Preliminary investigations into the interactions of herbicides with aqueous humic substances

The binding of the herbicides to aqueous humic substances was investigated using the herbicide mecoprop and a reference humic substance ‘Hohlohsee’. Initial results from 2D fluorescence spectrometry, infra-red spectroscopy and Curie-point pyrolysis gas chromatography/infra-red spectroscopy tentatively characterise the nature of the interaction. ©1997 SCI     

Assessing the distribution pattern of otters in four rivers of the Indian Himalayan biodiversity hotspot

1. The Eurasian otter (Lutra lutra), smooth‐coated otter (Lutrogale perspicillata), and Asian small‐clawed otter (Aonyx cinereus) have all been reported previously from the Indian state of Uttarakhand. However, little information is available about their current distribution in a mountainous region that is subject to increasing human‐induced stressors (such as hydropower plants, pollution, sand and boulder mining, destructive fishing techniques, poaching).
2. Owing to important roles played by these otters in structuring riverine food webs (particularly taking account of their roles as top carnivores), it is critical that they receive suitable protection in the face of projected temperature rise, change in precipitation patterns, and associated river flows in this Himalayan biodiversity hotspot. This study assesses otter distribution in four rivers of Uttarakhand as a basis for informing future conservation actions.
3. Field surveys were conducted (October 2018–January 2019) in reaches of the Kosi, Ramganga, Khoh, and Song rivers, supported by semi‐structured interviews (N = 379) conducted with members of local communities to collect qualitative data on views and perceptions of otter species. In addition, community‐based otter awareness camps were organized for local youths (N = 105), adults (N = 115), and schoolchildren (N = 256 covering 10 schools).
4. Habitat suitability maps were created using remote‐sensing data, survey findings, and a geographic information system to provide information about priority reaches of river to be targeted for future conservation efforts.
5. This study provides critical interdisciplinary baseline information to guide decision‐makers towards developing a targeted, otter‐specific conservation programme for this important Himalayan biodiversity hotspot.
6. The otter conservation education programmes conducted during this study resulted in a proposal to set up a community‐based conservation initiative to monitor and report otter sightings from the area, potentially representing a way forward for achieving simultaneous otter conservation and associated ecosystem benefits for local communities.

     

Analysis of viral load between different tissues and rate of progression of white spot syndrome virus (WSSV) in Penaeus monodon

At present the most common and most devastating disease of shrimp is caused by the white spot syndrome virus (WSSV), which has spread throughout the world mainly by different species of crustaceans carrying the virus. After experimental injection of Penaeus monodon with a known copy number of WSSV in the abdominal muscle, the rate of viral progression in different tissues at 12, 24, 36 and 48 hpi (hours post infection) was assessed using quantitative real‐time PCR. At 12 hpi the viral load was highest in haemocytes followed by pleopod, muscle and gills whereas at 48 hpi, the gills, the main target of WSSV, showed the highest viral load followed by pleopod, muscle and haemocytes. Viral copy number in the haemocytes was the lowest beyond 12 hpi indicating a remarkable reduction in the rate of viral replication in haemocytes compared with other tissues. The viral load in haemocytes, though increased again beyond 36 hpi, never surpassed the load in the other tissues. The real‐time PCR assay with its high sensitivity and wide dynamic range make it ideal for detecting low‐level WSSV infections that can occur in apparently healthy P. monodon.     

Hepatotoxicity associated with CCNU (lomustine) chemotherapy in dogs

One hundred seventy-nine tumor-bearing dogs were treated with 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) between 1995 and 2001. CCNU was given as a single dose of 50-110 mg/m2 body surface area PO. Treatment interval varied, but the minimal interval between CCNU doses was 3 weeks. After treatment, 11 dogs (6.1%) developed hepatic toxicity. The median number of CCNU doses and the median total cumulative CCNU dose were significantly higher in dogs that developed hepatic toxicity (4 doses; 350 mg/m2) than in dogs without hepatic damage (3 doses; 230 mg/m2). Median duration to detection of hepatic toxicity from the last dose of CCNU was 11 weeks (range 2-49 weeks). Common biochemical abnormalities were abnormally high serum liver enzyme activities and hypoalbuminemia. Six dogs with CCNU-associated hepatic toxicity had ascites, and 3 dogs had concurrent pleural effusion. Serum concentrations of bile acids were abnormally high in 4 of 5 dogs tested. Percutaneous ultrasound-guided liver biopsies were performed in 10 dogs, and findings were nonspecific and chronic in nature. Seven dogs were euthanized because of progressive liver failure, and their median survival from diagnosis of liver disease was 9 weeks. Three dogs died of other causes and 1 dog of unknown cause. Although clinical signs resolved in 3 dogs, biochemical abnormalities and histopathologic lesions persisted 4 to 38 months from the time of diagnosis of liver disease. Our findings suggest that CCNU can cause delayed, cumulative dose-related, chronic hepatotoxicity that is irreversible and can be fatal.     

Protective immune responses of recombinant VP2 subunit antigen of infectious bursal disease virus in chickens

Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease and poses a huge threat to poultry industry. The risks associated with conventional attenuated viral vaccines make it indispensable to probe into the development of novel and rationally designed subunit vaccines which are safer as well as effective. VP2 is the major host-protective antigen found in IBDV capsid. It encompasses different independent epitopes responsible for the induction of neutralizing antibody. Here, we report the efficacy of the immunodominant fragment of VP2 which induces both humoral and cellular immunity against infectious bursal disease. A 366bp fragment (52-417bp) of the VP2 gene from an IBDV field isolate was amplified and expressed in Escherichia coli as a 21kDa recombinant protein. The efficacy of rVP2(52-417) antigen was compared with two commercial IBDV whole virus vaccine strains. The rVP2(52-417) induced significantly high antibody titres in chicken compared to commercial vaccines and the anti-rVP2(52-417) sera showed reactivity with viral antigens from both commercial strains (P<0.0001) and field isolates. Also, the chicken splenocytes from rVP2(52-417) immunized group showed a significantly high proliferation (P<0.01) compared to other groups, which implies that the rVP2(52-417) fragment contains immunogenic epitopes capable of eliciting both B and T cell responses. Further, rVP2(52-417) conferred 100% protection against vIBDV challenge in the immunized chickens which was significantly higher (P<0.001) compared to 55-60% protection by commercial vaccine strains. Hence, the study confirms the efficacy of the immunodominant VP2 fragment that could be used as a potent vaccine against IBDV infection in chicken.     

The role of nuclear technologies in the diagnosis and control of livestock diseases—a review

Nuclear and nuclear-related technologies have played an important role in animal health, particularly in relation to disease diagnosis and characterization of pathogenic organisms. This review focuses primarily on how and where nuclear technologies, both non-isotopic and isotopic methods, have made their impact in the past and where it might be expected they could have an impact in the future. The review outlines the extensive use of radiation attenuation in attempts to create vaccines for a multiplicity of pathogenic organisms and how the technology is being re-examined in the light of recent advances in irradiation techniques and cryopreservation/lyophilization that might obviate some of the problems of maintenance of viable, attenuate vaccines and their transport and use in the field. This approach could be used for a number of parasitic diseases where vaccination has been problematic and where investigations into the development of molecular vaccines have still failed to deliver satisfactory candidates for generating protective immune responses. Irradiation of antigens or serum samples also has its uses in diagnosis, especially when the samples need to be transported across international boundaries, or when handling the pathogens in question when carrying out a test presents serious health hazards to laboratory personnel. The present-day extensive use of enzyme immunoassays and molecular methods (e.g., polymerase chain reaction) for diagnosis and characterization of animal pathogens has its origins in the use of isotope-labeled antigens and antibodies. These isotopic techniques that included the use of ⁷⁵Se, ³²P, ¹²⁵I, and ³⁵S isotopes enabled a level of sensitivity and specificity that was hitherto unrealized, and it is prescient to remind ourselves of just how successful these technologies were, in spite of their infrequent use nowadays. Finally, the review looks at the potential for stable isotope analysis for a variety of applications??in the tracking of animal migrations, where the migrant are potential carriers of transboundary animal diseases, and where it would be useful to determine the origins of the carrier, e.g., Highly Pathogenic Avian Influenza and its dissemination by wild water fowl. Other applications could be in monitoring sequestered microbial culture (e.g., rinderpest virus) where in the case of accidental or deliberate release of infective culture it would be possible to identify the laboratory from which the isolate originated.     

Controlled elimination of clathrin heavy-chain expression in DT40 lymphocytes

We exploited the high rate of homologous recombination shown by the chicken B cell line DT40 to inactivate the endogenous alleles for clathrin heavy chain and replace them with human clathrin complementary DNA under the control of a tetracycline-regulatable promoter. Clathrin repression perturbed the activities of Akt-mediated and mitogen-activated protein kinase-mediated signaling pathways and induced apoptosis; this finding suggests that in DT40 cells clathrin helps to maintain the integrity of antiapoptotic survival pathways. We also describe a variant cell line in which these signaling pathways were unaffected by clathrin down-regulation. This variant cell line did not undergo apoptosis in the absence of clathrin and was used to examine the effects of clathrin depletion on membrane-trafficking pathways. Receptor-mediated and fluid-phase endocytosis were both substantially inhibited, and transferrin-receptor recycling was modestly inhibited. Surprisingly, clathrin removal did not affect the morphology or biochemical composition of lysosomes.     

Effect of early age woody and herbaceous competition control on wood properties of loblolly pine

Early age competition control has been reported to significantly improve the growth and yield of plantation grown loblolly pine. The objective of this paper is to understand the changes in wood properties: basal area weighted whole disk SG, earlywood SG (EWSG), latewood SG (LWSG) and latewood percent (LWP) of 14 year-old trees which received early age herbaceous and hardwood competition control, using data collected from 13-sites across 4-physiographic regions in the southeastern USA. The study was laid out in a randomized complete block design and had four levels of weed control (no weed control; woody vegetation control; herbaceous vegetation control; and woody and herbaceous vegetation control), with four blocks at each site. Increment cores 12 mm in diameter were collected at breast height (1.37 m) from 9-trees in each plot and ring-by-ring SG, EWSG, LWSG and LWP measured using a X-ray densitometer. Whole disk basal area weighted SG and LWP were determined for each tree and used for analysis. A reduction in whole disk SG of 0.039 and 0.0014 and LWP of 7.38% and 3.62% was observed for trees which received total weed control compared to no weed control, for lower and upper Coastal Plain sites, respectively. For trees receiving total weed control compared to no weed control, it was observed that the diameter of the juvenile core increased by 20% on average across all physiographic regions. However, no change in the length of the juvenile period was observed among treatments other than the regional differences.     

Protein-lipid interactions in zein films investigated by surface plasmon resonance

Experiments on the adsorption of alpha-zein (characterized by SDS-PAGE) from aqueous ethanol and 2-propanol solutions onto hydrophilic and hydrophobic surfaces are reported. Zein adsorption onto self-assembled monolayers (SAMs) was detected by surface plasmon resonance (SPR). Gold substrates were prepared by thermal evaporation on glass slides. Gold-coated surfaces were modified by depositing SAMs of either a long-chain carboxylic acid terminated thiol [COOH(CH2)(10)SH] or a methyl-terminated alkanethiol [CH3(CH2)(7)SH]. Experimental measurements indicated that zein interacted with both hydrophilic and hydrophobic surfaces. Zein concentration affected the thickness of bound zein layers. The estimated thickness of the zein monolayer deposited on hydrophilic surfaces was 4.7 nm. Zein monolayer thickness on hydrophobic surfaces was estimated at 4.6 nm. The topography of zein layers was examined by atomic force microscopy (AFM) after solvent was evaporated. Surface features of zein deposits depended on the adsorbing surface. On hydrophilic surfaces, roughness values were high and distinct ring-shaped structures were observed. On hydrophobic surfaces, zein formed a uniform and featureless coverage.