INTRODUCTION

Buffalo play an important part in the agricultural economies of many developing countries, providing milk, meat, and draught power. The global population of buffalo is estimated to reach 199 million (1). The buffalo is a valuable cattle resource in many South Asian and Mediterranean countries (2, 3). Several factors influence the start of puberty and, as a result, the age at first calving (4). These include management, genotype, and climate. However, live body weight is one of the most important factors influencing puberty onset (4). Both Egyptian (5) and Mediterranean buffaloes (6) demonstrated an anticipation of puberty, with daily weight gains of 0.45 and 0.55 kg after weaning, respectively.

In relationship between the puberty onset in buffalo young and various hormonal and biochemical parameters has been explored (7). Researchers have identified metabolic indicators such as glucose, total protein, triglyceride and cholesterol that could potentially serve as signals of puberty in heifers (8).

The current study was aimed to determine the relationship between age of puberty in Iraqi-buffalo with body weight and some parameters biochemical.

This study was conducted in Sharqat city (Salah Alden Province) during the period from April 2022 to January 2023. This experiment included 18 Iraqi river heifers buffalos divided to three groups (first group which involved six buffalo’s heifers at age of 12 months, second group involved six buffalos aged 18 months and third group involved six buffalos aged 22 months). Estrus detection was done through visual observation in the early morning and evening (Estrus signs were monitored constantly for each females two time a day  07:00 Am and 06:00 Pm for 30 minutes) according (9). The body length and heart girth are the components to calculate the body weight using the formula (10) BW = Heart Girth Multiply Heart Girth Multiply Body Length Division 300. Body condition score (BCS) were scale monthly using the five point visual BCS technique (0 = extremely thin to 5 = extremely fat cows), two observations (lateral and back views) (11). Monthly blood samples (10 ml) from each animal were taken via jugular venipuncture at 7.00 AM. The samples were left to clot for 20 minutes at room temperature, after that they kept in refrigerator at (5°C) for 24 hrs. Serum decanted following immediate centrifugation of the samples (3000 cycle per 15 minutes) and kept under -20°C till analyzed. Blood concentration of cholesterol, glucose and total protein and triglyceride were estimated by spectrophotometer using special kit. Statistical analysis was carried out using the GraphPad Prism Software to detect significant differences at P‼0.05 (12).

Throwing the visual examination to detect estrus in Iraqi buffalo, the results of the present study showed that in the first group (12 months) three of the animals reveled the signs of estrus at first time at the age of18 months, one at 19 months, and two at 20 months, while in the second group, 3 animals showed the signs of estrus at first time at the age of 19 months and one at 20, 21, and 22 months, while the animals of the third group were all adults at the start of experiment.

The results of the study appears that the age of puberty in Iraqi buffaloes ranges between 18-22 months, this agrees with 18 months of age in Brazilian buffaloes, 20.7 months in Mediterranean Italian buffaloes (13, 14) Surti Buffalo Heifers (20.00 ± 2.12 months). Other study found that the average age of puberty for (riverine x swamp) crossbred buffalo was 22.93±1.6 months (15). Buffalo can reach puberty at the age of 20–24 mo; while, the results of the study differed with (16) in Nilli Ravi buffalo (32.5 months) and (17) in Surti buffalo (45.5 months of age), and (18) in India buffalo (31 months), (19) in Egypt River buffalo (15–17 months) This defenses may attribute to differences in genetic, nutrition, management and climatic factors. (20) reported that the female buffalo reproductive performance, including age at first mating, age at first calving, calving interval, and service per conception, is impacted by both genetic and non-genetic factors such as nutrition, management, and climate (21).The age at which buffalo reach puberty can vary greatly, ranging from 18 to 46 months, and this variance can be attributed to a variety of factors, including genotype, diet, management, social environment, climate, year or season of birth, and diseases. Balanced food and good management can help with growth and early sexual maturity (22). Puberty develops early (between 4 and 6.5 months of age) as a result of a high plan of feeding (23). Nutritional regimens that promote a relatively high rate of body weight gain (1 kg/day) are associated with increased adiposity and circulating concentrations of the metabolic hormones leptin, insulin, and insulin-like growth factor 1 (IGF1) when compared to regimens that promote slower growth (0.5 kg/day; 24). (25) Predicted that the positive metabolic profile caused by higher body weight growth would cause changes in the reproductive neuroendocrine system, leading in enhanced pulsatile release of GnRH and LH, which are necessary for first ovulation. A study found that buffalo heifers born in the spring experienced puberty for 404.3 ±3.9 days, while those born in the autumn did not (26). Recent studies (27) show a strong correlation between environmental changes and the onset and duration of puberty in animals and humans needs 543.2 ± 19.5 days.

Body weight and body condition score

The results of this study showed that the highest weight of the experimental animals was in the third group (338.09 ± 22.91 kg), compared to the second and first group (295.08 ± 33.15 and 219.48 ± 38.86 kg), and the weight of the buffalo in the second group was significantly higher P> 0.05 compared to the first group (219.48 ± 38.86 kg), (Table 1). The results of this study agreed with (28) who recorded 359.6 ± 13.3 kg. At puberty age and dissimilar with the result of (14) Surti Buffalo Heifers (200.00±1.62 kg) and (29) recorded 364±12.5 kg at first estrus in buffalo. in Nili-Ravi buffaloes (550 kg), also (30) The beginning of the first estrus in Nili-Ravi buffalo heifers was observed to occur at weights of the body ranging from 388 to 431 kg. A study showed that heifers fed an individual roughage-concentrate-based diet to provide a weight increase of 0.4 kg/day reached puberty at an average age of 946 days (31.53 months) and a body weight of 380.67 kg (31). This difference may perhaps be related to the type of breed and/ or nutritional status (32). Body condition scour compactness reached its highest level in the third and second groups (3.5, 3.5) compared with the first group (2.5). This result was comparable with (33) who recorded 3.06 BCS at puburtal age in cattle, while it was disagree whit (34) who report 4.05 BCS in cattle, this difference in result may due to different species, nutritional system or location area. (35)It has been observed that several factors influence puberty, in both direct and indirect ways. These characteristics include genetics and breed, weight and rate of weight gain, body composition, nutrition and feed, and specific environmental or social situations like as seasons, photoperiods, and the presence or absence of bulls. The body condition score (33) is one approach to assess the nutritional status of cattle. Body condition is frequently significantly connected with individual fitness, including survival and reproductive potential. Thus, evaluating animal status is critical in many ecological studies and as a wildlife management tool. For example, in disease ecology research, information on changes in body condition can show harmful effects of parasites on hosts or suggest hosts' relative susceptibility to infection (36). Adipose tissue mass and weight are thought to play animportant role in regulating puberty onset (37). Short-term dietary restrictions in prepubertal ruminants diminish the expression of the leptin adipose gene as well as its secretion (38). In this instance, nutritional restrictions will inhibit LH secretion. However, administering leptin will restore LH secretion. The effect demonstrates a positive correlation between LH and leptin secretion (38). Furthermore, during puberty, cows' leptin concentration and gene expression rise, as does their IGF-I level and weight (39).

Table 1: Body weight and BCS in pre, at and post puberty in Iraqi buffalo

Table 1: Body weight and BCS in pre, at and post puberty in Iraqi buffalo

Different small letters mean significant differences (P≤0.05)

Biochemical Parameters

Changes in blood biochemical components were often thought to be the cause of reproductive difficulties rather than the effect of them (40). The results of the current study, as shown in Table No. 2, showed that there was no significant difference in the values or levels of total protein (6.58 ± 6.0, 6.30 ± 5.8, 6.94 ± 6.5 in different groups respectively) and cholesterol (66.26 ± 1.9, 66.26 ± 1.9, 63.55 ± 1.58 in different groups respectively) between the experimental groups. The concentration of total protein in the present study was comparable with (41) who report 7.33 ± 0.64 in normal cycle buffalo and with (42) who recorded 6.46 ± 0.28 in Graded Murrah buffaloes and (43) in Murrah Buffaloes (6 ± 0.20). In contrast (44) recorded high concentration of total protein in buffalo (8.8 ± 0.2) after 18 months old. The present study had showed that there is no correlation between the level of total protein and age at puberty, this result in coincided with other work (45) attributed the significant differences in the serum protein concentration between pubertal and mature Angoni cattle. This difference may due to difference species. The concentration of cholesterol in the present study was comparable with (42) who reported 83.67 ± 8.81 in Graded Murrah buffaloes while it disagree with who reported 191 ± 5.21 in normal cyclic buffalo in Egypt. Changes between studies could be attributed to changes in breeds, environments, and levels of nutrition. Cholesterol is a steroid and a precursor to steroid hormones, including reproductive hormones (46). Furthermore, lipids constitute a rich energy supply necessary for oocyte maturation (47). Estrogen impacts lipid metabolism by lipogenesis, which leads to increased cholesterol synthesis. Triglycerides level was significantly higher P<0.05 in both the first and third groups (38.12 ± 2.29 g/L, 37.20 ± 4.50 g/dl), respectively, and compared with the second group (33.25 ± 10.16). These result was higher than recorded by (43) in Murrah buffaloes (30.25 ± 0.31). Triglyceride concentration in the present experiment exhibited curvilinear profiles relative to the onset of puberty with significant decreasing associated with puberty approaching, these results reversed with those obtained by Nazifi et al. (48) who recognized a significant decreases in the triglyceride concentration with goat age progression.The highest level of glucose was reached in the third group (65.95 ± 12.79) compared with first and second group (44.74 ± 9.25 and 52.32 ± 16.5 mmol/l respectively). Plasma glucose (GLU) is an essential supply of energy for several cells. GLU in plasma is generally maintained through the breakdown of dietary carbohydrates and a relatively sophisticated endogenous synthesis pathway (48). Endogenous glucose synthesis is caused by glycogenolysis (the breakdown of glycogen in the liver into glucose) as well as gluconeogenesis. Maintaining normal plasma glucose involves a fine balance of glucose availability and utilization (49, 50).Glucose is one of the important nutrients controlling reproduction and cyclicity in farm animals, and it was formerly thought to be the cause of poor fertility and non-cyclicity. The concentration of glucose in the present study was disagreed with (44) who found decreased glucose level with increasing age (41.5 ± 0.7 at 24 months age) and who found 102.42±7.45 in normal cyclic buffalo. These differences may due to different nutritional system or geographic area in deferent study. Plasma glucose proceeds primarily from gluconeogenesis and gastrointestinal absorption. In ruminants, little glucose is absorbed from the gut, so the vast majority of it is synthesized in the liver via gluconeogenesis, which uses other substrates such as ruminal volatile fatty acids produced by dietary glucose fermentation in the rumen (51; 52).

Table 2: levels of biochemical parameters in pre, at and post puberty in Iraqi buffalo

Different small letters between column means significant differences in (P< 0.05)

Correlation between body weight and age of puberty

The results of the current study, as shown in the figure (1), showed that body weight has a great relationship with the age of puberty, when the animal reach the suitable weight early the animal reach puberty earlier. This result was agreed with (53, 54) who reported that the body weight and growth are the major factors controlling age at puberty. Environmental factors, dietary quality, body condition score, and pre- and post-weaning weight increase all contribute to a lower age at puberty (55). Post-weaning diet plays an important influence in determining age at puberty. Sexual maturity in dairy heifers is governed by body weight rather than age (56). Under-feeding and uneven feed composition is the major causes of slow growth rates in field circumstances, delaying maturity (57). A study discovered that providing a high plane of nutrition (bypassing protein and fat) resulted in much more weight gain and a lower age at puberty, maturity (23 months), and first calving (14). Monteiro et al. (58) found that growth rate and body weight gain have a greater impact on puberty age. These findings were consistent with earlier investigations, which reported an age at puberty of 22-24 months and an age at first calving of 32-34 months (59). Heifers' puberty ages are directly related to their weight and physical condition. According to Romano et al. (60) who consumed more calories and gained more weight per day reached puberty earlier. Similarly, in the current investigation, pubertal heifers were heavier and younger than non-pubertal animals. In Bos taurus beef heifers, puberty is defined as the age at which the animal has reached around 60% of its mature weight (60; 61). This rate is around 65% for Bosindicus heifers (58).

Figure 1: Relationship between body weight and age of puberty in Iraqi buffalo

The age of puberty in Iraqi buffalo are about 18-20 months in Salah-Aldin province. The B.W and body condition score have high correlation with age of puberty in buffalo. The age of puberty have significant effect on the levels of glucose and Triglyceride especially.

Acknowledgments

The authors thank the faculty members of the Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Baghdad, Iraq.

Novelty statements

The novelty of the study is its focus on the relationship between the age of puberty in Iraqi buffalo with body weight and some biochemical parameters..

Author’s contributions

These authors each contributed equally.

Conflict of interest

The authors declare that they have no competing interests.

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