Temporal changes in circulating levels of plasma interleukin-8 during peripartum period in Murrah buffaloes (Bubalus bubalis) under subtropical climate

The objective of this study was to elucidate the changes in circulating levels of plasma interleukin-8 (IL-8) during peripartum period in Murrah buffaloes (Bubalus bubalis). IL-8 was estimated in blood plasma of healthy peripartum Murrah buffaloes (n = 6) on days ±30, ±15, ±5, ±3, ±1 and 0 pre- and postpartum with respect to the day of parturition (day 0) in each of the two different seasons (hot–humid and spring). The mean microclimate Temperature–Humidity Index (THI) during spring season was significantly lower (p < 0.01) than the corresponding THI in hot–humid season. In both the seasons, plasma IL-8 remained lower in prepartum period (≤46.56 ± 14.08 pg/ml during spring and ≤ 73.18 ± 18.56 pg/ml during hot–humid season) than in the postpartum period (≥51.41 ± 13.82 pg/ml during spring and ≥ 84.13 ± 16.97 pg/ml in hot humid season). During spring, the IL-8 levels were significantly higher (P < 0.05) on days +5 and +15 postpartum in comparison to the IL-8 levels on days −30, −5, and −3 prepartum. During hot–humid season, IL-8 level was significantly higher (P < 0.05) on day +30 as compared to the IL-8 levels on days −30 and −5 prepartum. The correlation between IL-8 and mean microclimate THI was significant (r = 0.25, P < 0.01). From the results, it is concluded that peripartum period in buffaloes is associated with an inflammatory response leading to significantly higher plasma IL-8 during parturition and postpartum period than in the pre-partum period.     

Ribotyping of Indian Isolates of <Emphasis Type="Italic">Pasteurella multocida</Emphasis> Based on 16S and 23S rRNA Genes

The applicability of ribotyping based on 16S and 23S rRNA was evaluated for molecular epidemiological studies. Forty-eight isolates of Pasteurella multocida isolated from different hosts and geographical locations and one reference isolate were ribotyped. Only four ribotypes were found. All the isolates including reference isolate from wild carnivores had the same ribotype, though they had different serotypes. The isolate from a tiger had one band in addition to the bands present in the major ribotype. The isolates from lions represented two ribotypes; of these ribotypes, one (r2) had an additional band of 3.6 kbp, which was absent in all other ribotypes. The second ribotype (r4) from a lion had one band missing (6 kbp) that was present in the other ribotypes. These isolates were further typed using ERIC-PCR and REP-PCR. With ERIC-PCR and REP-PCR, higher D values of 0.83 and 0.89 were obtained. The current study revealed that ribotyping is not a very efficient typing tool for use in molecular epidemiology for differentiation of isolates.     

Organic zinc and copper supplementation on antioxidant protective mechanism and their correlation with sperm functional characteristics in goats

Trace minerals feeding had significant effects on sperm production and fertility with better absorption and proper utilization within the body for optimum reproductive function. Several studies have shown that more influenced trace elements in the diets of animals are copper (Cu) and zinc (Zn). Bucks showing deficiency of this mineral might affect the quality of semen production which in turn would affect the fertility. This experiment was thus designed to test the effects of organic Cu and Zn supplementation on antioxidants enzyme activities and sperm functional attributes in fresh semen of bucks. Forty bucks (n = 40, Aged 5 months) were assigned to ten groups of four animals in each group, supplemented (for a period of 8 months) with different levels of organic Zn: 20 mg (T2), 40 mg (T3) and 60 mg (T4), organic Cu: 12.5 mg (T5), 25 mg (T6), 37.5 mg (T7) and combined organic Zn and Cu: 20 + 12.5 mg (T8), 40 + 25 mg (T9), 60 + 37.5 mg (T10), respectively, per kg dry matter and no additional mineral diet (control; T1). One hundred and sixty semen samples were collected through electro‐ejaculator and analysed for sperm quantity, quality, acrosome intactness and plasma membrane integrity and correlated with the catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase enzyme activities in seminal plasma. The results indicated organic Cu and zinc supplemented bucks produced more sperm cells, had higher sperm concentrations, maintained higher (p < .01) sperm livability, plasma membrane and acrosome integrities, more motility and velocity. The increased antioxidant enzyme activities, reduced oxidative stress and lowered lipid peroxidation were positively correlated (p < .05) with the sperm functional attributes. In conclusion, organic Cu and Zn supplement to male goats showed protective roles against oxidative damage and maintained better fresh semen characteristics.     

Identification of immunoreactive polypeptides of Babesia equi parasite during immunization

This study was carried out to identify immunoreactive polypeptides in Babesia equi merozoite antigen. Three fractions of killed B. equi merozoite antigen viz.; whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS) antigens were prepared from the parasite infected erythrocytes. These antigenic preparations along with ghost antigen from non-infected erythrocytes were fractionated on 12% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with sera showing high antibody titres. On SDS-PAGE, 16 polypeptides with molecular weight (Mr) in the range of 112-17kDa were obtained from the WM and CM antigens. But only six polypeptides were detected (96.5-28kDa) in the HSS antigen. On immunoblotting with high titred serum collected from donkeys following two immunizations with a killed B. equi merozoite immunogen, 11 polypeptides were observed in the WM and CM antigens (Mr 112-18kDa). Of these, four polypeptides (Mr 112, 45, 33 and 18kDa) were identified as most immunoreactive. Besides these, a 28kDa was observed as strong immunoreactive protein in WM and CM antigens. The HSS antigen showed only six polypeptides and one peptide (28kDa) was identified as immunoreactive. When high titred serum collected from immunized donkeys following challenge with B. equi infected blood and was used for immunoblotting, the protein profile of WM and CM antigens remained the same. However, three additional polypeptides (Mr 81, 54.5 and 39kDa) were detected in HSS antigen.     

Sonographic characteristics of goat testis on water bath based ultrasonography

Assessing the health of the testes in domestic animals is an important aspect of the breeding soundness examination and selection. The aim of the present study was to develop a simple method for scanning and to establish ultrasonographically the gross anatomic structures of the goat testes. Six adult male goats were examined to study the sonographic appearance of normal testes and epididymides using a water bath based ultrasound scanning technique. The ultrasonographic examinations were done using a 5–9 MHz/60 mm (7.5 MHz) linear-array transducer and a B-mode scanner. The ultrasonographic examination was performed in goats after standardizing the procedure on six testes collected from slaughter house. Results showed that in live goats when the probe was placed directly over the scrotum it gave distorted and unclear image. In water bath method the entire scrotum was dipped into a container filled with water and linear probe was used to observe the sonographic features of the testis. Each testis was viewed vertically, resulting in longitudinal image which was frozen, measured and printed through a thermal printer. The results of the ultrasonogram revealed that the testicular parenchyma was homogenous and moderately echogenic throughout. The diameter (mean±se) of the right and left testes was 4.47±0.14 and 4.42±0.07 cm respectively and no significant difference was observed between the testes. The mediastinum testis was a 1.50±0.22 cm wide linear structure of greater echogenicity than the testicular parenchyma when viewed in the transverse plane and nearly circular echogenic “spot” in the midline of the testis when viewed horizontally. The head and tail of the epididymides were easily identified on all the testes, but the epididymal body and ductus deferens were difficult to identify consistently. The tail of the epididymis was easily identified on the distal end of the testis with sonolucent tubules and appeared sonographically as a ‘peaked cap’ upon the testicular parenchyma. The diameter (mean±se) of the tail of right and left epididymis was 2.11±0.18 and 1.92±0.06 cm and no significant difference was observed between epididymides. The vascular pampiniform plexus (1.42±0.18 cm) was easily identified on the proximal end of the testes. The tunics of the testes appeared as a bright echogenic line. Inter-testicular septum appeared between testes as a hyperechoic line. It is concluded that ultrasonography permits a noninvasive evaluation of the internal structure of the scrotum and testes and water bath based sonographic examination may prove to be a valuable simple diagnostic methodology for evaluating physiopathologic conditions of goat testes and can be employed as a routine investigative method during breeding soundness and clinical examination.     

Effect of Supplementation of Taurine or Trehalose in Extender on Immunolocalization of Tyrosine Phosphoproteins in Buffalo and Cattle (Karan Fries) Cryopreserved Spermatozoa

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post‐thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris‐citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm ) or trehalose (100 mm ), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT‐154 (anti‐phosphotyrosine antibody) and FITC‐conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post‐thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing–thawing process.     

Larval and pupal toxicity effects of Plectranthus amboinicus, Bacillus sphaericus and predatory copepods for the control of the dengue vector, Aedes aegypti

The efficacy of copepod Mesocyclops aspericornis (Daday) combined with the larvicide Bacillus sphaericus (Bs) and a plant extract of Plectranthus amboinicus leaf abstract (PALE), used jointly and singly, was studied against Aedes aegypti in the laboratory. P. amboinicus leaf extract of 20, 40, 60, 80 and 100 ppm caused significant mortality against Ae. aegypti larvae. The LC₅₀ and LC₉₀ values for I to IV instars larvae and pupae were 26.12, 35.36, 45.76, 52.32 and 63.82 ppm, respectively. The LC₉₀ values for I to IV instars larvae and pupae were 82.53, 92.65, 108.06, 119.47 and 131.71 ppm, respectively. Under laboratory conditions, copepods treatment produced 7.9% predatory efficiency against 1st instar larvae of Ae. aegypti, at a copepod:larvae ratio of 1:10. When copepod treatment was combined with PALE this was increased to 8.7. The treatment of copepods combined with Bs and PALE yielded a better and more sustainable result (9.6%) than the agents used individually. This predation efficiency may be caused by detrimental effects of the P. amboinicus active principle compound (carvacrol) on the mosquito larvae. Our results suggest that the combined application of microbial insecticide (Bs), copepods and P. amboinicus leaf extract may be used to control Aedes populations.     

Rhizobacteria‐mediated resistance against the blackeye cowpea mosaic strain of bean common mosaic virus in cowpea (Vigna unguiculata)

BACKGROUND: The present study investigated the effect of seven Bacillus‐species plant‐growth‐promoting rhizobacteria (PGPR) seed treatments on the induction of disease resistance in cowpea against mosaic disease caused by the blackeye cowpea mosaic strain of bean common mosaic virus (BCMV). RESULTS: Initially, although all PGPR strains recorded significant enhancement of seed germination and seedling vigour, GBO3 and T4 strains were very promising. In general, all strains gave reduced BCMV incidence compared with the non‐bacterised control, both under screen‐house and under field conditions. Cowpea seeds treated with Bacillus pumilus (T4) and Bacillus subtilis (GBO3) strains offered protection of 42 and 41% against BCMV under screen‐house conditions. Under field conditions, strain GBO3 offered 34% protection against BCMV. The protection offered by PGPR strains against BCMV was evaluated by indirect enzyme‐linked immunosorbent assay (ELISA), with lowest immunoreactive values recorded in cowpea seeds treated with strains GBO3 and T4 in comparison with the non‐bacterised control. In addition, it was observed that strain combination worked better in inducing resistance than individual strains. Cowpea seeds treated with a combination of strains GBO3 + T4 registered the highest protection against BCMV. CONCLUSION: PGPR strains were effective in protecting cowpea plants against BCMV under both screen‐house and field conditions by inducing resistance against the virus. Thus, it is proposed that PGPR strains, particularly GBO3, could be potential inducers against BCMV and growth enhancers in cowpea. Copyright © 2009 Society of Chemical Industry     

Studies on immunocompetence status in two turkey varieties in India

(1) Two hundred and twenty-seven adult turkeys of both sexes, of two varieties (104 Black and 123 White) were used to evaluate their immunocompetence status and body weights. (2) Response to sheep red blood cells (SRBC) (humoral immunity) was measured by Haemagglutination (HA) test 5 days post immunisation (dpi) and expressed as log2 values. Mercaptoethanol resistant (MER) antibodies representing IgG were determined by Mercaptoethanol HA test and Mercaptoethanol sensitive (MES) antibodies, representing IgM as the difference in total HA titre and IgG. Serum lysozyme concentrations were estimated by 'Lysoplate assay' and expressed in log2 values. (3) Least squares analysis of variance revealed that the White variety had higher adult body weight (4.788 +/- 0.040 kg) than the Black (3.774 +/- 0.044 kg). Sexual dimorphism was apparent and meals were heavier than females in both varieties. The interaction effect of variety and sex on body weight was also significant. (4) Least squares means for immunological traits, namely, total anti-SRBC antibodies, MER, MES titres and serum lysozyme were 7.161 +/- 0.189, 0.801 +/- 0.071, 6.362 +/- 0.160 and 1.766 +/- 0.043 microg/ml, respectively. The Black variety had a higher MES antibody titre than the White. (5) Sex had an effect on all the immunological traits except on MER titres. Females generally had higher anti-SRBC, MER and MES titres and serum lysozyme. The variety x sex interaction effect was significant for MES titres and serum lysozyme. White males had the lowest MES titres.