Polarized epithelium-sperm co-culture system reveals stimulatory factors for the secretion of mouse epididymal quiescin sulfhydryl oxidase 1

Spermatozoa acquire fertilization ability through post-translational modifications. These membrane surface alterations occur in various segments of the epididymis. Quiescin sulfhydryl oxidases, which catalyze thiol-oxidation reactions, are involved in disulfide bond formation, which is essential for sperm maturation, upon transition and migration in the epididymis. Using castration and azoospermia transgenic mouse models, in the present study, we showed that quiescin sulfhydryl oxidase 1 (QSOX1) protein expression and secretion are positively correlated with the presence of testosterone and sperm cells. A two-dimensional in vitro epithelium-sperm co-culture system provided further evidence in support of the notion that both testosterone and its dominant metabolite, 5α-dihydrotestosterone, promote epididymal QSOX1 secretion. We also demonstrated that immature caput spermatozoa, but not mature cauda sperm cells, exhibited great potential to stimulate QSOX1 secretion in vitro, suggesting that sperm maturation is a key regulatory factor for mouse epididymal QSOX1 secretion. Proteomic analysis identified 582 secretory proteins from the co-culture supernatant, of which 258 were sperm-specific and 154 were of epididymal epithelium-origin. Gene Ontology analysis indicated that these secreted proteins exhibit functions known to facilitate sperm membrane organization, cellular activity, and sperm-egg recognition. Taken together, our data demonstrated that testosterone and sperm maturation status are key regulators of mouse epididymal QSOX1 protein expression and secretion.     

Insecticidal properties of some allethrin isomers used in coil formulations against mosquitoes

Previous studies have been made on the relative efficacy of the eight possible stereoisomers of allethrolonyl chrysanthemate. Some of these compounds and their mixtures have been classified according to their knockdown activity against Aedes aegypti. Two techniques were used, the contact method recommended by WHO and the method described by Katsuda and Ogami to study the effect on mosquitoes of the smoke from burning coils. Results show that on Aedes aegypti, the relative knockdown activity of the various stereoisomers of allethrin was about the same with both methods. The d trans chrysanthemate of d allethrolone (S-Bioallethrin) was the most effective.     

Cystoscopy in cattle--a valuable additional tool for clinical examination

Based on the findings of physical examination and on laboratory findings the urinary bladder of 23 cows was examined endoscopically in order to investigate the application of cystoscopy in cattle. The endoscopic findings of all examined cows were compared with the findings of physical examination and the results of macroscopic and microscopic urinalysis and the bacteriological culture of the urine. By physical examination only 3 cows were diagnosed to have urinary tract disease, whereas all other cows were suspected of having an urinary tract disease. Bacteriological culture of the urine revealed Corynebacterium renale and Escherichia coli infection in 18 cows, while the remaining 5 cows were negative. By cystoscopy catarrhal cystitis was diagnosed in 2 cases, haemorrhagic cystitis in 5 cases, and fibrinous-purulent and fibrinous-haemorrhagic cystitis in 13 cases. Three cows showed no pathological changes of the urinary bladder mucosa by endoscopic examination. Cystoscopy facilitates diagnosis through the direct visualisation of mucosal lesions and makes it possible to give a more accurate prognosis based upon the findings.     

Differentiation of Actinobacillus pleuropneumoniae by PCR-REA based on sequence variability of the apxIVA gene and by ribotyping

During the period of 2001-2003, a total of 591 Actinobacillus pleuropneumoniae field isolates from the Czech Republic were serotyped with a high occurrence of cross-reactions. The cross-reactions were observed in 416 isolates. Most frequently, in 401 isolates (67.9%), cross-reactions with antisera specific for serotypes 9, 11, and/or 1 were observed. Two additional molecular methods, ribotyping and restriction analysis of PCR amplified apxIVA gene (PCR-REA), were therefore used for detailed characterisation of A. pleuropneumoniae. In this subsequent analysis, reference strains representing serotypes 1-12 and 25 field isolates showing the most frequent serotype cross-reactions were examined. PCR-REA enabled all reference strains to be distinguished except for the strains of serotypes 9 and 11. Ribotyping distinguished all reference strains except two pairs of serotypes: 3 versus 6, and 9 versus 11, respectively. Field isolates with serotype cross-reactivity 9, 11, and/or 1 could not be differentiated by either of these methods.     

Pathophysiological reactions of the pulmonary circulation in airway and lung diseases

Hypoxic pulmonary vasoconstriction (HPV) is a physiological phenomenon in healthy lungs and a regulatory mechanism in alveolar hypoventilation by which blood flow is diverted from poorly ventilated areas to better ventilated alveolar regions. Maintained alveolar hypoxia (related either to respiratory diseases or to high altitude) results in a persistent activation of HPV. The latter mechanism is involved in the development of pulmonary vasoconstriction and right heart failure (Cor pulmonale) in chronic airway diseases. Compared to other species, especially the bovine and the porcine lung is highly susceptible to develop pulmonary hypertension because of anatomical and physiological pecularities. Consequently, therapeutic support of the heart is indicated in cattle and pigs suffering from respiratory diseases. In contrast, non-selective vasodilators are not recommended because they override the compensatory effect of hypoxic vasoconstriction and impair pulmonary gas exchange.     

Comparison of 13 single-round and nested PCR assays targeting IS900, ISMav2, f57 and locus 255 for detection of Mycobacterium avium subsp. paratuberculosis

For molecular biological detection of Mycobacterium avium subsp. paratuberculosis (MAP), PCR methods with primers targeting different regions specific for MAP are used worldwide. However, some uncertainties exist concerning the specificity of certain target regions and the sensitivity. To identify the methods which are best suited for diagnostics, 8 single-round and 5 nested PCR systems including 12 different primer pairs based on IS900 (9 x), ISMav2 (1x), f57 (1x), and locus 255 (1x) sequences were compared regarding their analytical sensitivity and specificity under similar PCR conditions. Reference strains and field isolates of 17 Mycobacterium species and subspecies, 16 different non-mycobacterial bovine pathogens and commensals were included in this study. Single-round PCR resulted in a detection limit of 100 fg to 1 pg, and nested PCR in 10 fg or below. Depending on the specific primer sequences targeting IS900, false positive results occurred with one of the five single-round and two of the four nested PCR systems. This also applied to the single-round PCR based on ISMav2 and the nested PCR based on f57. A high number of non-specific products were primarily detected for the single-round PCR assay based on ISMav2, but also for a single-round PCR targeting the IS900 and the locus 255. In conclusion, stringent selection of IS900-specific primers ensures that IS900 remains a favourite target sequence for amplification of MAP specific loci. The studied PCR systems based on f57, and locus 255 can also be recommended. Revision of ISMav2 primers is necessary. Single-round PCR systems are very reliable. Nested PCR assays were occasionally disturbed by contaminations, thus bearing a risk for routine diagnostics.     

Local Usage of Tiger Parts and Its Role in Tiger Killing in the Bangladesh Sundarbans

This article explored the local medicinal and traditional values of tiger parts and associated beliefs, and its link to the commercial trade in the Bangladesh Sundarbans. Using semi-structured qualitative interviews with 139 respondents, we found that the local use of, and belief in, the medicinal values of tiger parts is widespread and that virtually all parts of the tiger are used. Some of the local uses of tiger parts were unique in both the way and the purpose of use. For example, the soil of tiger pugmark was consumed by the women as a means of contraception. We established that local usage may be a significant threat to the tiger population of south Asia as it motivates stray tiger killing for collecting tiger parts for both local use and commercial demand, and provides the opportunities for poachers and the commercial trade to flourish.     

Prioritizing community behaviors to improve wild dog management in peri-urban areas

Wild dogs contribute to a range of negative impacts in Australian peri-urban regions. We identified and prioritized 13 behaviors that peri-urban residents could engage in to reduce the impacts of wild dogs in their communities. Key-informant interviews and stakeholder focus groups were used to develop a list of wild dog-management behaviors. On-line surveys with wild dog experts (n = 10) and peri-urban residents (n = 302) were used to create a Behavior Prioritization Matrix (BPM), which ranked the behaviors in terms of projected impact, based on: (a) expert estimates of effectiveness, (b) current adoption levels (penetration), and (c) likelihood of future adoption. BPM analysis indicated that increasing community reporting of wild dogs and their impacts would produce the greatest overall benefits for wild dog management. Behaviors rated as highly effective by experts (e.g., baiting and trapping) had low projected impact given that they were unlikely to be adopted by most residents.