Molecular epidemiology of a reproductive tract-associated colibacillosis outbreak in a layer breeder flock associated with atypical avian pathogenic Escherichia coli.

The molecular epidemiology of 70 Escherichia coli isolates from an infection outbreak in a layer breeder flock was examined by pulsed-field gel electrophoresis and for a range of virulence factors by polymerase chain reaction. Pulsed-field gel electrophoresis showed 35 of 45 isolates from eight disease cases were associated with a single clonal group that was the exclusive strain associated with reproductive tract. A second unrelated group was found in environmental isolates and healthy birds. The remaining isolates were unrelated to each other or either clonal group. Polymerase chain reaction virulotyping indicated the "epidemic" clonal group contains virulence factors including iss, sfa, tsh, iucC, ibeA, and sitA associated with avian pathogenic E. coli plus several virulence factors more normally associated with human urinary tract infection. Significantly, the "epidemic" clone was also found in an environmental sample, suggesting it may have been transmitted to the flock via the environment.

Observations on salpingitis, peritonitis and salpingoperitonitis in a layer breeder flock.

A flock of 13,951 hens and 1379 cockerels was monitored from 26 to 58 weeks of age for the complex of salpingitis, peritonitis and salpingoperitonitis (sps). Two hundred and forty-three hens (78 per cent of the hens that died) were examined postmortem, and sps was recognised by gross examination for inflammatory exudate, in the body cavity or oviduct in 111 (46 per cent) of them. Salpingoperitonitis was the most common form, followed by salpingitis and then peritonitis. There were acute and chronic cases in all three conditions, but only in peritonitis were acute cases more common than chronic cases. Seventeen birds that had died of sps were cultured for aerobic bacteria within 12 hours of death. Escherichia coli was recovered from a variety of tissues from all of them, and other bacteria, including staphylococci, Mannheimia haemolytica and Streptococcus bovis, were isolated from a few carcases, either alone or together with E coli. Relatively few isolations of E coli were made from normal hens cultured 48, 72 and 96 hours after death.

Gasless endoscopic anterior lumbar interbody fusion utilizing the B. E.R.G. approach.

BACKGROUND: Several authors have reported success using a gas-mediated transperitoneal approach for lumbar interbody fusion. However, this approach has not been shown to reliably and predictably address segments above L4-5. METHODS: The B.E.R.G. approach was attempted in 202 patients who required anterior lumbar interbody fusion (ALIF). Of those, 168 were completed successfully without conversion to an open procedure. The anterior retroperitoneal approach required no gas insufflation. The gasless environment allowed for the use of standard anterior instrumentation and a variety of fusion grafts and devices. RESULTS: Mean hospital stay was 1.95 days, with 73% of patients discharged in <47 h following surgery. Clinical results from the first 50 patients, with a minimum 2-year follow-up, include a 92% fusion rate and 78% of patients reporting significant pain relief of greater than 50%. CONCLUSIONS: The B.E.R.G. approach offers significant technical advantages over the standard gas-mediated transperitoneal approach for ALIF. The clinical results are similar to those reported for open approaches and the gas-mediated transperitoneal approach.

Minimally invasive 360 degrees instrumented lumbar fusion.

A retrospective preliminary study was undertaken of combined minimally invasive instrumented lumbar fusion utilizing the BERG (balloon-assisted endoscopic retroperitoneal gasless) approach anteriorly, and a posterior small-incision approach with translaminar screw fixation and posterolateral fusion. The study aimed to quantify the clinical and radiological results using this combined technique. The traditional minimally invasive approach to the anterior lumbar spine involves gas insufflation and provides reliable access only to L5-S1 and in some cases L4-5. A gas-mediated approach yields many technical drawbacks to performing spinal surgery. A minimally invasive posterior approach involving suprafascial pedicle screw instrumentation has been developed, but without wide-spread use. Translaminar facet fixation may be a viable alternative to transpedicular fixation in a 360 degrees instrumented fusion model. Past studies have shown open 360 degrees instrumented lumbar fusion yields high arthrodesis rates. The study examined the cases of 46 patients who underwent successful 360 degrees instrumented lumbar fusion using a combined minimally invasive approach. Anterior lumbar interbody fusion (ALIF) at one or two levels was performed through the BERG approach; a gasless retroperitoneal approach to the lumbar spine allowing the use of standard anterior instrumentation. Posteriorly, all patients underwent successful decompression, translaminar fixation, and posterolateral fusion at one or two levels through one small (2.5-5.0 cm) incision. Results showed mean hospital stay of 2.02 days; mean combined blood loss was 255 cc; and mean pain relief was 56%, with 75.5% of patients reporting good, excellent, or total pain relief. Forty-two of 46 patients (93.2%) achieved a solid fusion 24 months after surgery. A total of 47% of all patients working prior to surgery returned to work following surgery. The study showed that minimally invasive 360 degrees instrumented lumbar fusion, when performed utilizing these approaches, yields a high rate of solid arthrodesis (93.3%), good pain relief, short hospital stays, low blood losses, accelerated rehabilitation, and a quick return to the workforce. The BERG approach offers technical advantages over the traditional gas-mediated laparoscopic approach to the anterior lumbar spine.

The comparison of an aqueous preparation of tilmicosin with tylosin in the treatment of Mycoplasma gallisepticum infection of turkey poults.

A virulent strain of Mycoplasma gallisepticum (MG) was used to infect groups of 40 2-day-old poults kept in separate pens of 10 each. Of the six groups, three were treated with separate concentrations of tilmicosin, one was treated with tylosin, one remained untreated, and a final group was not infected and not treated. Mortality, clinical signs, and gross lesions were significantly less (P < 0.001) in the uninfected and infected medicated groups than in the infected unmedicated group. Also, the mean body weight gain of poults surviving to the end of the experiment was greater (P < 0.005) in the uninfected and infected medicated groups. MG was not recovered from the uninfected birds, and, among the infected poults, it was recovered from significantly fewer (P < 0.05) poults in the medicated groups. Serologic results were negative for the uninfected group, and there were fewer positive reactors for the infected medicated than the infected unmedicated group. In consideration of these results, tilmicosin should prove to be a useful addition to the antimicrobials in the treatment of MG infection in poults.

Experimental infection of chickens with Campylobacter jejuni: strains differ in their capacity to colonize the intestine.

Groups of broiler and layer type chickens (25 to 63 d.o.) were inoculated per os with separate isolates of 10 strains of Campylobacter jejuni. Nine of the 10 strains were originally isolated from chickens and one from a dog. The dog strain and five of the chicken isolates could be isolated after inoculation, but four strains were not recovered from cloacal swabs for up to 4 to 16 days after inoculation. However, it was possible to isolate C. jejuni from these birds, from cloacal swabs, when they were inoculated with organisms which had been previously shown to colonize other birds.

In vitro and in vivo comparisons of valnemulin, tiamulin, tylosin, enrofloxacin, and lincomycin/spectinomycin against Mycoplasma gallisepticum.

The minimum inhibitory concentrations (MICs) for valnemulin, tiamulin, enrofloxacin, tylosin, and lincomycin/spectinomycin were determined for a virulent strain of Mycoplasma gallispeticum (MG). At the initial reading, the lowest MICs were seen with valnemulin and tiamulin, followed by tylosin, enrofloxacin, and a relatively high MIC for lincomycin/spectinomycin. At the final reading, at 14 days, a similar pattern was obtained, with valnemulin giving the lowest MIC (< 0.008 mg/ml). The same strain of MG was used to infect groups of 20 2-day-old chicks in two separate experiments. In both, several concentrations of valnemulin and tiamulin and one each of tylosin and enrofloxacin were administered to separate groups in the drinking water. In the second experiment, one group of chicks was given lincomycin/spectinomycin. Each experiment had one infected unmedicated group and an uninfected unmedicated group. Mortality, clinical signs, and gross lesions, in both experiments, were significantly less (P < 0.001) in the uninfected and infected medicated groups (except for the two lowest dosages of valnemulin, lincomycin, and spectinomycin) than in the infected unmedicated groups. Also, the mean body weight gain was greater in the uninfected and infected medicated groups. Among the infected birds, MG was recovered from fewer chicks in the infected medicated groups except for the lowest two dosages of valnemulin. Serologic results were negative for the uninfected groups, and there were fewer positive reactors for the infected medicated groups except for the group treated with lincomycin/spectinomycin. Valnemulin should prove to be a useful addition to the antimicrobials in the control of MG infection in chickens.

The impact of non-endotoxin LAL-reactive materials on Limulus amebocyte lysate analyses.

Limulus amebocyte lysate (LAL) is activated by bacterial endotoxins and certain glucans (beta-D-glucan, LAL-RM). The potential for conflicting inter-laboratory results for LAL tests exists because commercial LAL reagents are highly variable in response to LAL-reactive glucans. The nature of beta-D-glucan activation of LAL and means for rendering LAL non-responsive to glucan are reviewed to provide a background for resolving conflicting data. Kinetic LAL methods are particularly useful for screening materials potentially contaminated with glucan. The presence of beta-D-glucan in parenterals is uncommon and is likely limited to products exposed to microbial or cellulosic materials. A scheme is suggested for identifying LAL-reactive glucans and for LAL release-testing without glucan interference.

The minimum inhibitory concentration of tilmicosin and tylosin for mycoplasma gallisepticum and Mycoplasma synoviae and a comparison of their efficacy in the control of Mycoplasma gallisepticum infection in broiler chicks.

The minimum inhibitory concentrations of tylosin tartrate and a new macrolid antimicrobial agent, tilmicosin, were assessed for six strains of Mycoplasma gallisepticum (MG) and three strains of Mycoplasma synoviae (MS) in vitro by the microbroth method. For four of the strains of MG, tilmicosin showed a slightly lower minimum inhibitory concentration (MIC) than did tylosin at both the initial reading (when pH 7.0 is first seen in the dilutions under test) and the final reading at 14 days of incubation. For one of the remaining strains, the MIC for tilmicosin was equal to or less than that for tylosin at the initial reading but greater at the final reading. For the other strain, the MIC for tilmicosin was greater than for tylosin, and for both of them the MICs were very much higher than for other strains. For the three strains of MS, there was little difference between the two drugs for one strain whereas the MIC for tilmicosin was slightly less for the other two groups. Groups of 30 chicks were infected with a virulent strain of MG and treated with either tylosin (0.5 g/liter) or tilmicosin (at concentrations of 0.125, 0.25 or 0.5 g/liter). One infected group was untreated and another group was uninfected and untreated. Clinical signs, mainly depression and nervous signs, were seen in two to five birds in the infected treated groups. In contrast, in the infected untreated group, 16 of 30 birds showed clinical signs. Mortality was significantly less in the infected treated groups compared with the infected untreated group (P < 0.001), and following infection there were significantly (P < 0.05) greater weight gains in the infected medicated groups. At necropsy the prevalence of gross lesions of the airsac walls was similar in all the infected medicated groups and was less than that for the infected unmedicated group. For the group on tylosin, MG was recovered from five chicks during life and from six dead chicks. The corresponding figures for the group receiving the lowest dose of tilmicosin were four for each; however, the organism was not recovered from the groups on the higher doses of tilmicosin either during life or from dead chicks. Serological results were negative for all groups except the infected untreated group, in which all three birds that were tested were positive.

A survey of the viral flora of two commercial Pekin duck flocks.

Ducklings on a problem farm which showed persistent and unacceptably high mortality yielded a larger range and greater number of viruses than did ducklings from a second flock, in which mortality was of a power and acceptable level. Reoviruses were the viruses most frequently isolated from young birds from both farms, but for longer at the problem site. ELAs (Embryo Lethal Agents), named since they caused high mortality in chick embryos, but could not otherwise be characterized, were recovered frequently and throughout the growth cycle of the problem flock, but not at all in the other flock. Lentogenic Newcastle disease virus was detected at all ages on the problem farm but less often than ELAs. The faeces of birds on the problem farm yielded rota-like viruses, corona-like viruses and adeno-like viruses, and on the farm with normal mortality, Egg Drop Syndrome-76 virus and adenovirus. Detection techniques included culture on chick embryos and chick embryo liver cells, and electron microscopy (EM). Inoculation of whole eggs was particularly valuable and more successful than cell culture for virus recovery. EM was most useful for direct examination of faecal preparations and confirmation of the viral type.

A model for testing the efficacy of enrofloxacin (Baytril) administered to turkey hens in the control of Mycoplasma iowae infection in eggs and embryos.

In a preliminary experiment, a field infection with Mycoplasma iowae was simulated by inoculating turkey eggs with various doses of two strains of M. iowae immediately before incubation. The strain and dose chosen for further study were those that best multiplied and resulted in infection of embryos from which the organism could be isolated after 25 days of incubation. Ten turkey hens free from infection with mycoplasmae were housed in isolation. The hens were given enrofloxacin in the drinking water at a concentration of 50 ppm on 3 successive days, on two occasions at intervals of 14 days. Within 48 hours of lay, their eggs were each inoculated with 0.1 ml of the selected strain and dose (10(5) colony-forming units/ml) of M. iowae. M. iowae was recovered from almost all eggs laid by hens before the initial medication but not from any of the eggs laid for several days after each period of medication. Thereafter, the organism could be recovered from a high proportion of inoculated eggs. The treatment of infected turkey laying flocks with enrofloxacin at strategic periods might be helpful in the control of this Mycoplasma by limiting both vertical and horizontal transmission.

Effect of duration of incubation of Mycoplasma gallisepticum cultures on the sensitivity and specificity of antigens for ELISA and microimmunofluorescence tests.

In the production of Mycoplasma gallisepticum (Mg) antigens for ELISA and microimmunofluorescence (MIF) tests, two strains of Mg, S6 and PG31, were grown in broth culture and harvested at intervals from 18 to 138 h. The effect of duration of incubation of culture on antigen sensitivity and specificity was assessed using homologous sera (against Mg S6), and sera prepared against Mycoplasma synoviae (Ms), and against mycoplasma media. It was found, in both Mg S6 and PG31 ELISAs, that in the period 18 to 70 h of incubation, sensitivity declined with homologous sera, but little thereafter. A more rapid and greater decline occurred with PG31 antigen than S6. The antigens showed a lack of specificity since antisera to Ms gave positive reactions. These were not influenced by duration of incubation of Mg culture. In the Mg S6 ELISA lack of specificity was also shown by positive reactions with sera against media components. These increased in the first 42 h of cultivation. In the MIF test there was no loss of sensitivity with Mg S6 antigen with increasing duration of incubation of culture but it occurred with PG31 antigen. The non-specific reactions with antisera to Ms increased with increased duration of incubation of both strains of Mg. This work demonstrates that the most sensitive antigens are produced by harvesting organisms early in culture, although this does not eliminate the Ms cross reactions.

A comparison of the efficacy of danofloxacin and tylosin in the control of Mycoplasma gallisepticum infection in broiler chicks.

Groups of chicks were infected with a virulent strain of Mycoplasma gallisepticum (MG) and treated with either danofloxacin or tylosin while one infected group was left untreated and a further group was uninfected and untreated. Control of clinical signs and mortality was better with danofloxacin than tylosin and there was significantly (P < 0.05) greater weight gain with danofloxacin at 21 days after infection. However at necropsy the prevalence of lesions of the airsac walls was similar in both groups. MG was recovered from fewer live chicks for the first week following treatment with danofloxacin, but at 2 weeks and at necropsy, at the termination of the experiment, it was recovered from a similar proportion of birds in both treated groups. This was reflected also in the serological results at the end of the trial.

Demonstration of sites of latency of infectious laryngotracheitis virus using the polymerase chain reaction.

Mature laying chickens were inoculated intratracheally with a field strain of infectious laryngotracheitis (ILT) virus. Tracheal swabs were collected regularly from all birds for virus culture. At various times post-inoculation, pairs of birds were killed and tissues removed for detection of virus products using conventional tissue homogenization and culture, organ culture, indirect immunofluorescence (IF) and also the polymerase chain reaction (PCR). The latter was used to detect a DNA sequence from the ILT virus thymidine kinase gene. Following inoculation the birds developed mild respiratory disease with clinical signs characteristic of ILT from 3 to 10 days post-inoculation. Trachea and turbinate tissues were virus-positive as determined by virus isolation, organ culture, IF and PCR on day 4 post-inoculation. After recovery from the acute phase, virus shedding initially ceased, then intermittent, low level shedding was recorded for five of the six remaining birds. In an attempt to locate sites of latency, pairs of birds were sampled at 31, 46 and 61 days post-inoculation. Virus was not detected in upper respiratory tract or ocular tissues by conventional techniques, or in the trigeminal, proximal and distal ganglia. All tissues were also negative by PCR, except for the trigeminal ganglia of five of the six birds. All PCR-positive birds had previously shed ILT virus intermittently between days 19 and 59 post-inoculation. As we did not detect viral DNA in any of the other tissues sampled from clinically recovered birds, we conclude that the trigeminal ganglion is the main site of latency of ILT virus.

Pathogenicity of latent infectious laryngotracheitis virus in chickens.

Groups of specific pathogen-free chickens were inoculated with the same dose of a field strain of infectious laryngotracheitis virus that had either been isolated from tracheal swabs taken from infected birds during acute phase shedding, or that had been isolated during an episode of virus shedding after a latent period of 12 to 17 weeks. Birds in both groups developed characteristic clinical signs of ILT including difficulty in breathing, rales and sneezing. Thus, ILT virus shed after a latent period is capable of causing disease in susceptible birds similar to that seen in the field.

The production of Mycoplasma iowae infection of turkey poults suitable for monitoring antimicrobials.

In an attempt to produce a persistent infection with M. iowae (Mi) three separate trials were conducted using strain B 11/80, a virulent strain, strain M 012-118, a recent isolate of unknown virulence, and strain Iowae 695 (I 695), the type strain. In each trial groups of 2-day-old poults were infected via the oesophagus, trachea, cloaca and directly into the lungs. Isolation during life was attempted from the oropharynx and the cloaca, and at necropsy at the end of the experiment (21 days after infection) from the trachea, lungs and airsacs, and the brain. The highest proportion of isolations were made at necropsy from the lungs and air sacs, and trachea, from birds infected with B 11/80 or M 012-118 via the lungs or B 11/80 given via the trachea. During life the proportion of isolations was lower than at necropsy but highest, overall, with B 11/80 given via the lungs or trachea and isolated from the oropharynx, or administered via the cloaca and isolated form this site. Strain I 695 was rarely isolated whatever the route of infection. There were few recoveries following infection via the oesophagus with any strain and no mycoplasmas were isolated from the brain. For the production of MI infection suitable for monitoring antimicrobials in young poults we would recommend infection with a pathogenic strain of the organism directly into the lungs.

Survey of field outbreaks of avian infectious laryngotracheitis in England and Wales.

Field outbreaks of infectious laryngotracheitis in commercial chicken flocks in England and Wales between 1985 and 1988 were investigated. Material from 49 outbreaks was submitted to Liverpool University, and virus was isolated from 17 of them. The results of a questionnaire on each outbreak are described. Generally, the disease was of moderate severity, and mainly affected laying flocks; it occurred in birds of a wide age range but most of the outbreaks were in birds between 10 and 20 weeks of age. The disease was not seen more frequently at any particular time of the year, and there was no evidence of a common source of infection. Three of the affected flocks had recently been moved and were beginning to lay; these stresses may have caused the re-excretion of latent virus.

A comparison of Baytril, Tylosin and Tiamulin in the control of Mycoplasma iowae infection of turkey poults.

Comparison was made of the protection afforded by Baytril, Tylosin and Tiamulin for turkey poults infected with Mycoplasma iowae (MI) by injection of the organism into the lungs. Poults were infected at 2 days of age and treatment commenced 3 days later by including the appropriate drug in the drinking water. Following treatment with Baytril at 50 mug/ml for 5 days, MI was not isolated at 7, 14 and 19 days after infection from the oropharynx or cloaca, during life, but was recovered from five of 23 poults at necropsy. With the same concentration of Baytril, but for only 3 days, isolations of MI were made at 19 days post infection from the oropharynx and cloaca, from 2 of 23 live birds, but at necropsy at this time from 19/23 birds. From poults given Tylosin or Tiamulin (at 500 and 250 mug/ml respectively for 5 days) MI was isolated from a proportion of birds on all occasions during life, after infection, and from 19/21 and 18/21, respectively, at necropsy.

Latency and reactivation of infectious laryngotracheitis vaccine virus.

Latency and reactivation of a commercial infectious laryngotracheitis virus vaccine were demonstrated in live chickens. Virus was re-isolated at intervals between seven and fourteen weeks post-vaccination and this may be of epizootiological significance.