Characterization of carboxylesterase in skin mucus of Cirrhinus mrigala and its assessment as biomarker of organophosphate exposure

Presence of carboxylesterase (CbE) activity in the skin mucus of Cirrhinus mrigala was investigated. CbE activity in skin mucus showed higher substrate preference for α-naphthyl acetate over p-nitrophenyl acetate. Four CbE isozymes—CbE-1, CbE-2, CbE-3, and CbE-4 were observed in skin mucus during zymography. The isozyme CbE-4 was characterized as typical serine esterase, whereas CbE-1, CbE-2, and CbE-3 were identified as sulphhydryl group-dependent serine esterases. In vitro treatment of skin mucus with the organophosphorus insecticide, Nuvan^® showed strong inhibition of CbE activity. In vivo exposure of the fish to sublethal test concentrations (5 and 15 mg/l) of the insecticide also revealed significant inhibition of CbE activity in mucus. After the cessation of exposure, CbE activity recovered to its control level during the recovery periods. Thus, CbE activity in skin mucus could be considered a biomarker of the organophosphorus insecticide exposure to fish and a useful tool in monitoring environmental toxicity.     

Seroprevalence of Actinobacillus (Haemophilus) pleuropneumoniae serotype-1 infection in swine herds in Quebec

Seroprevalence of Actinobacillus (Haemophilus) pleuropneumoniae serotype-1 infection was evaluated in pigs on 7 farms in Quebec. Commercial cross-bred herds A to G, ranging from 110 to 235 sows and infected with A pleuropneumoniae serotype-1 were selected. Five pigs/litter were selected at random and were identified (group 1). Blood samples were obtained from group-1 pigs at 2 to 4, 14, 28, 42, and 56 days of age. Blood also was obtained from group-1 pigs remaining in the postweaning unit at 70 days of age, and from 20 to 40 sows 1 to 3 times. To determine prevalence of seropositive pigs in all age groups for the entire study period in herds C to G, blood samples were obtained from 20 pigs/age group (group 2) selected at random at 28, 42, and 56 days of age at each visit. Group-1 pigs were included when they reached 28, 42, and 56 days of age. Pigs were serologically monitored in herds A and B for 3 months and in herds C to G for 5 to 6 months. Serologic status of pigs at 2 to 4 days of age was not statistically associated with status at 42 days (P = 0.6293) and at 56 days (P = 0.3098) of age for the same pigs. Therefore, seronegative pigs 2 to 4 days old did not seroconvert earlier than did those with detectable maternal antibodies at 2 to 4 days old. Only about 50% of the 70-day-old pigs were seropositive at 56 days. Seemingly, pigs seroconverted late in the postweaning period.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of heat treatment on the surface antigens of Haemophilus pleuropneumoniae

Coagglutination and ring precipitation tests were used to study the effect of heat on the surface antigens of Haemophilus pleuropneumoniae strains employing the reference strains belonging to serotypes 1 to 7 and field isolates belonging to serotypes 1, 2, 3, 5, 6 and 7. By immunising rabbits with formalin-fixed whole-cell suspension, antibodies were obtained which sensitised Cowan I Staphylococcus aureus to coagglutinate antigen preparations which had not been heated, or heated at 56 degrees C, or boiled or autoclaved. Similar positive reactions were obtained with the ring precipitation test. Heating the cultures at 56 degrees C for one hour was best for exposing the most potent serotype-specific antigens in all the strains studied. All the reference strains and most of the field isolates possessed the thermostable type specific antigens which could withstand autoclaving for one hour. However, many field isolates belonging to serotype 1 did not possess this antigen. The apparent antigenic heterogeneity of serotype 1 strains based on the presence or absence of these thermostable antigens could be valuable in epidemiological investigations. It was shown that most potent serotype-specific antigens are present as freely diffusible material on the surface layer of the bacterial cells, which could easily be removed by washing in saline solution. Well washed bacterial cells devoid of surface materials are poor antigens. It is recommended that test strains should not be heated above 56 degrees C for serotyping because higher temperatures are liable to destroy the capsular antigen of some strains and render the culture untypeable.     

Pain-related cerebral potentials in response to acute painful electrical stimulation in sheep

Objective To examine associations between late event-related cerebral potential amplitudes and behavioural responses to noxious electrical stimulation as an indicator of acute pain in sheep.
Design Analysis of variance for the effects of stimulus intensity on the behaviour and event-related cerebral potential variables.
Procedure Ninety-six brief constant current electrical pulse trains were presented to the front left leg of eight sheep at four intensities (2.5 to 10 mA) in a randomised order. An event-related cerebral potential and a graded flinch response were recorded for each stimulus and the 24 event-related cerebral potentials at each intensity were averaged to produce a mean waveform. Various components of this waveform were analysed and changes in these measures and the sheep's flinch response, as stimulus intensity increased, were determined.
Results Both the flinch response and some event-related cerebral potential components, that is, peak amplitudes 114 {N1}, 187 {P1}, 318 {P2} and 230 {Pm} ms after stimulus onset, were significantly affected as stimulus intensity increased.
Conclusion These corresponding behavioural and event-related cerebral potential changes indicate the usefulness of using changes in the event-related cerebral potential to measure acute pain in sheep.     

Sperm Motion Characteristics of Garole Rams Raised for a Prolonged Period in a Semi-arid Tropical Environment

The Garole is a prolific but less well known and rare breed of small sheep found in the hot and humid Sunderban region of West Bengal. An ability to breed throughout the year and to graze in knee-deep water, resistance to foot rot and a strong mothering instinct are some of the special features of this breed. Garole rams could provide germplasm to incorporate prolificacy traits by artificial insemination of the nonprolific sheep breeds found in abundance in the semi-arid and arid tropical climates of India. The aim of the present study was to evaluate the semen production by Garole rams maintained in a semi-arid climate for three years and to objectively assess their semen quality by a computer-assisted sperm analysis technique. The donor rams were randomly selected each year from the original flock procured from their natural habitat or from the offspring born at the Institute farm. Semen was collected weekly for three weeks each autumn for three consecutive years from 8 rams each year. The overall means (SD) of the traits that did not differ significantly with age or year were volume, concentration, curvilinear velocity, average path velocity, amplitude of lateral head displacement, beat frequency, motility and the percentages of rapid motile sperms and of slow motile sperms. The age of the rams had a significant effect (p<0.05) on the straight-line velocity but this was not significantly affected by the length of exposure to the semi-arid climate. However, the age and year had significant effects (p<0.05) on linearity, straightness and the percentage of medium motile sperms. It was concluded that Garole rams are capable of producing good-quality semen even after a prolonged period of exposure to a semi-arid tropical climate.     

The efficacy of diazinon impregnated ear tags against buffalo fly and resulting weight gains and diazinon residues in meat and milk

Ear tags impregnated with 20% diazinon were evaluated for their efficacy against the buffalo fly (Haematobia irritans exigua) on beef cattle in southern Queensland. Buffalo fly numbers and weight changes were recorded and diazinon residues in tissues of beef cattle and milk from lactating dairy cattle were assayed at different time intervals after tagging. In 2 efficacy trials conducted over 19 and 20 weeks, the mean numbers of buffalo fly on cattle each fitted with ear tags were 1 to 9 and 0 to 16, respectively, in trials 1 and 2, compared with 44 to 345 and 26 to 306 per head on untreated herds, respectively, despite regular spraying of the untreated herd in trial 1 with cyper-methrin to reduce fly burdens. Percentage buffalo fly control was 96.7 to 99.5% and 89.3 to 100% in the 2 trials. Cattle fitted with ear tags gained an average of 94 kg body weight after 5 months compared with 61 kg in the untreated herd, a net increase of 60% in treated animals compared with 28% in the untreated herd. Mean diazinon residue concentrations in the fat of perianal tissue biopsies were 0.02 to 0.03 mg/kg 1 to 8 weeks after tagging. Mean diazinon residue concentrations in the butterfat of milk from lactating dairy cattle were 0.01 to 0.04 mg/kg after tagging.     

Behaviour of Polyphenol Oxidase, IAA Oxidase and Catalase in Germinating Rice in Relation to Salt Tolerance

Activities of the enzymes polyphenol oxidase, indole-3-acetic acid (IAA) oxidase and catalase were compared from endosperms and embryoaxes of germinating rice ( Oryza sativa ) seeds of two sets of cultivars differing in salt tolerance, raised under increasing levels of NaCl salinity. Salt sensitive cvs. Ratna and Jaya were characterized by higher polyphenol oxidase activity in embryoaxes under salinisation as compared to salt tolerant cvs. CSR-1 and CSR-3. In endosperms of salt sensitive cultivars the enzyme activity decreased under salinisation whereas in tolerants it increased due to salinity. In vitro studies using desalted enzyme extracts showed a continuous increase in polyphenol oxidase activity with increasing concentration of NaCl (0–1000 mM) in the reaction medium. IAA oxidase activity increased due to salinity in embryoaxes of both sets of cultivars, the effect was greater in sensitive cultivars than tolerants. NaCl when incorporated in the reaction medium caused decrease in IAA oxidase activity. Activity of catalase decreased with increase in salinity with greater decrease in sensitive cultivars than tolerants. Results suggest possible different behaviours of oxidative enzymes in germinating seeds of rice cultivars differing in salt tolerance and that salt tolerance ability is associated with lower polyphenol oxidase and IAA oxidase activities and higher catalase activity in embryoaxes under salinisation.     

Identification of the gene homologous to HSV major DNA binding protein in the BHV-1 genome

By means of Southern blot hybridisation using a cloned herpes simplex virus (HSV) major DNA binding protein (MDBP) gene as probe, the putative MDBP gene of BHV-1 was located within the Hind III G fragment which mapped between 0.352 and 0.381 map units of the BHV-1 genome. Moreover, an antiserum raised to HSV MDBP precipitated a 120kD polypeptide in a radio-immunoprecipitation test.     

Serologic studies of Actinobacillus (Haemophilus) pleuropneumoniae strains of serotype-3 and their antigenic relationships with other A pleuropneumoniae serotypes in swine

Serotype-3 strains of Actinobacillus (Haemophilus) pleuropneumoniae possess epitopes shared with almost all other Actinobacillus serotypes. Common epitopes detected in particulate antigens were heat-labile and heat-stable and were of minor nature. Additional cross-reactive epitopes were detected in soluble and particulate antigens prepared from strains of serotypes 3, 6, and 8. Cross-reactions occurring between serotype-3 antigens and those of other serotypes were of 2 types: one associated mainly with 2-mercaptoethanol (2-ME)-sensitive IgM antibodies and the other associated mainly with 2-ME-resistant IgG antibodies. The cross-reactivity between serotypes 3, 6, and 8 was associated mainly with IgG antibodies, as shown by the results of 2-ME tube agglutination, 2-ME-indirect hemagglutination, and coagglutination tests. None of the tests was entirely satisfactory for serotyping serotype-3 isolates, mainly as a result of overlapping of type-specific antigenic determinants of serotypes 3, 6, and 8 in different combinations in the same strain. A combination of tests, using particulate and soluble antigens, may be necessary for typing of serotype-3 isolates.