Mitochondrial sequence based characterization and morphometric assessment of Diara buffalo population

Singh Karan Veer,Purohit Hitesh,Singh Ramesh Kumar 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.7

Objective: The present study is aimed at phenotypic characterization and mitochondrial d-loop analysis of indigenous “Diara” buffalo population, which are mostly confined to the villages on the South and North Gangetic marshy plains in the Bihar state of India. These buffaloes are well adapted and are best suited for ploughing and puddling the wet fields meant for paddy cultivation. Methods: Biometric data on 172 buffaloes were collected using a standard flexible tape measure. Animals are medium in size; the typical morphometric features are long head with a broad forehead and moderately long and erect ears. Genomic DNA was isolated from unrelated animals. The mtDNA d‐loop 358‐bp sequence data was generated and compared with 338 sequences belonging to riverine and swamp buffaloes. Results: Based on the mitochondrial d-loop analysis the Diara buffaloes were grouped along with the haplotypes reported for riverine buffalo. Sequence analysis revealed the presence of 7 mitochondrial D loop haplotypes with haplotype diversity of 0.9643. Five of the haplotypes were shared with established swamp breeds and with Buffalo population of Orissa in India. Conclusion: Morphometric analyses clearly shows distinguishing features like long and broad forehead which may be useful in identification. The germplasm of Diara buffalo is much adapted to the marshy banks of river Ganga and its tributaries. It constitutes a valuable genetic resource which needs to be conserved on priority basis. Objective: The present study is aimed at phenotypic characterization and mitochondrial d-loop analysis of indigenous “Diara” buffalo population, which are mostly confined to the villages on the South and North Gangetic marshy plains in the Bihar state of India. These buffaloes are well adapted and are best suited for ploughing and puddling the wet fields meant for paddy cultivation.Methods: Biometric data on 172 buffaloes were collected using a standard flexible tape measure. Animals are medium in size; the typical morphometric features are long head with a broad forehead and moderately long and erect ears. Genomic DNA was isolated from unrelated animals. The mtDNA d‐loop 358‐bp sequence data was generated and compared with 338 sequences belonging to riverine and swamp buffaloes.Results: Based on the mitochondrial d-loop analysis the Diara buffaloes were grouped along with the haplotypes reported for riverine buffalo. Sequence analysis revealed the presence of 7 mitochondrial D loop haplotypes with haplotype diversity of 0.9643. Five of the haplotypes were shared with established swamp breeds and with Buffalo population of Orissa in India.Conclusion: Morphometric analyses clearly shows distinguishing features like long and broad forehead which may be useful in identification. The germplasm of Diara buffalo is much adapted to the marshy banks of river Ganga and its tributaries. It constitutes a valuable genetic resource which needs to be conserved on priority basis.

Nutritional Management for Buffalo Production

Sarwar, M.,Khan, M.A.,Nisa, M.,Bhatti, S.A.,Shahzad, M.A. Asian Australasian Association of Animal Productio 2009 Animal Bioscience Vol.22 No.7

The buffalo (Bubalus bubalis) is an important contributor to milk, meat, power, fuel and leather production in many developing countries. Buffaloes can be categorized into Asian and Mediterranean buffaloes. Asian buffalo includes two subspecies known as Riverine and Swamp types. Riverine (water buffalo) and Swamp buffaloes possess different genetics (50 vs. 48 chromosomes, respectively), morphology (body frame, body weight, horn shape and skin color) and behavior (wallowing in mud or water) and thus, are reared and used for different purposes. Low per head milk yield, poor reproductive performance (seasonal breeding behavior, anestrous, and longer calving interval) and low growth rate in buffaloes have been attributed to insufficient supply of nutrients. In many parts of Asia, where the buffalo is an integral part of the food chain and rural economy, irregular and inadequate availability of quality feedstuffs and their utilization are hampering the performance of this unique animal. Balanced nutrition and better management can enhance buffalo productivity. Many efforts have been made in the last few decades to improve nutrient supply and utilization in buffaloes. Recent research on locally available feed resources such as crop residues, and industrial by-products, dietary addition of micronutrients, use of performance modifiers and use of ruminally protected fat and protein sources have shown significant potential to improve growth, milk yield and reproductive performance of buffaloes. However, a number of issues, including establishment of nutrient requirements for dairy and beef, development of buffalo calf feeding systems, nutritional management of metabolic and reproductive anomalies, and understanding and exploitation of the buffalo gut ecosystem, need to be addressed. Extensive coordinated research and extension efforts are required for improved buffalo nutrition in developing countries.

A Y-linked SNP in SRY Gene Differentiates Chinese Indigenous Swamp Buffalo and Introduced River Buffalo

Zhang, Yi,Sun, Dongxiao,Yu, Ying,Zhang, Yuan Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.9

The complete coding region sequence of the SRY gene in Chinese swamp buffalo was determined by PCR product sequencing. Comparison of swamp and river buffalo SRY gene sequences revealed a single nucleotide polymorphism (SNP, C/G) at the 202 bp site of the coding region. Further, a total of 124 male domestic buffaloes were genotyped at this SNP site using the PCR-SSCP method, and it was found that all Chinese indigenous swamp buffaloes had a guanine (G) at this site, while introduced river buffaloes and crossbred buffaloes showed a cytosine (C). Our findings suggested that this Y-linked SNP displayed type-specific alleles differentiating swamp and river buffaloes, and could be used as an effective marker to detect crossbreeding of swamp buffaloes with introduced river buffaloes in native buffalo populations, and thereby assess genetic diversity status and make proper conservation decisions for indigenous swamp buffaloes. In addition, this SNP can be potentially applied in the study of Asian water buffalo phylogeny from a male perspective.

Effect of Feeding Transgenic Cottonseed vis-à-vis Non-transgenic Cottonseed on Haematobiochemical Constituents in Lactating Murrah Buffaloes

Singh, Maha,Tiwari, D.P.,Kumar, Anil,Kumar, M. Ravi Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.12

An experiment was conducted to investigate the effect of feeding transgenic cottonseed (Bt.) vis-a-vis non-transgenic (non-Bt.) cottonseed on blood biochemical constituents in lactating Murrah buffaloes. Twenty Murrah buffaloes in mid-lactation were divided into 2 groups of 10 each. Animals of group I were fed with 39.5% non-transgenic cottonseed in concentrate mixture while the same percentage of transgenic (Bt.) cottonseed was included in the concentrate mixture fed to the animals of group II. Animals of both groups were fed with concentrate mixture to support their milk production requirements. Each buffalo was also offered 20 kg mixed green fodder (oats and berseem) and wheat straw ad libitum. The experimental feeding trial lasted for 35 days. There was no significant difference in the dry matter intake between the two groups of buffaloes. All the buffaloes gained body weight, however, the differences were non significant. Total erythrocyte count, hemoglobin content and packed cell volume were $9.27{\pm}0.70${\times}10^6/{\mu}l$, $13.01{\pm}0.60gdl$ and $34.87{\pm}1.47%$, respectively in group I with the corresponding figures of $8.88{\pm}0.33$, $12.99{\pm}0.52$ and $31.08{\pm}1.52$ in group II. The values of total erythrocyte count, haemoglobin content and packed cell volume did not differ significantly between the two groups of buffaloes. The concentration of plasma glucose, serum total proteins, albumin, globulin, triglycerides and high density lipoprotein were non significantly higher in buffaloes fed non-transgenic cottonseed than in buffaloes fed transgenic cottonseed. The cholesterol concentration was significantly (p<0.01) higher in buffaloes of group I ($136.84{\pm}8.40mg/dl$) than in buffaloes of group II ($105.20{\pm}1.85mg/dl$). The serum alkaline phosphotase, glutamic-oxaloacetate transaminase and glutamic-pyruate transaminase activities did not differ significantly between two groups of buffaloes. However, serum glutamic-pyruate transaminase activity was considerably high in buffaloes fed nontransgenic cottonseed as compared to buffaloes fed transgenic cottonseed. Bt. proteins in serum samples of animals of group II were not detected after 35 days of feeding trial. It was concluded that transgenic cottonseed and non-transgenic cottonseed have similar nutritional value without any adverse effects on health status of buffaloes as assessed from haematobiochemical constituents.

Lysozyme Activity in Buffalo Milk: Effect of Lactation Period, Parity, Mastitis, Season in India, pH and Milk Processing Heat Treatment

Priyadarshini, Subhadra,Kansal, Vinod K. Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.6

Lysozyme activity in buffalo milk in relation to the period of lactation, parity of animal, weather conditions and udder infections was studied. Effect of storage and heat processing of milk on lysozyme activity was determined. Lysozyme activity was higher in buffalo milk than in cow milk. Buffalo colostrum showed lysozyme activity 5 times of that in mature milk. Lysozyme activity in buffalo milk was not influenced by the parity of animal and the stage of lactation, however, it increased during extreme whether conditions (winter and summer). Lysozyme in both cow and buffalo milk exhibited maximum activity at pH 7.4. Buffalo milk lysozyme was fully stable while the cow milk lysozyme was partly inactivated by pasteurization (low temperature-long time as well as high temperature-short time treatments). Lysozyme in buffalo milk was more stable than in cow milk during storage and heat treatment. A 10 to 50-fold increase in milk lysozyme activity was observed in mastitic cows. An assay of lysozyme activity in milk can be used to diagnose mastitis in cattle but not in buffaloes. Some buffaloes exhibited 1000 fold greater lysozyme activity and moderately raised somatic cell count in milk, but there was no sign of mastitis in these animals. A possible role of milk lysozyme in prevention of mastitis in buffaloes is discussed.

Comparative Follicular Dynamics in Superovulated Crossbred Cows and Water Buffaloes

Manik, R.S.,Singla, S.K.,Madan, M.L. Asian Australasian Association of Animal Productio 1998 Animal Bioscience Vol.11 No.1

To understand the caused for poor response to superovulation in water buffalo compared to crossbred cows, follicular events, before start of superovulation, during superovulation and after superovulation were compared. Follicular development was monitored a day before start of superovulation, daily upto superestrus and on the day of flushing. A real time B mode diagnostic instrument equipped with a linear array, 5 MHz transducer was used in five crossbred cows and five Murrah buffaloes. Crossbred cows yielded significantly (p < 0.01) higher number of corpora lutea than buffaloes (21 vs 10). The mean number of small size (2 to 5 mm); medium size (6 to 9 mm) and large size $({\geq}10mm)$ follicles, a day before start of superovulation were almost similar or even slightly higher in buffalo. Though initial shift in the mean number of follicles was higher in buffalo than cow, yet, from Day 2 to Day 3 of the treatment, the average increase in medium (3.2 vs 1.2) and large size (5.0 vs 2.0) follicles was higher in cows than buffaloes. The mean number of medium and large size follicles was 9.8 and 14.4 in cows and 6.4 and 7.6 in buffaloes. On the day of flushing, the number of large size follicle was more in buffaloes than cows, indicating the ovulation problem in this species. The major conclusion from this investigation was that, a day before start of superovulatory treatment, the number of small and medium size follicles was slightly higher in buffaloes, even then superovulatory response was better in cows, due to shift, recruitment and passage of follicles from smaller size to larger size from Day 2 of treatment. Ovulation problem in buffaloes was also responsible for lower superovulatory responses as revealed by the presence of higher number of large size follicles on the day of flushing.

Influence of Varying Ruminally Degradable to Undegradable Protein Ratio on Nutrient Intake, Milk Yield, Nitrogen Balance, Conception Rate and Days Open in Early Lactating Nili-Ravi Buffaloes (Bubalus bubalis)

Nisa, Mahr-un,Javaid, A.,Shahzad, M. Aasif,Sarwar, M. Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.9

Twenty four early lactating Nili-Ravi buffaloes, eight in each group, were used in a Randomized Complete Block Design to evaluate the influence of varying ruminally degradable protein (RDP) to ruminally undegradable protein (RUP) ratio on feed intake, digestibility, N balance, milk yield and its composition, conception rate and days open. Three experimental diets were formulated to contain RDP:RUP of 50:50, 66:34 and 82:18 and were denoted as HRUP, MRUP and LRUP, respectively. Dry matter (DM) intake was higher (p<0.05) in buffaloes fed HRUP diet than in those fed MRUP and LRUP diets. Dry matter digestibility was higher (p<0.05) in buffaloes fed LRUP diet than in those fed HRUP and MRUP diets. Linear increase was observed in DM digestibility with increasing RDP:RUP while Neutral detergent fiber digestibility remained unaltered in buffaloes fed HRUP and MRUP diets, however, it was higher than in those fed LRUP diet. Crude protein digestibility remained unaltered across all treatments. Milk and 4 percent fat corrected milk (4% FCM) yield was higher (p<0.05) in buffaloes fed HRUP diet than those fed MRUP and LRUP diets. Linear decrease in milk yield was observed with increased RDP:RUP. Milk protein and fat yields were higher (p<0.05) in animals fed HRUP diet than those fed MRUP and LRUP diets. Milk protein percent in animals fed HRUP diet was higher than in those fed LRUP diet, whereas it did not differ with those fed MRUP diet. Percent of fat, total solids, solid not fat and lactose remained unaltered across all diets. Nitrogen balance was higher in buffaloes fed HRUP diet than in those fed other diets. Increasing the RDP:RUP resulted in a linear decrease in N balance. The blood urea nitrogen and milk urea nitrogen were lower (p<0.05) in buffaloes fed HRUP diet than those fed MRUP and LRUP diets. The blood pH remained unaltered across all treatments. Days open did not differ significantly. Conception rate was higher in buffaloes fed HRUP diet than those fed MRUP and LRUP diets. The findings of the present study indicate that feeding high (50% of the total crude protein) ruminally undegradable protein diet not only increased nutrient intake and milk yield but also improved conception rate in early lactating buffaloes.

Genetic Variation and Phylogenetic Relationships of Indian Buffaloes of Uttar Pradesh

Joshi, Jyoti,Salar, R.K.,Banerjee, Priyanka,Upasna, S.,Tantia, M.S.,Vijh, R.K. Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.9

India possesses a total buffalo population of 105 million out of which 26.1% inhabit Uttar Pradesh. The buffalo of Uttar Pradesh are described as nondescript or local buffaloes. Currently, there is no report about the genetic diversity, phylogenetic relationship and matrilineal genetic structure of these buffaloes. To determine the origin and genetic diversity of UP buffaloes, we sequenced and analysed the mitochondrial DNA D-loop sequences in 259 samples from entire Uttar Pradesh. One hundred nine haplotypes were identified in UP buffaloes that were defined by 96 polymorphic sites. We implemented neutrality tests to assess signatures of recent historical demographic events like Tajima's D test and Fu's Fs test. The phylogenetic studies revealed that there was no geographic differentiation and UP buffaloes had a single maternal lineage while buffaloes of Eastern UP were distinctive from rest of the UP buffaloes.

A Review of Recent Developments in Buffalo Reproduction - A Review

Warriach, H.M.,McGill, D.M.,Bush, R.D.,Wynn, P.C.,Chohan, K.R. Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.3

The buffalo is an important livestock resource in several countries of South Asia and the Mediterranean regions. However, reproductive efficiency is compromised due to known problems of biological and management origins, such as lack of animal selection and poor nutrition. Under optimal conditions puberty is attained at 15 to 18 months in river buffalo, 21 to 24 months in swamp buffalo and is influenced by genotype, nutrition, management and climate. However, under field conditions these values deteriorate up to a significant extant. To improve reproductive efficiency, several protocols of oestrus and ovulation synchronization have been adopted from their use in commercial cattle production. These protocols yield encouraging pregnancy rates of (30% to 50%), which are comparable to those achieved in buffaloes bred at natural oestrus. The use of sexed semen in buffalo heifers also showed promising pregnancy rates (50%) when compared with conventional non-sexed semen. Assisted reproductive technologies have been transferred and adapted to buffalo but the efficiency of these technologies are low. However, these latest technologies offer the opportunity to accelerate the genetic gain in the buffalo industry after improving the technology and reducing its cost. Most buffaloes are kept under the small holder farming system in developing countries. Hence, future research should focus on simple, adoptable and impact-oriented approaches which identify the factors determining low fertility and oestrus behaviour in this species. Furthermore, role of kisspeptin needs to be explored in buffalo.

Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells

Feng, Wanyou,Chen, Shibei,Do, Dagiang,Liu, Qinyou,Deng, Yanfei,Lei, Xiaocan,Luo, Chan,Huang, Ben,Shi, Deshun Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.10

Isolation and culture of spermatogonial stem cells (SSCs) are attractive for production of genetic modified offspring. In the present study, buffalo spermatogonial stem-like cells were isolated, cultured and expression pattern of different germ cell marker genes were determined. To recover spermatogonia, testes from age 3 to 7 months of buffalo were decapsulated, and seminiferous tubules were enzymatically dissociated. Two types of cells, immature sertoli cell and type A spermatogonia were observed in buffalo testes in this stage. Germ cell marker genes, OCT3/4 (Pou5f1), THY-1, c-kit, PGP9.5 (UCHL-1) and Dolichos biflorus agglutinin, were determined to be expressed both in mRNA and protein level by reverse transcription polymerase chain reaction and immunostaining in buffalo testes and buffalo spermatogonial stem-like cells, respectively. In the following, when the isolated buffalo buffalo spermatogonial stem-like cells were cultured in the medium supplemented 2.5% fetal bovine serum and 40 ng/mL glial cell-derived neurotrophic factor medium, SSCs proliferation efficiency and colony number were significantly improved than those of other groups (p<0.05). These findings may help in isolation and establishing long term in vitro culture system for buffalo spermatogonial stem-like cells, and accelerating the generation of genetic modified buffaloes.