High doses of highly pathogenic avian influenza virus in chicken meat are required to infect ferrets

High pathogenicity avian influenza viruses (HPAIV) have caused fatal infections in mammals through consumption of infected bird carcasses or meat, but scarce information exists on the dose of virus required and the diversity of HPAIV subtypes involved. Ferrets were exposed to different HPAIV (H5 and H7 subtypes) through consumption of infected chicken meat. The dose of virus needed to infect ferrets through consumption was much higher than via respiratory exposure and varied with the virus strain. In addition, H5N1 HPAIV produced higher titers in the meat of infected chickens and more easily infected ferrets than the H7N3 or H7N7 HPAIV.     

Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

Origin, course and distribution of the hypoglossal nerve in the New Zealand rabbit (Oryctolagus cuniculus L)

With 8 figures SUMMARY: This study aimed at revealing the origin, course and distribution of the hypoglossal nerve in 20 adult male New Zealand rabbits. In all the animals dissected, the hypoglossal nerve arose from the ventrolateral side of the medulla oblongata with two main roots and gave off a descending branch to the ansa cervicalis before reaching the division of the common carotid artery. This branch was not seen on the right side of only one case. At the lateral aspect of the hyoglossus muscle, the nerve then divided into the lateral and medial main branches, sent branches to the styloglossus, hyoglossus, genioglossus and geniohyoideus muscles and terminated in the intrinsic tongue muscles. A communicating branch was observed between the hypoglossal and accessory nerves in the right side of one animal and between the hypoglossal nerve and the ganglion nodosum in the right retropharyngeal area of another animal. An additional branch was observed innervating the stylohyoideus muscle in one animal only. A lateral lingual-hypoglossal communication was also seen between the lateral branch of the hypoglossal nerve and terminal branches of the lingual nerve.     

In vitro inhibition of Citrobacter freundii, a red-leg syndrome associated pathogen in raniculture, by indigenous Lactococcus lactis CRL 1584

Red-leg syndrome (RLS) is one of the main infectious diseases that cause economic losses in Lithobates catesbeianus hatcheries, Citrobacter freundii being an etiological agent. Treatment or prevention with therapeutics or chemicals results in modifications of the indigenous microbiota, development of antibiotic resistance, presence of their residues in food and enhancement of production costs. Thus, probiotics could be used as an alternative therapy. Lactic acid bacteria are part of the indigenous microbiota of healthy frogs and can prevent pathogen colonization by different mechanisms, including the production of antagonistic substances. In this work, the evaluation and characterization of the inhibition of C. freundii CFb by Lactococcus lactis subsp. lactis CRL 1584, a potentially probiotic candidate, were carried out. This strain produced lactic acid, H(2)O(2) and bacteriocin in static and shaken conditions and inhibited pathogen growth in associative cultures, with an earlier inhibition under agitated conditions. The elimination of each of the antimicrobial metabolites partially abolished the inhibition of the pathogen, suggesting that the inhibitory effect could be attributed to a combined action of the three antagonistic molecules. Electron microphotographs revealed the damage caused by L. lactis CRL 1584 supernatants to C. freundii cells. The addition of pure lactic acid, H(2)O(2) and bacteriocin to the culture media showed that each metabolite caused different morphological modifications in C. freundii, in agreement with the effect on viable cell counts. The results support the possibility that L. lactis CRL 1584 might be considered as a probiotic to be used in the prevention of RLS in raniculture.     

Resistance to cypermethrin, deltamethrin and chlorpyriphos in populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from small farms of the State of São Paulo, Brazil

This study was undertaken to determine the flea diversity on urban dogs and cats in Australia in 2009-2010. A total of 2530 fleas were recovered from 291 animals (151 dogs, 69 cats and 71 uncategorised dogs or cats) from veterinary clinics across five states of Australia. The majority of specimens were from coastal areas. The cat flea (Ctenocephalides felis felis) was the most frequent flea species identified (98.8%, 2500/2530). The only other flea species identified was the stickfast flea (Echidnophaga gallinacea) from Western Australia. Sequencing of the cytochrome oxidase subunit II mtDNA revealed a single haplotype across Australia within a subset of C. f. felis (n=19). Our study demonstrated dominance and haplotype homogeneity of C. f. felis on dogs and cats. Although Ctenocephalides canis was recovered from a feral fox, it was not identified from the sample of fleas analysed. This suggests that, under current conditions, it is unlikely that foxes are reservoirs of C. canis for domestic dogs or cats residing in coastal Australia, as previously speculated.     

Use of a gonadotrophin-releasing hormone vaccine in headshaking horses

The aim of this study was to investigate the use of a gonadotrophin-releasing hormone (GnRH) vaccine in the treatment of headshaking in horses. Fifteen geldings received two doses of the GnRH vaccine four weeks apart. Serum was collected before and after vaccination to measure concentrations of luteinising hormone (LH) (10 horses) and follicle-stimulating hormone (FSH) (six horses). Owners recorded the frequency of seven common headshaking behaviours using a visual analogue scale (VAS) before vaccination and at two, four, eight, 12, 16 and 20 weeks after vaccination. Serum LH (P=0.008) and FSH (P=0.03) concentrations decreased significantly following vaccination. Although approximately one-third of the owners reported a subjective improvement in headshaking, serial scoring did not indicate a reduction in headshaking behaviours following vaccination with a commercial GnRH vaccine. Vaccination reactions were observed in four of 15 horses (27 per cent), including one case of severe, presumed immune-mediated, myositis.     

Subcutaneous electronic identification in cattle: a field study

The use of injectable transponders in cattle for identification purposes, for up to 30 months, was investigated. Passive electronic transponders, encapsulated in either polymer or glass, were injected subcutaneously into either the ear base or the earlobe of 652 calves in three populations. The animals were clinically examined weekly, and transponder signalling was checked immediately before and after injection, after two weeks and after about eight months. About 10 per cent of animals in one population were also checked after 20 and 30 months. No severe clinical or visible pathological changes were observed, and the calves' welfare was not apparently affected by the procedure. None of the transponders migrated from their injection sites. Eight months after injection, a signal was detected from 98.2 per cent of transponders injected in the ear base and from 90.5 per cent of those in the earlobe. At 20 and 30 months after injection, 10.4 per cent and 2.6 per cent of transponders, respectively, had ceased to signal. Thus, most transponders in the calves' ear base demonstrated functionality for up to 30 months.