Both A and B grade cumulus-oocyte complexes (COCs) aspirated from cattle ovaries at slaughter were matured in vitro under normal (38.5 °C) and elevated temperatures (41 °C). Maturation competence based on cumulus expansion, COC diameter, and nuclear maturation were compared with and without antioxidant supplementation incorporating 7.5 µM retinol, 1 nM melatonin, and 1.5 µg/mL zinc chloride in an oocyte maturation medium. Heat stress significantly reduced cumulus expansion by approximately 20%, while only retinol could bring a significant (P ≤ 0.05) increase. Heat stress also decreased expansion of A (approximately 50%) and B (approximately 40%) grade COC diameter. All antioxidants significantly increased COC diameter at 38.5 °C in grade A COCs, but only retinol could significantly increase grade B COCs. At 41 °C, only retinol in grade A COCs significantly enhanced diameter. Elevated temperature also decreased the metaphase II stage of nuclear maturation by approximately 75%, and no antioxidant was protective, except retinol, which was only marginally so. Retinol (7.5 µM) was further supplemented in maturation and culture medium for in vitro embryonic development at 38.5 °C and demonstrated significantly higher (P ≤ 0.01) maturation, fertilization, and a 2-4 cell cleavage rate. Retinol supplementation not only showed better maturation results, but also was a better antioxidant in overcoming the deleterious effects of elevated temperature.
Domesticated fowls, pigeons and turkey birds were screened for avipoxvirus infection from different areas in Jammu region. Based on typical pox lesions the overall occurrence in fowl was found to be 18.52%, 17.03% in pigeons and 57.14% in turkeys. Mortality recorded in chicks was 41.96%, 45.36% in squabs, 100% in poults, and 20.00% in adult turkeys. Both cutaneous and diphtheritic forms of the disease was observed of which the latter was particularly prevalent in young birds. One sample of putative fowlpox virus (FWPV) from skin lesions of a fowl, and two samples of putative pigeonpox virus (PGPV) from skin and diphtheritic lesions each were inoculated on chorio-allantoic membrane (CAM) of 10-12 days old chicken embryonated eggs. A confirmatory diagnosis was made by PCR amplification of a highly conserved P4b gene locus detected in tissue samples from skin, diphtheritic membrane and virus inoculated CAM yielding a predicted 578 bp product. Phylogenetic analysis based on the same P4b gene locus revealed FWPV and turkeypox virus (TKPV) to be 99% related and belonging to clade 1, while PGPV was found to belong to clade 2. All three isolates illustrate considerable heterogeneity within the conserved P4b gene locus. The study indicates that the closely related FWPV and TKPV isolates may have the potential of cross infection between fowls and turkeys and therefore cross transmission studies are suggested.
Transmission electron microscopy (TEM) was employed for describing skin and scab lesions in goats affected by orf virus and to demonstrate the parapoxvirus from clinical suspensions by negative staining and ORFV confirmation by immunogold electron microscopy. All samples were confirmed as parapoxvirus by semi-nested PCR amplification of partial gene encoding for the B2L envelope protein. Skin lesions were characterized by ballooning degeneration and loss of desmosomes of the spinosum cells, cytolysis and vesicle formation. Nuclear changes included chromatin margination and increase in electron density. Cytoplasmic changes were typical of cell swelling, vacuolation and the presence of uniform, moderately electron dense viroplasm, situated in the perinuclear region. Various intracellular forms including immature virions (IV), mature virions (IMV) and wrapped virions (WV) were observed in the cytoplasm. All these forms of ORFV were observed morphologically akin to vaccinia virus (VACV). Negative staining of clinical samples and viral suspensions showed typical parapoxvirus morphology with a characteristic criss-cross tubular surface
NOCARDIOSIS has a worldwide distribution and has been reported in various species of mammals, fish and birds (Beaman 1984, Bolon and others 1989, Megid and others 1990, Panigraphy and Senne 1991, Boiron and others 1993, Biberstein and Hirsh 1999, Chen and others 2000. Among birds, the disease has been observed in parrots, birds of prey, passeriformes, black crakes (Limnocorax flavirostra) and mynahs (Cooper 1973, Panigraphy and Senne 1991, Baumgartner and others 1994, Bacciarini and others 1999. The disease is of zoonotic importance, and it has been estimated that more than 1000 human cases of the disease are diagnosed each year in the USA (Schaal 1992). The frequency of nocardial diseases has increased over the past 20 years (Grange 2002). To the authors' knowledge there have been no reports of the occurrence and pathology of nocardiosis in pigeons. This short communication describes an outbreak of nocardiosis in domestic pigeons (Columba livia).Sixty-seven of 250 pigeons (26·8 per cent) 12 to 18 months of age died over a period of one year, singly or a few at a time, in a private pigeonry in Srinagar, in the Kashmir valley of India. The birds were reared for racing. The pigeons had also been procured from Delhi and from the Jammu region of Jammu and Kashmir state. They were maintained on grains of maize (Zea mays), bajra (Pennisetum typhoideum), wheat (Triticum sativum) and gram (Cicer arietinum), depending on their availability. The affected pigeons had lameness, drooping wings, depression, gasping and emaciation. External examination of the birds revealed yellowish nodules on the wing joints, humerus, radius, ulna, hock, tarsal and metatarsal joints, and on the skin. The nodules were sometimes enucleable, having a hard consistency. Removal of the skin of dead birds revealed the presence of nodules in the skin, mostly in the neck region. At postmortem examination, yellowish nodules were observed in the liver, lungs, air sacs, on the thoracic wall, pericardium and bones, with darkcentred, necrotic areas. Splenomegaly was additionally observed. Representative tissues from various organs, showing lesions were collected into 10 per cent formol saline; bones were processed after routine decalcification in formic acid-sodium citrate solution.Material collected from the liver, lungs and bone nodules was homogenised and cultured on blood agar plates containing 5 per cent sheep blood. These were incubated aerobically at 37°C for up to five to 10 days. Samples were also cultured on Sabouraud's dextrose agar. The characteristics of any colonies were noted, and the organisms were stained with Gram stain.After fixation, tissues were embedded in paraffin wax and 5 µm sections were cut and stained with Harris's haematoxylin and eosin (Luna 1968). Subsequently, sections were stained with Brown and Brenn Gram stain, the Ziehl-Neelsen and Fite methods for acid-fast organisms (Luna 1968) and the toludine blue-A rapid method (Humason 1979) for demonstrating mast cells. This was followed by the chrysoidin method (Harada 1957), f...
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