.5 years 46.06% (82/178), and cattle originated from outside of Boyolali district 33.33% (71/213). Conclusion: This study showed a high prevalence of fasciolosis in Ampel abbatoir, as shown in the correlation between the risk factors of breed, sex, body condition score (BCS), origin, and age. Because of the high prevalence of fasciolosis in the abattoirs, it is essential to continue performing epidemiology studies in more expansive areas. The subsequent plans are important to reduce the risk of fasciolosis as a threat to productive cattle husbandry and warrant its transmission to humans as a foodborne-zoonotic disease.

Association of pleomorphic adenoma gene 1 with body weight and measurement of Bali cattle (Bos javanicus).

Background and Aim: Pleomorphic adenoma gene 1 (PLAG1) encodes a multifunctional transcription factor that controls many genes and pathways and is associated with cattle body weight and measurements. This study aimed to evaluate the association between PLAG1 polymorphisms with body weight and measurements in Bali cattle. Materials and Methods: A total of 87 Bali cattle, consisting of 48 bulls and 39 heifers at the Breeding Center for Bali Cattle, were used as the population in this study. Cattle were 2 years old and kept semi-intensively in the pasture. Phenotype data consisting of body weight, withers height, body length, chest girth, waist height, and chest depth were measured. Birth weight data were obtained from birth records, and weight gain, adjusted weaning weight, and yearling weight were calculated using formulas. Blood samples were taken from the jugular vein as much as 5 mL, and genomic DNA was isolated using the salting-out method. Polymerase chain reaction (PCR) was performed to amplify three target polymorphisms, namely, g.48308 C>T, g.32212 (19 bp indel), and g.45233 T>C. The presence of a 19 bp indel was determined by direct observation of the PCR product on a 2% agarose gel. Two other polymorphisms were detected by PCR-restriction fragment length polymorphism using the restriction endonuclease enzymes SacII and BclI. PLAG1 genotype and phenotype associations were analyzed using a general linear model. Results: The results showed that two of the target polymorphisms in PLAG1 did not vary. The DD genotype indicated by 123 bp of PCR product was the only genotype identified for g.32212 19 bp indel, and TT genotype was the only genotype found for g.45233 T>C single-nucleotide polymorphism (SNP). Conversely, g.48308 C>T SNP was found to be polymorphic. In addition, the g.48308 C>T polymorphism of PLAG1 was significantly associated with body length of Bali cattle. Cattle with the CC genotype had a greater body length than the other two genotypes. Conclusion: The g.48308 C>T SNP in PLAG1 was associated with Bali cattle body length characteristics. This finding could be used as a basis for selecting Bali cattle based on body length characteristics.

Genomic structure of Bali cattle based on linkage disequilibrium and effective population size analyses using 50K single nucleotide polymorphisms data.

Background and aim: Bali Cattle (Bos j. javanicus) is a local breed originating in Indonesia, accounting for 32.3% of the total cattle population. To date, no studies of the genetic structure and demographic status of Bali cattle have been conducted, even though the breeding of Bali cattle has a long and unique history that is likely to have impacted its genetic diversity. Therefore, a study that used molecular breeding technologies to characterize the demography of Bali cattle would be timely. This study aimed to examine genome diversity in Bali cattle and estimate the linkage disequilibrium (LD) and effective population size (N e) values in the cattle population. Materials and Methods: In this study, we explored the population structure and genetic diversity of Bali cattle using genomic-level analyses. Our study primarily studied cattle that had been bred in livestock breeding centers since these breeds had subsequently spread throughout Indonesia. We focused on characterizing the genetic structure, determining the level of LD present, and estimating the N e of the Bali cattle population. The genomic data used for this study were obtained from DNA samples of 48 Bali cattle collected at the Breeding Center of Bali Cattle as well as 54 genomic samples from Bali cattle collected elsewhere in Indonesia that had been used in recent publications. This genomic dataset included exclusively 50K single nucleotide polymorphisms (SNP) array (Illumina Bovine 50SNP bead chip, Illumina, USA) data. Results: We found that the LD values of Bali cattle from the breeding center and those raised elsewhere were 0.48±0.43 and 0.39±0.40, respectively. Subsequently, the Ne value of Bali cattle from the breeding center and farmers was 151 and 96, respectively. Conclusion: Our results suggest that the selection program of the breeding center is beneficial for maintaining the genetic diversity of Bali cattle.

First Autochthonous Report on Cattle Babesia naoakii in Central Java, Indonesia, and Identification of Haemaphysalis bispinosa Ticks in the Investigated Area.

In tropical countries, clinical bovine babesiosis is a tick-borne disease primarily caused by Babesia bovis and Babesia bigemina. Here, we investigated 11 cattle with presumptive diagnosis of clinical babesiosis in Boyolali district, Central Java, Indonesia. The majority of the animals were anemic, as evidenced by lower hematocrit, hemoglobin concentration, and red blood cell counts than the normal ranges. Blood DNA was analyzed by a PCR assay targeting the 18S rRNA-ITS region of babesial origin, and the results confirmed that the cattle were infected with Babesia species. The sequencing and phylogenetic analyses demonstrated that the animals were infected with Babesia naoakii. This is the first report of B. naoakii in Indonesia and of B. naoakii-induced clinical bovine babesiosis outside of Sri Lanka. B. naoakii causes a persistent infection, as indicated by positive PCR results for serial blood samples of the circulatory system taken two weeks after treatment. Consequently, subclinical or newly recovered cattle may serve as potential intermediate hosts and infect ticks as definitive hosts to complete the life cycle. To identify potential tick vectors, we collected ticks from cattle, including 11 animals with clinical babesiosis. Based on the morphology and the mitochondrial cytochrome c oxidase subunit 1 (COX1) of collected ticks, we found that all of the collected ticks were Haemaphysalis bispinosa, identifying this tick species as a potential vector of B. naoakii in Indonesia. In this study, the evaluation of local farmers' awareness and practices regarding tick-borne diseases is presented, as disease prevention is also reliant on the implementation of strategies for vector control. Since livestock activities in Java represent the country's busiest animal trade, thereby the spread of disease to other regions is possible through anthropogenic factors. In conclusion, B. naoakii is a causative pathogen of clinical bovine babesiosis autochthonously occurred in this report and further research on B. naoakii-infection is required in other regions of the country. The prompt treatment of the disease seemed crucial for animal survival, which implies the necessity of early diagnosis and a sensitive detection method.

A Novel Multiplex-PCR Assay to Detect Three Non-Halal Meats Contained in Meatball using Mitochondrial 12S rRNA Gene.

The objective of this study was to detect three non-halal meat products consisted of dog, pork, and rat species in meatball using novel multiplex-PCR with 12S rRNA gene as target sites. A total of 33 self-made meatballs were used, and they were grouped into eleven types of meatball based on meat species origin contained in the meatballs. Each type consisted of three meatballs. Extraction of genomic DNA from the meatballs was used as a DNA template for simplex-, duplex-, and multiplex-PCR processes. The result of simplex-PCR, duplex-PCR, and multiplex-PCR showed that the 12S rRNA primer gene successfully amplified DNA for each species bovine, dog, pig, and rat, which are respectively indicated by 155, 244, 357, and 491 bp of DNA bands. In addition, multiplex-PCR with 12S rRNA gene primers can be uniquely and accurately used for detection bovine, dog, pig, and rat species on beef meatball in one reaction.

Association of the thyroid hormone responsive spot 14 alpha gene with growth-related traits in Korean native chicken.

OBJECTIVE: Thyroid hormone responsive spot 14 alpha (THRSP) has been used to investigate the regulation of de novo lipogenesis because the variation of THRSP mRNA content in the tissue affects directly the ability of that tissue to synthetize lipids. Also, this gene responds to thyroid hormone stimulation and high level of carbohydrate feeding or insulin-injection. This study was carried out to investigate variations within THRSP and their effects on body and carcass weights in Korean native chicken (KNC). METHODS: A total of 585 chickens which represent the five lines of KNC (Black, Gray-Brown, Red-Brown, White, and Yellow-Brown) were reared and body weight data were recorded every two weeks from hatch until 20 weeks of age. Polymerase chain reaction- restriction fragment length polymorphism, DNA chips for Agilent 2100 Bioanalyzer, and Fluidigm Genotyping Technology, were applied to genotype selected markers. A linear mixed-effect model was used to access association between these single nucleotide polymorphism (SNP) markers and growth-related traits. RESULTS: A total of 30 polymorphisms were investigated in THRSP. Of these, nine SNPs for loci were selected to perform association analyses. Significant associations were detected between g.-49G>T SNP with body weight at 20 weeks of age (BW20), g.451T>C SNP with growth at 10 to 12 weeks of age (GR10-12), and g.1432A>C SNP with growth at 14 to 16 weeks trait (GR14-16) and body weight at 18 weeks of age (BW18). Moreover, diplotype of the THRSP gene significantly affected body weight at 12 weeks of age (BW12) and GR10-12 traits. Diplotype of ht1/ht2 was favorable for BW12 and GR10-12 traits. CONCLUSION: These results suggest that THRSP can be regarded as a candidate gene for growth traits in KNC.

Identification of quantitative trait loci for the fatty acid composition in Korean native chicken.

OBJECTIVE: Fatty acid composition is one of the most important meat quality traits because it can contribute to functional, sensorial, and nutritional factors. In this study, quantitative trait locus (QTL) analyses for fatty acid composition traits were investigated in thigh and breast meat of Korean native chicken (KNC). METHODS: In total, 18 fatty acid composition traits were investigated from each meat sample using 83 parents, and 595 F1 chicks of 20 week old. Genotype assessment was performed using 171 informative DNA markers on 26 autosomes. The KNC linkage map was constructed by CRI-MAP software, which calculated genetic distances, with map orders between markers. The half-sib and full-sib QTL analyses were performed using GridQTL and SOLAR programs, respectively. RESULTS: In total, 30 QTLs (12 in the thigh and 18 in the breast meat) were detected by the half-sib analysis and 7 QTLs (3 in the thigh and 4 in the breast meat) were identified by the full-sib analysis. CONCLUSION: With further verification of the QTL regions using additional markers and positional candidate gene studies, these results can provide valuable information for determining causative mutations affecting the fatty acid composition of KNC meat. Moreover, these findings may aid in the selection of birds with favorable fatty acid composition traits.

Genome scan linkage analysis identifies quantitative trait loci affecting serum clinical-chemical traits in Korean native chicken.

Alterations in robustness- and health-related traits lead to physiological changes, such as changes in the serum clinical chemical parameters in individuals. Therefore, clinical-chemical traits can be used as biomarkers to examine the health status of chickens. The aim of the present study was to detect the quantitative trait loci (QTLs) influencing eight clinical-chemical traits (glucose, total protein, creatinine, high-density lipoprotein cholesterol, total cholesterol, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and α-amylase) in an F1 nuclear families comprising 83 F0 founders and 585 F1 progeny of Korean native chickens. Genotypic data on 135 DNA markers representing 26 autosomes have been generated for this resource pedigree. The total length of the map was 2729.4 cM. We used a multipoint variance component linkage approach to identify QTLs for the traits. A significant QTL affecting serum α-amylase levels was identified on chicken chromosome (GGA) 7 [logarithm of odds (LOD) = 3.02, P value = 1.92 × 10(-4)]. Additionally, we detected several suggestive linkage signals for the levels of total cholesterol, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and creatinine on GGA 4, 12, 13, and 15. In this study, serum α-amylase levels related significant QTL was mapped on GGA7 and cholesterol, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and creatinine traits related suggestive QTLs were detected on GGA4, 12, 13 and 15, respectively. Further verification and fine mapping of these identified QTLs can provide valuable information for understanding the variations of clinical chemical trait in chickens.

Variance Component Quantitative Trait Locus Analysis for Body Weight Traits in Purebred Korean Native Chicken.

Quantitative trait locus (QTL) is a particular region of the genome containing one or more genes associated with economically important quantitative traits. This study was conducted to identify QTL regions for body weight and growth traits in purebred Korean native chicken (KNC). F1 samples (n = 595) were genotyped using 127 microsatellite markers and 8 single nucleotide polymorphisms that covered 2,616.1 centi Morgan (cM) of map length for 26 autosomal linkage groups. Body weight traits were measured every 2 weeks from hatch to 20 weeks of age. Weight of half carcass was also collected together with growth rate. A multipoint variance component linkage approach was used to identify QTLs for the body weight traits. Two significant QTLs for growth were identified on chicken chromosome 3 (GGA3) for growth 16 to18 weeks (logarithm of the odds [LOD] = 3.24, Nominal p value = 0.0001) and GGA4 for growth 6 to 8 weeks (LOD = 2.88, Nominal p value = 0.0003). Additionally, one significant QTL and three suggestive QTLs were detected for body weight traits in KNC; significant QTL for body weight at 4 weeks (LOD = 2.52, nominal p value = 0.0007) and suggestive QTL for 8 weeks (LOD = 1.96, Nominal p value = 0.0027) were detected on GGA4; QTLs were also detected for two different body weight traits: body weight at 16 weeks on GGA3 and body weight at 18 weeks on GGA19. Additionally, two suggestive QTLs for carcass weight were detected at 0 and 70 cM on GGA19. In conclusion, the current study identified several significant and suggestive QTLs that affect growth related traits in a unique resource pedigree in purebred KNC. This information will contribute to improving the body weight traits in native chicken breeds, especially for the Asian native chicken breeds.