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Characterization of growth performance, meat quality and serum biochemical parameters in chickens suffering from tibial dyschondroplasia
Livestock Science 233 (2020) 103956
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Characterization of growth performance, meat quality and serum biochemical parameters in chickens suffering from tibial dyschondroplasia
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Qin-Qin Caoa, An-An Konga, Kun-Sheng Taoa, Shi-Hao Zhenga, Chao Tonga, Xue-Bing Wanga, Zong-Xi Tonga, Mujeeb Ur Rehmanb, Shu-Cheng Huanga,⁎ a b
College of Animal Science and Veterinary Medicine, Henan Agricultural University, 95# Wenhua Road, Jinshui District, Zhengzhou, Henan 450002, PR China Research Center of Avian Disease, College of Veterinary Medicine of Sichuan Agricultural University, Chengdu, Sichuan, PR China
ARTICLE INFO
ABSTRACT
Keywords: Drumstick weight Meat quality Leg problems Tibial dyschondroplasia Serum biochemistry
The present study was conducted to examine the clinical manifestations of broilers suffering from tibial dyschondroplasia (TD) on growth performance, drumstick parameters, breast meat quality, and serum biochemical parameters. A total of 200 Arbor Acre broiler chickens were randomly allocated into two groups: control (basal diet) and TD group (basal diet supplemented with 50 mg/kg of thiram). Each group consisted of 4 replicates (25 chickens per replicate). The experiment lasted for 14 days. Results showed that TD chickens exhibited abnormal foraging behavior and resting position that caused a reduction in feed intake, daily weight gain and final body weight. Compared with the control group, the weight and length of drumstick in TD chicken group was significantly decreased (p < 0.05) and its color was pale. Interestingly, drumstick related-parameters were statistically correlated with breast muscle quality in control and TD chickens (p < 0.01). In addition, breast meat quality (assessed by shear force and pH) in TD group chickens was affected. Furthermore, TD chickens had significantly elevated serum creatine kinase (CK) and aspartic acid aminotransferase (AST) activities (p < 0.05), but no significant change was observed in serum amylase, triglycerides, and glucose levels (p > 0.05). In conclusion, the TD has adverse effect on the overall growth performance, drumstick parameters, breast meat quality, and some serum biochemical parameters of broilers. Moreover, decreased breast muscle quality was strongly correlated with leg problems in the tibia.
1. Introduction With the improvement of people's living standards, more attention is paid to food safety and health. Several studies reported that increase in red meat consumption is associated with higher mortality rates and cardiovascular disease (Guasch-Ferré et al., 2019; Zheng et al., 2019). In view of nutrition and human health concerns, chicken is an economical and affordable meat product that has a higher protein content and lower total fat, and provides essential vitamins and minerals. Therefore, it becomes an ideal meat source for consumers (FAO, 2010; Naji et al., 2014). The majority of the poultry meat products that reaches the food market are produced with birds reared under intensive conditions, which led to the selection of birds with rapid growth and feed efficiency genotypes (Cavani et al, 2009; Funaro et al., 2014). However, several authors have shown that the selection for fast-growing broiler has resulted in a disproportionate development of broiler bones compared to body weight gain, which eventually lead to a higher incidence of leg ⁎
problems (Garcia et al., 2013; González-Cerón et al, 2015). Most of the noninfectious disorders such as tibial dyschondroplasia (TD) and tarsal varus-valgus deformities (VVD) result in abnormal gait and even lameness. Reduction of leg abnormalities has become a major challenge in meat-type chicken because they impair growth performance, reduce meat quality and compromise welfare status of birds (Bizeray et al, 2002; Garcia et al., 2013). TD is an intractable leg problem in fast growing meat-type chickens that results in bone deformation and leg lameness (Herzog et al., 2011; Huang et al., 2017). This tibiotarsal bone disorder leads to apparent locomotion problem with rising prevalence of 30% in broilers flock (Pelicia et al., 2012; Huang et al., 2018). However, the relationship between TD and meat quality of broiler chickens is not known. Based on this consideration, the objective of this study was to establish a TD chicken model for studying the effects of leg problems (TD) on growth performance, meat quality and several biochemical parameters in broiler chickens.
Corresponding author. E-mail address: [email protected] (S.-C. Huang).
https://doi.org/10.1016/j.livsci.2020.103956 Received 7 July 2019; Received in revised form 2 February 2020; Accepted 3 February 2020 Available online 04 February 2020 1871-1413/ © 2020 Elsevier B.V. All rights reserved.
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2. Materials and methods
and record the readings.
2.1. Animals and experimental protocol
2.5. Serum biochemistry analysis
All procedures, treatments, and animal care were strictly in compliance with the guidelines of the Institutional Animal Care and Use Committee of Henan Agricultural University (Permit No: 11-0085). All arbor acres (AA) chickens (regardless of gender; one-day-old; 51.96 ± 3.87 g) were obtained from a commercial hatchery (Xingda Poultry Industry Co. Ltd., Kaifeng, China). Thiram (tetramethylthiuram disulfide) (AR, purity ≥ 98%, #C10036460) was acquired from Macklin Biochemical Co., Ltd. (Shanghai, China). All chicks were randomly allocated into two groups: the control group and the thiram-induced TD group (n = 100/group, with 25 chicks per replicate and 4 replicates per group), and raised in singlelayer metal cages (size, 120 cm × 90 cm × 40 cm). Standard feed and water was provided ad libitum. The environmentally controlled facility was maintained with the recommended standard room temperature, relative humidity, daily light, good ventilation, and hygienic conditions as previously described (Huang et al., 2017). After 3 days of acclimation, and from the fourth day the control group was offered a standard diet and the thiram - induced TD group received the same standard diet further supplemented with 50 mg/kg of thiram (thiram was removed on day 7) for the purpose of constructing a broiler TD model to facilitate the study of the TD mechanism as previously reported (Rath et al., 2005; Huang et al., 2017). The experiment lasted for 14 days.
The blood samples were centrifuged at 3000 × g for 20 min for the separation of serum and stored at −20 °C until further analyses. Creatine kinase (CK), aspartic acid aminotransferase (AST), amylase (AMY), triglycerides (TG), glucose (GLU) serum levels were determined. These biochemical indicators were quantified through a veterinary reagent tray purchased from Chengdu Puli Taisheng Technology Co., Ltd and quantitative detection was performed on a portable full automated veterinary biochemistry analyzer (#SMT-120V, Seamaty Technology Co., Ltd.,Chengdu, China) according to the manufacturer instructions. 2.6. Statistical analysis Correlation analyses were performed using Pearson's test, and the differences between groups of shear force and pH values of breast muscle were analyzed and displayed by a pseudocolor scale (from green to red) respectively, which are shown in the form of a heatmap. In addition, comparisons between two groups were implemented using unpaired, two-tailed Student's t-tests with SPSS (#Version 17.0., Chicago, IL) or Prism statistical analysis software (#Version 7.00, GraphPad Software, Inc.). Biochemical parameters were repeated at least three times to increase the reproducibility of the findings. In addition, representative results are displayed in figures. Data are presented as the means ± S.D. P < 0.05 was considered to be statistically significant.
2.2. Clinical manifestations and production performance analysis The chicks were group-weighed on day 3, 7, 10, and 14, and the average daily weight gain (ADG), average daily feed intake (AFI), and feed conversion ratio (FCR) were calculated per group. During the experiment, the clinical manifestations and feeding behavior of broilers were observed, and mortality of broilers was recorded on a daily basis.
3. Results 3.1. Effects of broiler leg problems on clinical features AA broiler chicken is a meat - type broiler and the fast weight gain adds a burden on the legs. TD is one of the most prone leg problems in broilers with fast growth rates. The experiment used thiram to establish a broiler TD model. The pathological feature is indicated by the fact that the proximal proximal tibial growth plate was clearly widened (Fig. 1A), and the tibia bone mass was significantly reduced (Fig. 1B). Once a broiler has a leg problem, some representative clinical symptoms are observed from the 10th day of age (Fig. 1). The foraging behavior in the control group showed eating in a normal standing position. In contrast, the posture of the broiler with legs stretched forward under its chest during feeding was observed in the TD group (Fig. 1C). Additionally, the change of resting position was noticed and TD group broilers’ leg showed to one side for outreach, even split-leg-like poses compared with the control broilers (Fig. 1C).
2.3. Morphology of the tibial growth plates On day 7 and 14, three chicks per treatment cage were randomly selected and euthanized as previously described (Huang et al., 2017). The stripping of the tibial longitudinalmuscles were performed and the tibiae were retrieved. The weight of the tibia was determined using an electronic balance (sensitive to 0.1 mg, #FA1204N, Jinghai Instrument Co., Ltd. Shanghai, China). In addition, preparation of sagittal sections of the proximal tibial growth plates was performed to analyze the morphology of growth plates. 2.4. Observation and evaluation of drumsticks and breast muscles The chicken drumsticks were collected from the control and the TD group on days 7 and 14. The drumstick muscle color was then visually evaluated and the length and weight of drumstick were measured and counted using an Digital Calipers (#SATA91511, TATA Company, Shanghai, China) and electronic balance (sensitive to 0.1 mg, #FA1204N, Jinghai Instrument Co., Ltd. Shanghai, China) in the two experimental groups. The electrodes of the corrected benchtop precision pH meter (#PHS3C, Lei Magnetic, Shanghai Yidian Scientific Instrument Co., Ltd.) were inserted into the muscle samples of the breast muscle, and three different positions were repeatedly measured, and the average value was calculated as the measurement result. For shear force analysis, breast muscle samples (size 2.5 cm × 1.0 cm × 0.5 cm) without fascia, sarcolemma and obvious fat were collected, then put them into the labeled ziplock bag and put them into the water bath at 75 °C for 30–45 s, until the meat turns white. A digital display muscle tenderness meter (#CLM36, Northeast Agricultural University School of Engineering, China) to cut three points in the direction of the vertical pectoral muscle fibers
3.2. Effects of broiler TD on production performance To investigate the effects of TD on production performance, the BW, AFI, ADG, and FCR of broiler chicken were measured (Fig. 2). There were no significant differences in BW, AFI, ADG, and FCR between the control and the TD group from day 0 to day 3 of experiment. However, on days 4–7, 8–10 and 11–14, there was a significant inhibition of AFI (p < 0.0001, p = 0.0003, and p = 0.0015, respectively) and ADG (p < 0.0001, p = 0.0020, and p = 0.0001, respectively) of TD broilers when compared to the controls, which led to an increase in FCR (p < 0.0001, p = 0.3548, and p = 0.0070, respectively) (Fig. 2B–D). Similarly, the BW of broilers in TD group showed a very significant inhibition trend from day 4 to day 14 of experiment compared to the control group chickens and the broiler weight of the control group (491.65 g) was 1.408 fold greater than that of the TD group (349.07 g) on day 14 (Fig. 2A). 2
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Fig. 1. The change of foraging behavior and resting position in the TD group chickens. Representative morphological images of proximal tibias from control and TD chickens taken under the same conditions. Black arrow refers to TD lesion (TDL). (B) Quantitative analysis of tibia bone weight in control and TD groups from 7- and 14-day-old broiler chickens. (n = 12 chickens in each group from four independent experiments). There was a statistical difference of p value less than 0.05; Data represent means ± s.d. two-tailed unpaired t-test. (C) Representative figure of broiler's foraging behavior and resting position in control and TD groups from 10-day-old broiler chickens.
3.3. Effects of broiler TD on the growth of drumstick
3.4. Effects of broiler TD on the meat quality of the breast muscle
Correlation analysis showed that the weight and length of broiler drumstick were positively correlated with the BW (R2 = 0.8444, p < 0.001; R2 = 0.9134, p < 0.001, respectively) in Fig. 3A and B. Therefore, the inhibition of BW of broiler chicken negatively affect the growth of drumstick muscles. The broiler drumsticks were weighed and measured at the 7th and 14th day. The results showed that the weight and length of the broiler drumstick in the TD group were significantly reduced (p < 0.001 and p < 0.001, respectively). In addition, visually observation of the drumstick muscle color showed that the control meat was brighter than that of the TD group, and this difference was more obvious on day 14 (Fig. 3C and D).
To further determine whether TD in broiler chickens has an impact on breast muscle quality, we compared shear force and pH values of breast muscle from control and TD groups. As shown in Fig. 4A and B, shear force and pH values of breast muscle on day 7 were markedly decreased (p < 0.0017 and p = 0.0008, respectively) in TD compared to the control group. On day 14, there was a significant difference in pH values (p = 0.0170), no significant difference in shear force values (p < 0.0741) of breast muscle of the TD group compared to the control group. Moreover, heatmap analysis clearly showed that the shear force and pH values of breast muscle in the TD chicken group (green) was markedly different from that of the normal group (red), especially on day 7; and the shear force value of the breast muscles was increased
Fig. 2. The overall performance of broilers in TD group was lower than that in control group. The broiler body weights in control and TD groups were recorded from whole experimental time (n = 12). The data are expressed as the mean ± s.d. *p < 0.05, ⁎⁎⁎ p < 0.001, two-tailed unpaired t-test. (B-D) The overall performance of broilers in control and TD groups were counted from four different time periods (including 0−3 days, 4−7 days, 8−10 days, and 11−14 days, n = 4 treatments). There was a statistical difference of p value less than 0.05, two-tailed unpaired t-test. 3
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Fig. 3. Lower drumstick parameters in broilers affected by TD. (A-B) Evaluation of the correlation between the drumstick weight (n = 24) and its length (n = 24) and body weight (n = 12). The data are analyzed with Pearson's test. **p < 0.01. (C-D) Representative morphological images of drumstick muscle from control and TD chickens taken under the same conditions, and quantitative analysis of drumstick muscle weight and drumstick length in control and TD groups from 7- and 14-day-old broiler chickens (n = 24). There was a statistical difference of p value less than 0.05; Data represent means ± s.d. two-tailed unpaired t-test.
with age (Fig. 4C). Correlation analysis indicated that the shear force value of breast muscle were positively correlated with the weight and length of broiler drumstick and body weight (R2 = 0.902, R2 = 0.920, and R2 = 0.895, respectively), however, there were no significant correlation between the pH of the breast muscle and the weight and length of broiler drumstick and body weight (R2 = 0.126, R2 = 0.153, and R2 = 0.06, respectively) as shown in Fig. 4D.
3.5. Effects of TD on serum biochemical parameters Serum biochemical parameters showed that CK and AST activities were markedly elevated (p = 0.0068 and p = 0.0083, respectively) in TD group chickens on day 7. No significant differences were observed on day 14 (p = 0.1849 and p = 0.2218, respectively) (Table 1). On the other hand, there were a downward trend, but no significant, on days 7 and 14 in the levels of AMY (p = 0.5145 and p = 0.6959, respectively) and TG (p = 0.2727 and p = 0.6913, respectively) in the TD chickens. The GLU level had an upward trend, but also no significant on days 7 4
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Fig. 4. The meat quality of the breast muscle has changed in TD group chickens. (A-B) The shear force values and pH of the pectoral muscle changed when the chicken undergoes TD. Data represent means ± s.d. There was a statistical difference of p value less than 0.05, two-tailed unpaired t-test. (C)The changes in shear force values and pH of the pectoral muscle were shown in heatmaps using the indicated pseudocolor scale from 0 percent (green) to 100 percent (red), relative to average values. (D) Evaluation of the correlation between the BW and drumstick, breast related parameters. The data are analyzed with Pearson's test. **p < 0.01.
Table 1 Evaluation of the serum biochemical indicators in normal and TD chickens. Item
Day 7 Control group
TD group
P-value
Day 14 Control group
TD group
P-value
CK (U/L) AST (U/L) AMY (U/L) TG (mmol/L) Glucose (mmol/L)
784.00 ± 30.50 147.50 ± 12.43 1092.25 ± 133.56 0.38 ± 0.09 8.38 ± 0.50
1600.50 ± 199.46 228.75 ± 16.95 898.50 ± 118.76 0.26 ± 0.03 9.80 ± 0.58
0.007 0.008 0.320 0.273 0.113
1352.75 ± 239.79 151.75 ± 28.53 839.25 ± 184.81 0.57 ± 00.10 11.29 ± 1.01
2081.25 ± 423.26 192.50 ± 8.91 934.75 ± 141.57 0.17 ± 0.08 9.96 ± 1.63
0.185 0.222 0.696 0.691 0.513
Notes: Data represent means ± standard error of the means. (n = 4 chickens from three independent experiments). There was a statistical difference of p value less than 0.05, two-tailed unpaired t-test. CK, Creatine kinase; AST, Aspartic acid aminotransferase; AMY, Amylase; TG, Triglyceride; GLU, glucose.
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Fig. 5. Proposed model of broiler chicken suffered from TD plays a detrimental role in breast muscle quality. TD is one of many factors that can cause leg problems in broilers. Once a broiler suffers from TD, it may affect the quality of the breast muscle by limiting the movement of the broiler chicken. ADG, average daily weight gain; AFI, average daily feed intake; FCR, feed conversion ratio; BW, body weight; CK, Creatine kinase; AST, Aspartic acid aminotransferase.
related products and also considered to be a reliable parameter of meat quality noted by consumers (Owens et al., 2000; zhao et al., 2019). These findings further indicated that the leg problems of broiler chickens not only affected the drumstick muscle quality, but also reduced the growth performance. Meat quality is generally reflected by several parameters as pH, meat color and shear force. Among them, shear force is a valuable and intuitive indicator for establishing standards for guaranteeing muscle tenderness (Lonergan et al., 2003; Bowker, Eastridge et al., 2011). Moreover, muscle tenderness has been shown to influence purchasing decisions of consumers who are willing to pay more for guaranteed tender meat (Huffman et al., 1996; Miller et al., 2001). Le Bihan-Duval et al. (2008) had suggested that pH of meat is a relevant selection criterion to improve meat characteristics because the pH parameter can be strongly correlated to meat color, water-holding capacity, and texture. Therefore, shear force and pH values are important indicators for evaluating the quality of poultry meat (Mueller et al., 2018; Soglia et al., 2018). In the present study, it was observed that the shear force values and pH of breast muscle on 7th day were markedly lower in the TD group as compared to the control group (Fig. 4). In addition, correlation analysis also pointed out that the shear force values of breast muscle were positively correlated with the weight and length of broiler drumstick and body weight. These results indicated that the meat quality of breast muscles is reduced after leg problems in broilers. However, the effect of broiler leg disease on breast muscle flavor and trace elements, as well as their nutritional value need further study. Many serum biochemical parameters can reflect an animal's physiological status and nutrient metabolic changes, nutrient deposition and simultaneously can also be influenced by numerous factors (Wang et al., 2009; Jin et al., 2019). Serum creatine kinase activity is directly related to muscle volume and activity, and an increase in serum creatine kinase concentration elevated indicates that muscle cells are damaged or the muscles are stressed resulting from the enzyme releases into the bloodstream (Śmiecińska et al., 2011; Matshogo et al., 2018). AST is transaminases widely present in animal mitochondria and have extremely important functions in liver and muscle health (Wei et al., 2019). Our results showed that the activity of serum CK and AST were markedly elevated in broiler chickens with TD, indicating that the occurrence of broiler leg disease has an adverse effect on the breast muscles. The serum CK activity in broilers with TD is in agreement with previous report by Mcdonald et al. (1991) who indicated that a 29-yearold man with proximal leg weakness has a high serum level of creatine kinase suggesting a myopathy. Additionally, serum AMY, TG, and GLU levels are considered important indicators for evaluating pancreatic
and 14 (p = 0.1127 and p = 0.5133, respectively) in the TD chickens (Table 1). 4. Discussion Due to the fast growth rate, broilers are very susceptible to leg disease (Nääs, et al., 2010; Huang et al., 2017). Broilers once suffering from TD will cause leg weakness, motion reduction and even fractures, which leads to change in drinking and feeding behavior and meat quality (Herzog et al., 2011; Pelicia et al., 2012; Huang et al. 2019). The present study showed that TD in broilers can decrease the growth performance, inhibit drumstick growth, and affect the meat quality of breast muscle that are reflected by changes in shear force, pH and serum biochemical parameter levels, which suggest that broiler leg problems is detrimental to breast meat quality (Fig. 5). The feeding behavior of broilers is concerned, not only related to production performance, but also to assess the difficulty of obtaining feed for lame broilers (Week et al., 2000). Danbury et al. (2000) studied the walking ability of lame broilers and pointed out that trained birds selected more analgesic supplemented feed when the severity of the lameness increased, indicating that lame broiler might feel pain. In the present study, it was observed that the foraging behavior and resting posture of the broiler chicken suffered from TD changed significantly compared with the normal broiler chickens during the trial, which may be related to the injury of the tibial growth plate and the reduction of the tibial bone mass, causing the leg pain of the broiler chickens. Growth performance is an important parameter used to evaluate poultry production and can be affected by a variety of factors (Jin et al., 2019). Pain in the legs changes the feeding behavior of broilers, which may affect broiler performance. We next found that the production performance (main including AFI and ADG) of the TD group was significantly lower than the control group, which resulted in a the slow growth in broiler weight gain during the whole experimental period. These results are agreement with Yao et al. (2018), who reported that the daily weight, AFI, and ADG was significantly reduced in TD group as compared with the control group from days 7 to 14. Therefore, these decreased growth performance of the TD group chicken may be related to movement limitation in the legs. In addition, the growth performance of broilers is also related to the inhibition of drumstick related - parameters (including drumstick weight and drumstick length), and the drumstick parameters in the TD group chickens were significantly suppressed and drumstick muscle color was pale (Fig. 3). In our lives, meat color is one of the important characteristics used by consumers when they purchase meat and meat 6
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function, blood lipids and glucose levels, respectively. Serum AMY, TG, and GLU levels showed no differences between control group chickens and TD group chickens in this study. Based on the above results, it is further confirmed by serum biochemical parameters that the leg disease problem of broilers may have certain adverse effects on chicken meat.
s41598-017-09664-6. Huffman, K.L., Miller, M.F., Hoover, L.C., Wu, C.K., Brittin, H.C., Ramsey, C.B., 1996. Effect of beef tenderness on consumer satisfaction with steaks consumed in the home and restaurant. J. Anim. Sci. 74, 91–97. https://doi.org/10.2527/1996.74191x. Jin, S., Fan, X., Yang, L., He, T., Xu, Y., Chen, X., Liu, P., Geng, Z., 2019. Effects of rearing systems on growth performance, carcass yield, meat quality, lymphoid organ indices, and serum biochemistry of Wannan Yellow chickens. Anim. Sci. J. 2019, 1–7. https:// doi.org/10.1111/asj.13220. Le Bihan-Duval, E., Debut, M., Berri, C.M., Sellier, N., Santé-Lhoutellier, V., Jégo, Y., Beaumont, C., 2008. Chicken meat quality: genetic variability and relationship with growth and muscle characteristics. BMC Genet. 9, 53. https://doi.org/10.1186/14712156-9-53. Lonergan, S.M., Deeb, N., Fedler, C.A., Lamont, S.J., 2003. Breast meat quality and composition in unique chicken populations. Poult. Sci. 82, 1990–1994. https://doi. org/10.1093/ps/82.12.1990. Matshogo, T.B., Mlambo, V., Marume, U., Sebola, N., 2018. Growth performance, blood parameters, carcass characteristics and meat quality traits in Potchefstroom Koekoek chickens fed Lippia javanica leaf meal. Trop. Anim. Health Prod. 50, 1787–1795. https://doi.org/10.1007/s11250-018-1620-9. McDonald, T.D., Medori, R., Younger, D.S., Chang, H.W., Minetti, C., Uncini, A., Bonilla, E., Hays, A.P., Lovelace, R.E., 1991. Becker muscular dystrophy or spinal muscular atrophy?—Dystrophin studies resolve conflicting results of electromyography and muscle biopsy. Neuromuscul. Disord. 1, 195–200. https://doi.org/10.1016/09608966(91)90024-M. Miller, M.F., Carr, M.A., Ramsey, C.B., Crockett, K.L., Hoover, L.C., 2001. Consumer thresholds for establishing the value of beef tenderness. J. Anim. Sci. 79 (12), 3062–3068. https://doi.org/10.2527/2001.79123062x. Mueller, S., Kreuzer, M., Siegrist, M., Mannale, K., Messikommer, R.E., Gangnat, I.D.M., 2018. Carcass and meat quality of dual-purpose chickens (Lohmann Dual, Belgian Malines, Schweizerhuhn) in comparison to broiler and layer chicken types. Poult. Sci. 97, 3325–3336. https://doi.org/10.3382/ps/pey172. Nääs, I.D.A., Paz, I.C.D.L.A., Baracho, M.D.S., Menezes, A.G.D., Lima, K.A.O.D., Bueno, L.G.D.F., Neto, M.M., de Carvalho, V.C., Almeida, I.C.D.L., Souza, A.L.D., 2010. Assessing locomotion deficiency in broiler chicken. Scientia Agricola 67, 129–135. http://dx.doi.org/10.1590/S0103-90162010000200001. Naji, T.A., Amadou, I., Zhao, R.Y., Tang, X., Shi, Y.H., Le, G.W., 2014. Effects of phytosterol in feed on growth and related gene expression in muscles of broiler chickens. Trop. J. Pharm. Res. 13 (1), 9–16. http://dx.doi.org/10.4314/tjpr.v13i1.2. Owens, C.M., Hirschler, E.M., Martinez-Dawson, R., Sams, A.R., 2000. The characterization and incidence of pale, soft, exudative turkey meat in a commercial plant. Poult. Sci. 79, 553–558. https://doi.org/10.1093/ps/79.4.553. Pelicia, K., Aparecido Jr, I.M., Garcia, E.A., Molino, A.B., Santos, G.C., Berto, D.A., Vieira Filho, J.A.I.V., Murakami, E.S.M.V., Montenegro, A.T., Silva, A.M., 2012. Evaluation of a radiographic method to detect tibial dyschondroplasia lesions in broilers. Rev. Bras. Cienc. Avic. 14, 129–135. http://dx.doi.org/10.1590/S1516635X2012000200007. Rath, N.C., Richards, M.P., Huff, W.E., Huff, G.R., Balog, J.M., 2005. Changes in the tibial growth plates of chickens with thiram-induced dyschondroplasia. J. Comp. Pathol. 133, 41–52. https://doi.org/10.1016/j.jcpa.2005.01.005. Śmiecińska, K., Denaburski, J., Sobotka, W., 2011. Slaughter value, meat quality, creatine kinase activity and cortisol levels in the blood serum of growing-finishing pigs slaughtered immediately after transport and after a rest period. Pol. J. Vet. Sci. 14, 47–54. https://doi.org/10.2478/v10181-011-0007-x. Soglia, F., Baldi, G., Laghi, L., Mudalal, S., Petracci, M., 2018. Effect of white striping on turkey breast meat quality. Animal 12, 1–7. https://doi.org/10.1017/ S1751731117003469. Wang, J.P., Yoo, J.S., Kim, H.J., Lee, J.H., Kim, I.H., 2009. Nutrient digestibility, blood profiles and fecal microbiota are influenced by chitooligosaccharide supplementation of growing pigs. Livest. Sci. 125, 298–303. https://doi.org/10.1016/j.livsci.2009.05. 011. Weeks, C.A., Danbury, T.D., Davies, H.C., Hunt, P., Kestin, S.C., 2000. The behaviour of broiler chickens and its modification by lameness. Appl. Anim. Behav. Sci. 67, 111–125. http://dx.doi.org/10.1016/s0168-1591(99)00102-1. Wei, J., Guo, W., Yang, X., Chen, F., Fan, Q., Wang, H., Zhang, N., Diao, Q., 2019. Effects of dietary ramie level on growth performance, serum biochemical indices, and meat quality of Boer goats. Trop. Anim. Health Prod. 1–7. https://doi.org/10.1007/ s11250-019-01891-5. Yao, W., Zhang, H., Jiang, X., Mehmood, K., Iqbal, M., Li, A., Zhang, J., Wang, Y., Waqas, M., Shen, Y., Li, J., 2018. Effect of total flavonoids of rhizoma drynariae on tibial dyschondroplasia by regulating BMP-2 and Runx2 expression in chickens. Front. Pharmacol. 9, 1251. https://doi.org/10.3389/fphar.2018.01251. Zhao, Y.R., Chen, Y.P., Cheng, Y.F., Qu, H.M., Li, J., Wen, C., Zhou, Y.M., 2019. Effects of dietary phytosterols on growth performance, antioxidant status, and meat quality in Partridge Shank chickens. Poul. Sci. 1–7. https://doi.org/10.3382/ps/pez059. Zheng, Y., Li, Y., Satija, A., Pan, A., Sotos-Prieto, M., Rimm, E., Willett, W.C., Hu, F.B., 2019. Association of changes in red meat consumption with total and cause specific mortality among US women and men: two prospective cohort studies. B.M.J. 365, l2110. https://doi.org/10.1136/bmj.l2110.
5. Conclusion The data in the present study indicated that leg problems (TD) in broilers can decrease the growth performance, inhibit broiler drumstick muscle growth, and affect the meat quality of breast muscle, with increased biochemical parameter levels (including CK and AST) in serum, which suggest that broiler leg problems is detrimental to meat quality. Acknowledgments We are grateful to the financial support from the Outstanding Talents of Henan Agricultural University (No. 30500421) for supporting this work. Supplementary materials Supplementary material associated with this article can be found in the online version, at doi:10.1016/j.livsci.2020.103956. References Bizeray, D., Leterrier, C., Constantin, P., Picard, M., Faure, J.M., 2002. Sequential feeding can increase activity and improve gait score in meat-type chickens. Poult. Sci. 81, 1798–1806. https://doi.org/10.1093/ps/81.12.1798. Bowker, B.C., Eastridge, J.S., Solomon, M.B., 2011. Use of gelatin gels as a reference material for performance evaluation of meat shear force measurements. J. Food Sci. 76, S210–S216. https://doi.org/10.1111/j.1750-3841.2011.02100.x. Cavani, C., Petracci, M., Trocino, A., Xiccato, G., 2009. Advances in research on poultry and rabbit meat quality. Italian J. Anim. Sci. 8, 741–750. https://doi.org/10.4081/ ijas.2009.s2.741. Danbury, T.C., Weeks, C.A., Waterman-Pearson, A.E., Kestin, S.C., Chambers, J.P., 2000. Self-selection of the analgesic drug carprofen by lame broiler chickens. Veterinary Rec. 146, 307–311. http://dx.doi.org/10.1136/vr.146.11.307. FAO, 2010. Agribusiness Handbook: Poultry Meat and Eggs. Rome, Italy. http://www. eastagri.org/publications/pub_docs/6_Poultry_web.pdf. Funaro, A., Cardenia, V., Petracci, M., Rimini, S., Rodriguez-Estrada, M.T., Cavani, C., 2014. Comparison of meat quality characteristics and oxidative stability between conventional and free-range chickens. Poult. Sci. 93, 1511–1522. https://doi.org/10. 3382/ps.2013-03486. Garcia, A.F.Q.M., Murakami, A.E., do Amaral Duarte, C.R., Rojas, I.C.O., Picoli, K.P., Puzotti, M.M., 2013. Use of vitamin D3 and its metabolites in broiler chicken feed on performance, bone parameters and meat quality. Asian-Australas. J. Anim. Sci. 26, 408–415. https://doi.org/10.5713/ajas.2012.12455. González-Cerón, F., Rekaya, R., Aggrey, S.E., 2015. Genetic relationship between leg problems and bone quality traits in a random mating broiler population. Poult. Sci. 94, 1787–1790. https://doi.org/10.3382/ps/pev159. Guasch-Ferré, M., Satija, A., Blondin, S.A., Janiszewski, M., Emlen, E., O'Connor, L.E., Campbell, W.W., Hu, F.B., Willett, W.C., Stampfer, M.J., 2019. Meta-analysis of randomized controlled trials of red meat consumption in comparison with various comparison diets on cardiovascular risk factors. Circulation 139, 1828–1845. https:// doi.org/10.1161/CIRCULATIONAHA.118.035225. Herzog, A., Genin, O., Hasdai, A., Shinder, D., Pines, M., 2011. Hsp90 and angiogenesis in bone disorders—lessons from the avian growth plate. Am. J. Physiol. Regul. Integr. Comp. Physiol. 301 (1), R140–R147. https://doi.org/10.1152/ajpregu.00134.2011. Huang, S., Kong, A., Cao, Q., Tong, Z., Wang, X., 2019. The role of blood vessels in broiler chickens with tibial dyschondroplasia. Poult. Sci. 98, 6527–6532. http://dx.doi.org/ 10.3382/ps/pez497. Huang, S., Rehman, M.U., Qiu, G., Luo, H., Iqbal, M.K., Zhang, H., Mehmood, K., Li, J., 2018. Tibial dyschondroplasia is closely related to suppression of expression of hypoxia-inducible factors 1α, 2α, and 3α in chickens. J. Vet. Sci. 19, 107–115. https:// doi.org/10.4142/jvs.2018.19.1.107. Huang, S.C., Rehman, M.U., Lan, Y.F., Qiu, G., Zhang, H., Iqbal, M.K., Luo, H.Q., Mehmood, K., Zhang, L.H., Li, J.K., 2017. Tibial dyschondroplasia is highly associated with suppression of tibial angiogenesis through regulating the HIF-1α/VEGF/ VEGFR signaling pathway in chickens. Sci. Rep. 7, 9089. https://doi.org/10.1038/
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Characterization of growth performance, meat quality and serum biochemical parameters in chickens suffering from tibial dyschondroplasia
T
Qin-Qin Caoa, An-An Konga, Kun-Sheng Taoa, Shi-Hao Zhenga, Chao Tonga, Xue-Bing Wanga, Zong-Xi Tonga, Mujeeb Ur Rehmanb, Shu-Cheng Huanga,⁎ a b
College of Animal Science and Veterinary Medicine, Henan Agricultural University, 95# Wenhua Road, Jinshui District, Zhengzhou, Henan 450002, PR China Research Center of Avian Disease, College of Veterinary Medicine of Sichuan Agricultural University, Chengdu, Sichuan, PR China
ARTICLE INFO
ABSTRACT
Keywords: Drumstick weight Meat quality Leg problems Tibial dyschondroplasia Serum biochemistry
The present study was conducted to examine the clinical manifestations of broilers suffering from tibial dyschondroplasia (TD) on growth performance, drumstick parameters, breast meat quality, and serum biochemical parameters. A total of 200 Arbor Acre broiler chickens were randomly allocated into two groups: control (basal diet) and TD group (basal diet supplemented with 50 mg/kg of thiram). Each group consisted of 4 replicates (25 chickens per replicate). The experiment lasted for 14 days. Results showed that TD chickens exhibited abnormal foraging behavior and resting position that caused a reduction in feed intake, daily weight gain and final body weight. Compared with the control group, the weight and length of drumstick in TD chicken group was significantly decreased (p < 0.05) and its color was pale. Interestingly, drumstick related-parameters were statistically correlated with breast muscle quality in control and TD chickens (p < 0.01). In addition, breast meat quality (assessed by shear force and pH) in TD group chickens was affected. Furthermore, TD chickens had significantly elevated serum creatine kinase (CK) and aspartic acid aminotransferase (AST) activities (p < 0.05), but no significant change was observed in serum amylase, triglycerides, and glucose levels (p > 0.05). In conclusion, the TD has adverse effect on the overall growth performance, drumstick parameters, breast meat quality, and some serum biochemical parameters of broilers. Moreover, decreased breast muscle quality was strongly correlated with leg problems in the tibia.
1. Introduction With the improvement of people's living standards, more attention is paid to food safety and health. Several studies reported that increase in red meat consumption is associated with higher mortality rates and cardiovascular disease (Guasch-Ferré et al., 2019; Zheng et al., 2019). In view of nutrition and human health concerns, chicken is an economical and affordable meat product that has a higher protein content and lower total fat, and provides essential vitamins and minerals. Therefore, it becomes an ideal meat source for consumers (FAO, 2010; Naji et al., 2014). The majority of the poultry meat products that reaches the food market are produced with birds reared under intensive conditions, which led to the selection of birds with rapid growth and feed efficiency genotypes (Cavani et al, 2009; Funaro et al., 2014). However, several authors have shown that the selection for fast-growing broiler has resulted in a disproportionate development of broiler bones compared to body weight gain, which eventually lead to a higher incidence of leg ⁎
problems (Garcia et al., 2013; González-Cerón et al, 2015). Most of the noninfectious disorders such as tibial dyschondroplasia (TD) and tarsal varus-valgus deformities (VVD) result in abnormal gait and even lameness. Reduction of leg abnormalities has become a major challenge in meat-type chicken because they impair growth performance, reduce meat quality and compromise welfare status of birds (Bizeray et al, 2002; Garcia et al., 2013). TD is an intractable leg problem in fast growing meat-type chickens that results in bone deformation and leg lameness (Herzog et al., 2011; Huang et al., 2017). This tibiotarsal bone disorder leads to apparent locomotion problem with rising prevalence of 30% in broilers flock (Pelicia et al., 2012; Huang et al., 2018). However, the relationship between TD and meat quality of broiler chickens is not known. Based on this consideration, the objective of this study was to establish a TD chicken model for studying the effects of leg problems (TD) on growth performance, meat quality and several biochemical parameters in broiler chickens.
Corresponding author. E-mail address: [email protected] (S.-C. Huang).
https://doi.org/10.1016/j.livsci.2020.103956 Received 7 July 2019; Received in revised form 2 February 2020; Accepted 3 February 2020 Available online 04 February 2020 1871-1413/ © 2020 Elsevier B.V. All rights reserved.
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2. Materials and methods
and record the readings.
2.1. Animals and experimental protocol
2.5. Serum biochemistry analysis
All procedures, treatments, and animal care were strictly in compliance with the guidelines of the Institutional Animal Care and Use Committee of Henan Agricultural University (Permit No: 11-0085). All arbor acres (AA) chickens (regardless of gender; one-day-old; 51.96 ± 3.87 g) were obtained from a commercial hatchery (Xingda Poultry Industry Co. Ltd., Kaifeng, China). Thiram (tetramethylthiuram disulfide) (AR, purity ≥ 98%, #C10036460) was acquired from Macklin Biochemical Co., Ltd. (Shanghai, China). All chicks were randomly allocated into two groups: the control group and the thiram-induced TD group (n = 100/group, with 25 chicks per replicate and 4 replicates per group), and raised in singlelayer metal cages (size, 120 cm × 90 cm × 40 cm). Standard feed and water was provided ad libitum. The environmentally controlled facility was maintained with the recommended standard room temperature, relative humidity, daily light, good ventilation, and hygienic conditions as previously described (Huang et al., 2017). After 3 days of acclimation, and from the fourth day the control group was offered a standard diet and the thiram - induced TD group received the same standard diet further supplemented with 50 mg/kg of thiram (thiram was removed on day 7) for the purpose of constructing a broiler TD model to facilitate the study of the TD mechanism as previously reported (Rath et al., 2005; Huang et al., 2017). The experiment lasted for 14 days.
The blood samples were centrifuged at 3000 × g for 20 min for the separation of serum and stored at −20 °C until further analyses. Creatine kinase (CK), aspartic acid aminotransferase (AST), amylase (AMY), triglycerides (TG), glucose (GLU) serum levels were determined. These biochemical indicators were quantified through a veterinary reagent tray purchased from Chengdu Puli Taisheng Technology Co., Ltd and quantitative detection was performed on a portable full automated veterinary biochemistry analyzer (#SMT-120V, Seamaty Technology Co., Ltd.,Chengdu, China) according to the manufacturer instructions. 2.6. Statistical analysis Correlation analyses were performed using Pearson's test, and the differences between groups of shear force and pH values of breast muscle were analyzed and displayed by a pseudocolor scale (from green to red) respectively, which are shown in the form of a heatmap. In addition, comparisons between two groups were implemented using unpaired, two-tailed Student's t-tests with SPSS (#Version 17.0., Chicago, IL) or Prism statistical analysis software (#Version 7.00, GraphPad Software, Inc.). Biochemical parameters were repeated at least three times to increase the reproducibility of the findings. In addition, representative results are displayed in figures. Data are presented as the means ± S.D. P < 0.05 was considered to be statistically significant.
2.2. Clinical manifestations and production performance analysis The chicks were group-weighed on day 3, 7, 10, and 14, and the average daily weight gain (ADG), average daily feed intake (AFI), and feed conversion ratio (FCR) were calculated per group. During the experiment, the clinical manifestations and feeding behavior of broilers were observed, and mortality of broilers was recorded on a daily basis.
3. Results 3.1. Effects of broiler leg problems on clinical features AA broiler chicken is a meat - type broiler and the fast weight gain adds a burden on the legs. TD is one of the most prone leg problems in broilers with fast growth rates. The experiment used thiram to establish a broiler TD model. The pathological feature is indicated by the fact that the proximal proximal tibial growth plate was clearly widened (Fig. 1A), and the tibia bone mass was significantly reduced (Fig. 1B). Once a broiler has a leg problem, some representative clinical symptoms are observed from the 10th day of age (Fig. 1). The foraging behavior in the control group showed eating in a normal standing position. In contrast, the posture of the broiler with legs stretched forward under its chest during feeding was observed in the TD group (Fig. 1C). Additionally, the change of resting position was noticed and TD group broilers’ leg showed to one side for outreach, even split-leg-like poses compared with the control broilers (Fig. 1C).
2.3. Morphology of the tibial growth plates On day 7 and 14, three chicks per treatment cage were randomly selected and euthanized as previously described (Huang et al., 2017). The stripping of the tibial longitudinalmuscles were performed and the tibiae were retrieved. The weight of the tibia was determined using an electronic balance (sensitive to 0.1 mg, #FA1204N, Jinghai Instrument Co., Ltd. Shanghai, China). In addition, preparation of sagittal sections of the proximal tibial growth plates was performed to analyze the morphology of growth plates. 2.4. Observation and evaluation of drumsticks and breast muscles The chicken drumsticks were collected from the control and the TD group on days 7 and 14. The drumstick muscle color was then visually evaluated and the length and weight of drumstick were measured and counted using an Digital Calipers (#SATA91511, TATA Company, Shanghai, China) and electronic balance (sensitive to 0.1 mg, #FA1204N, Jinghai Instrument Co., Ltd. Shanghai, China) in the two experimental groups. The electrodes of the corrected benchtop precision pH meter (#PHS3C, Lei Magnetic, Shanghai Yidian Scientific Instrument Co., Ltd.) were inserted into the muscle samples of the breast muscle, and three different positions were repeatedly measured, and the average value was calculated as the measurement result. For shear force analysis, breast muscle samples (size 2.5 cm × 1.0 cm × 0.5 cm) without fascia, sarcolemma and obvious fat were collected, then put them into the labeled ziplock bag and put them into the water bath at 75 °C for 30–45 s, until the meat turns white. A digital display muscle tenderness meter (#CLM36, Northeast Agricultural University School of Engineering, China) to cut three points in the direction of the vertical pectoral muscle fibers
3.2. Effects of broiler TD on production performance To investigate the effects of TD on production performance, the BW, AFI, ADG, and FCR of broiler chicken were measured (Fig. 2). There were no significant differences in BW, AFI, ADG, and FCR between the control and the TD group from day 0 to day 3 of experiment. However, on days 4–7, 8–10 and 11–14, there was a significant inhibition of AFI (p < 0.0001, p = 0.0003, and p = 0.0015, respectively) and ADG (p < 0.0001, p = 0.0020, and p = 0.0001, respectively) of TD broilers when compared to the controls, which led to an increase in FCR (p < 0.0001, p = 0.3548, and p = 0.0070, respectively) (Fig. 2B–D). Similarly, the BW of broilers in TD group showed a very significant inhibition trend from day 4 to day 14 of experiment compared to the control group chickens and the broiler weight of the control group (491.65 g) was 1.408 fold greater than that of the TD group (349.07 g) on day 14 (Fig. 2A). 2
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Fig. 1. The change of foraging behavior and resting position in the TD group chickens. Representative morphological images of proximal tibias from control and TD chickens taken under the same conditions. Black arrow refers to TD lesion (TDL). (B) Quantitative analysis of tibia bone weight in control and TD groups from 7- and 14-day-old broiler chickens. (n = 12 chickens in each group from four independent experiments). There was a statistical difference of p value less than 0.05; Data represent means ± s.d. two-tailed unpaired t-test. (C) Representative figure of broiler's foraging behavior and resting position in control and TD groups from 10-day-old broiler chickens.
3.3. Effects of broiler TD on the growth of drumstick
3.4. Effects of broiler TD on the meat quality of the breast muscle
Correlation analysis showed that the weight and length of broiler drumstick were positively correlated with the BW (R2 = 0.8444, p < 0.001; R2 = 0.9134, p < 0.001, respectively) in Fig. 3A and B. Therefore, the inhibition of BW of broiler chicken negatively affect the growth of drumstick muscles. The broiler drumsticks were weighed and measured at the 7th and 14th day. The results showed that the weight and length of the broiler drumstick in the TD group were significantly reduced (p < 0.001 and p < 0.001, respectively). In addition, visually observation of the drumstick muscle color showed that the control meat was brighter than that of the TD group, and this difference was more obvious on day 14 (Fig. 3C and D).
To further determine whether TD in broiler chickens has an impact on breast muscle quality, we compared shear force and pH values of breast muscle from control and TD groups. As shown in Fig. 4A and B, shear force and pH values of breast muscle on day 7 were markedly decreased (p < 0.0017 and p = 0.0008, respectively) in TD compared to the control group. On day 14, there was a significant difference in pH values (p = 0.0170), no significant difference in shear force values (p < 0.0741) of breast muscle of the TD group compared to the control group. Moreover, heatmap analysis clearly showed that the shear force and pH values of breast muscle in the TD chicken group (green) was markedly different from that of the normal group (red), especially on day 7; and the shear force value of the breast muscles was increased
Fig. 2. The overall performance of broilers in TD group was lower than that in control group. The broiler body weights in control and TD groups were recorded from whole experimental time (n = 12). The data are expressed as the mean ± s.d. *p < 0.05, ⁎⁎⁎ p < 0.001, two-tailed unpaired t-test. (B-D) The overall performance of broilers in control and TD groups were counted from four different time periods (including 0−3 days, 4−7 days, 8−10 days, and 11−14 days, n = 4 treatments). There was a statistical difference of p value less than 0.05, two-tailed unpaired t-test. 3
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Fig. 3. Lower drumstick parameters in broilers affected by TD. (A-B) Evaluation of the correlation between the drumstick weight (n = 24) and its length (n = 24) and body weight (n = 12). The data are analyzed with Pearson's test. **p < 0.01. (C-D) Representative morphological images of drumstick muscle from control and TD chickens taken under the same conditions, and quantitative analysis of drumstick muscle weight and drumstick length in control and TD groups from 7- and 14-day-old broiler chickens (n = 24). There was a statistical difference of p value less than 0.05; Data represent means ± s.d. two-tailed unpaired t-test.
with age (Fig. 4C). Correlation analysis indicated that the shear force value of breast muscle were positively correlated with the weight and length of broiler drumstick and body weight (R2 = 0.902, R2 = 0.920, and R2 = 0.895, respectively), however, there were no significant correlation between the pH of the breast muscle and the weight and length of broiler drumstick and body weight (R2 = 0.126, R2 = 0.153, and R2 = 0.06, respectively) as shown in Fig. 4D.
3.5. Effects of TD on serum biochemical parameters Serum biochemical parameters showed that CK and AST activities were markedly elevated (p = 0.0068 and p = 0.0083, respectively) in TD group chickens on day 7. No significant differences were observed on day 14 (p = 0.1849 and p = 0.2218, respectively) (Table 1). On the other hand, there were a downward trend, but no significant, on days 7 and 14 in the levels of AMY (p = 0.5145 and p = 0.6959, respectively) and TG (p = 0.2727 and p = 0.6913, respectively) in the TD chickens. The GLU level had an upward trend, but also no significant on days 7 4
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Fig. 4. The meat quality of the breast muscle has changed in TD group chickens. (A-B) The shear force values and pH of the pectoral muscle changed when the chicken undergoes TD. Data represent means ± s.d. There was a statistical difference of p value less than 0.05, two-tailed unpaired t-test. (C)The changes in shear force values and pH of the pectoral muscle were shown in heatmaps using the indicated pseudocolor scale from 0 percent (green) to 100 percent (red), relative to average values. (D) Evaluation of the correlation between the BW and drumstick, breast related parameters. The data are analyzed with Pearson's test. **p < 0.01.
Table 1 Evaluation of the serum biochemical indicators in normal and TD chickens. Item
Day 7 Control group
TD group
P-value
Day 14 Control group
TD group
P-value
CK (U/L) AST (U/L) AMY (U/L) TG (mmol/L) Glucose (mmol/L)
784.00 ± 30.50 147.50 ± 12.43 1092.25 ± 133.56 0.38 ± 0.09 8.38 ± 0.50
1600.50 ± 199.46 228.75 ± 16.95 898.50 ± 118.76 0.26 ± 0.03 9.80 ± 0.58
0.007 0.008 0.320 0.273 0.113
1352.75 ± 239.79 151.75 ± 28.53 839.25 ± 184.81 0.57 ± 00.10 11.29 ± 1.01
2081.25 ± 423.26 192.50 ± 8.91 934.75 ± 141.57 0.17 ± 0.08 9.96 ± 1.63
0.185 0.222 0.696 0.691 0.513
Notes: Data represent means ± standard error of the means. (n = 4 chickens from three independent experiments). There was a statistical difference of p value less than 0.05, two-tailed unpaired t-test. CK, Creatine kinase; AST, Aspartic acid aminotransferase; AMY, Amylase; TG, Triglyceride; GLU, glucose.
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Fig. 5. Proposed model of broiler chicken suffered from TD plays a detrimental role in breast muscle quality. TD is one of many factors that can cause leg problems in broilers. Once a broiler suffers from TD, it may affect the quality of the breast muscle by limiting the movement of the broiler chicken. ADG, average daily weight gain; AFI, average daily feed intake; FCR, feed conversion ratio; BW, body weight; CK, Creatine kinase; AST, Aspartic acid aminotransferase.
related products and also considered to be a reliable parameter of meat quality noted by consumers (Owens et al., 2000; zhao et al., 2019). These findings further indicated that the leg problems of broiler chickens not only affected the drumstick muscle quality, but also reduced the growth performance. Meat quality is generally reflected by several parameters as pH, meat color and shear force. Among them, shear force is a valuable and intuitive indicator for establishing standards for guaranteeing muscle tenderness (Lonergan et al., 2003; Bowker, Eastridge et al., 2011). Moreover, muscle tenderness has been shown to influence purchasing decisions of consumers who are willing to pay more for guaranteed tender meat (Huffman et al., 1996; Miller et al., 2001). Le Bihan-Duval et al. (2008) had suggested that pH of meat is a relevant selection criterion to improve meat characteristics because the pH parameter can be strongly correlated to meat color, water-holding capacity, and texture. Therefore, shear force and pH values are important indicators for evaluating the quality of poultry meat (Mueller et al., 2018; Soglia et al., 2018). In the present study, it was observed that the shear force values and pH of breast muscle on 7th day were markedly lower in the TD group as compared to the control group (Fig. 4). In addition, correlation analysis also pointed out that the shear force values of breast muscle were positively correlated with the weight and length of broiler drumstick and body weight. These results indicated that the meat quality of breast muscles is reduced after leg problems in broilers. However, the effect of broiler leg disease on breast muscle flavor and trace elements, as well as their nutritional value need further study. Many serum biochemical parameters can reflect an animal's physiological status and nutrient metabolic changes, nutrient deposition and simultaneously can also be influenced by numerous factors (Wang et al., 2009; Jin et al., 2019). Serum creatine kinase activity is directly related to muscle volume and activity, and an increase in serum creatine kinase concentration elevated indicates that muscle cells are damaged or the muscles are stressed resulting from the enzyme releases into the bloodstream (Śmiecińska et al., 2011; Matshogo et al., 2018). AST is transaminases widely present in animal mitochondria and have extremely important functions in liver and muscle health (Wei et al., 2019). Our results showed that the activity of serum CK and AST were markedly elevated in broiler chickens with TD, indicating that the occurrence of broiler leg disease has an adverse effect on the breast muscles. The serum CK activity in broilers with TD is in agreement with previous report by Mcdonald et al. (1991) who indicated that a 29-yearold man with proximal leg weakness has a high serum level of creatine kinase suggesting a myopathy. Additionally, serum AMY, TG, and GLU levels are considered important indicators for evaluating pancreatic
and 14 (p = 0.1127 and p = 0.5133, respectively) in the TD chickens (Table 1). 4. Discussion Due to the fast growth rate, broilers are very susceptible to leg disease (Nääs, et al., 2010; Huang et al., 2017). Broilers once suffering from TD will cause leg weakness, motion reduction and even fractures, which leads to change in drinking and feeding behavior and meat quality (Herzog et al., 2011; Pelicia et al., 2012; Huang et al. 2019). The present study showed that TD in broilers can decrease the growth performance, inhibit drumstick growth, and affect the meat quality of breast muscle that are reflected by changes in shear force, pH and serum biochemical parameter levels, which suggest that broiler leg problems is detrimental to breast meat quality (Fig. 5). The feeding behavior of broilers is concerned, not only related to production performance, but also to assess the difficulty of obtaining feed for lame broilers (Week et al., 2000). Danbury et al. (2000) studied the walking ability of lame broilers and pointed out that trained birds selected more analgesic supplemented feed when the severity of the lameness increased, indicating that lame broiler might feel pain. In the present study, it was observed that the foraging behavior and resting posture of the broiler chicken suffered from TD changed significantly compared with the normal broiler chickens during the trial, which may be related to the injury of the tibial growth plate and the reduction of the tibial bone mass, causing the leg pain of the broiler chickens. Growth performance is an important parameter used to evaluate poultry production and can be affected by a variety of factors (Jin et al., 2019). Pain in the legs changes the feeding behavior of broilers, which may affect broiler performance. We next found that the production performance (main including AFI and ADG) of the TD group was significantly lower than the control group, which resulted in a the slow growth in broiler weight gain during the whole experimental period. These results are agreement with Yao et al. (2018), who reported that the daily weight, AFI, and ADG was significantly reduced in TD group as compared with the control group from days 7 to 14. Therefore, these decreased growth performance of the TD group chicken may be related to movement limitation in the legs. In addition, the growth performance of broilers is also related to the inhibition of drumstick related - parameters (including drumstick weight and drumstick length), and the drumstick parameters in the TD group chickens were significantly suppressed and drumstick muscle color was pale (Fig. 3). In our lives, meat color is one of the important characteristics used by consumers when they purchase meat and meat 6
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function, blood lipids and glucose levels, respectively. Serum AMY, TG, and GLU levels showed no differences between control group chickens and TD group chickens in this study. Based on the above results, it is further confirmed by serum biochemical parameters that the leg disease problem of broilers may have certain adverse effects on chicken meat.
s41598-017-09664-6. Huffman, K.L., Miller, M.F., Hoover, L.C., Wu, C.K., Brittin, H.C., Ramsey, C.B., 1996. Effect of beef tenderness on consumer satisfaction with steaks consumed in the home and restaurant. J. Anim. Sci. 74, 91–97. https://doi.org/10.2527/1996.74191x. Jin, S., Fan, X., Yang, L., He, T., Xu, Y., Chen, X., Liu, P., Geng, Z., 2019. Effects of rearing systems on growth performance, carcass yield, meat quality, lymphoid organ indices, and serum biochemistry of Wannan Yellow chickens. Anim. Sci. J. 2019, 1–7. https:// doi.org/10.1111/asj.13220. Le Bihan-Duval, E., Debut, M., Berri, C.M., Sellier, N., Santé-Lhoutellier, V., Jégo, Y., Beaumont, C., 2008. Chicken meat quality: genetic variability and relationship with growth and muscle characteristics. BMC Genet. 9, 53. https://doi.org/10.1186/14712156-9-53. Lonergan, S.M., Deeb, N., Fedler, C.A., Lamont, S.J., 2003. Breast meat quality and composition in unique chicken populations. Poult. Sci. 82, 1990–1994. https://doi. org/10.1093/ps/82.12.1990. Matshogo, T.B., Mlambo, V., Marume, U., Sebola, N., 2018. Growth performance, blood parameters, carcass characteristics and meat quality traits in Potchefstroom Koekoek chickens fed Lippia javanica leaf meal. Trop. Anim. Health Prod. 50, 1787–1795. https://doi.org/10.1007/s11250-018-1620-9. McDonald, T.D., Medori, R., Younger, D.S., Chang, H.W., Minetti, C., Uncini, A., Bonilla, E., Hays, A.P., Lovelace, R.E., 1991. Becker muscular dystrophy or spinal muscular atrophy?—Dystrophin studies resolve conflicting results of electromyography and muscle biopsy. Neuromuscul. Disord. 1, 195–200. https://doi.org/10.1016/09608966(91)90024-M. Miller, M.F., Carr, M.A., Ramsey, C.B., Crockett, K.L., Hoover, L.C., 2001. Consumer thresholds for establishing the value of beef tenderness. J. Anim. Sci. 79 (12), 3062–3068. https://doi.org/10.2527/2001.79123062x. Mueller, S., Kreuzer, M., Siegrist, M., Mannale, K., Messikommer, R.E., Gangnat, I.D.M., 2018. Carcass and meat quality of dual-purpose chickens (Lohmann Dual, Belgian Malines, Schweizerhuhn) in comparison to broiler and layer chicken types. Poult. Sci. 97, 3325–3336. https://doi.org/10.3382/ps/pey172. Nääs, I.D.A., Paz, I.C.D.L.A., Baracho, M.D.S., Menezes, A.G.D., Lima, K.A.O.D., Bueno, L.G.D.F., Neto, M.M., de Carvalho, V.C., Almeida, I.C.D.L., Souza, A.L.D., 2010. Assessing locomotion deficiency in broiler chicken. Scientia Agricola 67, 129–135. http://dx.doi.org/10.1590/S0103-90162010000200001. Naji, T.A., Amadou, I., Zhao, R.Y., Tang, X., Shi, Y.H., Le, G.W., 2014. Effects of phytosterol in feed on growth and related gene expression in muscles of broiler chickens. Trop. J. Pharm. Res. 13 (1), 9–16. http://dx.doi.org/10.4314/tjpr.v13i1.2. Owens, C.M., Hirschler, E.M., Martinez-Dawson, R., Sams, A.R., 2000. The characterization and incidence of pale, soft, exudative turkey meat in a commercial plant. Poult. Sci. 79, 553–558. https://doi.org/10.1093/ps/79.4.553. Pelicia, K., Aparecido Jr, I.M., Garcia, E.A., Molino, A.B., Santos, G.C., Berto, D.A., Vieira Filho, J.A.I.V., Murakami, E.S.M.V., Montenegro, A.T., Silva, A.M., 2012. Evaluation of a radiographic method to detect tibial dyschondroplasia lesions in broilers. Rev. Bras. Cienc. Avic. 14, 129–135. http://dx.doi.org/10.1590/S1516635X2012000200007. Rath, N.C., Richards, M.P., Huff, W.E., Huff, G.R., Balog, J.M., 2005. Changes in the tibial growth plates of chickens with thiram-induced dyschondroplasia. J. Comp. Pathol. 133, 41–52. https://doi.org/10.1016/j.jcpa.2005.01.005. Śmiecińska, K., Denaburski, J., Sobotka, W., 2011. Slaughter value, meat quality, creatine kinase activity and cortisol levels in the blood serum of growing-finishing pigs slaughtered immediately after transport and after a rest period. Pol. J. Vet. Sci. 14, 47–54. https://doi.org/10.2478/v10181-011-0007-x. Soglia, F., Baldi, G., Laghi, L., Mudalal, S., Petracci, M., 2018. Effect of white striping on turkey breast meat quality. Animal 12, 1–7. https://doi.org/10.1017/ S1751731117003469. Wang, J.P., Yoo, J.S., Kim, H.J., Lee, J.H., Kim, I.H., 2009. Nutrient digestibility, blood profiles and fecal microbiota are influenced by chitooligosaccharide supplementation of growing pigs. Livest. Sci. 125, 298–303. https://doi.org/10.1016/j.livsci.2009.05. 011. Weeks, C.A., Danbury, T.D., Davies, H.C., Hunt, P., Kestin, S.C., 2000. The behaviour of broiler chickens and its modification by lameness. Appl. Anim. Behav. Sci. 67, 111–125. http://dx.doi.org/10.1016/s0168-1591(99)00102-1. Wei, J., Guo, W., Yang, X., Chen, F., Fan, Q., Wang, H., Zhang, N., Diao, Q., 2019. Effects of dietary ramie level on growth performance, serum biochemical indices, and meat quality of Boer goats. Trop. Anim. Health Prod. 1–7. https://doi.org/10.1007/ s11250-019-01891-5. Yao, W., Zhang, H., Jiang, X., Mehmood, K., Iqbal, M., Li, A., Zhang, J., Wang, Y., Waqas, M., Shen, Y., Li, J., 2018. Effect of total flavonoids of rhizoma drynariae on tibial dyschondroplasia by regulating BMP-2 and Runx2 expression in chickens. Front. Pharmacol. 9, 1251. https://doi.org/10.3389/fphar.2018.01251. Zhao, Y.R., Chen, Y.P., Cheng, Y.F., Qu, H.M., Li, J., Wen, C., Zhou, Y.M., 2019. Effects of dietary phytosterols on growth performance, antioxidant status, and meat quality in Partridge Shank chickens. Poul. Sci. 1–7. https://doi.org/10.3382/ps/pez059. Zheng, Y., Li, Y., Satija, A., Pan, A., Sotos-Prieto, M., Rimm, E., Willett, W.C., Hu, F.B., 2019. Association of changes in red meat consumption with total and cause specific mortality among US women and men: two prospective cohort studies. B.M.J. 365, l2110. https://doi.org/10.1136/bmj.l2110.
5. Conclusion The data in the present study indicated that leg problems (TD) in broilers can decrease the growth performance, inhibit broiler drumstick muscle growth, and affect the meat quality of breast muscle, with increased biochemical parameter levels (including CK and AST) in serum, which suggest that broiler leg problems is detrimental to meat quality. Acknowledgments We are grateful to the financial support from the Outstanding Talents of Henan Agricultural University (No. 30500421) for supporting this work. Supplementary materials Supplementary material associated with this article can be found in the online version, at doi:10.1016/j.livsci.2020.103956. References Bizeray, D., Leterrier, C., Constantin, P., Picard, M., Faure, J.M., 2002. Sequential feeding can increase activity and improve gait score in meat-type chickens. Poult. Sci. 81, 1798–1806. https://doi.org/10.1093/ps/81.12.1798. Bowker, B.C., Eastridge, J.S., Solomon, M.B., 2011. Use of gelatin gels as a reference material for performance evaluation of meat shear force measurements. J. Food Sci. 76, S210–S216. https://doi.org/10.1111/j.1750-3841.2011.02100.x. Cavani, C., Petracci, M., Trocino, A., Xiccato, G., 2009. Advances in research on poultry and rabbit meat quality. Italian J. Anim. Sci. 8, 741–750. https://doi.org/10.4081/ ijas.2009.s2.741. Danbury, T.C., Weeks, C.A., Waterman-Pearson, A.E., Kestin, S.C., Chambers, J.P., 2000. Self-selection of the analgesic drug carprofen by lame broiler chickens. Veterinary Rec. 146, 307–311. http://dx.doi.org/10.1136/vr.146.11.307. FAO, 2010. Agribusiness Handbook: Poultry Meat and Eggs. Rome, Italy. http://www. eastagri.org/publications/pub_docs/6_Poultry_web.pdf. Funaro, A., Cardenia, V., Petracci, M., Rimini, S., Rodriguez-Estrada, M.T., Cavani, C., 2014. Comparison of meat quality characteristics and oxidative stability between conventional and free-range chickens. Poult. Sci. 93, 1511–1522. https://doi.org/10. 3382/ps.2013-03486. Garcia, A.F.Q.M., Murakami, A.E., do Amaral Duarte, C.R., Rojas, I.C.O., Picoli, K.P., Puzotti, M.M., 2013. Use of vitamin D3 and its metabolites in broiler chicken feed on performance, bone parameters and meat quality. Asian-Australas. J. Anim. Sci. 26, 408–415. https://doi.org/10.5713/ajas.2012.12455. González-Cerón, F., Rekaya, R., Aggrey, S.E., 2015. Genetic relationship between leg problems and bone quality traits in a random mating broiler population. Poult. Sci. 94, 1787–1790. https://doi.org/10.3382/ps/pev159. Guasch-Ferré, M., Satija, A., Blondin, S.A., Janiszewski, M., Emlen, E., O'Connor, L.E., Campbell, W.W., Hu, F.B., Willett, W.C., Stampfer, M.J., 2019. Meta-analysis of randomized controlled trials of red meat consumption in comparison with various comparison diets on cardiovascular risk factors. Circulation 139, 1828–1845. https:// doi.org/10.1161/CIRCULATIONAHA.118.035225. Herzog, A., Genin, O., Hasdai, A., Shinder, D., Pines, M., 2011. Hsp90 and angiogenesis in bone disorders—lessons from the avian growth plate. Am. J. Physiol. Regul. Integr. Comp. Physiol. 301 (1), R140–R147. https://doi.org/10.1152/ajpregu.00134.2011. Huang, S., Kong, A., Cao, Q., Tong, Z., Wang, X., 2019. The role of blood vessels in broiler chickens with tibial dyschondroplasia. Poult. Sci. 98, 6527–6532. http://dx.doi.org/ 10.3382/ps/pez497. Huang, S., Rehman, M.U., Qiu, G., Luo, H., Iqbal, M.K., Zhang, H., Mehmood, K., Li, J., 2018. Tibial dyschondroplasia is closely related to suppression of expression of hypoxia-inducible factors 1α, 2α, and 3α in chickens. J. Vet. Sci. 19, 107–115. https:// doi.org/10.4142/jvs.2018.19.1.107. Huang, S.C., Rehman, M.U., Lan, Y.F., Qiu, G., Zhang, H., Iqbal, M.K., Luo, H.Q., Mehmood, K., Zhang, L.H., Li, J.K., 2017. Tibial dyschondroplasia is highly associated with suppression of tibial angiogenesis through regulating the HIF-1α/VEGF/ VEGFR signaling pathway in chickens. Sci. Rep. 7, 9089. https://doi.org/10.1038/
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