Effect of Progesterone and Ionomycin on Domestic Cat Sperm Motility Patterns and Acrosome Reaction

This study was aimed at assessing the changes in sperm motion patterns and the percentage of acrosome reaction (AR) in domestic cat semen after treatment with either ionomycin or progesterone (P₄). Ten ejaculates were collected from five tomcats using an artificial vagina, and were diluted, centrifuged and resuspended in a capacitation medium. Samples were evaluated and divided into seven equal aliquots and, after 2 h at 25°C, were incubated for 30 min at 38°C in 5% CO₂ and then analyzed. Computer-assisted sperm analysis and a combination of three fluorescent probes were used to assess sperm plasma, acrosomal membrane integrity and mitochondrial transmembrane potential. Thirty minutes after the start of incubation, P₄ was added (10 μg/ml) to the P1 group. Groups P2 and P3 were supplemented with P₄ (10 and 20 μg/ml, respectively) only after 2 h of incubation, and groups I1 and I2 were supplemented with ionomycin (4 and 8 μ m , respectively) 2 h after incubation. Group E was supplemented with ethanol (0.6%) at 2 h after incubation and group C received no supplementation. Ionomycin and P₄ treatments led to a hyperactivation-like sperm motion and an increase (p < 0.05) in the percentage of AR. Although a higher (p < 0.05) percentage of AR was obtained in group I2 when compared with all P₄ groups, a decrease (p < 0.05) in total and progressive motility was observed in I2 group. As I1 group was similar to I2 to induce AR without diminishing sperm motility, we can conclude that ionomycin at 4 μ m seems to be more suitable to trigger AR in domestic cat sperm.     

Beacon-like immunoreactivity in the hypothalamus of domestic chick

Beacon-immunoreactive (B-ir) fibres and neurons in the hypothalamus of the domestic chick (Gallus domesticus) were studied using an immunohistochemical technique in order to verify the presence and elucidate the pattern of distribution of this novel peptide in an avian brain. B-ir neurons were seen in the n. supraopticus, pars ventralis and pars externus; n. magnocellularis preopticus, pars dorsalis, medialis and ventralis; n. preopticus periventricularis; n. suprachiasmaticus, pars medialis; n. ventrolateralis thalami. Only few B-ir cells were scattered in the most anterior part of the lateral hypothalamic area. B-ir fibres, appearing as thin punctuate structures, were seen mainly along the walls of the third ventricle and in the ventromedial hypothalamus. Labelled fibres and terminals were located in the external and internal zones of the anterior and posterior median eminence. In particular, fibre terminals were seen close to the capillary loops of the hypothalamo-hypophysial portal system. The anatomical data of the present study regarding the distribution of B-ir in the chick hypothalamus suggest that beacon may play a key role in the regulation of the neuroendocrine system by acting as a neuromodulator and/or neurotransmitter.     

A survey of mutations in the Cochliomyia hominivorax (Diptera: Calliphoridae) esterase E3 gene associated with organophosphate resistance and the molecular identification of mutant alleles

Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species.     

Bluetongue in The Netherlands; description of the first clinical cases and differential diagnosis. Common symptoms just a little different and in too many herds

For the first time Bluetongue (BT) has been diagnosed in the Netherlands. The clinical symptoms of BT on five farms during the first outbreak ever in the Netherlands are described. Fever and swollen sensitive coronets leading to reluctance to stand and walk were sometimes the first symptoms. Later lesions in the mouth occurred with foamy salivation and respiratory problems. In other cases a swollen head with swollen lips and foamy salivation were the first clinical signs. Also sudden death occurred. In the first sixteen confirmed cases morbidity and mortality were lower than described in outbreaks in other countries. Good collaboration between practitioners, specialists of the Animal Health Service (GD-Deventer), and specialists of the Food and Consumer Product Safety Authority (VWA) and CIDC-Lelystad (Wageningen UR) led to a rapid notification and ultimately confirmation of the suspected diagnosis BT.     

DNA diagnosis in veterinary medicine: II. Polymerase chain reaction (PCR)

DNA diagnostic has become very important for genetic and infectious diseases. With a new method, called the polymerase chain reaction (PCR), specific pieces of genomes from viruses, bacteria, parasites or even cells can be amplified more than hundred million fold in a very short time period. For the following reasons this opens a new dimension in diagnostics: (1) The technique is so sensitive that a single virus, bacterium, parasite or cell is sufficient to be detected provided part of its nucleic acid sequence is known. (2) The material does neither need to be fresh nor be stored under special conditions since the nucleic acids are much more stable than e.g. enzymes or other proteins. (3) The method is quicker than the usual immunodetection methods. (4) The polymerase chain reaction has been standardized recently so that it can be carried out in diagnostic laboratories. The purpose of this minireview is to give an introduction into the basic technique and to mention possible and already existing applications in veterinary medicine.     

The biochemical, morphological and virulence profiles of Bacillus anthracis isolated in the Kruger National Park

The biochemical, morphological and virulence profiles of 44 Bacillus anthracis isolates, obtained from various localities and carcass remains of wild animals in the Kruger National Park, South Africa, were examined. The morphological characteristics tested for included: the formation of capsules on bicarbonate agar, the motility of the vegetative organism, the presence of haemolysis on blood tryptose agar, the sensitivity of the vegetative organism to bacteriophage, the production of lecithinase on egg yolk agar, the liquefaction (hydrolysis) of gelatine and the capability of each isolate to produce mucoid colonies when grown on bicarbonate agar with horse serum in an atmosphere containing CO2. The API 50CHB and 20E systems were used to evaluate the biochemical activity of each isolate. The virulence of each isolate was determined by its LD50, using an inbred line of Balb/C mice. A clear pattern in the biochemical reactions emerged that appeared to be specific for each isolate. On the API 50CHB test strip, only 2 of the 44 isolates gave a 100% positive reaction to all 10 of the biochemical substances to which it was supposed to react, 9 gave positive results to 90%, 19 were positive to 80%, and 14 were positive to 70%. The reactions on the API 20E were completely different from what was expected, with only 1 of the biochemical activities (gelatinase production) showing a positive reaction to all the isolates. The virulence test indicated that 27/44 isolates could be regarded as highly virulent with a LD50 of less than 1,000 organisms, and the rest of the isolates as virulent with a LD 50 of 1,001-10,000 organisms.(ABSTRACT TRUNCATED AT 250 WORDS)     

A dermoid of the eye in a blue-fronted Amazon parrot (Amazona aestiva)

A corneo-conjunctival dermoid is reported in a blue-fronted Amazon parrot ( Amazona aestiva ). After laminar keratectomy, histology showed the epidermis with feather follicles and dermal connective tissue with lymph follicles and sebaceous glands.