Effect of Different Levels and Forms of Boron and Chromium on Egg Quality, Hatchability and Fertility Percentages in Laying Japanese Quail

In intensive poultry farming, feed represents the largest expense. Thus, the identification of more economical and viable alternative feed additives is crucial for the development of cost-effective poultry feeds.

To guarantee that animals and birds have the best possible health, productivity, and reproduction, mineral nutrition is an essential component of the animal feeding system. Minerals are needed in smaller amounts than other nutrients like protein and energy, but their imbalances and deficiencies quickly show up in changes to animal welfare and productivity. They are necessary to preserve normal health and productivity; however, in some circumstances, adding more supplements may promote greater development and egg production (Swain et al., 2021). Traditionally, inorganic minerals have been included in chicken diets due to their affordability and accessibility; nevertheless, because of their poor bioavailability, they are comparatively less beneficial than organic minerals (Virden et al., 2004).

Therefore, current research has investigated the impact of significant mineral Nanoparticles (NPs) on the health of animals, consumption of feed, and growth performance. Mineral NPs seem to offer more potential for use as mineral supplementations than inorganic mineral supplementation because of their improved absorption, capacity to avoid negative interactions with other minerals, and capability to support growth and physiological processes. This is particularly true at smaller doses than those recommended for their larger particles; Nano-sized particles have more possible than their conventional sources, so less is needed; and nano-minerals increase bioavailability because of their adsorbing power, surface area, surface activity, and high catalytic efficiency (Patra and Lalhriatpuii, 2020). Accordingly, inorganic chromium (Cr) and boron (B) minerals, in addition to their nanoparticles, have been added to diet in order to investigate their impact on the quality of eggs laid by Japanese quail.

Boron has positive impacts on both human and animal biological processes, including growth, immunity, energy metabolism, calcium metabolism, creation of bones, and functioning of the brain (Biţă et al., 2022). Kara (2022) found that, with the exception of quails fed diets comprising 0 and 80 mg/kg B, the eggshell weight percentage of quails fed diets including 20 mg/kg B was considerably greater than that of every other group of quails. The findings of El-saadany et al. (2017) referred that laying hens' diets increased significantly in Haugh unit when B level increased from 100 to 200 and finally 300 mg/kg. While Mutlu et al. (2021) reported that there was no discernible change in the weight of the albumen and yolk in laying quails when calcium tetraborate was used as the source of calcium in the B (300 mg/kg diet) group. Boron supplementation at different levels (100, 200 and 300 mg/kg diet) significantly increased fertility and hatchability of fertile eggs percentage for laying hens (El-saadany et al., 2017). The mechanism of boron action, which involves raising the concentration of steroid hormones like beta-estradiol and testosterone, may be the cause of this positive effect of boron on hatchability and fertility in the current results (Naghii and Samman, 1993).

Quail egg quality metrics were enhanced by the addition of 800 ppb of Nano-Cr chloride (Ahamed et al., 2018). Consistent with these findings, Malathi (2015) demonstrated that adding Nano-Cr to layers at 50, 100, 200, and 400 ppb/kg diet enhanced the quality of the eggs. Conversely, the outcomes acquired by Mousa et al. (2022) observed a reduction in albumen height and yolk weight in Golden montazah layer hens, but no effect on eggshell thickness when fed at a level of 400 mg Cr chloride /kg diet. The findings of Abdel-Mageed and Hassan (2012) indicated that laying Japanese quail raised in Egyptian summer settings can have their egg hatching negatively reduced by supplementing with Cr at a level of 1200 μg/kg feed. The percentages of hatchability and fertility in Bandarh laying hens were increased by supplementing them with 250, 500, 1000, and 1500 μg/kg of their food as Cr yeast (Maysa, 2011). Therefore, the primary goal of the current study was to assess how parameters of egg quality and percentages of fertility and hatchability in laying Japanese quails were affected by boron and chromium in their inorganic and Nano forms.  

MATERIALS AND METHODS

This study was conducted at the quail research laboratory of the Animal and Fish Production Department at the Faculty of Agriculture (Saba Basha), Alexandria University, for a period of around three months, from July 10 to October 2, 2022.

Preparation of Nanoparticles from boron and chromium minerals:

As a source of inorganic B, sodium borohydride, 96% PS (NaH4B), which has 28.57% available B, was utilized. It is manufactured by PANREAC QUIMICA SA, Barcelona, Espana. As a source of inorganic Cr, Milan-Italy, a commercial company, produced Cr (III) chloride, or Cr trichloride hexahydrate (ClCr₃.6H2O), which has 19.96% available Cr.  Using a ball mill (Fritsch Pulverisette 7, Germany) running at 1400 rpm, the Advanced Technology and New Materials Research Institute at SRTA City, New Borg El-Arab City, Alexandria, Egypt, synthesized B and Cr mineral Nanoparticles physically. Materials are ground into a very thin, Nano-sized powder in a ball mill for use in feeding cattle (Bakker et al., 1995; Koch, 1997). The mechanical milling process, which is an impact process with high energy, employs balls within containers and can be done in a variety of mills, usually planetary and shaker mills (Gorrasi and Sorrentino, 2015). In Nano-B (N-B) and Nano-Cr (N-Cr), the average particle size was 23.69 nm and 23.53 nm, respectively, whereas the average particle size for conventional B was 433.46 µm and for Cr it was 15.66 µm. The Central Laboratory at Alexandria University in Egypt used a scan electron microscope (SEM) to measure the size of the Nanoparticles.

Experimental design and bird’s management:

During a 12-week period, a total of 243 Japanese quail hens were randomly assigned to nine treatment groups. These groups were then further split into three replicates, each consisting of nine birds, consisting of six females and three males. After two weeks of monitoring, the quail hens were chosen based on their ability to produce more than 70% of eggs. All quails were raised in wire batteries with identical climatic and sanitary settings for the duration of the 12-week trial. One of the nine experimental meals plus the following treatments were given to the quails: The control group's quails (T1) were fed a basic meal devoid of additives, whereas the treatments T2 (B25) and T3 (B50) were provided the same basal diet enriched with 25 and 50 mg/kg feed, respectively, of inorganic boron (B). Treatments T4 (N-B25) and T5 (N-B50) were given in the basal diet supplemented with Nano-inorganic boron (N-B) at levels of 25 and 50 mg/kg feed, respectively. Treatments T6 (Cr25) and T7 (Cr50) were fed the basal diet supplemented by inorganic chromium (Cr) at levels of 25 and 50 mg/kg feed, respectively. Treatments T8 (N-Cr25) and T9 (N-Cr50) werefed the basal diet inclusion of Nano-inorganic chromium (N-Cr) at levels of 25 and 50 mg/kg feed, respectively. The nine trial meals were designed to be isocaloric and isonitrogenous, and they were balanced to retain the same amount of lysine and sulfur amino acids in accordance with the National Research Council's requirements for laying Japanese quail (NRC, 1994). The experimental meals' composition and computed chemical analysis are displayed in (Table 1). Water and feed were freely provided to the animals during the trial.

MEASUREMENTS:

Egg quality characteristics:

In order to determine the exterior and internal egg quality measures that followed, twenty eggs were randomly selected from each treatment on the final three days at 20weeks of age. Each egg was then weighed separately, approximately the average egg weight.

External egg quality characteristics:

As reported by (Novikoff and Gutteridge, 1949), the specific gravity was obtained using (Olsson, 1934) saline flotation technique. According to (Romanoff and Romanoff, 1963), the egg shape index was calculated by dividing the egg's transverse diameter by its length and multiplying the response by 100. Three points on the egg (the air cell, the equator, and the sharp end) were used to quantify the thickness of the eggshell without the shell membrane by micrometers. Each dried eggshell was weighed, and the percentage of eggshell weight was computed by dividing the eggshell weight by the egg weight and multiplying the result by 100.

Internal egg quality characteristics:

Using a tripod micrometer and a caliper with a venire scale, the yolk index was determined as a percentage by calculating the ratio of each person's height to the average of their width and length, as described by (Funk, 1948). After the egg yolk and albumen were separated, the yolk weight was divided by the egg weight, and the result was multiplied by 100 to determine the yolk percentage. A Roche yolk color fan as used to measure the color of the egg yolk. The method of  An et al. (1997) was used to determine albumen height, Haugh unit, and albumen height per egg weight value. The weight of albumen was measured as egg weight - (yolk weight + shell weight), and its percentage was computed by dividing the albumen weight by the egg weight and multiplying the result by 100.

Hatchability and fertility percentages:

All the laid eggs from each treatment were collected daily over 4 days at 20 weeks of age and were incubated at a temperature of 37.6 °C with 45-55% relative humidity for 14 days. They were then transferred to hatcher trays at last 3 days of incubation and were maintained at 37.2 °C and 80 % relative humidity until hatching. After being hatched, and non-hatched eggs were broken to determine the percentages of fertility and hatchability. Hatchability of total eggs (%) = (number of hatched chicks / numbers of set eggs) ×100. Hatchability of fertile eggs (%) = (number of hatched chicks / numbers of fertile set eggs) ×100.

Statistical analysis: Using the SPSS® (2020) statistical software program for windows version 27.0.10.1, one-way ANOVA was used to statistically assess the differences between the treatments. Duncan's Multiple Range Test (Duncan, 1955) was used to isolate the significant differences across treatment means.

RESULTS AND DISCUSSION

1. External egg quality:

The results of external egg quality parameters are shown in Table 2. The results showed that adding boron and chromium in the form of inorganic and Nanoparticles had no noticeable impact on the external features of eggs.

The findings of Mutlu et al. (2024) showed that the egg weight of Japanese quail was not significantly affected by the addition of 300 mg/kg of calcium tetraborate to the meal as a source of B. Similarly, the findings of Sizmaz et al. (2021) demonstrated that the addition of boric acid (120 ppm) had no effect on the egg weight during the course of the trial in laying hens, independent of the time periods studied. Inorganic, organic, or Nano Cr supplements at 200 or 400µg/kg did not significantly affect layer bird egg weight, according to Sathyabama and Jagadeeswaran's (2016) findings. These outcomes are consistent with several studies that found no statistically significant relationship between the various levels of B and specific gravity (Kara, 2022; Olgun et al., 2012). Similar to this, laying hens given a diet containing 20 parts per million of Cr did not exhibit any appreciable change in specific gravity (Uyanik et al., 2002).

The results obtained by Mutlu et al. (2021)reported that the boron supplementation (calcium tetraborate at 300 mg/kg) had no effect on the egg shape index of laying quails when compared to the control. A similar finding was documented by Sizmaz and Yildiz (2016), who observed that during the course of the study period, the addition of boric acid at a level of 120 mg/kg feed did not substantially alter the thickness of the eggshells in laying hens. Furthermore, Sizmaz et al. (2021)reported that administering boric acid (120 ppm) to laying hens did not significantly alter the egg shape index or shell thickness. Similarly, Mousa et al. (2022)reported that after 32 weeks of treatment, the egg shape index, shell thickness and eggshell weight of golden montazah layer chickens were unaffected by the addition of 400 mg of chromium chloride per kg of feed.

Our results are in agreement with those of Adarsh et al. (2021), who found that boron (40 ppm) had no effect on the shell weight of eggs (as a percentage of egg weight) during the first 12-week feeding trial in laying hens. Furthermore, laying Japanese quails' eggshell weight was not significantly impacted by diets augmented with boric acid at 200 or 400 parts per million (Ayasan et al., 2011).

1. Internal egg quality:

All internal measurements of egg quality like yolk weight (g or %), yolk color, Haugh unit, and absolute albumen weight showed no significant variations between the experimental groups of laying quails fed on varying forms or levels of boron and chromium (Table 3), with the exception of yolk index and relative albumen weight. These results were in line with those obtained by Mutlu et al. (2021), who found no differences in yolk weight and yolk color between the supplemented calcium tetraborate (300 mg/kg) group and the control group. Moreover, the research by Sizmaz and Yildiz (2016) found that supplementing laying hens' diets with boric acid at a dose of 120 mg/kg did not substantially alter egg yolk weight and Haugh unit during the course of the trial. In another work, laying hens fed feed enriched with 100, 200, and 300 mg boron/kg feed showed no discernible change in the yolk ratio, according to El-saadany et al. (2017).

For Cr, Abdalla et al. (2019) stated that yolk weight and yolk color of Gimmizah chickens fed low metabolizable energy or low metabolizable energy and crude protein diets supplied with Cr Methionine and Cr chloride by 1200 µg/kg diet were not significantly differed, but meanwhile the Haugh unit significantly increased in this study. However,  Abdallah et al. (2013)highlighted that the egg yolk percentage of golden Montazah hens increased when chromium doses were increased in their diet. In another study, Şahin et al. (2001) found that dietary Cr supplementation increased the yolk weight of Japanese quails.

Our outcomes revealed that there are notable variations across treatments in the yolk index (p = 0.020). The group given 25 mg of Nano-inorganic boron (N-B) reported the greatest yolk index (44.91%), whereas the group given 50 mg of N-B showed the lowest yolk index (37.22%). However, as compared with the control group, there were no appreciable changes in the bird groups that were fed a feed that contained 25 mg of Nano-chromium or 50 mg of either inorganic or Nano-boron, respectively. Notably, these groups showed a significant increase in the yolk index value. These findings are consistent with those of Şahin et al. (2001), who reported that diet Cr

supplementation improved the Japanese quail's yolk index. However, research of Mousa et al. (2022) revealed that adding 400 mg/kg of Cr to the diet had no influence on the yolk index over the 40-week period of Golden Montazah layer-hens. Furthermore, Abdalla et al. (2019) reported that there was no significant difference in the yolk index of Gimmizah chickens given low metabolizable energy or crude protein diets supplemented by Cr-methionine and Cr-chloride at a dose of 1200 µg/kg. Additionally, According to the results of Sizmaz and Yildiz (2016), supplementing laying hens with boric acid at a dose of 120 mg/kg feed for the whole study period did not substantially alter the egg yolk index.

In this study, with regard to relative albumen weight, a significant effect was noted (p = 0.026), and the group that received 25 mg of inorganic chromium (Cr) had the lowest albumen percentage (51.97%) in comparison to the other groups. This suggests that the composition of albumen was influenced by forms of boron and chromium. Other treatment levels were closer to the control in terms of albumen composition, as evidenced by the higher albumen weights of other groups. Similar to these results, Şahin et al. (2001)found that dietary Cr supplementation enhanced the albumen weight of Japanese quail. Still, neither the crude protein nor low metabolizable energy diets enriched with 1200 µg/kg of Cr Methionine and Cr Chloride significantly changed the albumen weight% of Gimmizah chickens (Abdalla et al., 2019).

1. Hatchability and fertility:

The table 4 presents the impact of two forms and levels of boron and chromium on the hatchability and fertility percentages of laying Japanese quails at 20 weeks of age. The results showed significant differences in hatchability of total eggs percentage across the treatments (p = 0.006). The worst values of hatchability percentages are seen in groups supplemented with 50 mg either of inorganic or Nano boron, as well as 25 mg of inorganic boron compared with other treatments which achieved the best values. These results are consistent with those of Bozkurt and Peşmen (2023), who demonstrated that in Ovo injection of sodium borate (0.5 mg) seemed to boost the hatching power of broiler chicks compared to control group and sodium borate (1 mg) groups. Furthermore, compared to the control value, the hatchability % for laying hens increased considerably when boron was added at several amounts 100, 200, and 300 mg/kg diet (El-saadany et al., 2017). Similarly, Maysa (2011) observed that the hatchability proportion of Bandarh laying hens was enhanced by supplementing with Cr at doses of 250, 500, 1000, and 1500 μg/kg food as Cr yeast.

In this study, there are no discernible variations in the hatchability of viable eggs or fertility percentages between treatments for boron and chromium. The findings of Rossi et al. (1993), who found that feeding broiler breeders two types of boron at levels of 0–250 mg/kg had no effect on fertility, were in line with this discovery. However, compared to the control value, the fertility % of laying hens increased dramatically when boron was added at several amounts 100, 200, and 300 mg/kg diet (El-saadany et al., 2017). Additionally, Maysa (2011) found that the proportion of fertility for Bandarh hens improved when they were supplemented with Cr at doses of 250, 500, 1000, and 1500 μg/kg feed as Cr yeast.

CONCLUSION

This study revealed that optimal doses and specific forms of boron and chromium may enhance certain egg quality traits like yolk index and relative albumen as well as hatchability, though excessive levels may have adverse effects.