A novel 57-kDa merozoite protein of Babesia gibsoni is a prospective antigen for diagnosis and serosurvey of canine babesiosis by enzyme-linked immunosorbent assay

We isolated a novel single copy gene encoding a 57-kDa merozoite protein of Babesia gibsoni (BgP57). The nucleotide sequence of the cDNA was 2387 bp with an open reading frame (ORF) of 1644 bp encoding a 57-kDa predicted polypeptide having 547 amino acid residues. The recombinant BgP57 (rBgP57) without a predicted signal peptide was expressed in Escherichia coli as a soluble glutathione S-transferase (GST) fusion protein. Western blotting showed that the corresponding native protein was 57-kDa, consistent with molecular weight of predicted mature polypeptide. An indirect enzyme-linked immunosorbent assay (ELISA) using the rBgP57 detected specific antibodies in the sequential sera from a dog experimentally infected with B. gibsoni. Moreover, the antigen did not cross-react with antibodies to B. canis sub-species and closely related apicomplexan parasites indicating that the rBgP57 was a specific antigen for B. gibsoni antibodies. The diagnostic performance of ELISA based on rBgP57 using 107 sera from B. gibsoni-naturally infected dogs was the same as the previously identified rBgP32 but performed better than the previously studied rBgP50. Although, seminested PCR detected higher proportions (82%) of positive samples than the ELISAs, the Mcnemar's chi-square test showed that there was no significant difference in relative effectiveness of rBgP57-ELISA and seminested PCR (chi(2)=2.70; P=0.1003) in identifying positive samples. The rBgP57-ELISA when used in combination with rBgP32-ELISA and rBgP50-ELISA appeared to improve sensitivity of the rBgP57-ELISA for detection of B. gibsoni antibodies. Overall, the rBgP57-ELISA and seminested PCR when used in combination, could improve epidemiological surveys and clinical diagnosis of B. gibsoni infection.     

Predictive models for biomass and carbon stocks estimation in Grewia optiva on degraded lands in western Himalaya

Grewia optiva Drummond is one of important agroforestry tree species grown by the farmers in the lower and mid-hills of western Himalaya. Different models viz., monomolicular, logistic, gompetz, allometric, rechards, chapman and linear were fitted to the relationship between total biomass and diameter at breast height (DBH) as independent variable. The adjusted R² values were more than 0.924 for all the seven models implying that all models are apparently equally efficient. Out of the six non-linear models, allometric model (Y = a × DBH ^b ) fulfils the validation criterion to the best possible extent and is thus considered as best performing. Biomass in different tree components was fitted to allometric models using DBH as explanatory variable, the adjusted R² for fitted functions varied from 0.872 to 0.965 for different biomass components. The t values for all the components were found non-significant (p > 0.05), thereby indicating that model is valid. Using the developed model, the estimated total biomass varied from 6.62 Mg ha^?1 in 4 year to 46.64 Mg ha^?1 in 23 year old plantation. MAI in biomass varied from 1.66–2.05 Mg ha^?1 yr^?1. The total biomass carbon stocks varied from 1.99 Mg ha^?1 in 4 year to 15.27 Mg ha^?1 in 23 year old plantation. Rate of carbon sequestration varied from 0.63–0.81 Mg ha^?1 yr^?1. Carbon storage in the soil up to 30 cm soil depth varied from 25.4 to 33.6 Mg ha^?1.     

Performance of plastic biofilter media with different configuration in a water recirculation system for the culture of Nile tilapia (Oreochromis niloticus)

Three kinds of locally available plastic biofilter media with different configurations (plastic rolls, PVC pipes and scrub pads) were evaluated for their efficiency in organic waste removal from the effluents of an intensive recirculating tilapia culture system. A set of three types of solid-removing filters consisting of screened sedimentation; upflow sand as well as plastic bead filtration accomplished the mechanical filtration. Values of critical metabolic wastes like total ammonia nitrogen (TAN) (0.92 ppm) and nitrite-nitrogen (NO₂-N) (0.22 ppm) were found to be well within the acceptable limits, while other water quality parameters in the culture water were also maintained within the normal range by the filtration system. Removal rates of 3.46 g TAN/m³ per day and 0.77 g NO₂-N/m³ per day, as well as TAN and NO₂-N removal efficiencies of 29.37 and 27.3% respectively, were established to be the best for the plastic-roll biofilter medium as compared to PVC-pipe and scrub-pad media. Percent removal of TAN and NO₂-N per pass of the biofilter (25.49 and 26.3% respectively) and the specific TAN and NO₂-N removal rates (43 and 9.6 mg/m² per day) of plastic rolls were also found to be superior to the other two biofilter media. Pieces of PVC pipes as biofilter medium is recommended to be used in the biofilters in view of their cheaper cost.     

Replacement of Artemia with Moina micrura in the rearing of freshwater shrimp larvae

The effect of an abrupt change in the live diet of shrimp larvae was investigated by replacing Artemia with Moina micrura. The control treatment consisted of feeding Artemia throughout the rearing period (regime A), while in the other treatments the onset of Moina feeding was arbitrarily chosen at larval stages iv (A3M), vi (A5M), viii (A7M) and x (A9M). No significant differences ( = 0.05) were observed among the treatments during larval production, mean stage development (MSD) and growth of postlarvae. The mean (SD) yields of postlarvae (PL) were 11.97 (1.98), 15.10 (2.92), 14.72(1.56), 13.51 (1.74) and 12.70 (1.40) PL l^–1 respectively for the feeding regimes A3M, A5M, A7M, A9M and A. Up to stage v, the ingestion rate in the Moina treatment was as low as 0.01–0.47 larva^–1 h^–1 compared with that in the Artemia treatment (0.29–1.77 larva^–1 h^–1). However, the ingestion of Moina increased from stage vi–vii onwards.     

Effects of supplementation of coated crystalline amino acids on growth performance and body composition of juvenile kuruma shrimp Marsupenaeus japonicus

Supplementation of crystalline amino acids (CAA) in shrimp diets is unsuccessful in most cases. In the present study, various pre‐coated CAA were developed to minimize the leaching losses from diets in water and assessed using kuruma shrimp Marsupenaeus japonicus juveniles. Experimental diets were supplemented with coated CAA or non‐coated CAA to simulate the dietary amino acid profile to that found in the whole body protein of shrimp. The four diets contained CAA coated with either carboxymethyl cellulose (CMC) (diet 1), zein (diet 2), k‐carrageenan (diet 3) or agar (diet 4), respectively. Another four diets contained CMC‐coated CAA with the addition of either zein (diet 5), casein‐gelatin (diet 6), zein and k‐carrageenan (diet 7), or casein‐gelatin and k‐carrageenan (diet 8), respectively. The 8 weeks feeding trial demonstrated that the diets containing coated CAA gave significantly higher weight gains than diets containing non‐coated CAA, with the highest value for the diet 6. The diets containing coated CAA also yielded higher feed conversion efficiency, protein efficiency ratio, and specific growth rate than diets containing non‐coated CAA. Leaching trials suggested that shrimp growth performance was improved partly due to the retardation or reduction of CAA leaching by coating them with specific binders.     

Capillary flow responses in a soil–plant system for modified subsurface precision irrigation

Water movement in a soil–plant system was evaluated based on capillary flow in a modified subsurface irrigation system that incorporates a plant-water measuring device. Water from a reservoir tank located underneath the plant pot was supplied to the root zone through a fibrous medium. Evapotranspiration was measured from the water uptake and evaluations were performed based on soil moisture distribution and mass balance. Potential evapotranspiration was used as a reference for the plant–water uptake. Data were obtained from a test plant provided with the modified subsurface irrigation system. The plant was grown in a phytotron under controlled air temperature and humidity, and a comparison was made for different levels of soil moisture condition. The experimental results confirmed the operational efficiency of the modified subsurface irrigation system for precision irrigation.     

Voluntary food intake variation in chickens on lysine-free diet is attributed to the plasma lysine concentration

1. Growing chickens decrease their voluntary food intake when they receive a diet deficient in a single essential amino acid. Our previous studies suggest that the decreased food intake was associated with some metabolic changes.

2. In order to reveal the involvement of plasma lysine fluctuations in the reduction of food intake, we examined whether maintaining the plasma lysine concentration in chickens on a lysine-free diet (the purified diet contained no lysine) restored the food intake to that of the control (lysine hydrochloride 11.9 g/kg) group.

3. Male egg-type chickens at 21 d of age were injected with lysine at doses of 0.1 g/ml one hour after presenting the lysine-free diet. This injection increased the plasma lysine concentration one hour later and kept it similar to that of the control group for the following 2 h. Chickens ate the lysine-free diet as much as the control diet when their plasma lysine concentration was kept at a similar level to the control group. Injection of saline or alanine (0.12 g, isonitrogenous to lysine 0.1 g) into the crop of chickens on the lysine-free diet did not bring about the variations of food intake and plasma lysine concentrations as observed in those with lysine.

4. These findings show that the food intake variation was attributed to the plasma lysine concentration in the chickens on the lysine-free diet.     

Fate of carbofuran in rice-fish model ecosystem —An international study

Work done in six nations by seven research groups on Carbofuran metabolism in rice-fish ecosystem is summarized. The following results have been obtained: (a) Soil acts as a sink for the pesticide and there is considerable metabolism in the soil; (b) There is no bioaccumulation/biomagnification by the fish used in the experiments, the residue levels in water being extremely low; and (c) The plants translocate the chemical and even at the time of harvest, the grains show the presence of the pesticide.