• Animal Bioscience
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• v.35 no.7
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• pp.949-954
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• 2022

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.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.7
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• pp.1060-1068
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• 2009

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.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.3
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• pp.353-360
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• 2012

Mastitis is a highly morbid disease that requires detection at the subclinical stage. Tropical countries like India mainly depend on milch buffaloes for milk. The present study was conducted to investigate whether the trace minerals viz. copper (Cu), iron (Fe), zinc (Zn), cobalt (Co) and manganese (Mn) and enzyme activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in riverine buffalo milk can be used as an indicator of subclinical mastitis (SCM) with the aim of developing suitable diagnostic kit for SCM. Trace elements and enzyme activity in milk were estimated with Atomic absorption Spectrophotometer, GBC 932 plus and biochemical methods, respectively. Somatic cell count (SCC) was done microscopically. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. A statistically significant (p<0.01) increase in SCC, Fe, Zn, Co and LDH occurred in SCM milk containing gram positive bacterial agents only. ALP was found to be elevated in milk infected by both gram positive and negative bacteria. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and $SCC\geq2{\times}10^5$ cells/ml of milk as the benchmark. Only ALP and Zn, the former being superior, were found to be suitable for diagnosis of SCM irrespective of etiological agents. LDH, Co and Fe can be introduced in the screening programs where Gram positive bacteria are omnipresent. It is recommended that both ALP and Zn be measured together in milk to diagnose buffalo SCM, irrespective of etiology.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.7
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• pp.910-913
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• 2004

Insulin-like growth factor binding protein-3 (IGFBP-3) gene is a structural gene associated with the growth and development of the animals. The present investigation was carried out to unravel nucleotide sequence and polymerase chain reactionrestriction fragment polymorphism (PCR-RFLP) of IGFBP-3 gene in buffalo. Genomic DNA was isolated from a total of 157 animals belonging to Murrah, Surti, Jaffarabadi and Nagpuri breeds of Indian riverine buffalo. A 655 bp of IGFBP-3 gene was amplified in all the breeds and amplicons were digested with Hae III, Taq I and Msp I restriction enzymes. On digestion with Hae III yielded single restriction pattern of 8 fragments of sizes 201, 165, 154, 56, 36, 19, 16 and 8 bp in all the animals studied. Similarly Taq I and Msp I also revealed single restriction pattern yielding fragments of sizes 240 and 415 bp and 145 and 510 bp, respectively. This shows nonpolymorphic nature of restriction sites in buffalo. Nucleotide sequencing of 587 bp of IGFBP-3 gene in Murrah buffalo was done and submitted to the GenBank (Accession No. AY304829). Nucleotide sequencing revealed an addition of 4 bases in the intronic region as compared to cattle.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1556-1560
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• 2006

Six breeds of riverine buffalo viz. Murrah, Mehsana, Jaffrabadi, Nagpuri, Nili-Ravi and Bhadawari were characterized using FAO-recommended cattle specific microsatellite markers. Among the total of twenty microsatellite markers screened to explore genomic variability of six buffalo breeds, only ten were polymorphic in nature. Four out of ten polymorphic microsatellite loci were rated as informative. The numbers of alleles detected ranged from 2 to 7, with a mean of $5.5{\pm}0.07$ per microsatellite marker. The most polymorphic marker was BM1818 with a total of 7 alleles present at this locus. One breed specific marker was found in each of Mehsana (BM1818) and Bhadawari (ILSTS030) and four were found in Jaffarabadi (BM1818, ILSTS030, ILSTS054 and ILSTS011). Genetic distance (Ds) between the Mehsana and Bhadawari breed was the maximum (0.29), followed by Murrah and Mehsana (0.27), and Nili-Ravi and Bhadawari (0.26). The lowest Ds was found between the Jaffrabadi and Nagpuri breeds which was only 0.05. The highest divergence time of 1318 years was established between Mehsana and Bhadawari breeds whereas it was found to be lowest (272 years) between the Jaffrabadi and Nagpuri breeds.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.993-999
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• 2010

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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• pp.626-635
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• 2009

An experiment was conducted to investigate the effect of Butylated Hydroxy Anisole (BHA), Butylated Hydroxy Toluene (BHT), Pentoxifylline (PTX), Theophylline (TPY) and Theobromine (TBR) on cold protection ability of Murrah buffalo semen at room ($22-25^{\circ}C$) and refrigerated temperature ($4-7^{\circ}C$). Each semen sample was divided into six parts of equal volume and sperm concentration; the first was kept as a control and the remaining five were treated with BHA, BHT, PTX, TPY or TBR. Sperm motility, abnormal spermatozoa, live-dead count, hypo-osmotic swelling and acrosomal integrity were studied at room and refrigerated temperature for various incubation periods viz.; 0, 4, 8, 12 and 24 h at room and 0, 12, 24, 36, 48, 60 and 72 h at refrigerated temperature. Significant improvement in sperm motility, live-dead count, hypo-osmotic swelling and acrosomal integrity were observed in BHT, PTX and TPY fortified extender at room and refrigerated temperature for various incubation periods. From the present study it could be concluded that cold protection ability of buffalo semen can be improved through the addition of BHT followed by PTX and TPY.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.6
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• pp.788-794
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• 2013

Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. ${\alpha}_1$-anti trypsin, ${\alpha}_1$-acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with $SCC{\geq}2{\times}10^5$ cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that ${\alpha}_1$-anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of ${\alpha}_1$-acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and $SCC{\geq}2{\times}10^5$ cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that ${\alpha}_1$-anti trypsin indicates SCM irrespective of etiology, whereas ${\alpha}_1$-acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a good indicator of SCM. It is recommended measuring both ${\alpha}_1$-anti trypsin and ${\alpha}_1$-acid glycoprotein in milk to diagnose SCM in buffalo irrespective of etiology.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.9
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• pp.1166-1173
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• 2010

Sixteen Mediterranean pluriparous buffaloes were subdivided into two uniform groups of eight animals. The average weight of the two groups at the start of the trial was 671.2 and 656.7 kg. The number of days from calving were 33.4 and 33.3, and the average milk production was 12.73 and 12.33 kg/d. The trial lasted for 114 days, and was divided into two sub-periods of 58 and 56 days. The two diets, administered ad libitum, had the same forage/concentrate ratio (53/47) but in their formulation the percentage of the two forages varied. Diet 1: alfalfa hay = 10%, maize silage = 43%, concentrate 1 = 47% (6.63 MJ/kg DM of net energy; 179.5 g/kg DM of crude protein). Diet 2: alfalfa hay = 20%, maize silage = 33%, concentrate 2 = 47%, (5.99 MJ/kg DM of net energy; 155.4 g/kg DM of crude protein). For the overall trial period (33-146 days in milk), the intake of dry matter was 17.23 kg/d for Group 1 and 17.29 kg/d for Group 2 and corresponded to 2.50 and 2.58% (p<0.01) of live weight. There was no significant difference between the average weight (689.7 and 669.4 kg) and the body condition score (6.49 and 6.42) of the two groups of buffaloes. Group 1 produced a greater quantity of milk (11.89 vs. 10.90 kg/d, p<0.10) of better quality both for its higher fat content (82.32 vs. 77.29 g/kg, p<0.10) and its protein content (47.36 and 46.38 g/kg). The milk produced by the buffaloes receiving Diet 1 had a better clotting ability, lower values of r (15.98 and 16.42 min) and K20 (1.66 and 1.75 min) and a higher value of A30 (54.45 and 52.73 mm). Taking into consideration the two sub-periods, milk production was significantly different only in the first sub-period (33-90 DIM), in favour of Group 1 (13.08 vs. 11.56 kg/d, p<0.05), while the positive effect of Diet 1 was cancelled out (10.71 and 10.24 kg/d) in the second part of the trial (91-146 DIM).

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.3
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• pp.335-340
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• 2012

The data used came from two trials undertaken under the same climatic conditions (spring-summer). In both trials pluriparious buffaloes were utilized similar in weight, body condition score, and milk production from the previous year. From the first trial the data used was from the sub-period 23-88 DIM provided by seven animals fed ad libitum with diet A (6.69 MJ/kg DM; 158.30 g/kg of crude protein) with a forage/concentrate ratio of 48/52. From the second trial the data used was from the sub-period 33-90 DIM provided by seven animals fed ad libitum with diet B (6.63 MJ/kg DM; 179.50 g/kg of crude protein) and by seven animals fed ad libitum with diet C (5.99 MJ/kg DM; 155.40 g/kg of crude protein), each of the diets had the same forage/concentrate ratio (53/47). A significant difference was found in milk production between group B and C (13.08 vs. 11.56 kg/d, p<0.05), an intermediate production (12.10 kg/d) was noted in group A. A significant difference was found between fat (76.58 vs. 69.24 g/kg, p<0.05), protein (46.14 vs. 43.16 g/kg, p<0.05) and casein (39.94 vs. 34.98 g/kg, p<0.05) of the milk of group B with respect to group A. The milk of group C gave fat values (71.80 g/kg), protein (45.52 g/kg) and casein (39.06 g/kg) statistically equal to those of group B. The milk of groups B and C, in respect to the milk of group A, gave values of $K_{20}$ (1.77, 1.82 vs. 3.68 min, p<0.05), statistically lower and values of $A_{30}$ (48.28, 47.27 vs. 40.64 mm, p<0.05) statistically higher. Two simple linear regressions were calculated where the independent variable (x) was the daily standardized milk production, the dependent variable (y) or the daily intake of net energy or crude protein. Equation 1) NE (MJ/d) = 74.4049+2.8308${\times}$kg of normalized milk; equation 2) CP (kg/d) = 1.4507+0.1085${\times}$kg of normalized milk, both the equations were significant (p<0.05) with determination coefficients of 0.58 and 0.50 respectively. For a production of normalized milk that varies from 9 to 13 kg, the respective energy-protein concentrations fluctuate from 6.09 to 6.78 MJ/kg DM and from 148.00 to 174.46 g/kg DM.