Dissipation pattern and risk assessment of flubendiamide on chili at different agro-climatic conditions in India.

Dissipation pattern and risk assessment of flubendiamide and its metabolite (desiodo flubendiamide) on chili were studied at four different agro-climatic locations of India at the standard and double dose at 60 and 120 g a.i. ha(-1) at 10 days interval. Quantification of residues was done on a high-performance liquid chromatograph (HPLC) with a photo diode array detector. The limit of quantification (LOQ) of this method was found to be 0.01 mg kg(-1) while limit of detection (LOD) being 0.003 mg kg(-1). Residues of flubendiamide were found to be below the determination limit in 15 days at both the dosages in all locations. Half-life of flubendiamide when applied at 60 and 120 g a.i. ha(-1) ranged from 0.85 to 1.80 and from 0.95 to 2.79 days, respectively. On the basis of data generated under the All India Network Project on Pesticide Residues, a preharvest interval (PHI) of 1 day has been recommended and the flubendiamide 480 SC has been registered for use on chili in India by the Central Insecticide Board and Registration Committee, Ministry of Agriculture, Government of India. The maximum residue limit (MRL) of flubendiamide on chili has been fixed by the Food Safety Standard Authority of India, Ministry of Health and Family Welfare, Government of India, as 0.02 µg g(-1) after its risk assessment.

Residual behavior and risk assessment of flubendiamide on tomato at different agro-climatic conditions in India.

Supervised field trials were conducted at four different agro-climatic zones in India to evaluate the dissipation pattern and risk assessment of flubendiamide on tomato. Flubendiamide 480 SC was sprayed on tomato at 48 and 96 g active ingredient (a.i.) ha(-1). Samples of tomato fruits were drawn at 0, 1, 3, 5, 7, 10, 15, and 20 days after treatment. Quantification of residues was done on a high-performance liquid chromatography (HPLC) device with a photo diode array detector. The limit of quantification (LOQ) of this method was found to be 0.01 mg kg(-1) while limit of detection (LOD) being 0.003 mg kg(-1). Residues of flubendiamide were found below the determination limit of 0.01 mg kg(-1) in 20 days at both the dosages in all the locations. The half-life of flubendiamide at an application rate of 48 g a.i. ha(-1) varied from 0.33 to 3.28 days and at 48-g a.i. ranged from 1.21 to 3.00 days. On the basis of data generated under the All India Network Project on Pesticide Residues, a preharvest interval (PHI) of 1 day has been recommended, and the flubendiamide 480 SC has been registered for its use on tomato by the Central Insecticide Board and Registration Committee, Ministry of Agriculture, Government of India. The maximum residue limit (MRL) of flubendiamide on tomato has been fixed by the Ministry of Health and Family Welfare, Government of India under Food Safety Standard Authority of India, as 0.07 µg g(-1) after its risk assessment.

Safety evaluation of flubendiamide and its metabolites on cabbage and persistence in soil in different agroclimatic zones of India.

Supervised field trials following good agricultural practices were conducted at the research farms of four agricultural universities located at four different agroclimatic zones of India to evaluate the persistence and dissipation of flubendiamide and its metabolite, des-iodo flubendiamide, on cabbage. Two spray applications of flubendiamide 480 SC of standard and double dose at the rate of 24 and 48 g a.i. ha(-1) were given to the crop at a 15-day interval, and the residues of flubendiamide 2 h after spray were found in the range of 0.107-0.33 and 0.20-0.49 mg kg(-1) at respective doses. Residue of des-iodo flubendiamide was not detected in any cabbage sample during study period. No residues were found in the soil samples collected from all treated fields after 15 days of application. On the basis of data generated under All India Network Project on Pesticide Residues, a preharvest interval (PHI) of 10 days has been recommended, and the flubendiamide 480 SC has been registered for its use on cabbage by Central Insecticide Board and Registration Committee, Ministry of Agriculture, Government of India. The maximum residue limit (MRL) of flubendiamide on cabbage has been fixed by the Ministry of Health and Family Welfare, Government of India, under Food Safety Standard Authority of India as 0.05 µg/g after its risk assessment.

Dissipation and persistence of flubendiamide and thiacloprid in/on tomato and soil.

Flubendiamide, a phthalic acid diamide protects the plants against a broad range of economically important lepidopterus pests and thiacloprid a second generation neonicotinoid is effective against the sucking insects, white flies and jassids. To estimate the residues of flubendiamide and thiacloprid on tomato, analytical methods were validated by conducting recovery studies, residues were quantified by using HPLC on C(18) column and PDA at 260 λ. Residues of flubendiamide declined below detectable level of 0.01 mg kg(-1) after 5 and 7 days of application at lower and higher dose with RL(50) of 0.72 and 1.32 days, respectively. Thiacloprid residues reached below its detectable level of 0.01 mg kg(-1) after 5 and 7 days of its lower and higher dose with RL(50) of 0.83 and 1.79 days, respectively.

Dissipation study of Quinalphos (25 EC) in/on brinjal and soil.

A field experiment was conducted at Agricultural Research Station, Durgapura, Jaipur during kharif 2008 to study the dissipation of Quinalphos (25 EC) in/on brinjal and soil, when sprayed at its recommended dose (375 g a.i. ha(-1)) and double of the recommended dose (750 g a.i. ha(-1)). The residue data revealed the magnitude of dissipation and persistence by calculating safety parameters like RL(50) and T(tol). The initial deposit of Quinalphos in brinjal at 375 and 750 g a.i. ha(-1) were recorded as 0.0866 and 0.1517 mg kg(-1), respectively which reached to below detectable level (0.01 mg kg(-1)) in 7 and 10 days at recommended (375 g a.i. ha(-1)) and double of the recommended dose (750 g a.i. ha(-1)), respectively. The residues, however, had a half life value (RL(50)) of 2 days for lower dose and 3 days for higher dose. Hence 6 and 9 days waiting period was suggested for recommended and double of the recommended dose, respectively. No residues were detected in soil in treated plots at both the treatment levels 30 days after the spray of insecticide to the crop.

Persistence of propineb (polymeric zinc 1,2-propylene bisdithiocarbamate) fungicide in onion and cropped soil.

To study the persistence of propineb in/on onion a field experiment was conducted during 2007 at Agricultural Research Station, Durgapura, Jaipur Propineb70WP @ 1,225 and 2,450 g a. i. ha(-1) dose was applied as foliar spray to the crop thrice at an interval of 10 days. After third spray, onion samples comprising of green leaves and immature bulb were collected at 0,1,3,5,7,10 and 15 days. The initial deposit was 2.32 and 4.89 ppm of propineb (on CS(2) basis) which persisted up to the 10 and 15 days at normal 1,225 g a.i. ha(-1) and higher 2,450 g a.i. ha(-1) dose of fungicide application. The half life values of propineb ranged from 3.27 to 4.60 days.

Persistence and dissipation kinetics of spiromesifen in chili and cotton.

In a field study carried out at three different locations, the dissipation of spiromesifen on cotton and chili was studied and its DT50, and DT99 were estimated at each location. Spiromesifen was sprayed on chili at 96 and 192 g a.i. ha(-1) and cotton at 120 and 240 g a.i. ha(-1). Samples of chili fruits were drawn at 0, 1, 3, 5, 7, 10, 15, 21, 30 days after treatment and that of cotton seed and lint at first picking and harvest. Soil samples were drawn 30 days after treatment from 0 to 15 and 15 to 30 cm layer. Quantification of residues was done on GC-MS in Selected Ion Monitoring (SIM) mode in mass range 271-274 m/z. The LOQ of this method was found 0.033 microg g(-1), LOD being 0.01 microg g(-1). The DT50 of spiromesifen when applied at recommended doses in chili fruits was found to be 2.18-2.40 days. Ninety-nine percent degradation was found to occur within 14.5-16.3 days after application. Residues of spiromesifen were not detected in cotton seed and lint samples at the first picking. In soil, no residues of spiromesifen were detectable 15 days after treatment.

Sequential development of histologic lesions and their relationship with bacterial isolation, fecal shedding, and immune responses during progressive stages of experimental infection of lambs with Mycobacterium avium subsp. paratuberculosis.

Understanding pathogenesis during progressive stages of infection by Mycobacterium avium subsp. paratuberculosis (MAP) and finding suitable methods for its diagnosis are key to the control of Johne's disease in animals. Paratuberculosis was experimentally produced in 20 crossbred lambs by oral administration of MAP to study the sequential development of lesions between 10 and 330 days postinfection and to assess commonly used diagnostic methods such as bacterial culture, lymphocyte stimulation test (LST), and enzyme-linked immunosorbent assay (ELISA) during progressive stages of infection. Histologic lesions were classified into four grades from grade 1 (least severe) to grade 4 (most severe) on the basis of location of granulomatous lesions in different regions and layers of intestines, their association with intestinal lymphoid tissues, pattern and distribution of lesions, types of cellular infiltration, and presence of acid-fast bacilli. It is evident that infection first establishes in lymphoid tissues of the small intestine, possibly at multiple sites, producing segmental lesions and from there spreads to lamina propria and local lymph nodes. Wide variability in the histologic lesions in relation to postinfection periods and initial tropism of MAP to the intestinal lymphoid tissues (Peyer's patches) suggests a differential susceptibility of young animals, possibly because of compositional phenotypic variation of Peyer's patches influencing subsequent course of infection. Histopathology was found to be a better indicator of paratuberculous infection than bacteriology in sheep. The LST (reflecting the cellular immune response) and ELISA (reflecting the humoral immune response) had overall sensitivities of 65% (11 of 17) and 42% (8 of 19), respectively, in sheep with different types of pathology but when employed together could detect about 88% of infected animals.

Cloning and sequencing of a 16 kDa outer membrane protein gene of Pasteurella multocida P52.

The outer membrane proteins (OMPs) of Pasteurella multocida are potential immunogens. The 16 kDa OMP of P. multocida P52, serotype B:2, was identified as one of the major immunodominant antigens. The gene omp16, encoding a 16 kDa outer membrane protein, was amplified, cloned into a pBluescript SK(-) vector and sequenced. Complete sequence homology was observed between the 16 kDa OMP gene of P. multocida P52 (serotypes B:2) and P. multocida T16 (somatic serotype 3,4). This gene was distributed among different serotypes of P. multocida and found to localize in a 6.0 kb HindII fragment of the P. multocida genome.

An enzyme-linked immunosorbent assay using immonoaffinity-purified antigen in the diagnosis of caprine paratuberculosis and its comparison with conventional ELISAs.

An enzyme-linked immunosorbent assay (APA-ELISA) using an immunoaffinity-purified antigen was developed and compared with the unabsorbed and absorbed ELISA procedures, using a crude antigenic preparation, for its efficacy in detecting antibodies in goat sera against Mycobacterium ovium paratuberculosis. Serum samples from 89 goats belonging to three different flocks, two with a history and evidence of paratuberculosis and one without it, were subjected to each ELISA, which had been standardized on known positive sera from goats experimentally infected with paratuberculosis. Faecal culture, faecal examination and histopathology were used as indicators of infection. The diagnostic sensitivities of the unabsorbed, absorbed and APA-ELISA were 81.8%, 77.3% and 77.3% and the specificities were 90.6%, 93.7% and 96.8%, respectively. The positive predictive values of APA-ELISA (94.4%) was the highest, followed by absorbed ELISA (80.9%) and unabsorbed ELISA (72.0%). The negative predictive values for APA-ELISA, absorbed ELISA and unabsorbed ELISA were 93.0%, 92.7% and 93.8%, respectively. The results indicated the value of APA-ELISA in avoiding the need to absorb individual test sera with Mycobacterium phlei and giving more consistent results than the absorbed ELISA. The APA-ELISA was also better than the other two procedures in terms of specificity and positive predictive values.

Identification and characterization of a gene encoding a 35-kDa protein from Mycobacterium avium subspecies paratuberculosis.

Mycobacterium avium subspecies paratuberculosis is the causative agent of Johne's disease, a chronic enteritis in ruminants. A gene homologous to that of 35-kDa antigen of Mycobacterium leprae was cloned and sequenced from Mycobacterium paratuberculosis. The database searches revealed 82.79% and 95.67% similarities of its nucleotide sequence, with those of immunodominant 35-kDa protein of M. leprae and M. avium, respectively.

Effect of buffalo follicular fluid treatment on follicle population and ovulation rate in guinea pigs.

A number of workers have studied the effect of follicular fluid (FF) on the secretion of follicular stimulating hormone (FSH) but little is known about its potential as a regulator of ovarian activity, including ovulation rate. This paper describes the effect of charcoal treated-buffalo follicular fluid (buFF) treatment on follicular growth and ovulation rate in guinea pigs. Eighteen guinea pigs in three groups of 6 each were given 0.2 ml buFF at 12 hr interval for 3 days at different stages of estrous cycle viz., early-luteal, mid-luteal or follicular phase. One control group received equal volume of saline. Estrus was monitored every morning and evening by inspection of the opening of vaginal membrane and its cytology. All animals were sacrificed at 24 hr after the onset of estrus. Both the ovaries were dissected out, weighed and number of ovulation points recorded. One ovary from each animal was processed for histological examination to determine the population of healthy and atretic follicles. In early-luteal and follicular phase-treated animals the onset of estrus was delayed (P < 0.01) and ovulation rate was not affected. However, estrus occurred at normal when the treatment was initiated at midluteal stage and 50% animals failed to ovulate in this group. The total follicle population at metestrus increased significantly in all treated animals because of increase in number of follicles of size class II (400 to < 600 microns diam.). Atresia was also declined due to treatment. These results demonstrated that the buFF contained some inhibitory substances that delayed the onset of estrus in guinea pigs.

Effect of passive immunization with buffalo follicular fluid antisera on ovarian activity in guinea pigs.

Ovarian activity and follicular populations were studied in guinea pigs (Cavia porcellus) following administration of antisera against buffalo follicular fluid (buFF). Antibodies were raised in rabbits and the titre tested by immunodiffusion assay. Fourteen guinea pigs cycling normally were randomized into two groups. Animals in Group I (n = 8) were treated (i.p.) with 0.5 ml antisera and in Group II (control, n = 6) with the same volume of normal rabbit serum at 12 h intervals on the 10th and 11th day of their oestrous cycle. They were sacrificed 24 h after onset of estrus when ovulation points were counted and ovaries processed for microscopical examination. Treatment with buFF-antisera increased ovulation rate (3.6 vs. 2.0; p < 0.01) but had no significant effect on the total number of follicles. However, the treatment reduced the percentages of atretic follicles in all size classes. These results indicated that the administration of a buFF-antisera produced in the rabbits increased ovulation rate in guinea pigs by reducing the incidence of atresia.

Invasive pulmonary pseudallesheriosis in a cross-breed calf.

Pulmonary pseudallescheriosis was diagnosed in a two-months old calf. Pneumonic lungs with yellow-white nodules on the surfaces revealed granulomatous lesions microscopically. Septate, pleomorphic hyphae were present in the central caseated core with a bright eosinophilic periphery surrounded by polymorphonuclear cells and macrophages followed by a zone of epithelioid cells admixed with lymphocytes and plasma cells. The fungal agent was demonstrated by Grocott's silver methenamine staining. On isolation, morphologically it was found to be indistinguishable from that of Pseudallescheria boydii. It appears to be first report of fatal mycotic pneumonia in a calf due to P. boydii. The emphasis is given for further detailed investigation on this aspect in veterinary mycopathology.

Subacute toxicity of Basalin in rats.

Basalin, a herbicide, was administered orally to male rats at doses ranging from 60 mg to 1.92 g/kg for 13 weeks. Oral LD50 of the compound was 1.65 g/kg. Toxic effects included hyperexcitability and tremors. The cumulative lethal dose (CLD50) at the end of week 13 was 135 mg/kg with a cumulative toxicity factor (CTF) of 12.22. At 1.92 g/kg, no animals survived to 13 weeks. At 60 and 120 mg/kg, there were no significant changes in body weight gain compared with the controls and a significant decrease in total leukocyte count (TLC), erythrocyte sedimentation rate (ESR) and Hb was observed. There was a decrease in spermatogenesis and infiltration of mononucleated cells in the liver.