Source: UNIVERSITY OF GEORGIA submitted to
INVESTIGATION INTO FACTORS AFFECTING HATCHABILITY AND CHICK QUALITY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0189157
Grant No.
(N/A)
Project No.
GEOV-0459
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2001
Project End Date
Sep 30, 2004
Grant Year
(N/A)
Project Director
Sander, J. E.
Recipient Organization
UNIVERSITY OF GEORGIA
110 RIVERBEND ROAD
ATHENS,GA 30602
Performing Department
COLLEGE OF VETERINARY MEDICINE
Non Technical Summary
Microorganisms commonly associated with poultry hatcheries and egg contamination have been shown to cause disease in birds. Resistance to chemical disinfectants has been shown to occur with some of these bacteria. Without control of bacterial growth in hatcheries, poor chick quality may prevent profitable poultry production, This project will look at the efficacy of potentiated and new disinfectant products in relation to the currently used standards.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3114010110050%
3083220116050%
Goals / Objectives
a) To evaluate the efficacy of disinfectants against various poultry bacteria isolated from infected chicks or environments. b) To determine the effect of yolk sac contamination by these bacteria on chick quality and broiler production.
Project Methods
Stainless steel slides are used as the carrier for the bacterial samples in an effort to simulate industrial surfaces. Each slide is placed in a glass petri and sterilized by autoclaving. Each bacterial isolate, obtained from poultry environments, are incubated at 37 C overnight resulting in a suspension of approximately 109/ml. An aliquot of 10 ul of each bacterial suspension is evenly spread on the surface of the metal slides and allowed to dry. Disinfectants are applied to the slides using a stainless steel air brush. A fine mist of disinfectant solution is applied for 8 seconds at a distance of 28 cm to the surface of the slide providing consistent application over the entire surface. Following the contact times of 0, 5, 10, and 15 minutes with the disinfectant solution, the slide is removed from the petri dish with sterile forceps, and excess disinfectant removed from the slide by capillary action to the surface of the sterile filter paper lining the inside of the petri dish. The carrier slides are then transferred to glass tubes containing 18 ml of fluid tryptose soy broth. The media with the inoculated and disinfected slides are incubated at 37 C for 48 h. At the end of the incubation period, culture tubes are visibly inspected for the presence or absence of visible bacterial growth. Triplicate samples of each bacteria-coated slide, are exposed in this manner to the disinfectants diluted according to manufacturers recommendations. Thirteen hundred hatching eggs from a resident broiler breeder flock are obtained. Each egg is inoculated on its surface with an avian bacterial culture by individual immersion into a nutrient broth containing 2 X 106 bacteria/ml and allowed to air dry. The eggs are randomly placed into three identical incubators. A commercial hatchery disinfectant mixed at the manufacturers recommended dilution rate is misted into one incubator. A second machine received a mist of the same compound combined with EDTA-tris or another disinfectant altogether for efficacy comparison. The third machine received deionized distilled water. All liquids are administered for a period of 60 seconds each hour at a pressure of 40 psi. Temperature and relative humidity will be set at 99.5 F and 50% respectively. Air samples are taken weekly during the incubation period and increased to daily during the hatching period. Also, during the hatching period, due to expectedly higher levels of aerosol bacteria in the incubators the rate of disinfectant misting will be increased to 90 seconds per hour. Upon hatching, all unhatched eggs are broken open and evaluated for cause of death. A selection of 1-day-old chicks from each treatment group will be weighed, killed, yolk sacs weighed and cultured. A selection of the hatching chicks are placed into floor pens by treatment to provide three replicate pens of 40 birds each. All feed is weighed and recorded before being offered ad libitum to the chicks. Daily mortality is recorded. At 1 week of age, 10 chicks from each pen will be selected, weighed, killed, yolk sacs weighed and cultured. Two-week body weight, feed conversion, and mortality data is recorded.

Progress 10/01/01 to 09/30/04

Outputs
The objective of this study was to compare the efficacy of 904 used alone and in conjunction with EDTA-Tris as incubator disinfectants against eggs surface contaminated with an isolate of P. aeruginosa obtained from a commercial poultry hatchery. After misting these compounds into the incubator, air samples taken revealed bacterial levels stayed relatively low until most of the chicks were hatched at day 21 of incubation. At this time, bacterial levels in the incubator treated with distilled water were signaficantly higher than the bacterial levels of the incubator receiving either disinfectant. There were no significant difference in weight loss % between the groups of weighed eggs indicating that the disinfectant treatment did not affect moisture loss during incubation. Residue breakout data for days 13 and 21 shows no difference between the 3 treatments for contaminated eggs, cracked or cull eggs, middle and late embryonic mortality, cull chicks, fertility, hatch of fertile, and hatchability. However, early embryonic mortality was higher and the percentage of pipped was significantly less with disinfectant treatment when compared with eggs exposed to DH2O during incubation. This resulted in overall hatchability and hatch of fertile which were unaffected by the disinfectant treatment. The 1-day of age chick weight was not affected by treatment; however, cultures taken from the yolk saks of 1-day-old chicks showed a significant lower incidence of bacterial contamination among the chicks when the incubator environment was treated with 904 compared to the yolk sacs of chicks exposed to either DH2O or 904 and EDTA. Pseudomonas was present in 87% of the chicks samples and was equally represented in all treatment groups. All treatments had a moderate incidence of retained yolk sacs ranging from 40-52%. Of the yolks retained, most were also contaminated with bacteria (>78%) none of which included Pseudomonas. The mortality rates remained low for all treatment groups and did not differ statistically per treatment. Postmortem findings throughout the study indicated the primary cause of death was failure to consume feed and water. In conclusion, the use of Biosentry 904 or 904 and EDTA as disinfectants in the hatchery reduced the contamination of aerosol bacterial levels in the hatchery on day 21 of incubation while not affecting hatchability, livability, body weight, or feed conversion. Biosentry 904 when used alone reduced yolk sak contamination at 1 day of age.

Impacts
The determination of efficacious disinfectants for use in commercial poultry hatcheries can improve the number of high quality chicks produced and could potentially result in a lower incidence of contamination of poultry carcasses with potential pathogens.

Publications

  • Walker, S.E., J.E. Sander and R.E. Wooley. The in vitro efficacy of hatchery disinfectants against field isolates of Pseudomonas aeruginosa. Avian Dis. 46"826-830. 2002.
  • Sander, Jean E., Charles L. Hofacre, I.Hsin cheng and Roger D. Wyatt. Investigation of resistance of bacteria from commercial poultry sources to commercial disinfectants. Avian Dis. 46:997-1000. 2002.


Progress 10/01/02 to 09/30/03

Outputs
The objective of this study is to compare the efficacy of 904 used alone and in conjunction with EDTA-Tris as incubator disinfectants, against eggs surface contaminated with an isolate of P. aeruginosa obtained from a commercial poultry hatchery. The experiment was designed to determine the effects of misting these compounds into the incubator on aerosol bacterial levels, egg moisture loss, hatchability, yolk sac contamination and retention, early chick mortality, two-week body weight, and feed conversion. The bacterial levels from air samples taken from each incubator stayed relatively low until most of the chicks were hatched at day 21 of incubation. At this time, the bacterial levels in the incubator treated with distilled water were significantly higher than the bacterial levels of the incubator receiving either disinfectant solution. There was no statistically significant difference in weight loss percent between the groups of weighed eggs at the time of transfer, indicating that the disinfectant treatment did not affect moisture loss during incubation. Residue breakout data for days 13 and 21 shows no differences between the 3 treatments for contaminated eggs, cracked or cull eggs, middle and late embryonic mortality, cull chicks, fertility, hatch of fertile, and hatchability. However, early embryonic mortality was higher and the percentage of pipped, including dead in shell chicks, was significantly less with disinfectant treatment when compared with eggs exposed to DH2O during incubation. This resulted in an overall hatchability and hatch of fertile which were unaffected by the disinfectant treatment. The 1-day of age chick weight was not affected by treatment; however, cultures taken from the yolk sacs of 1-day-old chicks showed a significantly lower incidence of bacterial contamination among the chicks when the incubator environment was treated with 904 compared to the yolk sacs of chicks exposed to either DH2O or 904 and EDTA. It is interesting to note that the addition of the EDTA to the 904 resulted in a significantly higher incidence of gram- negative bacteria. Pseudomonas was present in 87% of the chicks sampled and was equally represented in all treatment groups. Yolk sac weights increased in response to the incidence of bacterial contamination. Day 14 body weights and yolk sac weights were not significantly different between treatment groups. All treatments had a moderate incidence of retained yolk sacs ranging from 40-52%. Of the yolks that were retained most were also contaminated with bacteria (>78%) none of which included Pseudomonas. The mortality rates remained low for all treatment groups and did not differ statistically per treatment. Postmortem findings throughout the study indicated the primary cause of death was failure to consume feed and water. In conclusion, the use of either Biosentry 904 or 904 and EDTA as disinfectants in the hatchery reduced the contamination of aerosol bacterial levels in the hatchery on day 21 of incubation while not affecting hatchability, livability, body weight, or feed conversion. Biosentry 904 when used alone reduced yolk sac contamination at 1 day of age.

Impacts
The determination of efficacious disinfectants for use in commercial poultry hatcheries can improve the number of high quality chicks produced and could potentially result in a lower incidence of contamination of poultry carcasses with potential pathogens.

Publications

  • Walker, S.E., J.E. Sander, and R.E. Wooley. The in vitro efficacy of hatchery disinfectants against field isolates of Pseudomonas aeruginosa. Avian Dis. 46:826-830. 2002.
  • Sander, Jean E., Charles L. Hofacre, I.Hsin Cheng, and Roger D. Wyatt. Investigation of resistance of bacteria from commercial poultry sources to commercial disinfectants. Avian Dis. 46: 997-1000. 2002.
  • Walker, S.E., J.E. Sander, J.L. Cline, and J.S. Helton. Pseudomonas aeruginosa isolates associated with mortality in broiler. Avian Dis. 46:1045-1050. 2002.
  • Walker, S.E., J.E. Sander. Effect of Biosentry 904 and ethylenediaminetetraacetic acid-TRIS disinfecting during incubation of chicken eggs on microbial levels and productivity of poultry. Avian Dis. (Submitted), 2002.


Progress 01/01/01 to 12/31/01

Outputs
Several Pseudomonas aeruginosa isolates were submitted for evaluation. Two isolates used for this study were obtained from the two different hatcheries (E-00-1997 and E-00-1996) and three isolates were obtained from broiler flocks (E-00-1963, E-00-1964 both originating in Hatchery 1; and E-00-1965 originating in Hatchery 2). Case Report: High early mortality was evident in several flocks originating from two commercial broiler hatcheries in Alabama. The commercial poultry production company involved was using a quaternary ammonium disinfectant in the hatchery. To provide a true association between the bacteria present in the hatcheries and yolk sacs and the high mortality in the chicks, 140 1-day old SPF White Leghorn chicks were purchased and placed in 14 clean positive pressure isolation units with 10 chicks per unit. Five P. aeruginosa isolates from the field case were purified and propagated in tryptic soy broth (TSB). Two units contained non-injected controls; chicks in 2 other units were inoculated with PBS. The rest of the chicks were injected with either 10 or 100 bacteria for each of the 5 isolates. Disinfectant Susceptibility: These same 5 P. aeruginosa isolates were evaluated for susceptibility to a quaternary ammonium disinfectant. It has been shown that the addition of EDTA-Tris can potentiate the action of antibiotics and some disinfectants. This study looked at the possible synergistic affect of combining these two chemicals for use against these isolates. The method of disinfectant testing used was described in the original proposal. The quaternary ammonium compound was used at a dilution of 1:256. The EDTA-Tris was used prepared from a stock solution containing 5 mM/L sodium EDTA and 50mM/L Tris-HCl and adjusted to pH of 8. When the 2 compounds were combined a 1:1 ratio was used resulting in the quaternary ammonia concentration to be 1:512. Contact times of the disinfectant to the bacteria ranged from immediate removal or 0.05 min., to 5, 10, and 15 minutes. Synergy was indicated if the combination of compounds was more rapidly bacteriocidal than either compound alone. If the bacteriocidal rate of the combination was slower than for either compound alone it is antagonistic. If the bacteriocidal rate is as rapid as that for the more bacteriocidal compound, the combination was said to be indifferent. One million CFU/ml bacterial concentration was used to distinguish disinfectant efficacy. Synergy and antagonism occurred using the same compounds against closely related bacteria.

Impacts
Proper hatchery sanitation could realistically reduce chick quality associated losses by 1 percent per year. This could result in an estimated savings of $150,000 annually.

Publications

  • Walker, S.E., J.E. Sander, and R. E. Wooley. 2001. The in vitro efficacy of hatchery disinfectants against field isolates of Pseudomonas aeruginosa. Avian Dis. (Accepted).
  • Sander, J.E., C.L.Hofacre, I.H. Cheng, and R.D. Wyatt. 2001. Investigation of Resistance of Bacteria from Commercial Poultry Sources to Commercial Disinfectants. Avian Dis. (In press).
  • Walker, S.E., J.E. Sander, J.L. Cline, and J.S. Helton. 2001. Case Report - Pseudomonas aeruginosa contamination of a hatchery. Avian Dis. (Accepted).