Antibacterial effect of kitchen herbs against pathogenic multidrug-resistant E. coli isolates from calf diarrhoea.

Calf diarrhoea remains the biggest challenge both in the small and large farms. Infectious diarrhoea is associated with many pathogens, Escherichia coli being one, but majority are systematically treated with antibiotics. Since antimicrobial resistance (AMR) is a growing menace, the need to find alternative prophylactic solutions using popular kitchen herbs such as Trachyspermum ammi (carom seeds), Curcuma longa (turmeric) and cinnamon (Cinnamomum sp.) extracts is been investigated against virulent form of E. coli isolated from calf diarrhoea. The virulence factors identified in these isolates were ST (32.5%), LT (20%), eaeA (15%), stx1 (2.5%) and stx2 (5%) with the occurrence of the most common serogroups as O18 (15%) followed by O111 (12.5%). Highest resistance was seen with beta lactam + beta lactamase inhibitor (amoxicillin/clavulanic acid) followed by beta lactams (ampicillin, cefuroxime and cefepime). The zone of inhibition due to cinnamon (methanol) and carom seed (ethanol) extracts (500 to 250 µg/mL concentration) on E. coli bacteria was >19 mm, respectively. Turmeric, cinnamon and carom had the potency of inhibiting the pathogenic E. coli which maybe suggestive of its use in calf diets as prophylaxis against diarrhoea.

Effect of reducing energy intake during the dry period on milk production, udder health, and body condition score of Jersey crossbred cows in the tropical lower Gangetic region.

To find out the effect of reducing energy intake during dry period on milk production, udder health, and body condition, the experiment was conducted on 14 Jersey crossbred cows during whole dry period and continued up to 120 days of lactation. Reduction in energy intake was done during far-off period for each dry cow of treatment group as compared to control group. Statistically analyzed data revealed that overall significantly (P < 0.01) lower DMI and WI were recorded in control than treatment group. Overall significantly (P < 0.01) higher total milk production was found in treatment than control group. Overall significantly (P < 0.01) lower milk SCC, MCMT, pH, and EC were found in treatment than control group. Nonsignificant difference in milk fat, SNF, total solid, total protein, and fat:protein ratio was found. Overall significantly (P < 0.01) better quality milk (MBRT) was found in treatment than control groups. BCS during dry period and at calving was significantly (P < 0.01) different between groups. Significantly (P < 0.01) higher plasma NEFA concentration was estimated in control than treatment groups in all stages. No significant difference was found for plasma concentrations of glucose, urea, and total protein. The coefficients of correlation indicated significant (P < 0.01) correlation among BCS, milk production, milk SCC, MCMT, pH, and EC. It can be concluded that reducing energy intake during far-off dry period can lead to achieve optimum BCS at calving. Suitable BCS at calving was beneficial to get higher milk production with improved quality, better maintenance of udder health and body condition of Jersey crossbred cows at tropical lower Gangetic region.

Bovine Circadian Locomotor Output Cycles Kaput (CLOCK) and Clusterin (CLU) mRNA Quantitation in Ejaculated Crossbred Bull Spermatozoa.

Mammalian circadian locomotor output cycles kaput (CLOCK) gene encodes a transcription factor that affects both the persistence and the period of circadian rhythms. Earlier reports suggested that CLOCK gene might be associated with male infertility in human. Present investigation, for the first time, reports that CLOCK gene expresses differentially between good and poor quality crossbred bull semen. The relative expression of CLOCK was significantly (p < 0.05) higher among good quality bull semen than motility-impaired ones. Clusterins (CLU) are series of genes associated with a variety of physiological activities including spermatogenesis, apoptosis and degenerative disease conditions. In the present context, we also investigated that the expression of CLU gene was significantly (p < 0.05) higher among motility-impaired crossbred bull semen compared to the good quality one.

Readily usable strategies to control mastitis for production augmentation in dairy cattle: A review.

Mastitis in dairy cattle is the most common management disorder that causes higher economic losses by lowering production and quality of milk leads to substantial economical loss. The aim of this article was to review worldwide important advances in strategies to control mastitis for production augmentation in dairy cattle. Many scientists worked to identify effective strategies to control mastitis caused by Streptococcus agalactiae, Staphylococcus aureus, and others. It is necessary to identify mechanisms of infection, define clinical and subclinical states of disease, determine exposure time, and identify pathogen-specific characteristics. Evolvement of management strategies that incorporated hygienic procedures (animal, floor, and milkman), post milking standing period of animal and strategic use of antibiotic or herbal therapy at dry-off, nutritional supplementation, fly control, body condition score optimization, etc., resulted in widespread control of mastitis. The udder, teat of animal, scientific management of milking, automatic milking procedure, genetic selection are considered as important factors to control mastitis. As farm management changed, scientists were directed to redefine control of mastitis caused by opportunistic pathogens of environmental sources and have sought to explore management strategies which will maintain animal well-being in a judicial way. Although significant advances in mastitis management have been made changing herd structure, changing climatic scenario and more rigorous milk processing standards ensure that mastitis will remain important issue for future research.

Osteosynthesis for intra-articular calcaneal fractures.

PURPOSE: To correlate treatment results of intra-articular calcaneal fractures with their computed tomographic (CT) classification. METHODS: 36 men and 4 women with 48 intra-articular calcaneal fractures (8 bilateral) underwent open reduction and internal fixation with bone grafting via an extensile lateral approach. Based on 2-dimensional CT scans, the fractures were categorised using the Sanders classification. There were 16 type-II, 20 type-III, and 12 type-IV fractures. Radiographs and Maryland foot scores were used for evaluation of the results at a mean of 38 (range, 26-66) months. RESULTS: Anatomic reduction of the posterior calcaneal facet was achieved in 38 of 48 fractures. The Bohler and Gissane angles were restored to between 92 and 99% of normal, respectively. Despite this, the mean functional scores were 84 in type-II, 83 in type-III and 67 in type-IV fractures. CONCLUSION: Surgical results were superior in type-II and -III fractures. Type-IV fractures fared poorly, despite excellent restoration of calcaneal anatomy; subtalar arthrodesis should have been considered.

Molecular markers, BM1500 and UMN2008, are associated with post-thaw motility of bull sperm.

Cryopreservation is one of the most important aspects of frozen semen technology and livestock breeding. Uses of candidate molecular markers in selection strategies for male fertility are well recognized. The present investigation targeted two microsatellite markers (BM1500 and UMN 2008) for association with semen quality variables and freezing capacity in Frieswal (HF×Sahiwal) crossbred bulls of Indian origin. Of the different alleles at the polymorphic locus BM1500, the 136bp allele was associated with greater (P<0.05) post-thaw motility percentage (PTM) while the 134 allele was associated with less (P<0.05) PTM. The 134/134 genotype at the polymorphic locus, UMN2008 was associated with greater (P<0.05) post-thaw motility while there was no allele effect on PTM. When combined genotypes UMN2008/BM1500 were analyzed, the 134/134-136/136genotype had the greatest (P<0.05) association with PTM. The present study is an initial report on the potential use of these markers as male reproductive biomarkers for improving semen freezing capacity in bulls.

A SNP (g.358A > T) at intronic region of CD9 molecule of crossbred bulls may associate with spermatozoal motility.

The surface expression of CD9 (cluster-of-differentiation antigen-9) in sperms of certain mammalian species has been attributed to its fusion with the egg and thereby dictating the fertility of species. In the present study, we investigated the association of CD9 with crossbred bull sperm quality and quantity trait was analyzed using a total of 96 Frieswal (HF × Sahiwal) crossbred. A single nucleotide polymorphism (g.358A > T) in intron 6 was significantly associated with sperm concentration (P < 0.05) and motility percentage (P < 0.01). mRNA was extracted from good (progressive motility > 50%) and motility impaired (progressive motility < 50%) bull semen. The mRNA expression and seminal plasma protein concentration of CD9 was significantly (P < 0.05) higher among good quality bull semen than motility impaired ones. Our results thus may indicate that, mutation in the intronic region may be responsible for the instability of RNA and the subsequent functional protein expression.

Interactions of concanavalin A with glycoproteins. A quantitative precipitation study of concanavalin A with the soybean agglutinin.

Certain oligomannose-type glycopeptides have been previously shown to be bivalent for binding to concanavalin A and capable of precipitating the lectin by forming homogeneous cross-linked lattices [L. Bhattacharyya, M. I. Khan, and C.F. Brewer, Biochemistry, 27 (1988) 8762-8767]. In the present study, the effect of protein environment on the binding properties of an oligomannose-type oligosaccharide has been examined through quantitative precipitation analysis of the interactions of concanavalin A (Con A) with the soybean (Glycine max) agglutinin (SBA), which is a tetrameric glycoprotein possessing a single Man9-oligomannose chain per monomer. The results showed that SBA forms two different types of cross-linked complexes with tetrameric Con A, depending on the relative ratio of the two molecules in solution. At a concentration of one equivalent or less, SBA forms a 1:1 complex with Con A. At concentrations exceeding one equivalent, SBA forms a 2:1 complex with Con A. However, SBA forms only 1:1 cross-linked complexes with dimeric forms of Con A, such as acetyl- and succinyl-Con A. The results demonstrated that the total valency of the carbohydrate of SBA is a function of both the quaternary structure of Con A, as well as the relative ratio of SBA to Con A. In addition, the individual Man9-oligosaccharide, which as a glycopeptide is bivalent for binding to Con A, expresses univalency when present on the protein matrix of SBA.

Analysis of equilibrium dissociation and unfolding in denaturants of soybean agglutinin and two of its derivatives.

The equilibrium denaturation of tetrameric soybean agglutinin (SBA) in urea and guanidine hydrochloride (GdnHCl) has been examined by steady-state fluorescence and size-exclusion chromatography. The denaturation of SBA reveals two distinct and separable transitions: dissociation (native tetramer<-->tertiary monomer) and unfolding (tertiary monomer<-->unfolded monomer). The urea denaturation curves of N-dimethyl and acetyl derivatives of SBA are also similar to unmodified lectin but the midpoints, [D](1/2), are shifted to lower denaturant concentrations. The free energy of stabilization of tertiary structure (DeltaG(u,aq)) of SBA is estimated to be 4.5-4.6 kcal mol(-1), which shows a decrease by approximately 10-15% for both N-dimethyl SBA and acetyl-SBA. The free energy term (DeltaG(d, aq)) for the relative stability of the quaternary structure of SBA and its derivatives shows that the decrease in stability relative to SBA occurs by <10% for N-dimethyl SBA while for acetyl-SBA, this occurs by approximately 30%. However, the m values depicting the dependence of free energy on denaturant concentration for SBA and its derivatives are similar for dissociation as well as unfolding, which suggest similar denaturation pathways of unmodified and modified SBA.

Physical and chemical characterization of macroglobulin from marine turtle eggwhite.

Macroglobulin was purified from the eggwhite of the marine turtle (Caretta caretta Linn.) by gel filtration through Sephadex G-200 and Sepharose 4B. It was characterized by physical techniques including sedimentation velocity, diffusion and viscosity. Its molecular weight (Mr) was determined as 724,000 with four subunits of equal molecular weight. A large amount of water was hydrodynamically associated with the macroglobulin. It inhibited the activities of trypsin and papain and did not cross-react with human alpha 2-macroglobulin. Its amino acid composition was similar to that of human alpha 2-macroglobulin. Results suggest that turtle eggwhite macroglobulin is a homologous but distinct protein from human alpha 2-macroglobulin.

Motion characteristics of Murrah buffalo bull spermatozoa in various seasons and its relationship with functional integrity of the plasmallema.

Semen was collected from six adult (3.5-7-year-old) Murrah buffalo bulls at weekly intervals for 1 year and evaluated for routine parameters, motion characteristics, reactivity in hypoosmotic solution, and acrosomal and other morphological abnormalities of the spermatozoa. The overall motility (MOT), straight line velocity (VSL), curvilinear velocity (VCL), linearity (LIN), lateral head displacement (ALH) and average path velocity (VAP) were 66.85+/-2.79%, 26.58+/-0.24 and 107.07+/-1.47 microm/s, 26.91+/-0.01%, 11.19+/-0.09 and 61.78+/-2.79 microm/s, respectively. Significant seasonal variation was observed in sperm kinematics and hypoosmotic swelling (HOS) reactivity. Except for LIN, the mean values of sperm dynamics were higher during summer and rainy season and significantly lower in winter season. Sperm kinematics showed significant (P<0.01) positive correlation (r=0.25-0.60) with plasmallemal integrity. Ejaculates with less than 50% HOS-reactive spermatozoa had significantly lowered MOT, VSL, VCL and VAP as compared to the ejaculates with >50% HOS-positive spermatozoa. No significant difference was observed in sperm kinematics among the ejaculates having 50-70% and >70% HOS-reactive spermatozoa. The trend of motion dynamics of the spermatozoa with respect to HOS reactivity was similar in all the three seasons (summer, rainy and winter). The results indicate that ejaculates having more than 50% of HOS-reactive sperm show a higher magnitude of sperm kinematics compared to ejaculates having less than 50% HOS-positive spermatozoa.

Spinal stenosis: its diagnosis and management--a clinical and radiological study.

A clinico-radiological study conducted on 124 patients including 89 cases of spinal canal stenosis and 143 normal (control) individuals is presented and the various causes of stenosis of the spinal canal are reviewed. Evaluation of canal to body ratio using plain radiographs of lumbo-sacral spine based on the Jones and Thomson technique and myelography were found very useful in diagnosing spinal canal pathology preoperatively. Spinal stenosis is more common in males, predominantly in the fourth decade. Extensive decompression laminectomy in appropriately selected cases yielded satisfactory results, any associated prolapse intervertebral disc requiring excision of the herniated disc in addition to laminectomy.

Hypertensive primary intraventricular haemorrhage.

A case of hypertensive primary intraventricular haemorrhage is reported. The patient recovered with conservative management. To the best of our knowledge, there has been no previous report of this rare entity in Indian literature.

Evaluation of three bovine Y specific microsatellite loci as an alternative biomarkers for semen quality traits in crossbred bull.

Although some of the studies earlier reported that bovine semen parameters are associated with some candidate markers genes, but scanty of reports available regarding the effect of allelic variation in Y specific microsatellite markers on semen quality parameters in bulls. In the present study we have targeted three Y specific microsatellite markers (INRA126, INRA 189 and BM861) for their association ship analysis with some semen quality parameters among Frieswal (HF × Sahiwal) crossbred bulls of Indian origin. The polymorphic loci of INRA 126, bulls with 182 and 184 alleles had significantly (P<0.01) higher semen volume as compared to 186 allele, however, 186 allele showed significantly (P<0.01) higher concentration per ml of semen compared to 182 and 184. Interestingly our study also revealed that number of sperm/ejaculate is also significantly (P<0.05) higher in 184 allele compared to 182 and 186. Similarly, association analysis of INRA 189 major three alleles also revealed a significant difference in semen volume and concentration. Allele 89 and 96 having significantly (P<0.01) higher volume compared to 86, whereas allele 86 having significantly (P<0.01) higher concentration per volume of semen than 89 and 96. Again after association of two major alleles (160 and 164) of BM861 loci with semen parameters revealed no significant difference with any of the semen quality parameters chosen here. Therefore the present study may be for the first time revealed that the Y chromosomal microsatellite alleles are important male reproductive biomarkers for improving semen quality traits in bulls.

Differential expression of protamine 1 and 2 genes in mature spermatozoa of normal and motility impaired semen producing crossbred Frieswal (HF×Sahiwal) bulls.

Mature spermatozoa contain thousands of mRNA transcripts. These untranslated mRNA may perhaps serve as a "footprint" of spermatogenesis since many of them might directly or indirectly be involved in fertilization, early embryo cleavage, poor semen quality and fertility. In this study, we tried to isolate high-quality RNA from mature spermatozoa and to monitor the expression profile of protamine 1 (PRM1) and protamine 2 (PRM2) gene in ejaculated spermatozoa of normal (good, % initial progressive motility: 57.61±1.41, n=9) and motility impaired (poor, % initial progressive motility: 18.45±1.61, n=8) crossbred Frieswal (HF×Sahiwal) bulls semen using real time quantitative PCR. Semen samples were subjected to discontinuous (45:90) Percoll gradient centrifugation, specifically to eliminate damaged spermatozoa and contaminating somatic cells. Total RNA was extracted from sperm pellets and cDNA was synthesized. Furthermore, the absence of contamination of germ cells, epithelial cells and leucocytes in all the RNA extractions was tested by RT-PCR targeting specific molecular markers like KIT, CDH1 and CD4, respectively. The presence of transcripts like PRM1, PRM2, DAZL, and PPIA were demonstrated in ejaculated spermatozoa using appropriate PCR primers without RNA amplification. Expression of PRM1 and PRM2 genes were evaluated by real time quantitative PCR using TaqMan chemistry, where PPIA was used as internal control. The cDNA synthesized from normal buffalo testicular tissue was served as positive control. The good quality semen producing group showed significantly higher level of PRM1 mRNAs expression as compared to the poor quality semen producers (P<0.05) indicating putative role of the gene and semen quality parameters especially initial progressive motility. However, PRM2 transcript levels were not significantly different between the groups (P>0.05).

Effect of age on spermiogram of Holstein Friesian × Sahiwal crossbred bulls.

This study was conducted on 94 Frieswal (5/8 Holstein Friesian 3/8 Sahiwal) crossbred bulls of three different grades, categorized based on their semen freezability visualising Group 1 (consistently freezable semen producer bulls, N = 11), Group 2 (inconsistent freezable, N = 16) and Group 3 (Non freezable, N = 67). Each group was further divided into two classes that is young (up to 30 months) and adult (31 to 70 months) bulls depending upon their age. Sperm morphology was studied by using the eosin-nigrosin staining technique. Bulls age significantly (P < 0.01) affected semen quality and sperm morphology. In adult bulls, semen volume, mass activity and sperm concentration were 36%, 17.56% and 19.6%, respectively, higher than young. Initial progressive motility (%) and livability showed significant (P < 0.01) improvement with the advancement of age (43.37 ± 1.21 and 67.71 ± 1.11, respectively, in young; 53.02 ± 1.11 and 74.17 ± 1.03, respectively, in adult). In young bulls, sperm head, mid piece, tail abnormality and total abnormal sperm percent (12.38 ± 0.92, 4.87 ± 0.24, 11.01 ± 0.60 and 28.26 ± 1.34, respectively) were 1.85, 1.27, 1.20 and 1.44 folds higher than that of their mature stage (6.69 ± 0.64, 3.82 ± 0.32, 9.14 ± 0.64 and 19.66 ± 1.31, respectively). Significant reduction (P < 0.01) in micro cephalic sperm, free heads, bent mid piece, looped mid piece and proximal protoplasmic droplets were observed at mature age as compared with their younger stage. In bulls of consistent freezing category, abnormal sperm heads significantly decreased from 4.40 ± 0.31% to 3.28 ± 0.02% on maturity. Similarly, in inconsistent freezing grade bulls sperm head abnormality (9.28 ± 0.75% to 5.13 ± 1.20%) and total abnormal sperm percent (24.89 ± 1.43 to 18.73 ± 3.40) was decreased over the age. On the contrary, in non-freezing category bulls' sperm morphology did not show significant (P > 0.05) improvement with age advancement, rather some abnormalities like long slender head, under developed/deformed head, abaxial implantation of mid piece, double mid piece, stump tail and distal protoplasmic droplets tend to increased significantly (P < 0.05) with age of bulls. Results indicated that in potential Frieswal bulls semen quality and sperm morphology were improved from young to mature stage, where as, in poor quality (non-freezing) semen producer bulls neither the morphology nor the semen quality showed any improvement with maturity. It was recommended that crossbred bulls producing more than 25% morphologically abnormal sperms in young age (below 30 months) along with poor progressive motility (<50%) and low sperm concentration (<1000 million/ml) need immediate culling with out any expectation of further improvement in semen quality with age advancement.

Cross-linking activity of the 14-kilodalton beta-galactoside-specific vertebrate lectin with asialofetuin: comparison with several galactose-specific plant lectins.

We have previously shown that plant lectins with a wide range of carbohydrate binding specificities can bind and cross-link (precipitate) specific multiantennary oligosaccharides and glycopeptides [cf. Bhattacharyya, L., Fant, J., Lonn, H., & Brewer, C. F. (1990) Biochemistry 29, 7523-7530]. This leads to a new source of binding specificity: namely, the formation of homogeneous cross-linked lattices between lectins and carbohydrates. Recently, we have demonstrated the existence of highly ordered cross-linked lattices that form between the D-Man/D-Glc-specific plant lectin concanavalin A and the soybean agglutinin which is a tetrameric glycoprotein possessing a single Man9 oligomannose chain per monomer [Khan, M. I., Mandal, D. K., & Brewer, C. F. (1991) Carbohydr. Res. 213, 69-77]. In the present study, we have compared the ability of the 14-kDa beta-galactoside-specific lectin from calf spleen, a dimeric S-type animal lectin, and several galactose-specific plant lectins from Erythrina indica, Erythrina cristagalli, and Glycine max (soybean agglutinin) to form specific cross-linked complexes with asialofetuin (ASF), a 48-kDa monomeric glycoprotein, using quantitative precipitation analyses. The results show the formation of 1:9 and 1:3 stoichiometric cross-linked complexes (per monomer) of ASF to the 14-kDa lectin, depending on their relative ratio in solution. Evidence indicates that the three triantennary N-linked complex-type oligosaccharide chains of ASF mediate the cross-linking interactions and that each chain expresses either trivalency in the 1:9 cross-linked complex or univalency in the 1:3 complex.(ABSTRACT TRUNCATED AT 250 WORDS)

Interactions of concanavalin A with glycoproteins: formation of homogeneous glycoprotein-lectin cross-linked complexes in mixed precipitation systems.

We have previously demonstrated that the interactions between branched chain oligosaccharides and glycopeptides isolated from glycoproteins and glycolipids with specific lectins lead to the formation of homopolymeric carbohydrate-protein cross-linked complexes, even in the presence of mixtures of the carbohydrates or lectins [cf. Bhattacharyya, L., Fant, J., Lonn, H., & Brewer, C. F. (1990) Biochemistry 29, 7523-7530]. Recently, we have shown that highly ordered cross-linked lattices are formed between the tetrameric glycoprotein soybean agglutinin (SBA), which possesses a Man9 oligomannose chain per monomer, and the Glc/Man-specific plant lectin concanavalin A (Con A) [Khan, M. I., Mandal, D. K., & Brewer, C. F. (1991) Carbohydr. Res. 213, 69-77]. Using radiolabeling and quantitative precipitation techniques, we show in the present study that Con A binds and forms unique cross-linked complexes with four different glycoproteins having different numbers and types of carbohydrate chains as well as different quaternary structures. The glycoproteins include quail ovalbumin, Lotus tetragonolobus isolectin A (LTL-A), Erythrina cristagalli lectin (ECL), and Erythrina corallodendron lectin (EcorL). The results show that a preparation of quail ovalbumin containing either one Man7 or Man8 oligomannose chain per molecule forms a 1:2 cross-linked complex with tetrameric Con A, thereby demonstrating bivalency of the single carbohydrate chain(s) on the glycoprotein. Tetrameric LTL-A and dimeric ECL, which possess two xylose-containing carbohydrate chains per monomer, both form 1:2 and 1:1 cross-linked complexes (per monomer) of glycoprotein to lectin, depending on their relative ratios in solution. However, dimeric EcorL, which has the same carbohydrate structure and number of chains as ECL, forms only a 1:2 cross-linked complex with tetrameric Con A.(ABSTRACT TRUNCATED AT 250 WORDS)