PCR-sequence characterization of new adenoviruses found in reptiles and the first successful isolation of a lizard adenovirus

A consensus nested PCR was used to screen diagnostic samples from approximately 70 reptiles for the presence of adenoviruses (AdV) in the years 2006-2007. Classical virus isolation methods were also used with all samples. After adenoviruses were detected in a group of helodermatid lizards in a Danish zoo, a follow-up study was also carried out on lizards from this group (10 Mexican beaded lizards and 24 Gila monsters) over the period of a year. Adenoviruses were detected in a total of 26 lizards and snakes by PCR. The PCR amplicons from all positive animals were sequenced and the resulting polymerase gene sequences were used for phylogenetic analysis. Altogether six Agamid AdVs were amplified, with a minimal sequence variation between one another and between these and GenBank Agamid AdVs. The sequence obtained from one of the Gila monsters is identical with the GenBank Helodermatid AdV, while the sequences from the Mexican beaded lizards differ from this. In a snake collection we have detected a new AdV from an Asp viper. All of the above mentioned adenoviruses cluster in the Atadenovirus genus. However, the sequence from a new Varanid AdV detected in this study clusters outside this genus. On cell culture, viruses were isolated from three of the AdV positive helodermatid lizards (one Mexican beaded lizard and two Gila monsters) and identified as AdVs based on electron microscopy and PCR and sequencing using cell culture supernatant. This is the first report of the successful isolation of a lizard AdV.     

Type and frequency of contacts between Belgian pig herds

Knowledge of the frequency of direct and indirect contacts between pig herds is a requirement for understanding the potential between-herd transmission of pathogens. Our aim was to investigate the different contacts between Belgian pig herds. We obtained data by conducting a postal survey on 421 pig herds in August 2005 and by analysis of available information on livestock movements in the national identification-and-registration database (18-months period in 2004-2006). Direct contacts included transports of pigs by onto-farm, off-farm and between-farm movements. Indirect contacts included vehicles entering the herd and visitors entering the stables. The median number of direct contacts per herd made by onto-farm movements was 0.2/month (Q1: 0; Q3: 0.5). About 1.2% of herds had > or = 3 onto-farm movements/month. We used a zero-inflated negative-binomial regression model to describe differences in the number of onto-farm movements according to herd size and herd type. Piglet multipliers followed by finishing herds were predicted to have the most onto-farm movements. Farrow-to-finishing herds made less movements compared to breeding herds. A median of 3997 between-farm movements/month was made in Belgium; these mainly concerned piglets. The median number of origin herds during an 8-month period for between-farm movements was 4 (Q1: 2; Q3: 8). For a typical 1-month period, we constructed directed graphs of between-farm piglet and replacement stock movements, illustrating potential receivers and distributors of infection. Of these between-farm movements, many were made over a short distance (median straight-line distance 19 km (Q1: 8; Q3: 36)). The median number of vehicles entering a herd and visitors entering the stables was 8/month (Q1: 6; Q3: 13) and 3/month (Q1: 2; Q3: 6) respectively. The number of indirect contacts by vehicles and persons were associated with herd size (Spearman's r: 0.7 and 0.2), herd type and other factors. Skewness of both direct and indirect contacts, illustrated that there was a wide variety in contact structure between pig herds in Belgium. Infection control might benefit by accounting for this variation in contacts and by targeting 'high-risk' herds in case of animal-disease emergencies.     

The effect of intravenous administration of variable-dose midazolam after fixed-dose ketamine in healthy awake cats

The effects of intravenous administration of variable-dose midazolam and ketamine (3 mg/kg) were studied in twelve healthy unmedicated cats from time of administration until full recovery. A range of midazolam doses (0.0, 0.05, 0.5, 1.0, 2.0 and 5.0 mg/kg) was chosen, so that beneficial and/or detrimental effects could be documented and the therapeutic window for further study determined. One minute after administration of ketamine, all cats had assumed a lateral position, mostly with head up. Muscle tone was increased (100%), apneustic breathing pattern evident in 92% of cats, chewing without stimulation of the oropharyngeal area was observed in most cats (97%), but most cats did not salivate (87%). At 2.5 min after completion of ketamine injection and 1 min after administration of saline, a similar picture was observed, except that salivation was evident. All cats chewed or swallowed in response to a finger or laryngoscope placed in the oropharyngeal area and, while most cats were not aware of a noxious stimulus to the tail, some cats were aware of a noxious stimulus to the paw. Recovery from ketamine alone was rapid and smooth with cats rolling into sternal recumbency and then cautiously walking with ataxia. Recovery to walking without incoordination was also rapid (< 2 h) and no abnormal behavioural patterns were observed during recovery. Administration of midazolam after ketamine, had beneficial effects and the therapeutic window for midazolam was found to lie between 0.05 mg/kg and 0.5 mg/kg. Administration of any dose of midazolam after ketamine caused a greater proportion of cats to assume a laterally recumbent position with head down compared with ketamine alone, however, the time period of recumbency was only significantly longer with a midazolam dose of 2.0 mg/kg or above. Doses of midazolam of 0.5 mg/kg or above decreased muscle rigidity but did not affect salivation or respiratory pattern observed in cats which received ketamine alone. A significantly greater proportion of cats which received ketamine and midazolam 0.5 mg/kg or above did not swallow in response to a finger or a laryngoscope placed in the mouth compared with that which received ketamine alone. The length of time in which cats did not swallow was only significantly longer at midazolam doses of 1.0 mg/kg and above. At midazolam doses of 0.5 mg/kg or above, the proportion of cats without a nociceptive response to a tail or paw clamp was significantly greater than cats which received ketamine alone. The time period without nociceptive response, however, was not influenced by midazolam administration. The time taken for cats which received ketamine and midazolam 0.05 mg/kg or 0.5 mg/kg to assume sternal position, walk with ataxia, walk without ataxia, behave normally when approached or restrained and recover normal arousal state was not significantly different from cats which received ketamine alone. Ketamine and midazolam 5.0 mg/kg significantly prolonged all recovery times compared with ketamine alone. Unfortunately, a greater proportion of cats which received ketamine and midazolam 0.5 or 5.0 mg/kg exhibited detrimental behavioural effects. These were more likely to be adverse and included restlessness, vocalization and difficulty approaching and restraining cats. In this study, an     

Fenbendazole-related drug residues in milk from treated dairy cows

Oral administration of [¹⁴C] fenbendazole (FBZ) at a dose of 5.Omg/kg leads to the presence of radiolabel in the milk of lactating dairy cows. However, the maximum mean concentration of total FBZ equivalents quantitated to one-third of the recommended safe concentration in milk (1.67 μg/mL). The label is equally distributed to the fat and aqueous portions of the milk. The maximum level, in general, is attained approximately 24-36 h after drug administration, with the highest levels ranging from 24 to 48 h after administration. The residues rapidly deplete, attaining levels of 10-20ng/mL by day 5, and are essentially undetectable by radiolabel monitoring by day 6. Extraction of the milk by matrix solid phase dispersion indicated that the label was distributed between traces of the parent drug, FBZ, and predominantly, the FBZ sulphoxide (SO) and sulphone (SO₂) metabolites. No other radiolabelled peaks were observed. Based on these data the metabolites of FBZ, FBZ-sulphone and FBZ-sulphoxide, could be used as marker residues for monitoring the administration of FBZ to lactating dairy cows.     

CONGENITAL DISLOCATION OF THE DEEP DIGITAL FLEXOR TENDON ASSOCIATED WITH HYPOPLASIA OF THE SUSTENTACULUM TALI IN A THOROUGHBRED COLT

An 11-month-old Thoroughbred colt was presented with a hard swelling at the proximal third of the right 4th metatarsal bone. A medial dislocation of the deep digital flexor tendon ( flexor digitorum profundus ) was also observed on the same leg. On the plantaroproximal-plantarodistal projection of the calcaneus, there was flattening and shortening of the sustentaculum tali. The smooth bony proliferation at the proximal third of the right 4th metatarus was compatible with a chronic splint bone fracture. This report describes a medial deep digital flexor dislocation associated with hypoplasia of the sustentaculum tali .     

Comparative metabolism of R(-)-fenoprofen in rats and sheep

The chiral inversion of 2-arylpropionic acids occurs in many species. It is a unique reaction specific to this group of drugs. In this study R-(-)-fenoprofen (R-(-)-FPF) was used as a model compound to investigate metabolic chiral inversion in sheep in vivo and in vitro and to compare the data with the results obtained in rats. Metabolic inversion in sheep was 80%. The apparent mean values of Km and V _max of thioester formation were: 392 μm and 2.08 nmol/min/mg in sheep and 500 μm and 22 nmol/min/mg in rats. For hydroxylation, the apparent mean values were V_max: 0.02 nmol/min/mg in rats and 0.01 nmol/min/mg in sheep. There was no correlation between in vitro thioesterification and in vivo chiral inversion in sheep as compared to rats. In sheep most of the thioester formed underwent inversion (80%) while in rats, where in vitro thioesterification was greater, in vivo inversion was less (42%). In consequence, in rats other metabolic pathways for R(-)-FPF-CoA, such as incorporation into triacylglycerols and conjugation with amino acids, may be quantitatively more important     

Performance of sows fed high levels of nonstarch polysaccharides during gestation and lactation over three parities

The effect of feeding sows a starch diet or a diet with a high level of nonstarch polysaccharides (NSP) during gestation, lactation, or both gestation and lactation during the first three parities on reproductive performance, body weight, and backfat was studied. Four-hundred and forty-four postpuberal gilts were allotted to a 2 x 2 x 2 factorial experiment. Treatments were diet composition during gestation (including the weaning-to-estrus interval; G-Starch: 274 g/kg of starch and 123 g/kg of fermentable NSP or G-NSP: 86 g/kg of starch and 300 g/kg of fermentable NSP), diet composition during lactation (L-Starch: 293 g/kg of starch and 113 g/kg of fermentable NSP or L-NSP: 189 g/kg of starch and 216 g/kg of fermentable NSP) and group-housing system during gestation (free access stalls or electronic feeding). Both gestation diets were formulated to be isoenergetic. During lactation, sows were given free access to the lactation diets from d 6 after parturition onwards. Body weight and backfat gains during gestation were lower in sows fed the G-NSP diet than in those fed the G-starch diet (P < 0.001). The effects were more pronounced in the electronic feeding system than in the free access stalls. These results indicate an overestimation of the energy value of fermentable NSP. Body weight and backfat losses during lactation were less in sows fed the G-NSP diet during gestation than in those fed the G-starch diet (P < 0.05),which can be explained by a 0.4 kg/d higher (P < 0.001) feed intake during lactation of the sows fed the G-NSP diet. Sows fed the L-NSP diet lost more backfat during lactation than sows fed the L-starch diet (P < 0.05). The number of total piglets born and live-born piglets was 0.5 piglet higher in sows fed the G-NSP diet than in those fed the G-starch diet (P < 0.05). Lactation diet did not affect the number of total piglets born or live-born piglets. This study shows that, although high NSP diets negatively influence body weight and backfat thickness of the sows, it is possible to feed sows a diet with a high level of fermentable NSP diet during both gestation and lactation without negative effects on reproductive performance. Under the conditions of this study, feeding sows a diet with a high level of fermentable NSP during gestation and a high level of starch during lactation seems the most favorable feeding strategy.     

Comparison between Haemophilus parasuis infection in colostrums-deprived and sow-reared piglets

The aim of this study was to compare the development of Glasser's disease in sow-reared and colostrum-deprived piglets. Ninety piglets from a commercial pig farm in Spain were used. The farm was positive for Haemophilus parasuis. Fifty-two pigs were sow-reared (SR) and 38 were colostrum-deprived (CD) piglets. The animals were intratracheally inoculated with H. parasuis serovar 5 and sacrificed at 1, 2 and 3 days post-infection. To assess the development of disease, antibody titers, clinical signs, pathological lesions, microbiological isolation and PCR amplification were compared between the groups. Inoculation of SR pigs did not cause clinical signs or lesions of Glasser's disease. In SR pigs, H. parasuis isolation and specific PCR amplification from tissues showed a very low number of positive samples. In contrast, in CD pigs, inoculation resulted in the typical signs and lesions of Glasser's disease. Positive microbiological isolation and specific PCR products were obtained from the majority of the tissues tested, and no antibodies against H. parasuis were detected. The experimental infection using CD pigs describes a successful method to study this microorganism and confirms the important role that maternal antibodies play in protection against clinical signs and disease.     

Aldicarb poisoning of dogs and cats in Gauteng during 2003

A survey of aldicarb poisoning in companion animals was conducted by posting questionnaires to all private practitioners in Gauteng Province, South Africa. The survey was designed to determine the percentage of aldicarb cases seen, treatment regimen, clinical signs observed, proposals for preventative actions and more effective treatments. Other questions included duration of treatment, survival rate, cost to clients, post mortem findings and reasons for poisonings.     

Endocrine and ovarian changes in response to the ram effect in medroxyprogesterone acetate-primed Corriedale ewes during the breeding and nonbreeding season

Two experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4-6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17beta were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17beta concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17beta, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17beta levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17beta concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.