The research aimed to determine the impact of synbiotic: 6 g of prebiotic inulin and 5 g of probiotic Saccharomyces cerevisiae strain 1026 on calves' productivity and greenhouse gas (GHG) production. The research was conducted with 10 Holstein Friesian and Red Holstein (Bos taurus L.) crossbreed calves of mean age 33±6 days and initial body weight 73.4±12.75 kg. We added the synbiotic into the diet of five dairy crossbreed calves (SynG) and five calves in control group (CoG) received non-supplemented diet. The duration of the experiment was 56 days. The weight of calves and amount of methane (CH4) and carbon dioxide (CO2) in the rumen were determined on day 1, 28, and 56. On day 56, three calves from each group were slaughtered. Meat samples were assessed for some indicators of meat quality. The main methanogens were detected in the rumen fluid and feces. The weight gain during the whole experiment period of 56 days was higher in the SynG (62.6±13.75 kg) compared to CoG (36.8±7.98 kg) calves (p<0.01). There were no significant differences in the levels of protein (%), fat (unsaturated and saturated – %), and cholesterol (mg/100 g) in meat samples from both groups. At the end of the experiment, the amount of CH4 in calves' rumen in CoG was higher (Me=792.06 mg/m3, interquartile range [IQR] 755.06-873.59) compared to SynG (Me=675.41 mg/m3, IQR 653.46-700.50) group (p<0.01). The values for CO2 were also increased in CoG (Me=4251.28 mg/m3, IQR 4045.58-4426.25) compared to SynG (Me=3266.06 mg/m3, IQR 1358.98-4584.91) group (p=0.001). There were no significant differences in the calves' weight and certain methanogen species in rumen liquid and feces on the 56th day of the experiment. Significantly higher results in the parameter total prokaryotes (V3) (bacteria+archaea) in rumen fluid were in SynG, whereas significantly higher results in the parameter total methanogens Met630/803 in rumen fluid were in CoG, p<0.05. The main results showed that the synbiotic can increase the daily weight gain in calves and decrease the amount of GHG in rumen but does not impact different methanogen species in rumen liquid and feces and meat protein, fat, and cholesterol levels. Keywords: calves, greenhouse gases, inulin, productivity, Saccharomyces cerevisiae, symbiotic.

The study was undertaken to find out the changes in biochemical and hormonal profile during transition period in single- and twin-bearing Beetal goats. Beetal goats are reared in North India for milk and meat purposes, popularly called poor man's cow, serving as a source of livelihood. The present study was conducted on Beetal goats, maintained at a goat farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, for characterization of plasma biochemical and hormonal changes during transitional phase in pregnant goats carrying single and twin fetus. The animals selected were expected to have parturition at the same time, to avoid environmental variation effects, and divided into two groups: Group 1 (carrying single fetus, n=14) and Group 2 (twin fetus, n=6). Blood samples were collected from goats prepartum (–30 days and –15 days), 0 day, and postpartum +15 days and +30 days, and plasma was separated for the analysis of biochemical metabolites and hormones by commercially available kits. Analysis of data revealed a significant decrease in triglyceride and cholesterol concentration during postpartum days. A significant increase in alanine aminotransferase (ALT) concentration was observed at the day of parturition. Furthermore, there was a significant (p≤0.05) decrease in ALT and aspartate aminotransferase concentration in goats at the 15th day and 30th day after parturition in both groups. A significant (p≤0.05) lower concentration of total protein and globulins was observed during late pregnancy and on the day of parturition, with a subsequent elevation during postpartum period (15 days and 30 days). Progesterone concentration was highest at –30 days prepartum and decreased sharply at parturition and thereafter postpartum, both in single- and twin-bearing goats. Progesterone level was lower in twin-bearing goats as compared to single-bearing goats at 0 day and postpartum days. No significant changes were observed in sodium, potassium, and chloride concentration during transitional period in goats. Transition period blood metabolite profile changes due to physiological need of mother and fetus were more prominent in metabolites regulating energy requirements and in goats bearing twin fetus. Keywords: biochemical, enzymes, goats, hormones, transitional period.

Candida albicans is a dimorphic fungus that has both yeast and filamentous forms. It is part of the normal flora in the oral and genital areas of mammals. One factor for the pathogenicity of C. albicans is its ability to switch from yeast to hyphae. The hyphal form adheres and penetrates tissues more readily than the yeast form and produces biofilms that are associated with chronic infection. Biofilms are protective niches that enable microorganisms to be more resistant to antibiotic treatment, thus allowing for persistent infection. The first stage in the transition from yeast to hyphae involves the formation of a germ tube, and this transition is triggered by interactions with host cells. Germ tube formation is dependent on serum, pH, temperature, and quorum-sensing molecules (QSMs). Farnesol, which is a QSM in C. albicans, can prevent yeast to hyphae conversion and inhibits the growth of fungal biofilm. Lyticase is a synergistic enzyme complex that catalyzes yeast cell lysis by β-1,3-glucanase and is a highly specific alkaline protease that produces protoplasts or spheroplasts. This study investigated the effect of farnesol and lyticase on the formation of C. albicans biofilms. C. albicans ATCC 2091 was cultivated on liquid and solid Sabouraud media. The presence of C. albicans was confirmed using HiCrome Candida Agar chromogenic medium. Enzyme activities were assayed using a HiCandida Identification Kit. The morphology and densitometry parameters of C. albicans biofilms were considered in the presence of farnesol (Sigma-Aldrich, Germany), lyticase (from Arthrobacter luteus; Sigma-Aldrich, Germany), and farnesol–lyticase. This study shows that both farnesol and lyticase possess antifungal activity against C. albicans biofilms. A significant difference among treatment groups (p<0.05) was observed from strong biofilm production to medium and weak. Many studies have been devoted to the antimicrobial action of farnesol. Bacterial enzyme lyticase is also used to degrade fungal cell walls. Both molecules show substantial antifungal properties that are similar to the properties of modern antimycotics. The current study demonstrates that farnesol and lyticase can disrupt biofilm formation in C. albicans ATCC 2091, which is an effective biofilm producer. Keywords: antimycotic activity, biofilms, Candida albicans, enzyme activities, farnesol, hemolytic activity, HiCrome Candida Agar, lyticase, optical density, quorum sensing.

Antimicrobial resistance is a global health threat. This study investigated the prevalence of Escherichia coli in imported 1-day-old chicks, ducklings, and turkey poults. The liver, heart, lungs, and yolk sacs of 148 imported batches of 1-day-old flocks (chicks, 45; ducklings, 63; and turkey poults, 40) were bacteriologically examined for the presence of E. coli. We isolated 38 E. coli strains from 13.5%, 6.7%, and 5.4% of imported batches of 1-day-old chicks, ducklings, and turkey poults, respectively. They were serotyped as O91, O125, O145, O78, O44, O36, O169, O124, O15, O26, and untyped in the imported chicks; O91, O119, O145, O15, O169, and untyped in the imported ducklings; and O78, O28, O29, O168, O125, O158, and O115 in the imported turkey poults. The E. coli isolates were investigated for antibiotic resistance against 16 antibiotics using the disk diffusion method and were found resistant to cefotaxime (60.5%), nalidixic acid (44.7%), tetracycline (44.7%), and trimethoprim-sulfamethoxazole (42.1%). The distribution of extended-spectrum β-lactamase (ESBL) and ampC β-lactamase genes was blaTEM (52.6%), blaSHV (28.9%), blaCTX-M (39.5%), blaOXA-1 (13.1%), and ampC (28.9%). Imported 1-day-old poultry flocks may be a potential source for the dissemination of antibiotic-resistant E. coli and the ESBL genes in poultry production. Keywords: Escherichia coli, extended-spectrum β-lactamase, imported, multidrug resistance, poultry.

Himatanthus sucuuba (Spruce) Woodson (Apocynaceae) is a medicinal plant known as "Bellaco- Caspi" widely distributed in Loreto, Peru. In the Peruvian traditional medicine, the latex of the plant is used for the treatment of wounds, inflammation, ulcers, and other ailments. This study aims to evaluate the wound healing effect of the latex of H. sucuuba in BALB/C albino mice. Thirty BALB/C male mice were used for wound healing study. In the experimental procedures, wound skin incision was performed at 2.0 cm in length until subcutaneous on the paravertebral of each animal. Under locally anesthetized with procaine cream, the treatment was conducted. All the mice were divided into three groups, the control group (A), zinc oxide cream (B), and H. sucuuba latex (C). The entire surface of the wound was treated for all the groups, and the treatments were performed daily for 15 days. The experiments were stopped on days 1, 7, and 15, respectively. The histopathological study of tissues revealed significant changes in wound healing effect in H. sucuuba latex compared to the control and B groups. Consequently, the mice treated with latex showed a significant reduction in epithelialization time and collagen formation. Furthermore, the latex showed a dose-dependent significant reduction of inflammation in the first 24 h of treatment. BALB/C mice treated with the latex of H. sucuuba possess a wound healing effect that can scientifically prove the traditional use of the plant as a wound healing agent. Keywords: Apocynaceae, BALB/C mice, latex, pathology, regeneration, wound healing.

Global trend to remove the antibiotic growth promoter (AGP) from animals contributes to the exploration of successful measures to sustain production and reduces the intestinal diseases in the post-AGP era. Plant extracts, therefore, have been used to improve performance and intestinal health. Here, we conducted a study to evaluate the effects of neem (Azadirachta indica) leaf extracts (NLE) as alternatives to AGPs in broiler chickens. Sixty day-old broiler chicks were assigned to 12-floor pens, each stocked with five birds and divided into three treatment groups of four pens per treatment. The groups were: Negative control, basal diet without additives; positive control, basal diet with antibiotics and vitamins; and NLE treated group, basal diet supplemented with 0.1% aqua extract of neem leaves. Overall feed intake was significantly (p≤0.05) highest in the positive control. Higher body weight, higher dressing percentage, and lower feed conversion ratio were observed in birds treated with NLE compared to the negative control group (p≤0.05) but not the positive control group (p>0.05). There was no significant variation in hematology between different groups. Furthermore, the economic evaluation indicated that the NLE treatment was found cheaper than control and antibiotic treatment in cost-benefit analysis. We suggest NLE might be a cheaper alternative to antibiotics in broiler production as indicated by improved body weight and feed efficiency. Keywords: alternatives to antibiotics, broiler chickens, growth performance, neem leaf extracts, production cost.

Food of animal origin such as milk is vital for nutritional security and should be free of any antibiotic residues due to its public health significance. We designed a study aiming to determine the occurrence of antibiotic residues and in further levels of oxytetracycline residues in pooled raw milk samples from Palakkad district, Kerala. We collected pooled raw milk samples were collected from Alathur, Chittoor, and Palakkad blocks of Palakkad district, Kerala. A total of 215 samples were screened for antibiotic residues by microbial inhibition assay (MIA) and the positive samples were subjected to enzyme-linked immunosorbent assay (ELISA) to determine oxytetracycline residues, this was further confirmed using high-performance liquid chromatography (HPLC). We found that out of the 215 pooled raw milk samples screened for antibiotic residues using MIA, 22 samples (10.23%) were positive for antibiotic residues from Palakkad, Kerala. Out of these 22 samples, five (2.33%) were positive for oxytetracycline residues. We further calculated the mean concentration of oxytetracycline residues in these five samples and estimated it to be 201.00±41.25 ng/mL and 272.11±53.21 ng/mL using ELISA and HPLC, respectively. On analyzing these five samples, we found that four samples (1.86%) exceeded the maximum residue limits level of 100 ng/mL for oxytetracycline residues in milk as specified by Codex Alimentarius Commission/Food Safety and Standards Authority of India (FSSAI). This study revealed that the occurrence of oxytetracycline residues in pooled raw milk samples in the Palakkad district of Kerala. Hence, there is a need for surveillance and monitoring of antibiotic residues in milk due to its impact on public health to ensure consumer safety. Keywords: enzyme-linked immunosorbent assay, high-performance liquid chromatography, microbial inhibition assay, oxytetracycline residues, Palakkad, pooled milk.

This study aimed to determine the prevalence of layer flock tumor disease in Lower Egypt during the period of 2018- 2019 and to undertake molecular characterization and determine the genetic diversity of all identified viruses. Forty samples were collected from layer chicken located in six governorates of Lower Egypt during the period of 2018-2019. Samples were taken from tumors in different organs. Tumor tissues were identified by histopathological sectioning and then further confirmed by a reverse-transcription polymerase chain reaction. Finally, genetic evolution of Avian leukosis virus (ALV-J) gp85 gene was studied. All the study samples were negative for Marek's disease virus, reticuloendotheliosis virus, ALV (A,B,C and D) and 20 samples were positive for ALV-J in backyard in six governrates. Sequencing of ALV-J gp85 gene was performed for six representative samples (one from each governorate), and they were found to be genetically related to prototype virus HPRS-1003 (identity percentage: 91.2-91.8%), but they were from a different group that was similar to the AF88-USA strain (first detected in 2000) with specific mutations, and they differed from a strain that was previously isolated in Egypt in 2005, forming two different subgroups (I and II) that had mutations in the hr1domain (V128F, R136A) and hr2 domain (S197G, E202K). The ALV-J virus was the main cause of neoplastic disease in layer chickens from Lower Egypt in the period of 2018-2019. We found that the genetic evolution of ALV-J gp85 gene was related to prototype virus HPRS-1003 but in a different group with a specific mutation. Further studies are needed to evaluate the antigenicity and pathogenicity of recently detected ALV-J strains. Keywords: Avian leukosis (J), gp85 gene, Marek's disease, reticuloendotheliosis virus, tumor viruses.

Water is the most important nutrient for the production of healthy livestock. Water scarcity bottlenecks livestock production in arid and semi-arid regions, particularly during the dry season. This study aimed to assess water availability and quality for small ruminants, and to identify major challenges of meeting their water requirements in two major small ruminant production systems in Jordan. Transhumant and sedentary production systems in the Northern Badia region of Jordan were the focus of this study. A questionnaire was distributed to 120 sheep and goat farmers (62 transhumant farmers and 58 sedentary farmers) and a water quality assessment was completed. Results showed that the two production systems varied their water source use seasonally. Water provision was perceived as one of the major constraints for Bedouins, particularly during the dry season in transhumant production systems, when longer distances to water sources and the high costs of fetching water daily aggravated the problem. The mean distance and travel times to the boreholes were less in the sedentary system. Watering frequency was significantly lower in the transhumant system compared to the sedentary system (p<0.05). Although the values of water quality parameters complied with guidelines for livestock consumption, low water quality was the main concern expressed by some of the survey households. Technical support to properly design, manage, and rehabilitate surface water harvesting systems is required for the sustainable use of water resources in the study region. Moreover, systematic water quality monitoring is necessary to ensure its suitability for livestock use. Further investigations on the microbiological quality of water and its effect on the health and performance of livestock are recommended. Keywords: Jordan, livestock, water access, water quality, water scarcity.

This study aimed to assess fundamental biochemical values of healthy animals and to provide useful data on comparative physiologies of Testudines, being assessed the serum biochemistry profiles, and body and tail biometry of Trachemys scripta elegans and Trachemys dorbignyi bred in interaction in the semiarid region of the São Francisco river valley. Serum biochemistry variables (urea, creatinine, glucose, total serum protein, albumin, globulin, and albumin/globulin ratio), and biometry values of the body (mass [body mass (BM)], maximum curvilinear length [carapace length (CL)], and width [carapace width (CW)] of the carapace, maximum curvilinear length [plastron length (PL)], and width [plastron width PW] of the plastron), and the tail (total length of the tail [TLT], pre-cloacal tail length [PrCL], post-cloacal tail length [PoCL]) were measured after 24 h fasting. T. s. elegans displayed higher BM, CL, CW, PL, PW, AST, TP, albumin, and globulin values. T. dorbignyi displayed higher values of glucose, TLT, and PrCL. Variables aspartate aminotransferase (AST) and total protein (TP) in T. s. elegans and glucose in T. dorbignyi explained most of the variance between the species and could serve to distinguish them. We conclude that most of the differences between T. s. elegans and T. dorbignyi shall be explained by biometric variables, AST, TP, and glucose, which characterize interspecific differences. Our results point out terms of reference for these species bred in captivity in the semiarid region of Brazilian Northeastern region and serve as a model for the comparative intra- and inter-species physiology and as a base for the health assessment of these species. Keywords: albumin, biometry, creatinine, glucose, urea.

This paper presents the results of the studies on "Stimix Zoostim" and "Normosil" probiotics and their influence on fecal microflora, hematologic indicators, immunoresistance, nutrient digestibility, and growth intensity of mother-bonded calves. The calves of the control group were fed with their basic diet (BD) without "Stimix Zoostim" or "Normosil", whereas the calves of the experimental group were fed with their BD supplemented with "Stimix Zoostim" and "Normosil". For 10-20-day-old calves, the daily dose was 10 mL per head, whereas 21-90-day-old calves received 15 mL of probiotics per head per day. The calves of the experimental group were administered probiotics every day. Calves aged 10 to 60 days received probiotics with milk, and then at the age of 61 to 90 days, received probiotics with water. (This is a sequential process. At 60 days old, calf rearing with milk stops, and after that, we use probiotics added to water to rear them). Both groups were administered probiotics twice a day, specifically 50% of the daily dose at each feeding time. The results in this study proved that "Stimix Zoostim" and "Normosil" probiotics exhibit high probiotic activity and have a positive effect on the calves' fecal microflora. Feeding the calves with probiotics resulted in a significant increase in the number of normal flora, such as lactic acid bacteria and bifidobacteria, and a decrease in the amount of Escherichia. It also resulted in an increase in red blood cells, hemoglobin, and ?-globulins within the physiological range. In our study, we found that the phagocytic reaction in the blood serum of the experimental group was slightly enhanced, suggesting a high response of the body to infectious agents. Thus, the group of calves receiving probiotic "Stimix Zoostim" exhibited an exceedance of phagocytic activity by 4.8% and the group receiving "Normosil" by 4.4%, in comparison with the control group. The daily dose of 10 mL of probiotics per head for 10-20-day-old calves and 15 mL per head for 21-90-day-old calves also had a positive effect on nutrient digestibility, growth, and forage consumption. The economic benefit per animal was 149.23 and 157.0 rubles, respectively. Keywords: average gain, blood immunoresistance, forage consumption, hematologic indicators, mother-bonded calves, nutrient digestibility.

Rabbit hemorrhagic disease (RHD) is an economically important disorder of rabbits, where infection results in severe losses to the meat and fur industries. Our goal was to characterize the RHD virus (RHDV) strains currently circulating in different regions of Egypt. Fifty rabbits suspected of harboring RHDV from 15 Egyptian governorates were evaluated. Diseased rabbits were identified by clinical signs and postmortem lesions. RHDV was confirmed through hemagglutination assay (HA) and polymerase chain reaction (PCR). Partial sequencing of the VP60 gene was performed for genotyping. From 50 rabbits, we identified 16 cases of RHDV (32%) by HA and PCR, including seven males and nine females. We identified two distinct genotypes through sequencing of an amplified fragment of the virus VP 60 gene. One group is composed of those circulating primarily in upper Egypt, which is closely related to the classical G3-G5 virus strains, and the second group, circulating predominantly in lower Egypt, was more closely related to the RHDV2 variant. The overall nucleotide sequence identity ranged from 78.4% to 100%, and identity with the vaccine strains ranged from 78.8% to 91.1%. Our results constitute important documentation of RHDV strains currently circulating in Egypt. The findings suggest that there may be a limit to the effectiveness of currently applied vaccine strains as this formulation may not cover all circulating strains. A wider investigation that includes both domestic and wild rabbits will be needed to identify appropriate control measures for this disease. Keywords: Egypt, native rabbits, prevalence, rabbit hemorrhagic disease virus, VP60.

Feline lower urinary tract disease (FLUTD) is one of the common cat diseases. The aim of this study was to detect feline idiopathic cystitis (FIC) as a cause of FLUTD in Sleman Regency, which is a problem in the population. Seventy-three cats with FLUTD symptoms were used from seven veterinary practices in Sleman Regency. The logging of each cat's medical history, clinical examination, urinalysis, routine blood screening, and ultrasonography was conducted to diagnose the cause of FLUTD. The percentages of diseases causing FLUTD included FIC 21.9%, urolithiasis 57.5%, urinary tract infection (UTI) 16.4%, neoplasia 1.4%, trauma 1.4%, and nervous disorders 1.4%. FIC, one of the causes of FLUTD, is found in cats and has become a problem among the cat population in Sleman Regency, Yogyakarta, Indonesia. Various handling and preventive efforts should be undertaken against the disease. Keywords: cat, detection, feline idiopathic cystitis, feline lower urinary tract disease, Indonesia, Sleman.

Pork leanness and marbling are among the essential traits of consumer preference. To acquire knowledge about universal epigenetic regulations for improving breed selection, a meta-analysis of methylated DNA immunoprecipitation sequencing (MeDIP-seq) profiling data of mixed loin muscle types was performed in this study. MeDIP-seq profiling datasets of longissimus dorsi muscle and psoas major muscles from male and female pigs of Landrace and Tibetan breeds were preprocessed and aligned to the porcine genome. Analysis of differential methylated DNA regions (DMRs) between the breeds was performed by focusing on transcription start sites (TSSs) of known genes (–20,000-3000 bases from TSS). All associated genes were further reviewed for their functions and predicted for transcription factors (TF) possibly associated with their TSSs. When the methylation levels of DMRs in TSS regions of Landrace breed were compared to those of Tibetan breed, 10 DMRs were hypomethylated (Landrace < Tibetan), and 19 DMRs were hypermethylated (Landrace > Tibetan), accordingly (p≤0.001). According to the reviews about gene functions, all associated genes were pieces of evidence for their roles in a variety of muscle and lipid metabolisms. Prediction of the binding TFs revealed the six most abundant binding TFs to such DMRs-associated TSS (p≤0.0001) as follows: ZNF384, Foxd3, IRF1, KLF9, EWSR1-FLI1, HES5, and TFAP2A. Common DMRs-associated TSS between the lean-type and the marbled-type loin muscles were identified in this study. Interestingly, the genes associated with such regions were strongly evidenced for their possible roles on the muscle trait characteristics by which further novel research topics could be focused on them in the future. Keywords: differential DNA methylation, lean, loin muscle, marbled, methylated DNA immunoprecipitation sequencing, transcription start site.

The present study was performed to assess the association of single-nucleotide polymorphisms (SNPs) in the fatty acid-binding protein 4 (FABP4) gene with birth weight (BW), final weight (FW), and average daily gain (ADG) in three Egyptian sheep breeds. Genomic DNA was extracted from the blood samples of 50 male and female individuals representing Ossimi, Rahmani, and Barki sheep breeds. A 407 bp nucleotide (nt) segment from the first intron of FABP4 was amplified by polymerase chain reaction, sequenced, and analyzed in the different samples. Sequence analysis of the determined segment (407 bp) revealed four SNPs (all transition types) at nt position 372 (CP011894.1:g.57605471) A>G, nt position 211 (CP011894.1:g.57605632) A>G, nt position 143 (CP011894.1:g.57605700) T>C, and nt position 111 (CP011894.1:g.57605732) T>C. The allelic and genotypic frequencies for the identified SNPs in the sheep breeds were calculated. At nt positions 372 and 211, two alleles were identified (A and G). Only two genotypes were present at nt position 372 (AA and AG), while three genotypes were present at nt position 211 (AA, AG, and GG). Two alleles (T and C) and three identified genotypes (TT, TC, and CC) were detected at nt positions 143 and 111. Analysis of the results revealed that AA genotype at nt position 372 is associated with higher estimates for BW, FW, and ADG when compared to all the other genotypes. Very high correlation coefficients were found between the genotypes 143-TT and 111-TT and also between 143-TC and 111-TC. The genotypes 372-AG, 211-GG, 211-AA, 143-TT, 143-CC, 111-TT, 111-TC, and 111-CC were associated with negative effects on BW, FW, and ADG. The detection of four SNPs in a partial sequence of the Egyptian ovine FABP4 gene intron 1 reflected that this gene harbors substantial diversity. In addition, a novel SNP at nt position 372 (CP011894.1:g.57605471) A>G was associated with higher estimates for BW, FW, and ADG. Keywords: body weight, FABP4 gene, growth rate, sheep, single-nucleotide polymorphisms.

Direct observation, scanning electron microscopy (SEM) is a common method used for the observations of biofilms. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide) (EDC) fixation method has proven to be a valuable fixation method in the observation of these biofilms. Still, it entails a method of biofilm fixation that can damage slim structures, leading to the impossible observation of biofilm development. In contrast, alcian blue and lysine (ABL) fixation technique appears more glycocalyx of biofilm, fully preserved samples, which may provide much insight into the development of B. subtilis biofilms. Here, the evaluation of the fixation of ABL technique for the study of B. subtilis biofilms was carried out in situ, on Congo red agar. In doing so, the comparison to commonly use conventional EDC technique for sample fixation, and observation was carried out. Observations were based on SEM over 30 samples. Overall, ABL technique provided excellent observation of biofilms formed in situ, on Congo red agar, and revealed slime structures, which have not been observed, much in standard EDC fixation or earlier in other studies of these biofilms in B. subtilis. This study reported the appropriate use of ABL in the fixation technique for the preservation of biofilm of B. subtilis. Keywords: alcian blue, Bacillus subtilis, biofilm, Congo red agar, scanning electron microscopy.

In Indonesia, highly pathogenic avian influenza (HPAI) H5N1 outbreaks in poultry are still reported. The disease causes a decrease in egg production and an increase in mortality; this has an impact on the economic losses of farmers. Several studies have considered that ducks play a role in the HPAI endemicity in the country; however, little is known about whether or not the type of duck farming is associated with HPAI H5 virus infection, particularly within clade 2.3.2.1c, which has been predominantly found in poultry since 2014. A cross-sectional study was conducted to determine the HPAI seroprevalence for H5 subtype clade 2.3.2.1c in laying ducks that are kept intensively and nomadically and to determine the associated risk factors. Forty-nine duck farmers were randomly selected from ten sub-districts in Purbalingga District, Central Java, Indonesia; a cross-sectional study was implemented to collect field data. Based on an expected HPAI prevalence level of 10%, estimated accuracy of ± 5%, and 95% confidence interval (CI), the total sample size was calculated at 36 individuals. Samples must be multiplied by 7 to reduce bias; thus, 252 ducks were taken as samples in this study. Considering that the maintenance and duck handling were uniform and farmers complained that the effect of activity to take duck samples would reduce egg production, this study only took samples from 245 ducks (oropharyngeal swabs and serum). Those samples were taken from five birds on each farm. Hemagglutination inhibition tests examined the serum samples for HPAI H5 Clade 2.3.2.1c, and pool swab samples (five swabs in one viral media transport) were examined by real-time reverse transcription-polymerase chain reaction (qRT-PCR) test for influenza Type A and H5 subtype virus. Information regarding farm management was obtained using a questionnaire; face-to-face interviews were conducted with the duck farmers using native Javanese language. Serum and swabs from 245 ducks were collected in total. For individual birds, 54.69% (134/245) of serum samples were H5 seropositive. Seroprevalence among nomadic ducks was 59.28% (95% CI: 0.48-0.61), which was higher than among intensively farmed ducks (48.57%, 95% CI: 0.38-0.58). Farm-level seroprevalence was 50% (95% CI: 0.30-0.69) for nomadic ducks but only 28.57% (95% CI: 0.11-0.51) for intensively farmed ducks. The farm-level virus prevalence (proportion of flocks with at least one bird positive for influenza Type A) was 17.85% (95% CI: 0.07-0.35) for nomadic ducks and 4.76% (1/21) for intensively farmed ducks (95% CI: 0.008-0.23). All influenza Type A positive samples were negative for the H5 subtype, indicating that another HA subtype AI viruses might have been circulating in ducks in the study area. A relationship between duck farms that were H5 seropositive and their maintenance system was present; however, this relationship was not significant, the nomadic duck system detected 2 times higher H5-seropositive ducks than the intensive farming system (OR: 2.16, 95% CI: 0.33-14.31). This study found that the seroprevalence of HPAI in the duck population level in Purbalingga was 54.69% and demonstrated that the nomadic duck farming system was more likely to acquire HPAI H5 infection than the intensive farming duck system. Other risk factors should be further investigated as the diversity of the farming system is partially related to HPAI H5 infection. Keywords: avian influenza, ducks, farming system, highly pathogenic avian influenza H5, risk factors, seroprevalence.

Congestive heart failure (CHF) is a clinical disorder that results from cardiac dysfunction with subsequent fatal outcomes in most cases. Several diseases are incriminated in occurrence of CHF. Therefore, the aims of this study were to identify CHF etiology and associated clinical findings in 67 cows and to investigate the relationship between CHF and the other body organs using ultrasonographic examination. Sixty-seven cows affected by CHF admitted to the clinic with a history of loss of appetite, decrease in milk production, constipation, and brisket edema were thoroughly investigated clinically and ultrasonographically. In addition, ten apparently healthy cows were used as a control group. Clinically, cows with CHF manifested jugular engorgement and pulsation (88.1%), brisket and/or intermandibular edema (77.6%), and muffled heart sounds (76.1%). Based on the ultrasonographic examination, traumatic pericarditis (82.1%) was the most prevalent etiology of CHF. Extracardiac etiology of CHF identified were exudative pleurisy (10.4%) and mediastinal abscesses (7.5%). Hepatomegaly (88.1%) and pleural effusion (61.2%) were the most documented consequences. Both cardiac and extracardiac diseases could be associated with CHF in cattle. Ultrasonographic changes in liver and pleura secondary to CHF were the most common findings. Ultrasonography is a good tool for the diagnosis of cardiac and extracardiac etiologies of CHF in cattle. Keywords: cattle, congestive heart failure, mediastinal abscess, pericarditis, pleurisy.

The aims of this study were first to estimate calcification in the esophagus and abomasum of cows and second to quantify its appearance with increasing age using histopathological and immunohistochemical techniques. Esophageal and abomasal samples from 24 healthy cows (Bos taurus) were collected. Hematoxylin and eosin, alizarin red, and von Kossa stains were used for histopathological analysis. Histopathological changes were confirmed with immunohistochemical staining, followed by digital image analysis. Histological findings revealed the esophagus and abomasum wall comprised four fundamental layers, the mucosa, submucosa, muscularis, and serosa or adventitia. At 1 year old, calcification was beginning to appear as fine diffused points in mucosa, submucosa, and muscularis of both esophagus and abomasum, appearing as small spots at 2 years old. With advancing age in all animals, this calcification began to appear as medium spots spread throughout all wall layers of these organs at 3 years old. By 4 years old, calcification had evolved into large dark foci spread substantially throughout the tunica submucosa and tunica muscularis. Immunohistochemical results exhibited positive immunoreaction to calcium salts in the esophagus and abomasum layers in all animals, which increased with age. The current study concluded that calcification is a pathological event appearing spontaneously in various types of soft tissue, significantly increasing with age, either because of hypercalcemia and hyperphosphatemia or secondary to other diseases. Keywords: abomasum, alizarin red, calcification, esophagus, immunohistochemistry, von Kossa.

The purpose of the present study was to evaluate the effects of Beauveria bassiana (Hypocreales) inoculum on plant growth, volatile constituents, and tick repellency of the extracts of Tulbaghia violacea (Amaryllidaceae). Eight-week-old potted seedlings of T. violacea were each inoculated with conidia of B. bassiana (strain SM3) suspended at a concentration of 1×106 conidia mL–1. Tissue colonization by fungal conidia was assessed after 3 weeks. Plant growth, volatile constituents, and tick repellency were assessed after 12 weeks post-treatment. B. bassiana conidia successfully colonized leaf and root tissues of T. violacea. The growth of fungal hyphae out of the leaf and root sections occurred in 75% and 91.6% of plants, respectively. Inoculation of the plants with B. bassiana significantly (p<0.05) influenced root length and plant height but did not have substantial effects on weights and leaf number of T. violacea. While the fungus did not have significant effects on overall number of the volatile chemical constituents, significant variations in the quantity (area ratio) were observed in at least four compounds that were detected. In the tick repellency bioassay, high concentration (20 w/v%) of acetone extract from fungus-exposed plants produced the least repellent effect on Rhipicephalus appendiculatus larvae (Ixodidae), while at lower concentrations (5 w/v% and 10 w/v%) of acetone extracts of T. violacea, tick repellent activity of the extract of the fungus treatment was significantly improved and was comparable to commercial N,N-Diethyl-m-toluamide and the other treatments. Experimental fungal inoculation positively influenced plant growth in height and root length and tick (R. appendiculatus) repellency of acetone extracts of T. violacea at a concentration of 10 w/v% compared to the control treatment. Keywords: Beauveria bassiana, fungal endophyte, secondary metabolites, tick repellency, tick toxicity, Tulbaghia violacea.

Numerous studies have found benefits of omega-3 polyunsaturated fatty acids (PUFAs), namely, for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in dogs. The objective of the present study was to assess the efficacy of dietary inclusion of equal amounts of omega-3 FAs in phospholipid (PL) from krill meal to triglyceride structure from fish oil to increase the omega-3 FA profile in red blood cells (RBCs) in dogs. Ten adult Alaskan Huskies of both genders were supplemented with daily 1.7 g EPA and DHA from krill meal for 6 weeks, while another ten dogs received 1.7 g EPA and DHA from fish oil. FA and omega-3 index measurements of the two groups were taken after 0, 3, and 6 weeks for comparison. It was mainly the EPA levels that increased in the krill meal group (from 1.84% to 4.42%) compared to the fish oil group (from 1.90% to 2.46%) (p<0.001), which drove the group differences in the omega-3 index. This resulted in the krill meal group having a mean omega-3 index increase from 3.9 at baseline to 6.3%, which was significantly greater than the increase from 3.9% to 4.7% observed in the fish oil group (p<0.001). Concomitantly, omega-6 PUFAs, such as arachidonic acid and linoleic acid, were reduced in RBC membranes and the omega-6 to omega-3 ratio was significantly more reduced in the krill meal compared to the fish oil group. The results showed that krill meal supplementation was associated with a reduction of omega-6 PUFAs, which compensated for the increased omega-3 index, suggesting that PLs are efficient delivery molecules of omega-3 PUFAs. Keywords: dog, fish oil, krill meal, omega-3 index, phospholipids, triglycerides.

Dermatophytosis, a zoonotic disease, is caused by fungi of three main genera, namely, Micropsorum, Trichophyton, and Epidermophyton. Specific lesions of dermatophyte infections are localized in the face, legs, and/or tail. Skin lesions in infected animals demonstrate localized alopecia, erythema, and crust, which are more commonly known as ringworm. Factors that affect dermatophytosis include the dermatophyte species; virulence factors of the agent; and the immune status, age, and sex of the host. High levels of cortisol and pro-inflammatory cytokines have also been reported to play an important role in dermatophyte infection. This review aims to explore and understand factors that affect dermatophyte infection with an emphasis on the prevalence, clinical signs, pathogenesis, immune response, and the roles of cortisol and cytokines in companion animals infected by a dermatophyte.

This paper reports a retrospective study performed in 185 cats diagnosed with feline lower urinary tract disease (FLUTD). The analyzed population involved feline patients at the Veterinary Clinic of Universitas Gadjah Mada, Indonesia. This research aimed to evaluate the clinical indications and causes of FLUTD in the Yogyakarta cat population. The medical data of all feline patients were obtained to conduct this study. FLUTD diagnoses were based on physical examinations, urinalyses, ultrasound examinations, and bacterial cultures. Only cats with a complete examination were used in the study. The clinical signs were evaluated and accompanied by the results of laboratory tests in cats that showed symptoms of FLUTD. The medical history of all feline patients was analyzed thoroughly. Most of the feline's urine samples were collected by catheterization. The most commonly diagnosed of FLUTD in the Yogyakarta cat population were: feline idiopathic cystitis (FIC) (56%), urinary tract infection (UTI; 25%), urolithiasis (13%), urethral plugs (UP) (4.9%), and neoplasia (0.4%), respectively. The prevalence of UTI is higher than that reported in Europe or the US. Older cats more often show symptoms of UTI and neoplasia, whereas young cats more often show symptoms of FIC and UP. The prevalence of male cats experiencing FLUTD in Yogyakarta is much higher than female cats. The incidence rate of FLUTD cases in Yogyakarta is related to age and sex. The results of this study are similar to those of the previous research studies conducted in other countries. Keywords: cats, feline lower urinary tract disease, Yogyakarta.

The current phytobiotic industry is struggling to determine a proper sterilization method for an herbal formula that comprises multiple plant extracts. Hence, this study aims to investigate the effects of two sterilization methods of herbal formula on phytochemical compounds and antibacterial activity against mastitis-causing bacterial isolates. The herbal formula comprised the extracts of Piper betle leaves, Curcuma domestica, and Curcuma zanthorriza. We applied two sterilization methods – sterilization using 0.45 μM syringe filter and sterilization using an autoclave at 121°C for 15 min. After sterilization, we measured phytochemical contents (phenol and flavonoid) of the herbal formula. Using the disk diffusion method, the antibacterial susceptibility test of the sterilized herbal formula against mastitis-causing bacterial isolates was conducted. Tetracycline, erythromycin, and penicillin – common antibiotics for mastitis therapy in dairy farms – were used as standard antibiotics. Compared with autoclave sterilization, syringe filter sterilization resulted in less (p<0.05) phenolic and flavonoid contents. Against Escherichia coli and Staphylococcus aureus, the filter sterilized herbal formula (E. coli: 65.9%-73%; S. aureus: 6.2%-18.1%) markedly reduced the antibacterial activity than the autoclave-sterilized herbal formula (E. coli: 2.1%- 3%; S. aureus: 4.5%-10.7%). This study establishes that autoclave sterilization of the herbal formula is the best sterilization method that exerts minimal adverse effects on the phytochemical compounds and could sustain the antibacterial efficacy against mastitis-causing bacteria. Hence, the herbal formula could be used as an alternative treatment for bovine mastitis. Keywords: herbal extract, mastitis, phytobiotic, phytochemical compound, sterilization method.

Milk production is one of the main props for the national economy. One of the crucial problems in this industry is subclinical mastitis, which harms this industry that considered the backbone of the economy. It is an infectious and zoonotic disease; the infection can spread between dairy animals through milkers' hands, and milking machines, while the human infection occurs due to the consumption of apparently hygienic milk. Staphylococcus aureus is one of the main causative agents of clinical and subclinical mastitis. It is also considered one of the bacteria incriminated in food intoxication of humans due to its virulence factors as enterotoxins and toxic shock syndrome. The current study was designed to assess the prevalence of S. aureus and its enterotoxins, as well as, its other virulence factors in milk collected from cows that suffer from subclinical mastitis. Sixty cows were collected from different dairy farms located in Assiut Governorate, Egypt. These cows were subjected to the clinical examination of the udder and its lymph nodes before sampling. Milk samples were collected from clinically healthy udders. All the milk samples were examined by California mastitis test (CMT), polymerase chain reaction (PCR), and enzyme-linked immunosorbent assay (ELISA) for confirmation subclinical mastitis, presence of S. aureus and its enterotoxins genes and other virulence factors in the examined milk samples. The cows included in the current study had healthy udders. The sixty collected milk samples were tested by CMT. 48/60 (80.0%) were positive samples; from the 48 positive samples, 46 (95.83%) samples were confirmed positive by S. aureus 16s rRNA PCR assay. Multiplex PCRs confirmed the presence of staphylococcus enterotoxin gene C (sec) in one sample, staphylococcus enterotoxin gene D (sed) in 23 samples, while ELISA assay confirmed the presence of the same enterotoxin in only two samples. On the other hand, other groups of genes responsible for some other virulence factors of S. aureus like the extracellular thermostable nuclease (nuc) gene were found in 33 samples, while toxic shock syndrome (tsst) gene and methicillin restraint S. aureus (mecA) gene were not detected in this study. Subclinical mastitis is one of the hidden factors that adversely affect the health of both animals and humans. The milk is usually appeared good and may be consumed by humans especially children; however, it causes severe public health problems. In addition, the infected animals with this form of mastitis can spread the infection to other dairy animals and may be turned to a clinical case of contagious mastitis that may be ended by animal culling or death. S. aureus is one of the main causes of subclinical mastitis in cattle. In addition to extracellular thermostable nuclease (nuc) gene, staphylococcus enterotoxin gene C (sec) and staphylococcus enterotoxin gene D (sed) are the most common virulence genes confirmed in subclinical mastitis milk. These results highlighted the need to apply more hygienic measures in the dairy farms to avoid spreading the infection between animals to ensure the production of safe and healthy food to humans. Keywords: enterotoxins, mecA, nuc, polymerase chain reaction, Staphylococcus aureus, subclinical mastitis.

Vibrio bacteria are autochthonous inhabitants of aquatic and marine environments. Certain strains are responsible for important seafood-borne outbreaks in developed nations. The aim of this study was to investigate the occurrence of Vibrio spp. along the Algerian Mediterranean coast in several samples of two prominent wild and farmed marine fishes, Sparus aurata and Dicentrarchus labrax. A total of 350 S. aurata (n=280 farmed and n=70 wild) and 340 D. labrax (n=250 farmed and n=90 wild) were sampled among three different locations along the Algerian Mediterranean coastal area. The samples were processed for Vibrio research according to the ISO methods. Isolated colonies were identified utilizing biochemical tests and consecutively confirmed with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, combined with polymerase chain reaction (PCR) analysis when appropriate, or confirmed with PCR analysis alone. A total of 42 Vibrio spp. were detected only among the farmed fishes. Taking into account, all 690 fishes sampled, the incidence of Vibrio spp. was 6.08% (with peaks up to 7.92%) among the total number of farmed fishes. Overall, 25 strains were isolated from S. aurata and 17 strains were isolated from D. labrax. The isolated strains belonged to four different species and were represented as follows: Vibrio alginolyticus (n=20), Vibrio cholerae (n=15), Vibrio fluvialis (n=5), and Vibrio hollisae (n=2). The incidence of Vibrio was higher in places characterized by greater levels of anthropogenic contamination of seawater. Considering the growing production and consequent rising consumption of farmed fish in Algeria, the reported incidence of Vibrio and the presence of potentially pathogenic strains of Vibrio such as V. cholerae cause particular concern for food safety matters. Even if innovative and natural techniques are desired in aquaculture, proper hygiene and manufacturing practices are essential for the correct management of Vibrio infection risk in farmed fishes at both industrial and domestic levels. Keywords: Algeria, gilt head sea bream, Mediterranean sea, sea bass, Vibrio spp.

This study is aimed at characterizing the genetic and phylogenetic structure of Lakor goats as indigenous livestock from the Southwest Maluku Regency based on mitochondrial COI gene sequences. The genomes of 103 follicle samples from Lakor goats, collected from Lakor Island, were analyzed. The polymerase chain reaction was used to amplify 1548 bp of the mitochondrial COI gene using two primer pairs (COIA and COIB). Following sequencing, genetic variation and phylogenetic relationship were established using MEGA version X software. The results of multiple COI gene alignment of the total sequences identified four polymorphic nucleotides that function as genetic markers between individual animals within the Lakor goat population. These correspond to positions 228 (A-G), 519 (G-A), 900 (C-T), and 1266 (T-C). Phylogenetic signals based on the COI gene showed that Lakor goat breed is a monophyletic group or single clade with a bootstrap value of 100% by the neighbor-joining (NJ) and maximum likelihood (ML) evolutionary models. This data indicated that evolutionarily, the Lakor goat breed has a very close kinship with three goat breeds from China: The Meigu goat (KM 244714.1), Chinese Tibet (Capra hircus) (KJ 940969.1), and C. hircus (KP 677510.1). Phylogenetic information based on the cladistics system classified the Lakor goat as a single clade (monophyletic group). The low-genetic diversity within populations indicates that there has been an inbreeding depression occurring at a very high frequency. We conclude that the Lakor goat may be divided into a single clade or monophyletic group based on the COI gene sequence. Four nucleotides were identified that can be used as genetic markers among individual animals within the Lakor goat population, as well as C. hircus and others as derived from GenBank data. The Lakor goat population has a high level of inbreeding depression as a result of geographical isolation, which supports the formation of a monophyletic group with different genetic characteristics, and does not allow the introduction of males from other breeds. Phylogenetic signals indicated that Capra aegagrus (bezoar) is the ancestor of the native goats in Indonesia, including the Lakor goats. Keywords: artiodactyla, CO1 gene, Lakor goat, mitochondrial, phylogenetic study.

The current study was carried out to determine the prevalence, seasonality, and antimicrobial profile of thermotolerant Campylobacter isolated from broiler chickens in Batna, East Algeria, from June 2016 to June 2018. A total of 960 samples, including 480 cloacal swabs, 240 cecal contents, and 240 neck skin samples collected from 6 poultry farms and 12 slaughterhouses, were included in this study. After isolation and identification, susceptibility to seven antimicrobial agents was tested by the disk diffusion method. The seasonality of Campylobacter infection at broiler farms was statistically analyzed. The data showed that 65%, 55%, and 70% of the cloacal swab, neck skin, and cecal content samples were contaminated with thermotolerant Campylobacter strains, respectively (p<0.05). Among the isolated campylobacteria, Campylobacter jejuni was the predominant species (73.5%). Sampling season exhibited a significant impact on the prevalence of Campylobacter (p<0.01), with peak occurrence in summer. All of the isolates were susceptible to gentamicin and resistant to ampicillin and amoxicillin/clavulanic acid, while 83.3% of them were resistant to erythromycin. Interestingly, 16 different resistance profiles were noted, with the combination of "ampicillin, amoxicillin/clavulanic acid, chloramphenicol, erythromycin, and tetracycline" being the most common, identified in 20.7% of isolated strains. This study demonstrates the presence of a high contamination rate of multidrug-resistant Campylobacter in farms and slaughterhouses in East Algeria. These findings underscore the need to apply strict control measures to avoid any associated public health hazard among Algerian consumers. This initial finding of the contamination of poultry with this zoonotic pathogen in East Algeria suggests the value of periodic comprehensive evaluation of associated disease in poultry as well as in humans in this region. Keywords: antimicrobial resistance, broilers, poultry farms, slaughterhouses, thermotolerant Campylobacter.

The objective of this study was to assess a new lead system method to improve electrocardiographic measurement in horses. Twenty-two horses with an average age of 8.8±0.8 years were enrolled in this study. Horses were divided into two groups, consisting of a control group (n=11) and athlete group (n=11). Electrocardiography (ECG) and echocardiography were performed to provide information on the structure and function of the heart. Two lead systems, base apex and modified precordial leads, were used for the electrocardiogram to assess the cardiac electrophysiological functions. PR interval, QT interval, and QRS-T angle presented significant differences between the control and athlete groups when the modified precordial lead system was used. However, significant variations in the mean electrical axis were found when the base apex lead system was used. The modified precordial lead system resulted in more significant differences in cardiac electrophysiological parameters than the base apex lead system. In the athlete group, echocardiography showed cardiac adaptations such as increases in the left atrial and left ventricular dimensions and stroke volume and a decrease in heart rate in response to exercise and training. The observed differences in cardiac morphology and function between groups suggested differences in health performance in the athlete group. These data provided the first evidence that the modified precordial lead system improved statistical variation in ECG recording and provided the most reliable method for health screening in horses. Keywords: echocardiography, electrocardiography, horse, non-invasive cardiac evaluation.

This study aimed to investigate the effects of protein reduction with supplementation of limiting amino acids (AA, tryptophan, threonine, and valine) on growth performance, blood biochemical, immunity parameters, and carcass traits in broiler chickens. Three hundred one-day-old broiler chicks were randomly allotted into three treatment groups, with five replicates per treatment and 20 broiler chickens per replicate. The three experimental diets were formulated with different dietary crude protein (CP) %, (control [CON] and CON with 1% [CP-1%] or 2% [CP-2%] less CP units) during the starter, grower, and finisher phases. The CP of the experimental diets were 22, 21, and 20% for the starting period (day 1-14); 20, 19, and 18% CP for the growing period (day 15-28); and 18, 17, and 16% CP for the finishing period (day 29-35) in CON, CP-1%, and CP-2%, respectively. The low-CP diets (CP-1% and CP-2%) were supplemented with combined AA, threonine+tryptophan+valine, to meet the respective levels of the CON diet. The CP-2% group had greater (p<0.05; linear, p<0.05) final body weight and gain and better feed conversion ratio. The combined AA inclusion in the low-CP diet (CP-1% and CP-2%) increased (p<0.001; linear, p<0.001) carcass and breast meat yield as well as CP% of breast meat. The reduction of CP% with AA supplementation (CP-2%) decreased (p<0.05; linear, p<0.05) serum triglycerides, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, and alkaline phosphatase, while increased (p<0.01; linear, p<0.01) phagocytic activity and phagocytic index. The mRNA expression of splenic and cecal tonsil interleukin 4 and interferon gamma was upregulated (p<0.001; linear, p<0.001) in the low-CP diets supplemented with AA (CP-1% and CP-2%). Dietary supplementation with AA to low-protein diets improved (p<0.01; linear, p<0.01) the economic returns of broiler chickens. A reduction of dietary CP and increased synthetic AA such as threonine, valine, and tryptophan should be considered to improve performance, health, and immunity in broiler chickens. Keywords: amino acids, broiler chickens, immune-related genes, meat composition, performance.