Evaluation of the impact of formaldehyde fumigation during the hatching phase on contamination in the hatch cabinet and early performance in broiler chickens.

Commercial hatch cabinet environments promote replication of microorganisms. These pathogenic or apathogenic microorganisms may serve as pioneer colonizers of the gastrointestinal tract (GIT) of poultry. Some of these pioneer colonizers, such as Escherichia coli and Enterococcus spp., are opportunistic pathogens that lead to reduced performance in commercial poultry. Effective hatchery sanitation is imperative to limit contamination of naïve neonatal chicks and poults. Formaldehyde fumigation has been traditionally used to reduce the pathogen load in commercial hatch cabinets. To investigate potential alternatives to formaldehyde fumigation, models to mimic the microbial bloom in a laboratory setting must be utilized. The purpose of the present study was to evaluate the impact of a multispecies environmental challenge model (PM challenge) with and without formaldehyde fumigation during the hatching phase on early performance in broiler chicks. Three experiments were conducted to evaluate microbial contamination in the hatch cabinet environment (air samples, fluff samples), enteric colonization at day-of-hatch (DOH), and 7-day performance. In all experiments, significantly (P < 0.05) more gram-negative bacteria were recovered from the GIT at DOH in the PM challenge control group as compared to the nonchallenged control (NC) group and the formaldehyde-treated group (PM + F). There were no statistical differences in 7-day body weight gain or feed conversion ratio between the PM challenge control group, the NC group or the PM + F group. These data suggest this model could be utilized to evaluate alternatives to formaldehyde fumigation for controlling the microbial load during the hatching phase in a laboratory setting.

Development of an environmental contamination model to simulate the microbial bloom that occurs in commercial hatch cabinets.

Microbial blooms that emerge in commercial hatch cabinets consist of apathogenic and pathogenic microorganisms, including Escherichia coli, Enterococcus faecalis, and Aspergillus fumigatus. Objectives of the present study included the development of a multipathogen contamination model to mimic commercial conditions and optimization of sampling methods to quantify bacterial or fungal presence within the hatch cabinet. The pathogen challenge mix (PM) was recreated from select bacterial or fungal isolates recovered from an egg homogenate (EH) derived from the contents of infertile eggs and late embryonic mortalities. Isolates selected for PM included Enterococcus faecalis (∼108 CFU/egg), Staphylococcus aureus (∼107 CFU/egg), Staphylococcus chromogenes (∼107 CFU/egg), Aspergillus fumigatus (∼106 spores/egg), and 2 Escherichia coli (∼108 CFU/egg) isolates. Challenge (100 µL of PM or EH) was administered using a sterile loop to a 28 mm area on the blunt end of the eggshell at day 19 of embryogenesis (DOE). In 3 experiments, microbiological data were collected from environmental hatcher samples (open-agar plate method), fluff samples, postmortem whole-body chick rinse samples, and gastrointestinal tract (GIT) samples to evaluate select bacteria and fungi circulating within the hatch cabinet and colonization of GIT. Cumulative bacterial and fungal recovery from the PM hatching environment from DOE20 to hatch was higher than the nonchallenged group (NC) and EH group at ∼860 and ∼1,730 CFU, respectively. Bacterial recovery from GIT, fluff, and chick rinse samples were similar for the PM and EH group in Exp. 1. However, Aspergillus fumigatus recovery from fluff and chick rinse samples for the PM group was significantly (P < 0.001) higher than the NC and EH group. In Exp. 2 and 3, PM challenge significantly (P < 0.05) increased Gram-negative bacterial recovery from the GIT, fluff and chick rinse samples compared to both the NC and EH group. These data suggest this innovative multispecies environmental contamination model using PM could be utilized to evaluate strategies to mitigate microbial contamination in commercial hatch cabinets in a laboratory setting.

Research Note: Application of an Escherichia coli spray challenge model for neonatal broiler chickens.

Avian pathogenic Escherichia coli (E. coli) is an opportunistic pathogen often introduced to neonatal chicks during the hatching process. This commensal bacterium, particularly as a pioneer colonizer of the gastrointestinal tract, can have substantial implications in the rearing of poultry because of reduced flock performance. In order to mimic the effects of the natural bacterial bloom present during the hatch, a seeder challenge model was developed to expose neonatal chicks to virulent E. coli. On day 20 of embryogenesis, selected early hatched chicks (n = 18/hatcher) were briefly removed and sprayed challenged with saline (vehicle) or E. coli at 1 × 107 colony-forming unit (CFU)/chick (exp 1) and 2.5 × 107 CFU/chick (exp 2). These challenged chicks were returned to the hatcher to serve as seeders to transmit the pathogen to the indirect challenged or contact chicks (n = 195/hatcher). For two 7-d experiments, the efficacy of transmission was evaluated via enteric bacterial recovery, body weight gain (BWG), and mortality. For exp 1 and exp 2, significantly (P < 0.0001) more gram-negative bacteria were recovered from the seeder and contact gastrointestinal samples than the negative control samples on day of hatch. In addition, there was a reduction (P < 0.05) in 7-d BWG and significantly (P < 0.0001) higher mortality in the contact-challenged chicks than the negative control chicks in both exp 1 and exp 2. These data suggest that this challenge model could be used to evaluate different methods of controlling the bacterial bloom that occurs in the hatching environment.

Development of a wild-type Escherichia coli environmental bloom model to evaluate alternatives to formaldehyde fumigation in broiler chicken hatch cabinets.

Horizontal transmission of opportunistic Escherichia coli during hatch can have detrimental effects on early performance, particularly as pioneer colonizers. Commercially, formaldehyde is often applied in the United States to combat the bacterial bloom that occurs inside of the hatching environment. The purpose of these experiments was to develop a replicable E. coli horizontal challenge model to evaluate alternatives to formaldehyde sanitation applied to the hatching environment. In experiment 1, two trials were conducted for 2 wild-type (WT) E. coli isolates (isolate 1 [I1] or isolate 2 [I2]) to determine the appropriate in ovo challenge dose and day of embryogenesis (DOE) for challenge administration. In experiment 1 trial 1, the most appropriate inoculation dose and time point were determined to be 102 cfu/embryo on DOE 19. Experiment 1 trial 2 evaluated whether placement of seeder (direct-challenged) embryos with contact (indirect-challenged) embryos during hatch affected contact hatchability. Trial 2 showed no differences in hatchability between groups. A 7-day experiment (experiment 1 trial 2) was conducted to evaluate the effects of I1 or I2 on horizontal transmission, gram-negative bacterial (GNB) recovery from the gastrointestinal tract (GIT), and impact on BW gain (BWG). Compared with the negative control, seeder, and contact chicks challenged with I1 or I2, we observed increased (P < 0.05) GNB recovered from GIT on the day of hatch. There was a marked (P < 0.05) reduction in 7-day BWG between the I1 indirect-challenged group and the negative control group. To further validate the model, 2 7-day trials (experiment 2, experiment 3) were conducted to evaluate the effects of formaldehyde fumigation on coliform recovery from the hatching environment and on early performance using I1 for the challenge. Isolate 1 positive control hatchers had increased levels of circulating coliforms compared with the negative control and formaldehyde-treated hatchers, although there was no significant impact on performance induced by challenge or formaldehyde treatment in experiment 2 or experiment 3. These data provide a potential model for investigations related to horizontal transmission of WT E. coli at a low dose on DOE 19 to promote simulated commercially relevant bacterial blooms under laboratory conditions.

Development of a novel in ovo challenge model for virulent Escherichia coli strains.

During the hatching process, chicks are exposed to opportunistic and/or pathogenic organisms, such as virulent or avirulent Escherichia coli. Virulent E. coli strains have not been feasible for induction of neonatal colibacillosis via in ovo challenge due to high embryonic mortality. In this manuscript, we describe the addition and co-administration of the bacteriostatic antibiotic tetracycline to a virulent E. coli challenge culture, improving hatchability and livability of seeder chicks while allowing robust horizontal transmission in the hatching cabinet to contact chicks. Experiment 1 consisted of 3 trials. Experiment 1, trial 1 was conducted to determine an effective ratio of E. coli challenge and tetracycline dose to be utilized in the seeder model. Trials 2 and 3 were conducted to evaluate the transmission of E. coli from seeder to contact chicks. Experiment 2 consisted of 3 independent 7-D trials where body weight gain (BWG), mortality, and selected enteric bacterial recovery were evaluated. In trials 1 to 3, significantly (P < 0.05) more Gram-negative bacteria were recovered from whole gut samples (GIT) vs. negative controls on day of hatch, from both seeder and contact chicks. At day 7 in trial 1, contact chicks had significantly (P < 0.05) more Gram-negative bacteria recovered from the GIT than the negative control, but not in trials 2 and 3. Presumptive lactic acid bacterial recovery was elevated in contact and seeder chicks compared to the negative control in all 3 trials. Contact challenge caused a significant (P < 0.05) reduction in BWG in 2 out of 3 trials at day 7, and there was a significant (P < 0.05) increase in mortality as compared to the negative controls in all trials. These data suggest that co-administration of a virulent E. coli strain with tetracycline allows for hatch of direct challenged chicks and effective horizontal transmission to contact chicks during the hatching process, as evidenced by reduced day 7 performance and altered selected enteric bacterial recovery.

Characterization and evaluation of lactic acid bacteria candidates for intestinal epithelial permeability and Salmonella Typhimurium colonization in neonatal turkey poults.

The present study evaluated the microbiological properties of three probiotic candidate strains of lactic acid bacteria (LAB) (128; 131; CE11_2), their effect on intestinal epithelial permeability, and their ability to reduce intestinal colonization of Salmonella Typhimurium (ST) individually or as a batch culture in neonatal turkey poults. Isolates were characterized morphologically and identified using 16S rRNA sequence analyses. Each isolate was evaluated for tolerance and resistance to acidic pH, high osmotic NaCl concentrations, and bile salts in broth medium. In vitro assessment of antimicrobial activity against different enteropathogenic bacteria was determined using an overlay technique. In vitro intestinal permeability was evaluated using a stressed Caco-2 cell culture assay treated with/without the probiotic candidates. The in vivo effect of the selected LAB strains on ST cecal colonization was determined in two independent trials with neonatal turkey poults. The results obtained in this study demonstrate the tolerance of LAB candidates to pH 3, a NaCl concentration of 6.5%, and high bile salts (0.6%). All strains evaluated exhibited in vitro antibacterial activity against Salmonella Enteritidis, ST, and Campylobacter jejuni. Candidates 128 and 131 exhibited a coccus morphology and were identified as Enterococcus faecium, and bacterial strain CE11_2 exhibited clusters of cocci-shaped cells and was identified as Pediococcus parvulus. All three candidate probiotics significantly (P < 0.05) increased transepithelial electrical resistance (TEER) in Caco-2 cells following a 3-h incubation period with hydrogen peroxide compared to control and blank groups. The combination of all three candidates as a batch culture exhibited significant efficacy in controlling intestinal colonization of ST in neonatal turkey poults. Evaluation of the combination of these selected LAB strains according to performance and intestinal health parameters of chickens and turkeys are currently in process.

In ovo evaluation of FloraMax®-B11 on Marek's disease HVT vaccine protective efficacy, hatchability, microbiota composition, morphometric analysis, and Salmonella enteritidis infection in broiler chickens.

Three experiments were conducted to evaluate the effect of in ovo administration of FloraMax®-B11 (FM) on Marek's disease (MD) herpesvirus of turkeys (HVT) vaccine protective efficacy, hatchability, microbiota composition, morphometric analysis, and Salmonella enteritidis (SE) infection in chickens. Experiment 1 consisted of 3 trials. In trials 1 and 2, d 18 White Leghorn 15I5x71 embryos were randomly distributed in 4 groups: 1) HVT vaccinated in ovo and no Marek's disease virus (MDV) challenge; 2), HVT + FM vaccinated in ovo and no MDV challenge; 3) HVT vaccinated in ovo and challenge with virulent MDV (vMDV; strain 583A); and 4), HVT + FM vaccinated in ovo and challenge with vMDV. Trial 3 was designed exactly the same as Experiment 1 but chicks were challenged with very virulent MDV (vvMDV; strains Md5 and 612). Birds were monitored until 8 wk of age, and tested for MD incidence. Experiment 2 consisted of 3 trials. In each trial, d 18 broiler embryos were injected in ovo with either saline or FM to measure hatchability and gastrointestinal bacterial composition. In Experiment 3, d 18 broiler embryos were injected in ovo with either saline or FM. All chickens that hatched were orally gavaged with SE at hatch and kept for 7 d to monitor post-hatch BW. No significant difference (P > 0.05) between MD percentage in birds vaccinated with HVT alone or HVT + FM were observed in Experiment 1. In Experiment 2, probiotic did not negatively affect hatchability, but did reduce lactose positive Gram-negative bacteria. Further, increase in BW was associated with higher villi surface area in the ileum in chickens that received the probiotic as well as a significant reduction in the SE incidence in Experiment 3. These results suggest that in ovo administration of FM does not negatively impact the ability of HVT to protect against MD or hatchability of chickens, but improves BW during the first 7 d of life and decreases SE recovery in chickens.

Melorheostosis--an unusual cause of amputation.

A 24-year-old female developed, in infancy, progressive right upper and lower limb muscle and soft tissue contractures and had a diagnosis of melorheostosis made on X-ray and pathological specimens. At the age of 11 years she began to have pain in the right hip and lower limb and this later became the dominant feature. She ultimately required amputation through the right hip joint and prosthetic fitting. She now has independent mobility with her prosthesis and has had no recurrence of pain. Her right arm remains flexed, shortened and contracted, but some hand function is retained. A review of the medical literature is discussed.

Two case study reports of sleep apnoea in patients with paraplegia.

STUDY DESIGN: Case studies of sleep apnoea occurring in two patients with paraplegia. OBJECTIVE: To raise awareness of sleep apnoea in paraplegia. SETTING: Belfast, Northern Ireland. CASE REPORT: We report two patients with paraplegia, one who was having apparent episodes of loss of consciousness and the other daytime somnolence, who were found to have sleep apnoea. The first patient had been medically investigated extensively and a diagnosis of epilepsy was being considered. A joint consultation with the respective partners in each case revealed periods of night-time apnoea and led to sleep study investigations. CONCLUSION: Sleep apnoea is a treatable condition that can occur in patients with paraplegia who are not necessarily obese. Once diagnosed, resolution of symptoms can be rapid and can result in improved quality of life for patients.

Seasonal dynamics of phytoplankton and planktonic protozoan communities in a northern temperate humic lake: diversity in a dinoflagellate dominated system.

Species diversity and richness, and seasonal population dynamics of phytoplankton, planktonic protozoa, and bacterioplankton sampled from the epilimnion of Crystal Bog in 2000, were examined in order to test the hypothesis that these groups' diversity and abundance patterns might be linked. Crystal Bog, a humic lake in Vilas County, Wisconsin, is part of the North Temperate Lakes Long-Term Ecological Research Site. Phytoplankton and planktonic protozoa were identified and enumerated in a settling chamber with an inverted microscope. Bacterial cells were enumerated with the use of fluorescence 4', 6'-diamidino-2-phenylindole (DAPI)-staining procedures, and automated ribosomal intergenic spacer analysis (ARISA) was used to assess bacterioplankton diversity. Bacterial cell counts showed little seasonal variation and averaged 2.6 x 10(6) cells/mL over the ice-free season. Phytoplankton and planktonic protozoan numbers varied by up to two orders of magnitude and were most numerous in late spring and summer. Dinoflagellates largely dominated Crystal Bog throughout the ice-free period, specifically Peridiniopsis quadridens in the spring, Peridinium limbatum in summer, and Gymnodinium fuscum and P. quadridens in fall. Brief blooms of Cryptomonas, Dinobryon, and Synura occurred between periods of dinoflagellate domination. The dominant dinoflagellate, Peridinium limbatum, was calculated to have a growth rate of 0.065 day(-1) and a doubling time of 10.7 days. Heterotrophic nanoflagellates (HNFs) were a consistent component of the planktonic protozoa; seasonal patterns were determined for three genera of HNFs (Monosiga, Bicosoeca, and Desmarella moniliformis). Three genera of ciliates (Coleps, Strobilidium, and Strombidium) comprised the greater part of the planktonic protozoa in Crystal Bog. The number of species of planktonic protozoa was too low to calculate a diversity index. Shannon-Weaver diversity indices for phytoplankton and bacterioplankton in the epilimnion followed very similar seasonal patterns in this lake, supporting the hypothesis that in freshwaters, diversity patterns of these groups are linked.

The Northern Ireland troubles and limb loss: a retrospective study.

The Troubles in Northern Ireland have now lasted 34 years. Divisions and strife between the opposing loyalist and republican communities, and between the communities and the security forces, have led to thousands of deaths and injuries. The violence has often been indiscriminate injuring and killing totally innocent people. Staff at the Regional Disablement Services at Musgrave Park Hospital, Belfast have had the responsibility for helping to rehabilitate those who have suffered limb loss, both civilians and security forces personnel. In this study the authors present patient demographics for those survivors, referred for prosthetic fitting, who have sustained limb amputations as a result of the Troubles from 1969 to 2003, with the cause of injury, resulting levels of amputation, associated injuries, time to first prosthetic fitting and reason for any delay in fitting identified. One hundred and twenty-nine (129) patients sustained amputations, 110 male and 19 female with an age range at the time of injury from 7 to 60 years. Seventy-two (72) were civilian. Ninety-three (93) underwent immediate amputation, the most frequent level of amputation being trans-femoral. Delayed healing of deep wounds was the most common reason for delayed amputation; other causes were chronic osteomyelitis, malunited fractures and failed arthrodesis, often associated with chronic pain. Ninety-two (92) patients required amputation of one limb or part thereof, 35 required amputation of 2 limbs and 2 underwent triple amputation. Three (3) patients lost both hands. Sixty seven percent (67%) had other associated physical injuries. Thirty-two (32) patients had a delay of 6 months or more in fitting a prosthesis. The most common cause of injury was the car bomb.

Telemedicine--the way ahead for medicine in the developing world.

An e-mail link with the facility to send high-resolution digital images is a cheap and uncomplicated telemedicine method. The Swinfen Charitable Trust helped establish such a link in Patan Hospital Kathmandu, Nepal in March 2000. Over 12 months using this link 42 telemedicine referrals were sent to specialists throughout the world. Referrals were: 36% respiratory medicine; 21% neurology, 21% dermatology; 14% cardiology; 5% nephrology; and 3% radiology--28 had digital pictures attached, of which 96% were of high enough quality on which specialists were able to comment. Thirty-nine replies were received. The average time for a specialist reply was 2 days, and 45% were answered within 24 hours. All replies were judged by independent assessors to be helpful or very helpful for diagnosis, management and education. The assessors decided that in 50% of cases the advice if acted upon would have shortened hospital stay. This pilot study has shown that a low-cost telemedicine link is technically feasible and can be of significant benefit for diagnosis, management and education in a developing world setting.

Complex regional pain syndrome post mastectomy.

Complex regional pain syndrome includes the previously termed condition reflex sympathetic dystrophy. It is a chronic pain disorder diagnosed on the basis of symptoms and skin changes and is known to have a psychological element. It is a rare complication after surgery, especially mastectomy. We present two females who developed this syndrome after undergoing mastectomy for chronic mastalgia. These cases demonstrate that amputation of an organ for chronic pain can result in reflex sympathetic dystrophy developing in a nearby limb.

High-temperature, acid-hydrolyzed remains of Polytrichum (Musci, Polytrichaceae) resemble enigmatic Silurian-Devonian tubular microfossils.

Gametophytes and sporophyte components of two species of the evolutionarily early-divergent moss Polytrichum were separately subjected to high-temperature acid hydrolysis, and remains were examined by fluorescence microscopy and scanning electron microscopy. Remains included fragments of capsule, seta, leaves, stems, and calyptra. Cell walls of all remains were autofluorescent in violet and UV excitation, suggesting occurrence of resistant polyphenolic compounds. Calyptras of both species dissociated into smooth- walled, acutely branched filamentous associations of tubular cells with distinctively thickened cell junctions. In these aspects and measurements of wall dimensions made from SEMs, the hydrolysis-resistant Polytrichum calyptra remains were similar to several tubular microfossils described from Silurian and Lower Devonian deposits, whose provenance is unknown or ascribed to fungi. Our data suggest the possibility that at least some ancient tubular microfossils might have originated from Polytrichum-like early mosses. They add to increasing evidence that bryophytes left microfossil evidence for their presence millions of years earlier than is indicated by their macrofossil record.