The livestock sector contributed 53.2 percent of the agricultural value added and 11.4 percent to the national GDP (gross domestic product) during 2009-10 [1]. Livestock play a vital role in the rural economy; 30-35 million people are engaged in livestock raising in rural areas [2], deriving 30-40 percent of the household income from this sector [3]. Gross value addition of livestock increased to Rs. 1287 billion in 2008–09 as compared to Rs. 1052 billion in 2007–08, showing an increase of 22.3 percent. The value of livestock is 6.1 percent more than the combined value of major and minor crops [4].

Pakistan is endowed with rich livestock genetic resources that are well adapted to the local conditions. There are 15 breeds of cattle, 5 breeds of buffalo, 33 breeds of sheep, and 36 breeds of goat [5,6]. The population of cattle in Pakistan is 29.56 million, buffalo 27.33 million, sheep 26.49 million, and goats 53.79 million [5]. There are 177.247 million buffalo heads worldwide in 50 countries, of which 171 million (97%) are found in Asia, while 5.38 million (3%) are found in the rest of the world. Pakistan, with a population of 29.9 million (14%) is the second-highest buffalo-inhabited country in the world after India, which has a population of 98.7 million (56 % of the total world buffalo) [4]. Among dairy animal species, buffalo is certainly the second largest source of milk supply in the world and capable of yielding 75 million tons of milk [7]. The present investigation is thus designed to study the Azikheli buffalo breed with the following objective: to determine the morphological and morphometric characteristics of the Azikheli buffalo.

Buffalo Coat Color

Various coat colors of different breeds of buffalo have been reported in the literature. However, the coat color of the majority of buffalo is black, as in Nagpuri buffalo [8], Mehsana buffalo [9] of India, Nili Ravi buffalo, Kundi buffalo [10] of Pakistan, and Anatolian buffalo of Turkey [11]. Light brown, grey-brown, and black-to-brown colors have also been found in Chinese buffalo [12] and Manda buffalo of India [13]. The color of the Egyptian buffalo is blackish gray [14], whereas the color of the Bhadawari breed of India is copper [15]. Some buffalo have white markings on various parts of their body, like the face, legs, and switch of the tail, such as the Nagpuri buffalo [8] and the Nili Ravi buffalo [10].

Study Area

Swat Valley is situated at 34.4° and 35° North and 72° and 74.6° east. It covers an area of 8220 square kilometers, and the elevation above mean sea level varies from 600 meters in the south to 6000 meters in the north, with a human population of 12,49,572 [16]. There are two main rainy seasons: from the end of December to the end of April and from the end of July to mid-September, and the annual precipitation in Azikheil has a range of 1000–1750 mm per year. Khwazakhela Valley (Azikheil) of Swat District was selected as the study area, which is a central location in the original home tract of the Azikheli buffalo [Figure 1] [17].

Figure 1: Map of the Study Area and Azikheli Buffalo Home Tract. The Encircled Regions Represent The Sampling Areas. Source: [17].

Collection of Data

The data was collected based on the morphological characteristics of the Azikheli buffaloes.

Selection of Azikheli Buffalo Cows

On the basis of parity, thirty-six Azikheli buffalo cows, each from the first, second, and third parity (n = 108), were randomly selected from the Khwazakhela valley of District Swat, KPK, during the last month of pregnancy.

Selection of Azikheli Buffalo Bulls

A total of 27 Azikheli buffalo bulls were also randomly selected from the Khwazakhela valley of District Swat, KPK, for the morphological study, including their physical characteristics. Data on physical characteristics was collected as per the standard procedure of random sampling [18] from both Azikheli buffalo cows and buffalo bulls.

Physical Characteristics

Physical characteristics like the color of the coat, forehead, eyelashes, eyes, horn, muzzle, forelegs, hind legs, and hooves of each animal were recorded.

Statistical Analysis

Mean, standard error, and Student’s t-test were calculated for various comparisons. The chi-square test and analysis of variance were also performed using GraphPad Prism-5 (GraphPad Software, San Diego, CA).

Study Area

This study was carried out on 135 buffalo, of which 108 were buffalo and 27 were buffalo bulls from Khwazakhela valley (Azikheil) of District Swat, KPK, and Pakistan.

Physical Characteristics

Physical characteristics studied were color patterns of the coat, forehead, eyelashes, eyes, horns, muzzle, forelegs, hind legs, and hoof. 

Statistical Analysis

Statistical analyses were applied to analyze the data. Different tests of significance were applied to ascertain significant or non-significant differences among the variables under study. Where needed, the Chi-square test of significance, t-test, analysis of variance, and regression analysis of variance were applied. 

Color Pattern of Coat

Four distinct coat colors were observed in the Azikheli buffalo breed, namely brown (Figure 2 a, b), black (Figure 3 a, b), black and white (Figure 4 a, b), and pure white (Figure 5 a, b), both in buffalo and bulls (Table I). The prevailing higher percentage of coat color in buffalo was brown, followed by black and black and white. Bull also follows the same coat color pattern as in buffalo, with brown, black, and white in 4% (n = 1) in descending order of percentage. A completely white coat color was observed only in 4% (n = 5) bulls. Sex differences for the prevalence of brown coat color (χ2 (2) =0.07; P = 0.96), black (χ2 (2) =0.65; P = 0.72), black and white (χ2 (2) =0.35; P = 0.83), and white were statistically not significant (χ2 (2) =2.40; P = 0.30). 

Figure 2: Brown Coat Color (A) Azikheli Buffalo Cow (B) Azikheli Buffalo Bull.

Figure 3: Black Coat Color (A) Azikheli Buffalo Cow (B) Azikheli Buffalo Bull.

Figure 4: Black And White Coat Color (A) Azikheli Buffalo Cow (B) Azikheli Buffalo Bull.

Figure 5: White Coat Color (A) Azikheli Buffalo Cow (B) Azikheli Buffalo Bull.

                     Table 1: Coat Color Pattern of Azikheli Buffalo and Bull in Khwazakhela Valley of District Swat, KPK, Pakistan.

Color of the Forehead:

Forehead color was white in this breed (Figure 6 a); however, a white-spotted forehead (Figure b) and a white-colored forehead extended to the nose bridge (Figure 6 c) were also noted (Table II). Based on coat color and color of forehead, buffalo are locally termed "Sara Chargai, which means brown coat color with a completely white forehead, and are considered the most preferred. The other is “Sara Tikai,” which refers to a brown coat color with a white spot on the forehead. The term “Tora Chargai” is used for buffalo with a black coat and white forehead. Tora Tikai is a buffalo with a black coat and a white spot-on forehead. Out of 108 Azikheli buffalo, the forehead of 61% (n = 66) was completely white; the forehead of 30% (n = 32) was with a white spot; and the forehead of 9% (n = 10) was with a white color extended to the nose bridge. Among the 27 bulls, 55.56% (n = 15) were completely white, 29% (n = 8) had a white spot, and 14.81% (n = 4) had a white-colored forehead extended to the nose bridge (Table III). Sex differences for the prevalence of completely white color (χ2 (2) =0.41; P=0.81), white spotted (χ2 (2) =1.01; P=0.05), and white color forehead extended to nose bridge (χ2 (2) =0.058; P=0.97) were not statistically different. 

Figure 6: Spotting Pattern of Forehead of Azikheli Buffalo (A) Completely White (B) White Spotted (C) White Spotted Extended to Nose Bridge.

                      Table 2: Color Pattern of the Forehead, Eyelashes and Eyes in Azikheli Buffalo and Bull in Khwazakhela Valley of District Swat, KPK, Pakistan.

                   Table 3: Color Pattern of Horns and Muzzle in Azikheli Buffalo and Bull in Khwazakhela Valley of District Swat, KPK, Pakistan.

Shape and Color of Horns

Horns are flat laterally and directed backward and slightly upward without twisting. Upward turning is variable and gives a sickle or semi-sickle appearance to the horn. The majority of the horns observed in this breed were black (Figure 7 b); however, brown (Figure 7 a), brown with a black tip (Figure 7 c), and black with a brown tip (Figure 7 d) were also noted (Table III). Out of 108 buffalo, 34% (n = 37) were completely brown, 52% (n = 56) were with black horns, 9% (n = 10) were brown with black tip horns, and 4.63% (n = 5) were black with brown tip horns. Among 27 bulls, 29.63% (n = 8) of bulls were completely brown, 51.85% (n = 14) bulls had a black horn, 11.11% (n = 3) bulls had a brown horn with a black tip, and 7.41% (n = 2) bulls possessed a black horn with a brown tip (Table III). Sex of buffalo was not a significant source of variation for the prevalence of completely brown color (χ2 (2) =0.10; P = 0.95), black (χ2 (2) =0.02; P = 0.99), brown with black tip (χ2 (2) =0.04; P = 0.98), and black with brown tip (χ2 (2) =1.28; p = 0.53).

Figure 7: Colors of the Horns of Azikheli Buffalo (A) Completely Brown (B) Black (C) Brown with Black Tip (D) Black with Brown Tip.

Color of the Muzzle

The muzzle was mostly white (Figure 8 a) in this breed; however, black (Figure 8 b), black and white (Figure 8 c), and light black pigmented to whitish pigmented were also noted. Muzzle color of 55% (n=60) buffalo was white, 16% (n=17) buffalo had black muzzle, 15% (n=16) buffalo had black and white color muzzle, and 14% (n=15) buffalo had light black pigmented to whitish pigmented muzzle. Among 27 bulls, the muzzle color of 51.85% (n = 14) bulls was white, 18.51% (n = 5) bulls had a black color muzzle, 41.81% (n = 4) bulls had a black and white color muzzle, and 14.81% (n = 4) bulls had a light black pigmented to whitish pigmented color muzzle (Table 3). Sex differences for the distribution of white color (χ2 (2) =0.09; P=0.95), black (χ2 (2) =0.04; P=0.98), black and white (χ2 (2) 2 =0.95; P=0.62), and light black pigmented to whitish pigmented were not significant (χ2 (2) =0.43; P=0.80). 

Figure 8: Colors of the Muzzle of Azikheli Buffalo (A) White (B) Black (C) Black And White.

Shape and Color of the Forelegs

The legs are strong and well-built. The front legs are almost straight, whereas the hind legs have medium angularity. In this breed, most forelegs were white below the knee (Figure 9 a); however, completely white forelegs (Figure 9 b) and black and white forelegs (Figure 9 c) were also noted (Table IV). Out of 108 buffalo, 75% (n=81) had completely white forelegs below the knee, 10% (n=11) had black and white forelegs, and 15% (n=16) had black and white forelegs. Among the 27 bulls, 70.37% (n = 19) were white below the knee, 7.41% (n = 2) had completely white forelegs, and 22.22% (n = 6) had black and white forelegs (Table IV). No significant effect of buffalo sex was observed on the prevalence of white color below the knee (χ2 (2) =0.13; P = 0.93), completely white color of the foreleg (χ2 (2) =1.09; P = 0.58), and black and white color of the foreleg (χ2 (2) =0.99; P = 0). 

Figure 9: Colors of the Foreleg of the Azikheli Buffalo (A) White Bellow Knee   (B) Completely White (C) Black And White.

Color of Hind Legs

Most hind legs were white bellow hocks (Figure 10 a) in this breed. However, completely white hind legs (Figure 10 b) and black and white (Figure 10 c) hind legs were also noted. Sixty-six percent (n=71) of the 108 buffalo had white bellow hock hind legs, 15% (n=516) had completely white hind legs, and 19% (n=21) had black and white hind legs. Among the 27 bulls, 59.26% (n = 16) had white bellow hocks, 18.52% (n = 5) had completely white hind legs, and 22.22% (n = 6) had black and white hind legs (Table IV). Sex differences for the prevalence of white bellow hock (χ2 (2) =0.09; P=0.98), completely white (χ2 (2) =0.26; P=0.87), and black and white (χ2 (2) =0.41; P=0.81) were not significantly different.

Figure 10: Colors of the Hind Leg of Azikheli Buffalo (A) White Below Hock (B) Completely White (C) Black and White.

                     Table 4: Color Pattern of Hoof, Fore Legs and Hind Legs in Azikheli Buffalo and Bull in Khwazakhela Valley of District Swat, KPK, Pakistan.

Color of Hoof

The color of the hoof observed was mostly brown in this breed (Figure 11 a); however, black (Figure 11 b) and brown with black striation (Figure 11 c) were also noted. Out of 108 buffalo's hoofs, 60.19% (n = 65) were brown, 37.04% (n = 4) were black hoofs, and 2.77% (n = 3) were brown with black striation hoofs. Among the 27 bulls, 55.55% (n = 15) were brown, 40.74% (n = 11) were black, and 3.70% (n = 1) were brown with black striation hoof (Table IV). The sex of the buffalo was not a significant source of variation in the distribution of brownish color (χ2 (2) =0.21; P = 0.89), black (χ2 (2) =0.33; P = 0.85), and brown with black striation (χ2 (2) =4.00; P = 0.13). The color of the hoof in the Azikheli buffalo breed in Khwazakhela valley, district Swat.

Figure 11: Colors of the Hoof of Azikheli Buffalo (A) Brownish (B) Black (C) Brownish With Black Striation.

Azikheli buffalo is an indigenous breed of District Swat, KPK. Very little information is available on the phenotypic, productive, and reproductive characteristics of this Azikheli buffalo breed. The present study was conducted to investigate the physical characteristics of the Azikheli buffalo in District Swat, KPK. Physical characteristics include the color of the coat, eyelashes, muzzle, hooves, and legs. The coat color of domestic animals is an important characteristic used to distinguish breeds. Most of the riverine buffalo are black to ashy gray [11,19] whereas the skin color of swamp buffalo ranges from grey to completely black, with very few being white [19,20]. Dark coat color makes buffalo heat intolerant, rendering the animal to stay near the swamp [20]. The dominant brown coat color of Azikheli buffalo thus seems an adaptation to the mountainous environment and genetic in origin [19] because swamp facilities are not available to buffalo in Swat.

The present study on Azikheli buffalo and buffalo bulls in district Swat revealed that the majority of Azikheli buffalo and buffalo bulls are brown in coat color (Table.I), with no sex differences in these colors. Azikheli buffalo have a high percentage (62.04%) of brown coat color compared to a very low percentage (8%) among other Pakistani buffalo [19] and 8% in Nagpuri buffalo of India [21] where the dominant coat color is black. It was observed that the majority of Azikheli buffalo and buffalo bulls have black horns (52% in buffalo and 51.85% in bulls) with no sex differences. The color of the horns in other buffalo breeds in Pakistan, like Nili-Ravi and Kundi, is also black [19]. It was observed that the majority of Azikheli buffalo and bulls have a completely white forehead (61% in buffalo and 55.56% in bulls), white eyelashes (50% in buffalo and 44.4% in bulls, Table II), a white muzzle (55% in buffalo and 51.85% in bulls, Table III), and white forelegs below the knee joint (75% in buffalo and 70.37% in bulls).

Our current research revealed that on the basis of the color pattern of the coat and other body parts, Azikheli buffalo are locally termed "Sra Chargai” (brown coat color with completely white forehead) and are preferred to “Sara Tikai” (brown coat color with white spot-on forehead), “Tora Chargai” (black coat color and white forehead), and “Tora Tikai” (black coat color and white spot-on forehead). These local terminologies regarding the morphological description not only facilitate selection criteria but also denote the cultural association of the farming community with the breed. White markings on the forehead, face, muzzle, legs, tail, and wall eyes (eyes with a whitish iris) have also been reported in Nili-Ravi buffalo, hence the name Panch Kalyani due to the possession of these five white markings [19]. It was observed that the majority of Azikheli buffalo and bulls had white-colored forelegs below the knee joint (75% in buffalo and 70.37% bulls) and hind legs below the hock joint (66% in buffalo and 59.26% bulls; Table IV). Hooves of brownish color were 60.19 % in buffalo and 55.55% in bulls (Table IV), with no sex differences for these color patterns. Maqsood [19] reported that legs are white or of gray color from hoof to above the knee (stocking) in Swamp buffalo, and in Nili-Ravi buffalo, legs have white marking on the lower part of the legs. Sukla [22] observed that the color of the legs is like wheat straw in the Bhadawari buffalo, and there is white marking on the legs of the Mehsana buffalo. Soysal [11] reported white marks on the lower part of legs and black hooves in Anatolian water buffalo. On the other hand, Kolte and Sadekar [21] reported a higher percentage of buffalo with black legs (90%) but legs with white patches (10%) in Nagpuri buffalo. Different authors have observed different color patterns of legs in different buffalo breeds, making them distinct from one another. These morphological differences may be used as an identification mark for various breeds of buffalo.

According to the present study, we focused mainly on qualitative traits (phenotypic characteristics), which include the color of the coat and other body parts, eyelashes, muzzle, hooves, and legs of the Azikheli buffalo. Therefore, it has been concluded that the dominant coat color of Azikheli buffalo and bulls was brown. However, black, black and white, and white coat colors were also observed. Our research also elucidated the local names of the coat color among Azikhele Buffaloes, Sra Chargai, Sara Tikai, Tora Chargai, and Tora Tika, which will help in recruitment criteria but also represent the artistic linkage of the farming community to the breed existing in Pakistan.

Ethical Committee:

The ethical committee localized at the Faculty of Veterinary and Animal Sciences, Lasbela University of Agriculture, Water, and Marine Sciences, Uthal, Balochistan, Pakistan, assessed and examined various techniques involved in this research (Date: January 10, 2022, approval number: 60).

Peer-review: externally peer-reviewed

Statement of Interest:

All the authors of this manuscript hereby confirm no conflict of interest.

Funding and Acknowledgement

The authors express gratitude for the monetary assistance provided by the Department of Biological Sciences, Quaid e Aazam University, Pakistan.

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