• Title/Summary/Keyword: Alternative protein

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Post HCV Infection Due to MX Gene Stimulation Produced Post Treatment with Imported and Locally Produced Egyptian Biosimilar IFN

  • Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5635-5641
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    • 2015
  • Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

Anti-inflammatory Effect of Gagam-GongJin-dan in mouse peritoneal macrophages (마우스 복강대식세포에서 가감공진단(加減拱辰丹)의 항염증 효과)

  • Kim, Hong-Jun;Kim, Young-Sik;Mok, Ji-Ye;Jeong, Seung-Il;Hwang, Sung-Yeoun;Cho, Jung-Keun;Jang, Seon-Il
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.207-217
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    • 2011
  • Objectives : In a previous study, we have shown that Gagam-Gongjin-Dan(GGD) has an inhibitory effect on the ovalbumin-induced immune responses and a hepatoprotective effect on actaminophen-induced liver injury in Balb/c Mice. However, the possible anti-inflammatory effect of GGD extract for inflammatory mediators was not reported. Therefore, the purpose of this study was to investigate an inhibitory effects of GGD extract against lipopolysaccharides(LPS) induced inflammatory mediators in mouse peritoneal macrophages. Methods : GGD extract was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Accumulated nitrite, an oxidative product of nitric oxide(NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin $E_2(PGE_2)$, interleukin-$1{\beta}$(IL-$1{\beta}$), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) were measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2) were measured by Western blot analysis. Results : GGD extract (50-$400\;{\mu}g$/ml) per se had no cytotoxic effect in LPS-stimulated peritoneal macrophages. GGD extract dose-dependently reduced NO, $PGE_2$, IL-$1{\beta}$ and TNF-${\alpha}$ production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with GGD extract in a dose dependent manner. Conclusions : These results suggest that GGD extract has an anti-inflammatory effect against LPS-induced inflammatory mediators in peritoneal macrophages, these properties may contribute to inflammation disease care.

Subacute Oral Toxicity Evaluation of Freeze-Dried Powder of Locusta migratoria

  • Kwak, Kyu-Won;Kim, Sun Young;An, Kyu Sup;Kim, Yong-Soon;Park, Kwanho;Kim, Eunsun;Hwang, Jae Sam;Kim, Mi-Ae;Ryu, Hyeon Yeol;Yoon, Hyung Joo
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.795-812
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    • 2020
  • Novel food sources have enormous potential as nutritional supplements. For instance, edible insects are considered as an alternative food source due to their higher protein content; moreover, they are economically efficient reproducers and have high in nutritional value. In this study, we investigated the toxicity of the freeze-dried powder of Locusta migratoria (fdLM), known to contain rich proteins as well as fatty acids. The objective of the present study was to evaluate the subacute toxicity of fdLM in male and female Sprague-Dawley (SD) rats. The SD rats were divided into four groups based on the dosage of fdLM administered: dosage of 0 (vehicle control), 750, 1,500, and 3,000 mg/kg/day were administered for 28 days. Toxicological assessments including observations on food consumption, body and organ weights, clinical signs, mortality, ophthalmologic tests, urinalyses, hematologic tests, clinical chemistry tests, gross findings, and histopathology tests were performed. Clinical signs, urinalyses, hematology, serum biochemistry tests, and organ weight examinations revealed no fdLM-related toxicity. The no-observed-adverse-effect level for fdLM was higher than 3,000 mg/kg/day in rats of both sexes; therefore, fdLM, in conclusion, can be considered safe as an edible alternative human and animal food source material.

Screening of Complement-System Activating Polysaccharide from Edible Plants and Its Action Mode (식물성 식품재료로부터 보체계 활성화 다당의 검색 및 그 활성검토)

  • Shin, Kwang-Soon;Ra, Kyung-Soo;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.25 no.3
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    • pp.197-203
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    • 1993
  • Screenings were performed on edible plants to examine their complement-system activating ability (anti-complementary activity) by hemolytic complement assay $(TCH_{50})$. Among 38 kinds of plant extracts, 5 kinds showed relatively strong anti-complementary activity which decreased $TCH_{50}$ more than 60% comparison with control and the order of activity was Zingiber officinale>Colocasia antiquorum>Capsella bursapastoris>Ginkgo biloba>Alium monanthum in $1000{\mu}g/ml$. The anti-complementary activity of ZR-1 prepared from the root of Zingiber officinale which was showed the most potent activity, did not change by pronase treatment, but decreased greatly by periodate oxidation. These results indicate that not protein moiety but carbohydrate moiety in ZR-1 fraction may also contribute to the anti-complementary activity. Also, the anti-complementary activity of ZR-1 was reduced partially in the absence of the $Ca^{2+}$ ion. When crossed immunoelectrophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the ZR-1 in $Ca^{2+}$ free condition, a cleavage of C3 precipitin line was observed. Furthermore this polysaccharide fraction considerably inhibited $ACH_{50}$. These results also indicate that the mode of complement activation by polysaccharide from Zingiber officinale is via not only the classical pathway but also the alternative pathway.

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Production of Keratinolytic Protease by Bacillus pumilus RS7 and Feather Hydrolysate As a Source of Amino Acids (Bacillus pumilus RS7에 의한 난분해성 케라틴 분해효소의 생산 및 아미노산 공급원으로서 우모 분해산물)

  • Woo, Eun-Ok;Kim, Min-Ju;Son, Hyeng-Sik;Ryu, Eun-Youn;Jeong, Seong-Yun;Son, Hong-Joo;Lee, Sang-Joon;Park, Geun-Tae
    • Journal of Environmental Science International
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    • v.16 no.10
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    • pp.1203-1208
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    • 2007
  • Feathers are produced in huge quantities as a waste product at commercial poultry processing plants. Since feathers are almost pure keratin protein, feather wastes represent an alternative to more expensive dietary ingredients for animal feedstuffs. Generally they become feather meal used as animal feed after undergoing physical and chemical treatments. These processes require significant energy and also cause environmental pollutions. Therefore, biodegradation of feather by microorganisms represents an alternative method to prevent environment contamination. The aim of this study was to investigate cultural conditions affecting keratinolytic protease production by Bacillus pumilus RS7. We also assessed the nutritive value of microbial and alkaline feather hydrolysates, The composition of optimal medium for the keratinolytic protease was fructose 0.05%, yeast extract 0.3%, NaCl 0.05%, K2HPO4 0.03%, KH2PO4 0.04% and MgCl2 6H2O 0.01%, respectively. The optimal temperature and initial pH was $30^{\circ}C$ and 9.0, respectively. The keratinolytic protease production under optimal condition reached a maximum after 18 h of cultivation. Total amino acid content of feather hydrolysates treated by NaOH and B. pumilius RS7 was $113.8\;{\mu}g/ml$ and $504.9\;{\mu}g/ml$, respectively. Essential amino acid content of feather hydrolysates treated by NaOH and B. pumilius RS7 was $47.2\;{\mu}g/ml$ and $334.0\;{\mu}g/ml$, respectively. Thus, feather hydrolysates have the potential for utilization as an ingredient in animal feed.

A chemical conjugate between HER2-targeting antibody fragment and Pseudomonas exotoxin A fragment demonstrates cytotoxic effects on HER2-expressing breast cancer cells

  • Lee, Sunju;Park, Sangsu;Nguyen, Minh Tan;Lee, Eunyoung;Kim, Julee;Baek, Sangki;Kim, Chong Jai;Jang, Yeon Jin;Choe, Han
    • BMB Reports
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    • v.52 no.8
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    • pp.496-501
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    • 2019
  • Conventionally, immunotoxins have been produced as a single polypeptide from fused genes of an antibody fragment and a toxin. In this study, we adopted a unique approach of chemical conjugation of a toxin protein and an antibody fragment. The two genes were separately expressed in Escherichia coli and purified to high levels of purity. The two purified proteins were conjugated using a chemical linker. The advantage of this approach is its ability to overcome the problem of low recombinant immunotoxin production observed in some immunotoxins. Another advantage is that various combinations of immunotoxins can be prepared with fewer efforts, because the chemical conjugation of components is relatively simpler than the processes involved in cloning, expression, and purification of multiple immunotoxins. As a proof of concept, the scFv of trastuzumab and the PE24 fragment of Pseudomonas exotoxin A were separately produced using E. coli and then chemically crosslinked. The new immunotoxin was tested on four breast cancer cell lines variably expressing HER2. The chemically crosslinked immunotoxin exhibited cytotoxicity in proportion to the expression level of HER2. In conclusion, the present study revealed an alternative method of generating an immunotoxin that could effectively reduce the viability of HER2-expressing breast cancer cells. These results suggest the effectiveness of this method of immunotoxin crosslinking as a suitable alternative for producing immunotoxins.

Effect of feeding a diet comprised of various corn silages inclusion with peanut vine or wheat straw on performance, digestion, serum parameters and meat nutrients in finishing beef cattle

  • Zhang, Hongrui;Zhang, Liyang;Xue, Xiao;Zhang, Xiaoxia;Wang, Hongyi;Gao, Tengyun;Phillips, Clive
    • Animal Bioscience
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    • v.35 no.1
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    • pp.29-38
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    • 2022
  • Objective: The objective of this study was to compare the feeding value, meat nutrients and associative effects of a diet comprised of various corn silages inclusion with peanut vine or wheat straw in finishing beef cattle. Methods: One hundred and eighty Simmental crossbred beef steers were blocked and assigned to the follow treatments: i) whole plant corn silage-based diet (control, WPCS), ii) mixed forages-based diet (replacing a portion of corn silage with wheat straw, WPCSW), iii) corn stalklage-based diet (CS), and iv) sweet corn stalklage-based diet (SCS). Each group consisted of 5 repeated pens with 9 steers/pen. The diets were formulated to be isonitrogenous and isoenergetic with same forage to concentrate ratio. Experimental diets were fed for 90 d. Results: The effective ruminal degradability of dry matter and crude protein were highest for WPCS diet (p<0.05), for neutral detergent fiber was highest in SCS diet (p<0.05). The average daily gain was greater for cattle offered the WPCS diet, intermediate with WPCSW and SCS and lowest with CS (p<0.001). The concentration of non-esterified fatty acid in serum was higher for steers fed with CS and SCS diets than those offered WPCS and WPCSW steers (p<0.001). The treatments did not affect the general nutritional contents and amino acids composition of Longissimus dorsi of steers (p>0.05). Conclusion: The corn silage-based diet exhibited the highest feeding value. The sweet corn stalklage and wheat straw as an alternative to corn silage offered to beef cattle had limited influence on feeding value and meat nutrients. However, the value of a corn stalklage-based diet was relatively poor. To sum up, when the high quality forage resources, such as corn silage, are in short supply, or the growth rate of beef cattle decreases in the later finishing period, the sweet stalklage and wheat straw could be used as a cheaper alternative in feedlot cattle diet without sharp reducing economic benefits.

Effect of Bacillus amyloliquefaciens-based Direct-fed Microbial on Performance, Nutrient Utilization, Intestinal Morphology and Cecal Microflora in Broiler Chickens

  • Lei, Xinjian;Piao, Xiangshu;Ru, Yingjun;Zhang, Hongyu;Peron, Alexandre;Zhang, Huifang
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.2
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    • pp.239-246
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    • 2015
  • The present study was conducted to evaluate the effect of the dietary supplementation of Bacillus amyloliquefaciens-based direct-fed microbial (DFM) on growth performance, nutrient utilization, intestinal morphology and cecal microflora in broiler chickens. A total of two hundred and eighty eight 1-d-old Arbor Acres male broilers were randomly allocated to one of four experimental treatments in a completely randomized design. Each treatment was fed to eight replicate cages, with nine birds per cage. Dietary treatments were composed of an antibiotic-free basal diet (control), and the basal diet supplemented with either 15 mg/kg of virginiamycin as antibiotic growth promoter (AGP), 30 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 30) or 60 mg/kg of Bacillus amyloliquefaciens-based DFM (DFM 60). Experimental diets were fed in two phases: starter (d 1 to 21) and finisher (d 22 to 42). Growth performance, nutrient utilization, morphological parameters of the small intestine and cecal microbial populations were measured at the end of the starter (d 21) and finisher (d 42) phases. During the starter phase, DFM and virginiamycin supplementation improved the feed conversion ratio (FCR; p<0.01) compared with the control group. For the finisher phase and the overall experiment (d 1 to 42) broilers fed diets with the DFM had better body weight gain (BWG) and FCR than that of control (p<0.05). Supplementation of virginiamycin and DFM significantly increased the total tract apparent digestibility of crude protein (CP), dry matter (DM) and gross energy during both starter and finisher phases (p<0.05) compared with the control group. On d 21, villus height, crypt depth and villus height to crypt depth ratio of duodenum, jejunum, and ileum were significantly increased for the birds fed with the DFM diets as compared with the control group (p<0.05). The DFM 30, DFM 60, and AGP groups decreased the Escherichia coli population in cecum at d 21 and d 42 compared with control group (p<0.01). In addition, the population of Lactobacillus was increased in DFM 30 and DFM 60 groups as compared with control and AGP groups (p<0.01). It can be concluded that Bacillus amyloliquefaciens-based DFM could be an alternative to the use of AGPs in broilers diets based on plant protein.

A Baseline Study on the Choice of Optimal Screening Test Items among Workers with Abnormal Liver Function Tests on Workers' Periodic Health Examination (근로자 건강진단시 간기능 이상자의 정밀검사항목 개선을 위한 조사연구)

  • Cheong, Hae-Kwan;Lim, Hyun-Sul;Kim, Gyu-Hoi
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.4 s.48
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    • pp.747-761
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    • 1994
  • Workers' periodic health examination is the main tools used to manage the health problems of most workers in Korea. The most common health problem found in workers' periodic health examination is liver disorder. Liver disorder is also one of the most common health problems in general population and one of the leading causes of mortality in adult population. Regulation proposed by government (No. 207, Ministry of Labor, 1992) defines the criteria for selection of workers with the liver dysfunction for further evaluative examination and the examination items used for diagnosis of the workers with liver dysfunction. This study was designed to evaluate the proficiency of each examination items presently defined in Regulation and propose the optimal examination items for detection of the liver disorders found by workers' periodic health examination. Study subjects are 186 workers with abnormal liver function tests in screening examination of workers' periodic health examination. Questionnaire survey including past history of liver disorder, drinking history, height and weight was done. Physical examination by physician, routine test items defined by Regulation (SGOT, SGPT, $\gamma$-GTP, protein, albumin, total and direct bilirubin, alkaline phosphatase, $\alpha$-feto protein, HBsAg and anti-HBs), anti-HCV antibody test and liver ultrasonography were done. Results are as follows; 1. Result of evaluative examination utilizing only the items defined in Regulation was; There were 75 workers with suspected live. disorder(40.3%), 63 with no liver dysfunction (33.9%), 13 with suspected hepatitis B(7.0%), 10 workers with hepatitis B(5.4%), 10 workers with hepatitis B carrier state(5.4%), 10 with alcoholic liver disorders(5.4%), 5 with fatty liver(2.7%). When alternative diagnostic criteria applying additional examination items (drinking history, body mass index, anti-HCV antibody and ultrasonography) diagnosability of liver disorder was increased. When all four items were included, final results were; 23 workers (17.8%) with hepatitis B (10 carriers, 13 suspects and 10 hepatitis B), 10 (5.4%) with hepatitis C(4 carriers, 5 suspects and 1 hepatitis C), 13(7.0%) with alcoholic liver disorder, 45(24.2%) with fatty liver (40 suspects, 5 fatty liver), 410%) with suspected liver disorders and 44 (23.7%) with normal liver. 2. Of examination items defined by Regulation, only SGOT, SGPT, $\gamma$-GTP and HBsAg were significantly different in abnormal rate and mean value, and all other laboratory findings did not showed significant difference between two groups. Drinking history, body mass index and anti-HCV antibody test which are the items that authors included in this study, also showed significant difference between two groups. Utilization of body mass index (BMI) for abnormal liver function group in diagnosis of fatty liver had high specificity (97.6%) but sensitivity (22.3%) was low. Therefore we suggest that SGOT, SGPT, $\gamma$-GTP, HBsAg, alcohol drinking history, BMI and anti-HCV Ab were useful for diagnosis of liver disorders among worker's periodic health examination.

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Growth Performance, Carcass and Meat Characteristics of Black Goat Kids Fed Sesame Hulls and Prosopis juliflora Pods

  • Abdullah, Abdullah Y.;Obeidat, Belal S.;Muwalla, Marwan M.;Matarneh, Sulaiman K.;Ishmais, Majdi A. Abu
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1217-1226
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    • 2011
  • Finding alternative feeds, such as sesame hulls and Prosopis juliflora species can attenuate difficulties of feed shortage and reduce the cost of animal feed in arid and semi-arid countries. Thirty-two Black male kids with similar initial weights (BW = $16.7{\pm}0.80\;kg$) and $120{\pm}5\;d$ of age, were used to evaluate the effect of replacing barley grains and soybean meal with Prosopis juliflora pods (PJP) and sesame hulls (SH) on growth performance, digestibility and carcass and meat characteristics. Kids were equally divided into four dietary treatment groups for an 84-d fattening period. Treatment diets had similar crude protein (CP) and metabolizable energy (ME). The treatment groups were: (T1) no PJP nor SH, (T2) 10% PJP and 20% SH, (T3) 15% PJP and 15% SH, and (T4) 20% PJP and 10% SH. A tendency was detected (p<0.08) for dry matter (DM), crude protein (CP), organic matter (OM) and neutral detergent fiber (NDF) intakes that were greater for T2 than T3 and T4 while T1 was not different from all other treatment groups. Ether extract (EE) intake was the greatest (p<0.05) for T2 and the lowest for T1. Acid detergent fiber (ADF) intake was greater (p<0.05) for T2 than T1 while T3 and T4 were intermediate. Final live weight, average daily gain (ADG) and feed conversion ratio (FCR) were comparable among different treatment groups. Digestibility of DM, OM, CP, NDF and ADF were similar among all treatment groups, however, EE digestibility was the lowest (p<0.05) for T1 when compared to other treatments. In addition, nitrogen intake, nitrogen in urine and retained and retention percentages were similar among all treatment groups. However, N loss in feces was higher (p<0.05) for T2 than T3 and T4 while T1 was intermediate. No differences were observed among treatment groups with respect to fasting live weight, hot and cold carcass weights, dressing-out percentages, mesenteric fat, visceral organs, carcass cuts percentages and carcass linear dimensions. No differences were also observed for dissected loin, leg, rack and shoulder tissues except in the total bone % for loin cuts and in the meat to bone ratio for rack cuts. T3 has the greatest total bone % and the lowest meat to bone ratio when compared to all other treatment groups. No differences were observed between treatment groups in all quality characteristics of the longissimus muscle. The present study demonstrates the potential of using PJP and SH for growing kids without adverse effects on growth performance, carcass characteristics and meat quality.