Detection of chromogranin A in the adrenal gland extracts of different animal species by an enzyme-linked immunosorbent assay using Thomsen-Friedenreich antigen-specific Amaranthus caudatus lectin

The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen-Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.     

Analyses of factors affecting dry matter intake of lactating dairy cows

An experiment was conducted to analyze feed, climate and animal factors affecting dry matter intake (DMI) in lactating dairy cows. Sixteen lactating Holstein cows, with parity from 1 to 6, were assigned to a feeding trial for 2 years, comprising 31 lactations. The animals were fed Italian ryegrass silage, oat hay, alfalfa hay, beet pulp and three types of concentrate. The data, pooled and classified by stage of lactation, season of lactation and parity were analyzed by stepwise multiple regression to determine the nature and extent of factors affecting DMI. A total of 45 prediction equations for DMI were derived. Energy‐corrected milk yield or milk yield was selected as the primary factor of DMI in all the equations in which the ratio of contribution (R²) varied from 0.26 to 0.67. The dietary concentration of organic cell wall, crude fiber, crude protein, organic b fraction, forage to concentrate ratio, average ambient temperature and temperature–humidity index were selected as the secondary factors affecting DMI for pooled data, late lactation (251–350 days of lactation), summer (June–August), spring (March–May), ≥4th lactation, autumn (September–November) and 3rd lactation, respectively, and improved R² up to 0.77. Except for an impact of bodyweight in several equations, feed and climatic factors significantly improved prediction equations effectively for data classified in different ways. To estimate DMI accurately in lactating dairy cows, feed and climatic factors should be considered for specific conditions.     

Serosurvey of five viruses in chickens on smallholdings in Bangladesh

A serologic survey was undertaken in chickens in smallholdings in Bangladesh for avian influenza A virus (AIV), egg drop syndrome '76 virus (EDS'76V), infectious bronchitis virus (IBV), Newcastle disease virus (NDV) and reovirus (RV) in three phases: January 2002-May 2003, September 2003-August 2004, and August 2005-March 2006. Four hundred thirty-six sera collected in the 2nd phase, 295 in the first phase, 755 in the 1st plus 2nd phases and 295 in the 1st phase were investigated for AIV, EDS'76V, IBV and RV, respectively, using enzyme linked immunosorbent assays. All 854 sera collected in the three phases were screened for NDV using hemagglutination inhibition test. In chickens 20% were seropositive to AIV, 3% to EDS'76V, 74% to IBV, 88% to NDV, and 47% to RV. The seroprevalence in flocks was 23% to AIV, 6% to EDS'76V, 79% to IBV, 89% to NDV and 56% to RV. Twenty-five percent chickens had > or = 10log(2)HI titers to NDV.     

Investigation on association and expression of ESR2 as a candidate gene for boar sperm quality and fertility

ESR2 is involved in oestrogen-related apoptosis in cell cycle spermatogenesis but their effects have not yet confirmed in pig. Therefore, this study was aimed to investigate the association of ESR2 polymorphism with sperm quality and boar fertility traits and to analyse the ESR2 mRNA and protein expressions in boar reproductive tissues. DNA samples from 203 Pietrain (PI) and 100 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility (MOT), semen volume (VOL), plasma droplet rate (PDR) and abnormal spermatozoa rate (ASR)] and fertility [non-return rate (NRR) and number of piglet born alive (NBA)] traits were available. A SNP in coding region of ESR2 g.35547A>G in exon 5 was associated with MOT and PDR in the PI and with SCON, VOL, MOT and PDR in PIHA population. For mRNA and protein expression study, a total of six boars were divided into two groups with group I (G-I) and group II (G-II) where G-I characterized for relatively a better sperm quality according to the mean of two groups. mRNA expression was higher in brain and testis than that in all parts of epididymis. Both qRT-PCR and western blot analysis revealed that the ESR2 gene expression and protein expression were significantly higher in testis collected from G-II compared with that of G-I boars. Moreover, ESR2 protein localization in germ cell, Leydig and Sertoli cells, epithelial cells and spermatozoa was remarkable, which indicated the important role of ESR2 in spermatogenesis process. These results might shed new light on the roles of ESR2 in spermatogenesis as candidate for boar fertility, but still the lack of association across populations should be considered.     

Survivability and causes of loss of broody-hen chicks on smallholder households in Bangladesh

We determined the flock sizes and rates of loss caused by different factors in broody-hen chicks (BHC) up to 60 days of age on 600 randomly selected smallholdings in Bangladesh. The smallholders were beneficiaries of a village poultry production chain called ‘Smallholder Livestock Development Project-2’ (SLDP-2) which was undertaken with the financial assistance of the Danish International Development Agency (DANIDA). For estimating survival time of BHC, we observed chicks in 80 smallholdings. SLDP-2 aims at ameliorating poverty among women by poultry rearing at village level; in total, 104,000 key rearers, constituting 96% of all of the beneficiaries of the SLDP-2 area, were enrolled in 26 upazilas (a lower administrative unit of Bangladesh). A key rearer is a smallholder who rears at least five ‘Sonali’ (RIR × Fayoumi) and some indigenous (desi) chickens in a semi-scavenging system. Sonali chickens are supplied from the development project, and have higher egg production while the broodiness of the desi hens is exploited to get chicks hatched for future stocks; thus, the chicks hatched and reared to 60 days old at key rearers’ households are called BHC. In this study 32% of the smallholders had BHC each month. At the beginning of a month, the median number of chicks in a flock was 8, and the mean survival time was 50.5 days. Incidence rates of loss of BHC from disease, predation, selling and slaughtering were 0.102, 0.086, 0.009 and 0.002 per chick-month at risk, respectively. The major predators were crows, mongooses and eagles with incidence rates of loss being 0.018, 0.016 and 0.010 per chick-month at risk, respectively. Colibacillosis (both single and mixed infections) contributed to the death of 21% of dead BHC collected; Newcastle disease and salmonellosis contributed to the next highest (14 and 12%) proportional mortalities.     

Regional Specialisation of the Ganglion Cell Density in the Retina of the Native Duck (Anas platyrhynchos) of Bangladesh

In this study, retinal whole‐mount specimens were prepared and stained with 0.1% cresyl violet for the ganglion cell study in the native duck (Anas platyrhynchos). The total number, distribution and size of these cells were determined in different retinal regions. The mean total number of ganglion cells was 1 598 501. The retinal area centralis had the highest ganglion cell density with 11 200 cells/mm². Number of ganglion cell bodies was the highest in temporal area, followed by dorsal, nasal and ventral areas. Ganglion cell size ranged from 5.25 to 80 μm². In the temporal and nasal region, most of the cells were ranged from 15 to 25 μm², and in the dorsal and ventral region, most of the cells were ranged from 12 to 25 μm². There was a marked trend for the retinal ganglion cell size to increase as the population density decrease towards the periphery. A population of small ganglion cells persisted into the central area just above the optic disc and the largest soma area was in the ventral zone of the retina. Thus, the specialisation of ganglion cell densities and their sizes support the notion that the conduction of visual information towards the brain from all regions of the retina is not uniform, and the central area is the fine quality area for vision in native duck.     

Efficacy of strain RB51 vaccine in protecting infection and vertical transmission against Brucella abortus in Sprague-Dawley rats

Immunizing animals in the wild against Brucella (B.) abortus is essential to control bovine brucellosis because cattle can get the disease through close contact with infected wildlife. The aim of this experiment was to evaluate the effectiveness of the B. abortus strain RB51 vaccine in protecting infection as well as vertical transmission in Sprague-Dawley (SD) rats against B. abortus biotype 1. Virgin female SD rats (n = 48) two months of age were divided into two groups: one group (n = 24) received RB51 vaccine intraperitoneally with 3 × 10¹⁰ colony forming units (CFU) and the other group (n = 24) was used as non-vaccinated control. Non-vaccinated and RB51-vaccinated rats were challenged with 1.5 × 10⁹ CFU of virulent B. abortus biotype 1 six weeks after vaccination. Three weeks after challenge, all rats were bred. Verification of RB51-vaccine induced protection in SD rats was determined by bacteriological, serological and molecular screening of maternal and fetal tissues at necropsy. The RB51 vaccine elicited 81.25% protection in SD rats against infection with B. abortus biotype 1. Offspring from rats vaccinated with RB51 had a decreased (p < 0.05) prevalence of vertical transmission of B. abortus biotype 1 compared to the offspring from non-vaccinated rats (20.23% and 87.50%, respectively). This is the first report of RB51 vaccination efficacy against the vertical transmission of B. abortus in the SD rat model.