The Effects of Different Levels of Tomato Juice in Drinking Water on the Egg Quality, Performance, and Hematological Parameters of Babcock White Laying HensABSTRACT
This study investigates the use of tomato juice (TJ) to assess its effects on egg quality, laying hen performance, and various blood parameters. A total of 128 Babcock white laying hens, aged 50 weeks, were randomly divided into four groups (n=32), each consisting of four subgroups with eight hens. The experimental groups received tomato juice supplementation in their drinking water at concentrations of 0%, 1%, 2.5%, and 5% for four weeks. Fecal samples were collected on days 15 and 30 to evaluate total aerobic bacteria, Lactobacillus spp., Enterococcus spp., Salmonella spp., and Clostridium perfringens counts. The results showed that the 5% tomato juice group significantly increased egg weight (65.95 g/hen/day) and egg mass (56.52 g/hen/day) compared to the control group (62.63 g/hen/day and 45.35 g/hen/day, respectively). Additionally, the 5% TJ group showed improvements in egg quality, with a higher Haugh unit (80.77) and yolk color (11.21) compared to the control group (75.65 and 10.13, respectively). In conclusion, tomato juice supplementation positively affected laying hen performance, egg quality, and blood Vitamin E levels without adverse effects. Additionally, TJ supplementation appeared to enhance intestinal microbiota balance. These findings suggest that tomato juice, when added to drinking water at different concentrations, can improve both egg quality and intestinal health in older laying hens. Furthermore, the unchanged Enterococcus counts in the SBM agar indicate that tomato juice supplementation did not significantly impact this bacterial population, suggesting it may not affect all gut microbiota uniformly. However, the reduction in Lactobacillus spp. in the 5% TJ group suggests a selective influence on intestinal microbiota, which could be important for gut health management in laying hens.
Keywords:
Blood parameters; egg yield; gut microbiota; poultry performance; tomato supplementation
INTRODUCTION
In the 19th century, synthetic antibiotics were widely used in poultry production to maintain health and productivity. However, recent discoveries have highlighted the potential of plant-based extracts as alternatives to synthetic antibiotics, offering benefits without compromising the performance of poultry farms (Gultepe et al., 2020). Numerous herbal and phytogenic products, including essential oils, powders, and extracts, have shown positive effects on the performance of laying hens (Ahmadi & Rahimi, 2011). These herbal extracts often possess antioxidant properties that help neutralize free radicals, protecting hens from stress-related health issues (Tatli et al., 2015). For example, peppermint oil has been shown to enhance the immune system and performance of laying hens, and to improve growth, immunity, and feed conversion ratios of broilers (Durrani & Chand, 2008).
Tomatoes (Lycopersicon esculentum Mill.) are particularly important vegetables, ranking second only to potatoes in global production. In 2022, global tomato production reached 186 million tons, with major producers including China, India, Turkey, the United States, Egypt, Italy, Mexico, Brazil, Nigeria, and Spain (FAOSTAT, 2023). Although most tomatoes are sold fresh, a significant portion is processed into products such as sauces, ketchup, tomato paste, canned tomatoes, and tomato juice. Tomato by-products, such as pomace, seeds, and peels, comprise 5% to 13% of the total fruit mass (Del Valle et al., 2007).
Tomatoes are rich in antioxidants, including carotenoids, vitamin C, and flavonoids, as well as phytochemicals like lycopene (Bohm et al., 1995). Lycopene, the red pigment in tomatoes, is a potent natural antioxidant capable of neutralizing free radicals. Its singlet oxygen quenching ability is ten times greater than that of α-tocopherol (Di Mascio et al., 1989). Consumption of tomato extracts has been linked to reductions in total cholesterol, triglycerides, and blood sugar levels, making it valuable for reducing the risk of cardiovascular conditions such as atherosclerosis (Duttaroy, 2011).
Tomato waste in the diets of laying hens has been shown to reduce egg yolk cholesterol and improve yolk color (Mahata et al., 2016). It is well known that the microorganisms in the digestive system are influenced by the consumption of herbal sources (Armstrong, 1996). In modern poultry production, vegetable-based additives are increasingly used to promote beneficial gut flora, replacing previously used antibiotics to boost yield (Markowiak & Slizewska, 2018). For instance, Mountzouris et al. (2007) demonstrated that probiotics delivered through drinking water were as effective as those included in feed at positively influencing broiler chicken intestinal flora.
Despite the known benefits of tomatoes and their by-products in poultry diets, the use of tomato juice in poultry drinking water has not been extensively studied, particularly in relation to its effects on egg production, egg quality, and gut health. While previous research has focused on tomato waste and extracts added to feeds, there is limited information on the impact of liquid tomato supplementation, such as juice, delivered via water. This method may offer enhanced bioavailability and rapid metabolism. This is a critical gap, as drinking water supplementation provides a more direct and efficient route for nutrient uptake compared to feed-based approaches, allowing for quicker physiological responses in hens.
This study aims to fill this research gap by exploring the effects of tomato juice supplementation through drinking water on the performance, egg characteristics, hematological parameters, and intestinal microbiota of laying hens. This method provides a unique approach to delivering antioxidants and other beneficial compounds present in tomatoes, with the potential to improve egg quality and hen health while enhancing intestinal microbiota balance. Furthermore, this research seeks to provide new insights into the bioavailability of tomato-derived nutrients when administered in liquid form, contributing to the growing body of literature on plant-based supplements as alternatives to synthetic antibiotics in poultry production.
MATERIALS AND METHODS
Ethical Approval
The entire experimental procedure was conducted at the Veterinary Research Laboratory of Afyon Kocatepe University, Turkey, with the approval of the ethics committee (Approval No: AKÜHADYEK-127-16, dated 08/11/2016).
Poultry Management and Experimental Design
A total of 128 fifty-week-old Babcock white laying hens were used in this study. The hens were randomly divided into four groups based on the level of tomato juice supplementation in their drinking water. Each group was further subdivided into 4 subgroups (n = 32), with each subgroup consisting of 8 hens. Freshly squeezed tomato juice was added to the drinking water at concentrations of 0%, 1%, 2.5%, and 5% for a duration of four weeks. The tomatoes were washed thoroughly, and the juice was extracted using a commercial juicer. The juice was then filtered to remove seeds and peels and stored in sterilized containers at 4°C to maintain freshness until use. Tomato juice was administered fresh every 24 hours to prevent spoilage or fermentation. A lighting schedule of 16 hours of light and 8 hours of darkness was maintained throughout the study. Feed was provided ad libitum, and all groups, including the control, received a basal diet formulated to meet the nutritional needs of laying hens, as specified by the NRC (1994) (Table 1).
Data Collection and Analysis
At the beginning and end of the study, all hens were weighed to determine their live body weights. Daily records of total egg production and mortality were kept, while feed consumption was measured weekly. The feed conversion ratio (FCR) and egg mass were calculated based on total feed consumption and egg production. Eggs were weighed weekly, and after the 2nd and 4th weeks, eggs were taken to a laboratory to analyze quality parameters such as egg weight, yolk color index, albumen index, yolk index, eggshell thickness, and Haugh unit. The Haugh unit was calculated using albumen height (Digital Caliper, CD-15CP, Mitutoyo Ltd., UK), following the method of Haugh (1937). The color of the egg yolk was assessed using the Improved Roche Yolk Color Fan (Yolk-Fan™, DSM Nutritional Products AG, Kaiseraugst, Switzerland), which compares yolk color with a 15-band color scale. Albumen and yolk indices were calculated according to the methods described by Tilki & Saatci (2004).
At the end of the trial, blood samples were randomly collected from 3 hens per subgroup. Blood samples were drawn directly from the heart, and blood cell counts of full blood were performed using a compact blood analyzer. Parameters measured included lymphocytes (LC), neutrophils (NC), total leukocyte count (TLC), red blood cell count (RBC), and monocytes (MC). Additional hematological parameters included mean corpuscular volume (MCV), hemoglobin concentration (He), mean corpuscular hemoglobin concentration (MCHC), and mean platelet volume (MPV).
For serum analysis, samples were centrifuged, and the supernatants were stored under appropriate conditions until analysis. An ELISA analyzer was used to measure high density lipoprotein (HDL), low density lipoprotein (LDL), glucose, total protein (TPRO), alanine aminotransferase (ALT), total cholesterol (CHO), aspartate aminotransferase (AST), alkaline phosphatase (ALP), phosphorus, calcium, Vitamin E, total oxidant capacity (TOS), total antioxidant capacity (TAC), Gamma Glutamyl Transpeptidase (GGT), and Immunoglobulin G (IgG). Serum analyses were performed using colorimetric methods and relevant commercial kits, following the manufacturer’s guidelines for each parameter.
Microbiological Analysis
Fecal samples were collected from four hens per subgroup on the 15th and 30th days of the trial. Samples were randomly selected and collected in sterile fecal containers before being transported to the diagnostic laboratory of Afyon Kocatepe University for microbiological analysis. On fecal sampling days, plastic covers were placed beneath each cage and monitored frequently. Observations continued until a sufficient quantity of sample material was collected from each representative group.
Fecal samples (1g) were weighed and diluted in sterile saline solution at a ratio of 1:10. A total of 12 sterile Falcon tubes (15 mL) were used to prepare the dilutions. The agars for microbiological culturing were prepared under standardized conditions to assess total aerobic bacteria, Lactobacillus spp., Enterococcus spp., and the presence of Salmonella spp. and Clostridium perfringens. Further details of the microbiological procedures are presented in Table 2.
To ensure accurate results, a pilot study determined the appropriate dilution rates for each bacterial sample based on their multiplication rates. Dilutions were as follows: for PCA, 10-8 and 10-10; for MRS, 10-6 and 10-8; for SBM, 10-6 and 10-8; for PBLS, 10-6 and 10-8; and for Clostridium perfringens, 10-6 and 10-8, following the methodology of Siriken et al. (2003). Samples were incubated under either aerobic or anaerobic conditions, depending on the bacterial species, at appropriate environmental temperatures. Bacterial colonies were then counted, and the data were recorded digitally for further analysis.
Statistical Analysis
All data were analyzed using a randomized block cage design, with each cage treated as the experimental unit to account for variability between cages. The PROC GLIMMIX procedure was used to analyze performance parameters, such as feed consumption, hen day egg production (HDEP), feed conversion ratio (FCR), egg weight, and egg mass, with repeated measures over time. Tomato juice supplementation, sampling week, and their interactions were considered as fixed effects, while the cage was treated as a random effect. Different covariance structures were tested, and the best structure for each parameter was selected based on Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC). Tukey-Kramer adjustments were applied to control for Type I errors in post-hoc comparisons.
For single-point data such as egg traits, serum biochemical parameters, and fecal microbial counts, the same PROC GLIMMIX model was applied without considering time. Linear, quadratic, and cubic contrasts were used to explore trends across the different tomato juice supplementation levels (0%, 1%, 2.5%, and 5%). SAS PROC IML was used to calculate contrast coefficients, allowing for comparisons between treatment levels.
Data are presented as pooled SEM ± least-square means, with statistical significance set at p≤0.05. Tendencies for significance were noted for p-values between 0.05 and 0.15.
RESULTS AND DISCUSSION
In the present study, feed consumption increased linearly with the dose of tomato juice supplementation. Across the experimental groups, egg production rose over time compared to the control group. The experimental groups exhibited significantly higher (p<0.05) egg weight and egg mass values as compared to the control group. However, no significant differences were observed in feed conversion ratio (FCR) and body weight among the groups (Table 3).
Water consumption was highest in the group supplemented with 2.5% tomato juice (Table 4). However, these observations only reflect group-level records, since statistical analysis was not feasible, as individual subgroups could not be recorded.
Previous studies have reported mixed results regarding the effects of tomato-based supplements on egg production and feed intake. Akdemir et al. (2012) found that increasing tomato powder concentration in hen diets led to greater egg production, feed consumption, and egg weight, along with improved FCR. In contrast, Roberta et al. (2007) noted a decrease in egg yield with more than 5% tomato meal in the diet. Salajegheh et al. (2014) observed that up to 15% tomato powder could be included in laying hen diets, with 5% and 10% concentrations enhancing egg yield and mass, though FCR worsened at 15%.
The increased feed consumption in our study with tomato juice supplementation might be attributed to the high glutamic acid content in tomatoes. Glutamic acid, an excitatory neurotransmitter, enhances appetite by improving palatability. Safamehr et al. (2011) reported glutamic acid levels in tomatoes ranging from 282 to 3632 mg/L, with red tomatoes containing the highest concentrations.
Higher egg weight and mass in the experimental groups may be related to the elevated lycopene content in tomato juice. Nguyen and Schwartz (1998) found that tomato juice contains 5.0-11.6 mg/100g of lycopene, compared to 0.9-4.2 mg/100g in raw tomatoes. Lycopene’s strong antioxidant properties could contribute to improved egg quality.
Regarding egg quality parameters, the Haugh unit and yolk color values significantly increased (p<0.05) in the 5% tomato juice group, though eggshell thickness was reduced. The enhanced yolk color is likely due to the lycopene content. Yilmaz et al. (2010) similarly observed that tomato powder darkened the yolk color. The reduced eggshell thickness in the 5% group might be attributed to the relatively low calcium content in tomato juice. Coïc et al. (1961) highlighted the importance of calcium for eggshell formation, suggesting that sodium and potassium in tomato juice might impair calcium absorption.
Biochemical parameters revealed a linear increase in total antioxidant capacity (TAC) and calcium levels in the experimental groups, while total protein values increased quadratically. Vitamin E levels were significantly higher in the 1% tomato juice group (p<0.05), and total oxidative status (TOS) values increased cubically. Other parameters, including IgG, HDL, GGT, cholesterol, and glucose, were not significantly affected (p>0.05) (Table 6). The observed increase in serum calcium levels in the experimental groups may indicate reduced calcium absorption, potentially due to high sodium and potassium levels in tomato juice, as noted by Kemi et al. (2007) and Sellmeyer et al. (2002).
The study’s findings align with previous research on the antioxidant effects of lycopene, which may enhance egg quality and biochemical parameters by scavenging free radicals and reducing oxidative stress.
This study investigated the effects of tomato juice supplementation on the hematological parameters of older laying hens (Table 7). Compared to the control group, hens receiving 1%, 2.5%, and 5% tomato juice supplementation showed significant differences in lymphocyte counts (p<0.05) and some other hematological parameters. Notably, 2.5% tomato juice supplementation positively influenced mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC). The results suggest that tomato juice supplementation may have limited but potentially beneficial effects on specific hematological parameters.
Olugbenga et al. (2022) found that most hematological parameters and serum enzymes were unaffected by tomato supplementation. Similarly, our study showed limited but positive effects on lymphocyte counts, MCV, and MCHC, without significant changes in other blood markers. This suggests that while tomato supplementation can enhance certain blood parameters, its overall impact on poultry health is beneficial and safe, with no observed adverse effects.
Microbiological results indicated no significant effect of tomato juice supplementation on Enterococcus spp. levels in SBM agar. However, significant changes were observed in MRS and PCA counts. At the second measurement, PCA values were significantly lower in the 1% and 2.5% tomato juice groups compared to the control group (p<0.01), while MRS counts were lower in the 2.5% group (p<0.05), but increased in the 5% group (p<0.05) (Table 8). The reduction in PCA values may be attributed to the antioxidant properties of lycopene, which can inhibit aerobic bacterial growth.
The suppression of Salmonella spp. in this study may be related to the low pH of tomato juice (4.10-4.60), which could inhibit bacterial growth. McClane et al. (2013) also noted that tomato products could reduce the growth of harmful bacteria such as Cl. perfringens. At the study’s end, the control group had the highest levels of Cl. perfringens, while the 5% tomato juice group had the lowest levels.
CONCLUSION
In conclusion, tomato juice supplementation in the drinking water of laying hens may lead to improvements in performance parameters such as egg production and quality without negatively affecting egg characteristics. While positive effects were observed with lower doses, higher doses did not show the same level of benefit, particularly with respect to eggshell thickness and microbial counts. These findings suggest the possible application of tomato juice as a dietary supplement in older laying hens and highlight the need for further research to elucidate its underlying mechanisms and long-term effects on production and gut health.
ACKNOWLEDGEMENTS
The authors would like to thank KUHAM (Kocatepe Livestock Research and Application Center, Afyon Kocatepe University, Turkey) for their support during this study.
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FUNDING
This work was supported by the Commission of Research Projects (BAPK) of AKU University (Grant no. 17 Kariyer 190) for the microbiological component of this study.
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DATA AVAILABILITY STATEMENT
Data is available upon request.
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DISCLAIMER/PUBLISHER’S NOTE
The published papers’ statements, opinions, and data are those of the individual author(s) and contributor(s). The editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions, or products referred to in the content.
Data is available upon request.
