Molecular identification of blast resistance genes in rice landraces from northeastern India

Rice blast disease caused by the fungus Magnaporthe oryzae is one of the most devastating diseases causing huge losses worldwide. In the present study, major blast resistance genes were investigated in landraces originating from northeastern India. Based on phenotypic evaluation, 288 landraces were classified into three distinct groups: resistant (75), moderately resistant (127) and susceptible (86). The genetic frequencies of the 18 major blast resistance genes were between 6.2% and 27.4%, with only two genotypes possessing a maximum of nine blast resistance genes. The cluster and population structure analysis grouped the landraces into two groups. Through principal coordinate analysis, the scatter plots partitioned the resistant and moderately resistant landraces into different groups. Analysis of molecular variance showed maximum (96%) diversity within populations and least (4%) diversity between populations. Association analysis identified six markers, CRG4_2, RM72, tk59-2, pi21_79-3, RM1233 and RM6648, that are significantly associated with blast disease and explained a phenotypic variance of 1.1–6.5%. The associated genes could be used in marker-assisted rice breeding programmes for gene pyramiding to develop rice varietal resistance against blast disease. The present study represents a valuable blast resistance genetic resource that could be used for identification of new R genes, donors for blast resistance, and genomic studies.     

Frequent human‐poultry interactions and low prevalence of Salmonella in backyard chicken flocks in Massachusetts

The backyard chicken (BYC) movement in the USA has increased human contact with poultry and subsequently, human contact with the pathogen Salmonella. However, to date, there have been few studies assessing prevalence of Salmonella in backyard flocks, despite the known public health risk this zoonotic bacterium poses. The objective of this study was to characterize human‐BYC interactions and assess the prevalence of Salmonella among BYC flocks. We interviewed 50 BYC owners using a structured questionnaire to determine flock and household characteristics that facilitate contact with BYC and that may be associated with Salmonella in the BYC environment. Composite faecal material, cloacal swabs and dust samples from 53 flocks housed on 50 residential properties in the Greater Boston, Massachusetts area were tested for Salmonella using standard culture techniques and confirmed using Matrix‐Assisted Laser Desorption/Ionization‐Time of Flight Mass Spectrometer. Microbroth dilution and whole genome sequencing were used to determine phenotypic and genotypic resistance profiles, respectively, and sequence results were used to determine multilocus sequencing type. No owners self‐reported a diagnosis of salmonellosis in the household. Over 75% of a subset of owners reported that they and their children consider BYC pets. This perception is evident in how owners reported interacting with their birds. Salmonella enterica subspecies enterica serotype Kentucky ST152 (serogroup C)—a strain not commonly associated with human infection—was confirmed in one flock, or 2% of tested flocks, and demonstrated resistance to tetracycline and streptomycin. We detected Salmonella at low prevalence in BYC. Further study of the health effects of exposure to zoonotic gastrointestinal pathogens such as Salmonella among families with BYC is warranted.     

Expression of cytokines and respiratory burst activity of milk cells in response to Azadirachta indica during bovine mastitis

The immuno therapeutic potential of hydro-methanolic extract of Azadirachta indica (A. indica) was studied during bovine clinical mastitis (CM). The somatic cell count (SCC), total bacterial count (TBC), milk differential leukocyte count (DLC), hydrogen peroxide (H₂O₂), superoxide anion (O₂ ⁻) production and interleukin- 2 (IL-2) and gamma interferon (IFN-γ) cytokines expression were studied before and after intramammary infusion of A. indica extract in diseased cows. The results revealed that A. indica treatment significantly (P < 0.05) decreased the SCC, TBC, milk neutrophil percent and significantly (P < 0.05) enhanced milk lymphocyte percent, H₂O₂ and O₂ ⁻ production by milk cells. The IL-2 and IFN-γ were expressed in normal healthy cows and diseased cows after A. indica treatment, whereas both the cytokines could not be expressed in cows treated with antibiotic and in untreated diseased cows. The results of the present study indicated anti inflammatory, antibacterial and immunomodulatory potential of the herb, these activities could be due to the presence of bioactive principle in the extract. This is a preliminary trial indicated beneficial effect of the herb against bovine mastitis it can be developed as an alternative therapy where the use of antibiotics is normally restricted.     

Acclimation Induced Responses of SDH Activity of Tilapia (Oreochromis mossambicus) Following Introduction in a New Pond Habitat

Responses of the major mitochondrial respiratory enzyme succinate dehydrogenase (SDH) activity in gill and muscle tissue of tilapia (Oreochromis mossambicus) were investigated upon their transfer to a new pond habitat. Sixty healthy adult fish were procured from each of the three ponds used and were distributed in three cages in equal numbers; the cages were placed in each of the other two ponds as well as into original rearing pond. Four fish were removed from each cage every week and were sacrificed for the assay of SDH activity in gills and muscles. Transfer of fish from a stress condition to a benign environment and vice versa resulted in an increase and decrease in SDH activity of fish, respectively. The intensity of increase upon transfer was again dependent upon the amount of stress in the new habitat. In general, the SDH activity of the test fish strongly correlated with the ammonia concentration (P?<?0.05) of water regardless of habitats. Time required for the introduced fish to acclimatize with new environmental conditions was dependent upon the environmental milieu of both its initial and final place of rearing and hence their physiological states. Differences in water quality, especially ammonia concentration, between its original place of rearing and the place of transfer were found to be responsible for the differences in acclimation period of two or three weeks. Relatively less difference in ammonia (2.68 for gills and 3.20 for muscles) between the initial place of rearing and new habitat resulted in acclimation of the fish one week earlier (second week) than the relatively wide difference (4.46 for gills and 5.62 for muscles) for third week. It may be reasonable to conclude that the varied responses of the mitochondrial respiratory enzyme of exogenously introduced fishes with the differences in the water quality—especially the ammonia concentration of ambient water between the original fish holding pond and growout pond—can be used to predict the time that will be required for the exogenously introduced fish to fully acclimatize with the new habitat.     

Latent heat flux estimation in clear sky days over Indian agroecosystems using noontime satellite remote sensing data

A simplified evaporative fraction (Λ) based single-source energy balance scheme was tested with moderate resolution (1 km) noontime satellite observations to evaluate clear sky latent heat flux (λE) estimates over diverse agricultural landscapes. This approach uses two-dimensional (2D) scatter between land surface temperature (LST) and albedo to determine Λ. The operational utility of this scheme was demonstrated for estimating regional evapotranspiration and consumptive water use during rabi (November to April) crop growing season to predict pre-harvest wheat yield (error within 15.9% of reported mean) using time series data. The existence of triangular relations between Λ and LAI (leaf area index) or NDVI (normalized difference vegetation index) was found with basal line (hypotenuse) linearly coupled with LAI or NDVI at low level of surface soil wetness. The analysis of diurnal course of in situ Λ proved the validity of constant-Λ hypothesis over pure, uniform, homogeneous crop canopies but showed irregular and wave-like patterns over heterogeneous, mixed crop canopies. The root mean square error (RMSE) of noontime and daytime average λE estimates with respect to in situ λE measurements were also smaller over homogeneous agricultural canopies (41 and 23 W m⁻²) with correlation coefficients (r) 0.94 and 0.96, respectively, from 135 clear sky datasets as compared to RMSE over heterogeneous ones (59 and 28 W m⁻² with r = 0.66 and 0.82, respectively from 22 datasets). The intercomparison with another Λ based approach (LST–NDVI 2D scatter) showed the supremacy of Λ determined from LST–albedo 2D scatter. The efficiency of LST–NDVI scatter was better during the dry down or water limited phases of crop growth only. The uncertainties of λE estimates were attributed to errors in core radiation budget inputs, relative loss of conservativeness of Λ due to canopy heterogeneity, and the inherent limitations of the single-source approach. There is further scope to reduce present λE uncertainties by combining the new findings on Λ (LST–albedo scatter)–NDVI triangular relations, diurnal Λ and two-source radiation budget.     

Sequence and evolutionary analysis of ribosomal DNA from Huanglongbing (HLB) isolates of Western India

Citrus greening (Huanglongbing, HLB) is a widespread and economically important citrus disease all over the world. The disease is caused by a phloem-limited fastidious gram negative bacterium, “Candidatus Liberibacter spp.” which belongs to the alpha-proteobacteria group classified on the basis of its 16SrDNA sequence. Although the pathogen has been classified under three distinct groups, viz. Asian, African and American isolates, nothing is known about the status and the molecular variabilities among the Indian HLB isolates collected from different citrus cultivars grown in India. Five different HLB isolates showing variable symptoms based on their severity of infection on different citrus, viz. Mosambi, Rangpur lime, Cleopatra mandarin, acid lime and rough lemon, were studied by PCR amplification, sequence and evolutionary analysis of their 16S and 16S/23S rDNA intergenic regions. Analysis of the 16S/23S rDNA intergenic region separated all five Indian isolates from existing African isolates but failed to differentiate among Asian, American and Indian isolates. However, further analysis of complete 16S rDNA clearly indicated that Indian isolates fall within the Asian HLB group. Overall, our results suggest that all the five Indian HLB isolates taken for the current analysis belong to the Candidatus Liberibacter asiaticus strain, which showed distinct sequence variabilities and produced noticeable symptoms on the citrus trees. These results provide a robust framework for understanding how differences in pathogenicity among various HLB isolates may be related to evolutionary history.     

Effect of supplementation of vitamin E,copper and zinc on the in vitro phagocytic activity and lymphocyte proliferation index of peripartum Sahiwal (Bos indicus) cows

To study the effect of vitamin E (VE), copper (Cu) and zinc (Zn) supplementation on the in vitro phagocytic activity (PA) and lymphocyte proliferation response (LPR) of blood neutrophils and lymphocytes, thirty Sahiwal pregnant cows (six in each group) in their late gestation at 30 days before the expected date of calving were selected from the NDRI experimental herd and supplemented with various micronutrients from 30 days before calving to 45 days after calving. Cows were supplemented individually with VE (1000 IU/cow/day), Cu (20 ppm/cow/day) and Zn (80 ppm/cow/day) and also with a combination of VE, Cu and Zn to study cumulative effect of all micronutrients. One group without any supplementation acted as a control. Blood neutrophils and lymphocytes were isolated and studied for their PA and LPR. Supplementation of micronutrients like VE, Cu, Zn and a combination of all these nutrients significantly (p < 0.01) increased the PA of experimental cows as compared to control (unsupplemented) cows during the pre‐partum period. During post‐partum, all the micronutrients (VE, Cu, Zn and their combination) showed a significant (p < 0.01) increase in the PA of experimental cows as compared to control cows. Of all the groups, significant (p < 0.01) and maximum PA was observed in the combination group followed by Zn‐supplemented group during both the pre‐ and post‐partum period. A significant (p < 0.01) increase in LPR of B lymphocytes was observed in combination‐supplemented group during the pre‐partum period and during both the pre‐ and post‐partum period in the Cu‐supplemented group.     

Assessment of DNA Damage during In Vitro Development of Buffalo (Bubalus bubalis) Embryos: Effect of Cysteamine

Comet assay was used in the present study to examine DNA damage to buffalo oocytes and embryos during in vitro culture. Embryos were produced in vitro from oocytes obtained from slaughterhouse ovaries in presence of cysteamine (IVM and IVC media supplemented with 50 and 100 μm , respectively) or in its absence (controls). Compared to controls, cysteamine supplementation increased (p < 0.01) cleavage rate and proportion of oocytes that developed to 8‐ to 16‐cell stage. The incidence of DNA damage was lower (p < 0.01) in cysteamine group than that in controls at 8‐ to 16‐ (19.3 ± 4.24 vs 72.0 ± 5.22%) but not in 2‐cell stage embryos (11.7 ± 5.63 vs 20.8 ± 5.49%) or in mature oocytes (5.3 ± 3.43 vs 10.3 ± 4.73%). The tail length, which indicates magnitude of DNA damage, was shorter (p < 0.01) in cysteamine group than in controls in mature oocytes (25.5 ± 0.5 vs 36.0 ± 0.71 pixels) and 8‐ to 16‐cell stage (49.2 ± 1.64 vs 152.7 ± 1.28 pixels) but not in 2‐cell stage embryos (36.3 ± 1.54 vs 36.4 ± 0.75 pixels). Also, exposure of oocytes/embryos to UV radiation or H₂O₂ caused extensive DNA damage. In conclusion, these results suggest that oocytes/embryos suffer from DNA damage during progress of in vitro culture, which can be partly ameliorated by cysteamine supplementation of culture media.     

Antibacterial spectrum of Hypericum hookerianum.

The antibacterial activity of the chloroform, acetone and methanol extracts of Hypericum hookerianum leaves and stems was evaluated. All extracts showed antibacterial activity against six different gram-positive and gram-negative bacteria, the methanol extracts exhibiting the maximum inhibitory activity at 400 microg/ml.     

In vitro phagocytic activity of milk neutrophils during lactation cycle in Murrah buffaloes of different parity

Milk samples were collected from 34 lactating Murrah buffaloes on days 0, 5, 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, 270 and day 300 after calving. Milk somatic cell counts (SCC) were highest in multiparous buffaloes . Milk SCC were significantly lower in buffaloes of third and fourth parity during early lactation and than increased significantly (p < 0.01) by the end of lactation. Milk neutrophils were significantly lower in all the buffaloes during early lactation, but increased significantly (p < 0.01) afterwards. Milk lymphocytes were significantly higher during early lactation, but decreased significantly (p < 0.01) by the end of lactation. Phagocytic activity (PA) was highest in day 1 colostrum and then decreased significantly (p < 0.01) by the fourth milking in buffaloes of second, third and fourth parity . Phagocytic index (PI) was also highest in colostrums of primiparous buffaloes. Irrespective of parity, maximum PA and PI was observed during mid lactation. In terms of in vitro phagocytic activity, early lactation is the most critical period followed by late and mid lactation.