Reduction of Airborne Microorganisms by Filtering Recycled Air in a Chick Hatcher1,2 JOHN S. A V E N S , CAREY L . QUARLES AND DIANE J. FAGERBERG
Department of Animal Sciences, Colorado State University, Fort Collins, Colorado 80523 (Received for publication July 4, 1974)
ABSTRACT An experimental chick hatcher designed to filter recycled ventilation air was tested for its effectiveness in reducing the number of viable airborne microorganisms. Chicks in a filtered hatcher and a control hatcher (no filter) were artificially contaminated with Serratia marcescens as ventilation air was recycled in the hatchers for twelve hours. The number of viable S. marcescens particles in the filtered air of the conditioning chamber was less than detectable. The number of viable airborne S. marcescens particles in the hatching chamber of the filtered hatcher indicated a reduction of greater than 90 percent over the number in the unfiltered hatcher. The filter was effective in reducing the number of airborne particles carrying viable S. marcescens organisms in the hatcher. POULTRY SCIENCE 54: 479-482, 1975
INTRODUCTION ONTAMINATION or infection of hatched chicks may be attributed to microorganisms on or in some hatching eggs and on surfaces within the hatcher. Microorganisms on chick down or other particles prevalent at hatching time may become airborne and distributed throughout the hatcher by air movement caused by ventilation, potentially contaminating or infecting all chicks in the machine. These microorganisms may be pathogenic causing morbidity or mortality of the hatched chicks. Magwood (1964) observed a direct relationship between airborne microorganisms and the amount of contamination of various hatchery surfaces. Reduction in the number of airborne particles transmitting microorganisms should decrease contamination and infection of hatched chicks. An experimental chick hatcher was designed (Robbins Incubator Co.) to achieve this reduction. Ventilation air leaving the hatching chamber was filtered and recirculated in a positive pressure system. Many of the viable
C
1. Published with the approval of the Director of the Colorado State University Experiment Station as Scientific Series Paper No. 1982. 2. This research was supported in part by Robbins Incubator Company, Project 6535.
airborne particles could adhere to the filter and be eliminated from circulation in the hatching chamber. MATERIALS AND METHODS Hatcher Design. Two similar experimental hatchers designed and built by Robbins Incubator Company, one filtered and one control (no filter) were simultaneously tested. Each hatcher was designed with a conditioning chamber and hatching chamber, separated by a filter in the filtered hatcher (Figures 1 and 2). The filter frame was refilled with new dry filter medium before each experiment. Four 90-egg capacity trays were in the hatching chambers. Eggs. Fertile eggs were obtained from caged Single Comb White Leghorn hens artificially inseminated twice each week. Seven hundred-twenty eggs were incubated in a Robbins Hatch-O-Matic incubator. At 18.5 days 360 eggs were transferred to the filtered hatcher and 360 to the control hatcher. Artificial Contamination. The effectiveness of the filter in reducing viable airborne particles was determined by artificial contamination of hatched chicks with S. marcescens. This organism was used because of its characteristic red pigmentation in colonial growth,
479
480
J. S. AVENS, C . L . QUARLES AND D . J . FAGERBERG
into sterile, plastic petri dishes containing folded paper tissue ("Kim-Wipes"). One petri dish containing the culture-saturated tissue was placed open in the center of each of the four hatching trays—5 ml. on bottom tray, 10 ml. on each middle tray and 15 ml. on the top tray. These open culture dishes were a source of airborne particle contamination since many of the hatched chicks walked through, fell in and/or pecked at the culture, thus contaminating their down.
FIG. 1. Experimental chick hatcher. Side and front panels have been removed to show blower, heater, humidifier, cooling coil and filter in conditioning compartment. Door to the hatching compartment is open. allowing easy identification. A twelve-hour culture of S. marcescens in Trypticase Soy Broth (BBL) containing 2 percent Yeast Extract (BBL) was placed in the hatchers after the majority of the chicks had pipped and 12 hours before air sampling. Specific volumes of the 12-hour culture were pipetted
Air Sampling and Enumeration. An Andersen Viable Sampler Model 10-000 (Andersen, 1958) (Figure 3), operated according to manufacturer's instructions (1971), was used to impinge air onto pertri dishes (Andersen 00201) each containing 27 ml. Tryptone Glucose Yeast Agar (BBL). Consecutive air samples were taken first from the conditioning chamber followed by the hatching chamber of the filtered and control hatchers. Sampling probes were inserted through ports in front of each chamber of the hatchers just prior to sampling. Each probe was made of unpainted copper tubing (1-1/16 inch I.D.)
•TEMPERATURE SENSORS
HATCHING COMPARTMENT-
- CONDITIONING COMPARTMENT (CONTAINS HEATERS, COOLING COILS, HUMIDIFIER, HUMIDISTAT, ETC.)
ILTER TEST HATCHER
FIG. 2. Experimental chick hatcher—schematic, front view.
481
FILTERING AIRBORNE MICROORGANISMS
TABLE 1.—Andersen Viable Sampler Serratia marcescens colony counts (viable particles per cubic foot of hatcher air): hatcher with filter and hatcher without filter, conditioning and hatching chambers
Experiment
Hatcher
1
Filtered Control
2
Filtered Control
3
Filtered Control
Chamber Conditioning Hatching 2 0 1500 3300 0 120 0 170
2 40 10 140
and its respective chambers are shown in Table 1.
FIG. 3. Andersen Viable Sampler in place at end of sampling probe. Sampling probe sealed in place around hatcher port.
with one 90° bend and was flange-gasketed for a friction-tight seal around the hatcher ports (Figure 3). The Andersen Viable Sampler was gasketed for a friction-tight connection to the end of the probe (Figure 3). One probe extended into the center of the conditioning chamber just above the floor; the other into the center of the hatching chamber below the bottom tray. The protruding end of each probe remained closed until air sampler was attached. Air sampling time was 30 seconds. Air flow through the sampler was one cubic foot (28.3 liters) per minute. After sampling, plates were incubated at 25-30° C. for 24 hours. Viable particle counts were determined by the "positive hole" method (Andersen Air Samplers, 1971). RESULTS Serratia marcescens colony counts (viable particles per cubic foot of air) for each hatcher
Experiment 1. Ninety-nine percent fewer viable S. marcescens particles per cubic foot of air were collected from the hatching chamber of the filtered hatcher compared to the control hatching chamber. No viable S. marcescens particles were found in the conditioning chamber of the filtered hatcher. Experiment 2. The filtered hatcher had 95.0 percent fewer viable S. marcescens particles per cubic foot of air in the hatching chamber than the unfiltered control hatcher. No viable S. marcescens particles were found in the conditioning chamber of the filtered hatcher. Experiment 3. The filtered hatcher had 92.9 percent fewer viable S. marcescens particles per cubic foot of air in the hatching chamber than the unfiltered control hatcher. No viable S. marcescens particles were found in the conditioning chamber of the filtered hatcher. DISCUSSION The filter effectively reduced the number of viable airborne particles in the chick hatcher during the hatching process. Filtration effectively reduced the number of S. marcescens-contaminated particles in both chambers of the filtered hatcher compared to the unfiltered control hatcher. It should now be determined if this reduction of viable airborne particles during hatch by filtration
482
J. S. AVENS, C. L. QUARLES AND D . J. FAGERBERG
would d e c r e a s e bird morbidity and mortality
during growth. REFERENCES Andersen, A. A., 1958. New sampler for the collection, sizing and enumeration of viable airborne particles. J. Bact. 76: 471-484.
Andersen Air Samplers, 1971. Instructions for operation and care of the Andersen Viable Sampler, pp. 1-3. 2000 Inc., Salt Lake City. Magwood, S. E., 1964. Studies in hatchery sanitation. 3. The effect of airborne bacterial population on contamination of egg and embryo surfaces. Poultry Sci. 43: 1567-1572.
Behavioral and Electrophysiological Consequences of Cycloheximide Administration in Chick Embryos JlNDRICH S E D L A C E K
Research Laboratory of Psychiatry, Faculty of General Medicine, Charles University, Prague, Czechoslovakia (Received for publication July 5, 1974) ABSTRACT The effect of cycloheximide administration upon the spontaneous motility, electroencephalogram (EEG) and infra slow potential oscillations (ISPO) of brain hemispheres and upon the tectal optic evoked responses was investigated in 15-day- and 20-day-old chick embryos. Cycloheximide evoked in 15-day-old embryos, within some hours after systemic administration, a full depression of EEG activity and an apparent inhibition of spontaneous motility. The ISPO's of brain hemispheres remained unchanged during the entire investigation period. The inhibitory effect of cycloheximide was manifested in 20-day-old embryos by a high degree of ISPO depression only, whereas the EEG activity, optic evoked responses and spontaneous motility did not change. The information collected suggested that the different types of electrical activity generated in the brain at different stages of development were in different relationship to the normal proteosynthesis in embryonic brain tissue. POULTRY SCIENCE 54: 482-487,
INTRODUCTION
R
ECENT investigations of some inhibitors of proteosynthesis (puromycin, cycloheximide, acetoxycycloheximide) have shown that they have a depressive effect on simple and complicated functions and processes in the nervous system (Paggi and Toschi, 1971; Barondes and Cohen, 1967, 1968). It has been shown in many cases that the effect of different drugs connected with proteosynthesis and amino acid metabolism depends on the developmental stage (Schade andPascoe, 1964; Sedlacek and Schade, 1969; Stastny, 1971). The purposes of this report were: first to investigate the effect of cycloheximide at two different stages of chick embryo development, and second to obtain some information on the dependence of EEG activity and infra slow potentials on the
1975
process of proteosynthesis in brain tissue. MATERIAL AND METHODS This study was performed on 130 White Leghorn chick embryos: 70 embryos on day 15, 60 embryos on day 20 of incubation. The fertile eggs were incubated in a commercial incubator with forced air circulation and with automatic turning of eggs several times a day (38° C , 60-70% relative air humidity). Registration of spontaneous motility (10 embryos) was carried out on intact eggs with an automated technique using three phonograph cartridges (Supraphon VK 311) as transducers (Kovach, 1970). Electrical activity (65 embryos) was recorded from the exposed surface of the right brain hemisphere on a 4-channel direct writing apparatus (4-EEG 1, USSR) with a frequency
Department of Animal Sciences, Colorado State University, Fort Collins, Colorado 80523 (Received for publication July 4, 1974)
ABSTRACT An experimental chick hatcher designed to filter recycled ventilation air was tested for its effectiveness in reducing the number of viable airborne microorganisms. Chicks in a filtered hatcher and a control hatcher (no filter) were artificially contaminated with Serratia marcescens as ventilation air was recycled in the hatchers for twelve hours. The number of viable S. marcescens particles in the filtered air of the conditioning chamber was less than detectable. The number of viable airborne S. marcescens particles in the hatching chamber of the filtered hatcher indicated a reduction of greater than 90 percent over the number in the unfiltered hatcher. The filter was effective in reducing the number of airborne particles carrying viable S. marcescens organisms in the hatcher. POULTRY SCIENCE 54: 479-482, 1975
INTRODUCTION ONTAMINATION or infection of hatched chicks may be attributed to microorganisms on or in some hatching eggs and on surfaces within the hatcher. Microorganisms on chick down or other particles prevalent at hatching time may become airborne and distributed throughout the hatcher by air movement caused by ventilation, potentially contaminating or infecting all chicks in the machine. These microorganisms may be pathogenic causing morbidity or mortality of the hatched chicks. Magwood (1964) observed a direct relationship between airborne microorganisms and the amount of contamination of various hatchery surfaces. Reduction in the number of airborne particles transmitting microorganisms should decrease contamination and infection of hatched chicks. An experimental chick hatcher was designed (Robbins Incubator Co.) to achieve this reduction. Ventilation air leaving the hatching chamber was filtered and recirculated in a positive pressure system. Many of the viable
C
1. Published with the approval of the Director of the Colorado State University Experiment Station as Scientific Series Paper No. 1982. 2. This research was supported in part by Robbins Incubator Company, Project 6535.
airborne particles could adhere to the filter and be eliminated from circulation in the hatching chamber. MATERIALS AND METHODS Hatcher Design. Two similar experimental hatchers designed and built by Robbins Incubator Company, one filtered and one control (no filter) were simultaneously tested. Each hatcher was designed with a conditioning chamber and hatching chamber, separated by a filter in the filtered hatcher (Figures 1 and 2). The filter frame was refilled with new dry filter medium before each experiment. Four 90-egg capacity trays were in the hatching chambers. Eggs. Fertile eggs were obtained from caged Single Comb White Leghorn hens artificially inseminated twice each week. Seven hundred-twenty eggs were incubated in a Robbins Hatch-O-Matic incubator. At 18.5 days 360 eggs were transferred to the filtered hatcher and 360 to the control hatcher. Artificial Contamination. The effectiveness of the filter in reducing viable airborne particles was determined by artificial contamination of hatched chicks with S. marcescens. This organism was used because of its characteristic red pigmentation in colonial growth,
479
480
J. S. AVENS, C . L . QUARLES AND D . J . FAGERBERG
into sterile, plastic petri dishes containing folded paper tissue ("Kim-Wipes"). One petri dish containing the culture-saturated tissue was placed open in the center of each of the four hatching trays—5 ml. on bottom tray, 10 ml. on each middle tray and 15 ml. on the top tray. These open culture dishes were a source of airborne particle contamination since many of the hatched chicks walked through, fell in and/or pecked at the culture, thus contaminating their down.
FIG. 1. Experimental chick hatcher. Side and front panels have been removed to show blower, heater, humidifier, cooling coil and filter in conditioning compartment. Door to the hatching compartment is open. allowing easy identification. A twelve-hour culture of S. marcescens in Trypticase Soy Broth (BBL) containing 2 percent Yeast Extract (BBL) was placed in the hatchers after the majority of the chicks had pipped and 12 hours before air sampling. Specific volumes of the 12-hour culture were pipetted
Air Sampling and Enumeration. An Andersen Viable Sampler Model 10-000 (Andersen, 1958) (Figure 3), operated according to manufacturer's instructions (1971), was used to impinge air onto pertri dishes (Andersen 00201) each containing 27 ml. Tryptone Glucose Yeast Agar (BBL). Consecutive air samples were taken first from the conditioning chamber followed by the hatching chamber of the filtered and control hatchers. Sampling probes were inserted through ports in front of each chamber of the hatchers just prior to sampling. Each probe was made of unpainted copper tubing (1-1/16 inch I.D.)
•TEMPERATURE SENSORS
HATCHING COMPARTMENT-
- CONDITIONING COMPARTMENT (CONTAINS HEATERS, COOLING COILS, HUMIDIFIER, HUMIDISTAT, ETC.)
ILTER TEST HATCHER
FIG. 2. Experimental chick hatcher—schematic, front view.
481
FILTERING AIRBORNE MICROORGANISMS
TABLE 1.—Andersen Viable Sampler Serratia marcescens colony counts (viable particles per cubic foot of hatcher air): hatcher with filter and hatcher without filter, conditioning and hatching chambers
Experiment
Hatcher
1
Filtered Control
2
Filtered Control
3
Filtered Control
Chamber Conditioning Hatching 2 0 1500 3300 0 120 0 170
2 40 10 140
and its respective chambers are shown in Table 1.
FIG. 3. Andersen Viable Sampler in place at end of sampling probe. Sampling probe sealed in place around hatcher port.
with one 90° bend and was flange-gasketed for a friction-tight seal around the hatcher ports (Figure 3). The Andersen Viable Sampler was gasketed for a friction-tight connection to the end of the probe (Figure 3). One probe extended into the center of the conditioning chamber just above the floor; the other into the center of the hatching chamber below the bottom tray. The protruding end of each probe remained closed until air sampler was attached. Air sampling time was 30 seconds. Air flow through the sampler was one cubic foot (28.3 liters) per minute. After sampling, plates were incubated at 25-30° C. for 24 hours. Viable particle counts were determined by the "positive hole" method (Andersen Air Samplers, 1971). RESULTS Serratia marcescens colony counts (viable particles per cubic foot of air) for each hatcher
Experiment 1. Ninety-nine percent fewer viable S. marcescens particles per cubic foot of air were collected from the hatching chamber of the filtered hatcher compared to the control hatching chamber. No viable S. marcescens particles were found in the conditioning chamber of the filtered hatcher. Experiment 2. The filtered hatcher had 95.0 percent fewer viable S. marcescens particles per cubic foot of air in the hatching chamber than the unfiltered control hatcher. No viable S. marcescens particles were found in the conditioning chamber of the filtered hatcher. Experiment 3. The filtered hatcher had 92.9 percent fewer viable S. marcescens particles per cubic foot of air in the hatching chamber than the unfiltered control hatcher. No viable S. marcescens particles were found in the conditioning chamber of the filtered hatcher. DISCUSSION The filter effectively reduced the number of viable airborne particles in the chick hatcher during the hatching process. Filtration effectively reduced the number of S. marcescens-contaminated particles in both chambers of the filtered hatcher compared to the unfiltered control hatcher. It should now be determined if this reduction of viable airborne particles during hatch by filtration
482
J. S. AVENS, C. L. QUARLES AND D . J. FAGERBERG
would d e c r e a s e bird morbidity and mortality
during growth. REFERENCES Andersen, A. A., 1958. New sampler for the collection, sizing and enumeration of viable airborne particles. J. Bact. 76: 471-484.
Andersen Air Samplers, 1971. Instructions for operation and care of the Andersen Viable Sampler, pp. 1-3. 2000 Inc., Salt Lake City. Magwood, S. E., 1964. Studies in hatchery sanitation. 3. The effect of airborne bacterial population on contamination of egg and embryo surfaces. Poultry Sci. 43: 1567-1572.
Behavioral and Electrophysiological Consequences of Cycloheximide Administration in Chick Embryos JlNDRICH S E D L A C E K
Research Laboratory of Psychiatry, Faculty of General Medicine, Charles University, Prague, Czechoslovakia (Received for publication July 5, 1974) ABSTRACT The effect of cycloheximide administration upon the spontaneous motility, electroencephalogram (EEG) and infra slow potential oscillations (ISPO) of brain hemispheres and upon the tectal optic evoked responses was investigated in 15-day- and 20-day-old chick embryos. Cycloheximide evoked in 15-day-old embryos, within some hours after systemic administration, a full depression of EEG activity and an apparent inhibition of spontaneous motility. The ISPO's of brain hemispheres remained unchanged during the entire investigation period. The inhibitory effect of cycloheximide was manifested in 20-day-old embryos by a high degree of ISPO depression only, whereas the EEG activity, optic evoked responses and spontaneous motility did not change. The information collected suggested that the different types of electrical activity generated in the brain at different stages of development were in different relationship to the normal proteosynthesis in embryonic brain tissue. POULTRY SCIENCE 54: 482-487,
INTRODUCTION
R
ECENT investigations of some inhibitors of proteosynthesis (puromycin, cycloheximide, acetoxycycloheximide) have shown that they have a depressive effect on simple and complicated functions and processes in the nervous system (Paggi and Toschi, 1971; Barondes and Cohen, 1967, 1968). It has been shown in many cases that the effect of different drugs connected with proteosynthesis and amino acid metabolism depends on the developmental stage (Schade andPascoe, 1964; Sedlacek and Schade, 1969; Stastny, 1971). The purposes of this report were: first to investigate the effect of cycloheximide at two different stages of chick embryo development, and second to obtain some information on the dependence of EEG activity and infra slow potentials on the
1975
process of proteosynthesis in brain tissue. MATERIAL AND METHODS This study was performed on 130 White Leghorn chick embryos: 70 embryos on day 15, 60 embryos on day 20 of incubation. The fertile eggs were incubated in a commercial incubator with forced air circulation and with automatic turning of eggs several times a day (38° C , 60-70% relative air humidity). Registration of spontaneous motility (10 embryos) was carried out on intact eggs with an automated technique using three phonograph cartridges (Supraphon VK 311) as transducers (Kovach, 1970). Electrical activity (65 embryos) was recorded from the exposed surface of the right brain hemisphere on a 4-channel direct writing apparatus (4-EEG 1, USSR) with a frequency
