ISOLATION, PARTIAL PURIFICATION AND IMMUNOBIOCHEMICAL CHARACTERIZATION OF INHIBIN FROM GOAT OVARIAN FOLLICULAR FLUID

In the present study Inhibin was purified from goat ovarian follicular fluid by Gel filtration chromatography on Sephacryl S-200. SDS PAGE of purified inhibin revealed the molecular weight of 55 kDa. The purified follicular fluid had been found to be immunoreactive by DID test and Western Blot analysis when treated against hyperimmune sera

PARTIAL PURIFICATION AND IMMUNO-BIOCHEMICAL CHARACTERISATION OF FERTILITY ASSOCIATED PROTEIN OF KARAN FRIES BULL SEMINAL PLASMA

The objective of the present study was detection, isolation, partial purification and immunobiochemical characterization of fertility associated protein in the seminal plasma of high prolific Karan fries bull. Seminal plasma of Karan Fries bull was partially purified by gel filtration chromatography and analyzed by 10% SDS-PAGE for their polypeptide profile. PAGE analysis revealed major band of 55 kDa, and 26 kDa. Hyperimmune serum was raised in rabbit against crude seminal plasma protein. Single precipitin line was observed in DID test when each of the partially purified 26 kDa and 55 kDa proteins were reacted with hyperimmune serum. These proteins were also found to be immunoreactive against hyperimmune serum in Western blot technique

Study on the effect of toxicity under highly arsenic prone zone in Nadia district of West Bengal in India

The present study was carried out on the basis of status of arsenic in soil, drinking water and plants, blood, urine and faeces of animals at arsenic prone zone. Within the ambit with the environment, the examination of animals was taken into consideration. They were screened and categorised on the degree of As toxicity. For field works animals were randomly selected from arsenic prone zone. The external manifestation indicated a complex syndrome and characteristic signs such as increased heart rate and respiratory rate, red urine, congested mucous membrane, anorexia, absence of ruminal motility, diarrhoea with blood, polyuria and unusual weight loss. The haematobiochemical changes such as low Hb level, decreased level of TEC, TLC and increased level ALT, AST, BUN and creatinine. Increased level of arsenic in urine, blood and faeces than the value of control animals could be the confirmatory indication of arsenic toxicity

Effect of micronutrient supplementation on hormonal profile of local goat and sheep breeds of West Bengal

The investigation was carried out to assess the influence of trace mineral supplementation on certain steroid and thyroid hormones that affect reproduction in Black Bengal goat and Garole sheep. Trace mineral feeding to local goat and sheep breeds showed positive influence. Rise of the serum trace minerals does not necessarily influence rising of serum steroid and thyroid hormone levels within short period

Molecular characterization of buffalo αs1-casein gene

The caseins (Alpha s1, Alpha s2, beta and kappa) comprise the major protein component of ruminant milk and are secreted in the form of stable calcium phosphate micelles. The present study was proposed to characterize αs1-casein gene in buffalo at the molecular level to determine complete αs1-casein cDNA sequence. Sequencing results were analyzed and aligned with available mRNA GenBank sequences of other domestic animals (Accession AAB34797, NM_001009795, X72221, AY344966, NM_181029, AY948385 and BC109618). The complete amino acid coding region of the buffalo αs1- casein was deduced from cloned cDNA generated from mammary tissue mRNA. Amino acid sequence comprises of 199 amino acid residues. When aligned with previously reported buffalo sequence (Accession-AY948385) it showed one amino acid (Glutamine) deletion at amino acid position 78

PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF IgG FROM SERUM OF YAK ( BOS GRUNNIENS )

In the present study Immunoglobulin G was purified from serum of Yak by gel filtration chromatography on Sephacryl S-200. SDS- PAGE of purified yak IgG showed four major polypeptides of 62.94 kDa, 53.29 kDa 29.22 and 28.21 kDa. The purified Immunoglobulin has been found to be immuno-reactive by DID test and Western Blot analysis when treated against hyper-immune sera

Immunochemical characterization of purified buffalo beta-lactoglobulin

Beta-lactoglobulin (β-lg) is a major whey protein found in cow milk and other ruminants, deer, bison, and buffalo. β-lg is considered as one of the main allergenic components in bovine milk and therefore the modification of β-lg is considered as a promising treatment for milk allergy. Thus the present study was conducted to assess the antigenicity of purified β-lg that can be exploited for detection of buffalo milk-protein intolerance. β-lg was purified from milk of Murrah buffalo of Haringhata Farm, West Bengal (India) by combination of gel filtration (using Sephacryl S-200) and anion-exchange chromatography (using DEAE-Sepharose). The molecular weight was derived as 18.05 kDa in 15 percent one-dimensional SDS-PAGE. Antigenicity (seroreactivity) of purified β-lg was evaluated by double immuno diffusion (DID), western blotting, dipstick ELISA and indirect ELISA using hyperimmune sera raised in rabbit against purified buffalo β-lg. In indirect ELISA, titer of rabbit serum against buffalo β-lg was observed to be in between 200-400. Therefore, the study revealed that purified buffalo β-lg is highly antigenic and a seroreactive protein and may be used for detection of buffalo milk proteinintolerance using rapid, user-friendly, cost effective sero-diagnostic technique/too

PARTIAL PURIFICATION AND IMMUNE-BIOCHEMICAL CHARACTERIZATION OF DOG SERUM IMMUNOGLOBULIN G

In the present study Immunoglobulin G was purified from serum of dog by gel filtration chromatography on Sephacryl S-200. SDS- PAGE analysis of purified dog IgG showed major polypeptides of 66 kDa, 52.40 kDa and 20.72 kDa. The purified Immunoglobulin has been found to be immune-reactive by DID test and Western Blot analysis when treated against hyperimmune sera which was raised in rabbit

Isolation and purification of beta-lactoglobulin from cow milk

Aim: The present study was undertaken to standardize a convenient method for isolation and purification of β-lactoglobulin (β-lg) from cow milk keeping its antigenicity intact, so that the purified β-lg can be used for detection of cow milk protein intolerance (CMPI). Materials and Methods: Raw milk was collected from Gir breed of cattle reared in Haringhata Farm, West Bengal. Milk was then converted to skimmed milk by removing fat globules and casein protein was removed by acidification to pH 4.6 by adding 3 M HCl. β-lg was isolated by gel filtration chromatography using Sephacryl S-200 from the supernatant whey protein fraction. Further, β-lg was purified by anion-exchange chromatography in diethylaminoethyl-sepharose. Molecular weight of the purified cattle β-lg was determined by 15 percent one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was analyzed by gel documentation system using standard molecular weight marker. Results: The molecular weight of the purified cattle β-lg was detected as 17.44 kDa. The isolated β-lg was almost in pure form as the molecular weight of purified β-lg monomer is 18kDa. Conclusion: The study revealed a simple and suitable method for isolation of β-lg from whey protein in pure form which may be used for detection of CMPI

Characterization of enterovirulent Escherichia coli isolated from yak calves died of diarrhoea in India

Shiga toxin producing (STEC), enteropathogenic (EPEC) and enterotoxigenic Escherichia coli (ETEC) were isolated from 29 diarrhoeic yak calves from two districts of Arunachal Pradesh, India (West Kameng and Tawang) during 2005 to 2011. The STEC (28) and EPEC (12) isolates belonged to 25 different O serogroups. Among the 28 STEC isolates, 8 (28.5%) isolates carried only stx1, 10 (35.7%) isolates were positive for both stx1 and stx2, 10 (35.7%) isolates carried only stx2. The stx variants such as stx1c, stx2c, stx2d and stx2e were detected in 7 (25%), 9 (32.1%), 1 (3.5%) and 4 (14.28%) isolates, respectively. Among the 12 EPEC isolates, 2 (4.65%) strains were ‘typical’ possessing bfpA gene. The ehxA and saa genes were present in 19 and 4 isolates, respectively. Among the 14 ETEC isolates, 6 (42.8%), 7 (50%), 5 (35.7%), 5 (35.7%), 7 (50%) and 6 (42.8%) isolates carried genes coding for STa, STb, LT, F5, F41 and EAST1, respectively. The present study detected LT, STb, F41 as the most prevalent type in yaks associated with diarrhoea. Further, the STEC, EPEC and ETEC isolates showed resistance against furazolidone (40%), nitrofurantoin (39%), nalidixic acid (38%), erythromycin (38%), kanamycin (38%), amikacin (37%). The dendrogram revealed clonal relationship among the isolates after RAPD/ERIC analysis. The study could not detect any specific RAPD/ERIC cluster associated with origin, serotypes or virotypes of the isolates. Thus, it was concluded that genetically diverse, multi-drug resistant enterovirulent E. coli were associated with death due to diarrhoea in yak calves in India.</jats:p