Detection of Babesia bigemina DNA in ticks by DNA hybridization using a nonradioactive probe generated by arbitrary PCR

The Dig-labeled probe specific to Babesia bigemina generated from monomorphic RAPD fragment of approximately 873 bp size amplified by a 10 mer CGGTGGCGAA, detected up to 100 ng of template DNA. This nonradioactive probe also detected B. bigemina in preparations of larval tick DNA from two of the five samples on dot-blot hybridization.     

High antigenic cross-reaction among the bacterial species responsible for diseases of cultured freshwater fishes and strategies to overcome it for specific serodiagnosis

Antigenic sharing among the most commonly bacterial pathogens such as Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens of Indian major carps has been studied using immunological reactions such as cross-agglutination, disc diffusion and indirect enzyme linked immunosorbent assay (ELISA). The data were analysed using statistical analysis (SAS), version 6.12. The results showed high antigenic similarities among the bacterial whole cells, whole cell lysates, somatic 'O' antigens, lipopolysaccharides (LPS) and extracellular products (ECP). However, few or no similarities were observed in ECP components of <20kD. The present study indicates a need to develop differential diagnostic methods based on serology by choosing the highly specific less cross-reactive ECP antigen.     

High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis

Toxoplasmosis, caused by Toxoplasma gondii, is a disease of economic importance in livestock, especially in sheep and goats, where it causes abortion. Although several serological tests are in use for diagnosis of infection, production of reliable reagents is a constraint. An 814 bp sequence coding for a truncated surface antigen surface antigen 1 (SAG1), a tachyzoite stage-specific protein, as well as a 657 bp sequence coding for granule protein 7 (GRA7), a dense granule protein were PCR amplified from the genomic DNA of T. gondii. The amplified products were ligated in pET-32b(+) and pET-32c(+) expression vectors, respectively and subsequently transformed into BL21(DE3)pLysS cells. A high-level expression of the histidine-tagged SAG1 and GRA7 fusion proteins were obtained after 7h of incubation. The recombinant proteins were purified using Ni-NTA column and were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis using reference positive sera from goat, rabbit and humans at 1:100 dilution. Subsequently, the diagnostic efficiency of the recombinant proteins, either individually or as a cocktail of the recombinant proteins, was assessed with 56 reference goat sera by enzyme-linked immunosorbent assay (ELISA). The immunoreactivity of the refolded SAG1 and GRA7 was evidenced by high OD values. The reactivity of the recombinant proteins as a cocktail preparation was more than that of individual proteins in ELISA and could detect accurately the infection in goats. This is the first report of serological detection of caprine toxoplasmosis by ELISA using a cocktail of recombinant Toxoplasma proteins.     

Nasal vascular hamartoma in a Domestic Shorthair cat

A nasal mass in a Domestic Shorthair cat was causing facial deformity, sneezing and intermittent epistaxis. Biopsy samples obtained previously had been non-diagnostic. Computed tomography images revealed an irregular, contrast-enhancing mass occupying a large portion of the righthand side of the nasal cavity. Previously described criteria for malignancy were not present. A ventral surgical approach combined with temporary, ipsilateral, common carotid arterial occlusion provided excellent access for debulking the lesion and collecting samples for histopathology. A nasal vascular hamartoma was diagnosed and clinical signs resolved postoperatively. This is the first documentation of this abnormality in the cat. Hamartomatous abnormalities should be included on the list of differential diagnoses for feline nasal mass lesions. The prognosis for hamartomatous lesions postoperatively is good, in keeping with their limited propensity for growth after maturity.     

Foliage nutrient concentration and lopping time of agroforestry tree species in the Western Himalayan region of India

Tree foliage is a nutrient-rich and important source of green fodder to sustain the livestock during the lean winter period of a season in the low and mid hill regions of Indian Himalaya. Knowledge of the variations in mineral nutrient composition of the foliage during an active growth season helps in determining optimum lopping time to harvest nutrient-rich fodder. Changes in the nutrient concentrations of Albizia stipulata and Ougenia oojeinensis foliage during the annual growth season were studied to find out an optimum lopping time. Two dissimilar trends in variation of nutrients in foliage with the advancement of growth season were observed. These were: (1) depletion/dilution of N, P, K and S and (2) accumulation of Ca, Mg and Na. The depletion was more from October onwards, and accumulation reached to its maximum in November-December. Therefore, the period from October to December, preferably November, can be regarded as optimum lopping time for A. stipulata. For O. oojeinensis, the corresponding time could be September to November, preferably October. Ougenia foliage contained higher concentration of N, P, Ca, Mg, and Na than that of Albizia. This revised version was published online in June 2006 with corrections to the Cover Date. This revised version was published online in June 2006 with corrections to the Cover Date.     

Changes in Plasma Concentrations of LH, FSH, Estradiol 17-β and Progesterone During Oestrus in Mithun (Bos frontalis)

The objective of the present study was to establish the changes in plasma concentrations of LH, FSH, estradiol 17-beta (E2) and progesterone (P4), as well as to understand their temporal relationships during oestrus in mithun (Bos frontalis). The experiment was conducted on 11 mithuns during third or fourth postpartum oestrous cycle. Since oestrus onset the jugular vein blood samples were collected every 2 h for 72 and 96 h, respectively from the animals without and with standing heat. The LH, FSH, E2 and P4 concentrations were estimated in plasma. The P4 concentration was fluctuated throughout the oestrus period and the average P4 concentration was found significantly (p<0.05) lower on the day of oestrus onset. The multiple rises in LH and FSH concentrations above the basal level in spike like fashion were observed throughout the oestrus period irrespective of the occurrence of standing heat. A significant (p<0.01) gradual increase in the average daily E2 concentration was observed till day 2 following oestrus onset irrespective of the occurrence of standing heat. A significant (p<0.05) simultaneous increase in LH, FSH and E2 concentrations and a transient increase in P4 concentration at approximately the time of standing heat onset were observed. During investigation a definite temporal coupling between LH and FSH rises was absent throughout the oestrus period. The results suggest that (1) the multiple short-duration low-amplitude LH and FSH surges during oestrus may be crucial for the final maturation of ovulatory follicle and subsequent ovulation in mithun; (2) a differential mechanism for controlling LH and FSH secretions probably exists in mithun.     

Goat serum, a substitute of bovine serum in cultivation of Babesia bovis.

Babesia bovis (a Mexican isolate) was cultivated in MASP culture system using goat serum in various concentrations as substitute of bovine serum. It was observed that 20% goat serum + 20% bovine serum + 60% Parker's medium 199 supported the growth of the parasite, which was maintained in this medium through 8 subcultures. The soluble exoantigen (vaccine) present in the culture supernatant is to be quantified and tested in vitro. Goat serum from slaughterhouses may be utilized for in vitro cultivation of the parasite and, expectedly, production of vaccine. This study may prove to be useful in reducing the cost of vaccine at least in tropical countries.     

Inorganic pyrophosphatase activity in water stressed excised rice leaves

Summary Changes in the inorganic pyrophosphatase (EC 3.6.1.1) activity and the chlorophyll and -amino nitrogen levels were studied in excised leaves of rice (Oryza sativa L. cv. Ratna) subjected to various degrees of water stress. Chlorophyll level decreased and a-amino nitrogen level increased in the turgid leaves floating on water; these changes were accelerated when the leaves were subjected to water stress (Figs. 2, 3). The decrease with time in the activity of alkaline inorganic pyrophosphatase and the increase with time in the activity of acid inorganic pyrophosphatase in water-floated turgid leaves were accelerated following water stress (Figs. 4, 5). There was an accelerated decline in the ratios of alkaline to acid inorganic pyrophosphatase activity in stressed compared to turgid leaves (Fig. 6). It was concluded that water stress enhances the senescence of excised rice leaves and that the changes in the activities of alkaline and acid inorganic pyrophosphatases can be taken as indicators of water stress in rice leaves.Work supported by grant from the Extra Mural Research Unit, Council of Scientific and Industrial Research, New Delhi to Dr. D. Mishra. Mr. S. Dwivedi is a University Grants Commission Junior Research Fellow