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Poult Sci 2007. 86:698-704
© 2007 Poultry Science Association
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METABOLISM AND NUTRITION

Improved Colostomy Technique and Excrement (Urine) Collection Device for Broilers and Broiler Breeder Hens

M. K. Manangi, F. D. Clark and C. N. Coon1

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701

1 Corresponding author: ccoon{at}uark.edu


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
In conducting nutritional experiments using chickens, scientists are limited in determining the urinary excretion of nutrients because of the difficulty of separating urine from feces. Our main objective was to improve the colostomy procedure for urine collection in broilers and both urine and egg collection for broiler breeder hens. Ketamine HCl (10 to 30 mg/kg i.m.) in combination with Xylazine (2 to 6 mg/kg i.m.) was used to anesthetize broilers (4 wk old) and broiler breeder hens (25 wk old). The colostomy technique involved: 1) transecting the distal colon at approximately 1.5 to 2 cm from the proximal cloaca and ligating the distal colonic segment with 3-0 absorbable surgical suture, 2) ligating the seromuscular coat of the colon to the peritoneal tissue at 3 points in a triangular shape using 4-0 silk suture, 3) ligating the mesentery at the skin level to prevent continued bleeding of the colostomy stoma, and 4) placing 3 sutures using a triangulation technique that consisted of the seromuscular aspect of the transected proximal colonic segment and the skin, and finally, 5) suturing all exteriorized edges of the transected proximal colonic segment after mucosal eversion to the skin with simple interrupted sutures using absorbable suture. For the purpose of urine or egg collection or both, the appropriate size of drainable pouch with a curved tail closure was used. Feces were collected on a tray. The colostomized broilers could be kept for several days and the colostomized broiler breeder hens for several months to collect urine, eggs (for breeder hens), and feces separately without the problem of cross-contamination.

Key Words: colostomy • broiler • broiler breeder hen • urine


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
In conducting metabolism studies using chickens, scientists are limited in determining the urinary excretion of nutrients because of the difficulty of separating urine from feces. However, some of these difficulties are circumvented by cannulation, catheterization, or surgical exteriorization of ureters and by using funnels designed to fit into an urodeum (Davis, 1927; Coulson and Hughes, 1930; Hester et al., 1940; Hart and Essex, 1942; Dixon and Wilkinson, 1957; Ainsworth, 1965; Buss et al., 1980) to separate urine from feces. Although all these procedures are useful for short-term collection, they have side effects such as increased urine flow, tendency for suture breakdown (Fussell, 1969), and fistulation into the cloaca, especially in ureteral exteriorization (Ainsworth, 1965; Tao et al., 1969), for which birds need to wear a harness to facilitate urine and feces collection. Colostomy, a procedure that involves surgically severing the colon just anterior to the coprodeum or rectum and exteriorizing, offers an opportunity to keep birds for long-term experiments to collect urine and feces separately without the problem of cross-contamination between urine and feces. Various surgical techniques have been reported in the literature. Rothchild (1947) anchored the anterior part of the colon to be exteriorized to the body wall by suturing the muscularis and serosa to the peritoneum and abdominal muscle, and the exteriorized edges to the adjacent abdominal muscle and skin. Dixon (1958), Colvin et al. (1966), and Paulson (1969) attached the colon only to the skin. Fussell (1969) reported suturing the peritoneum to serosa, muscle to serosa, everted mucosa to serosa, and muscle to skin. The exteriorized colon was everted using a rubber tube clamped into the lumen of the colon, and the edge of the colon was sutured to its own serosal wall (Fussell, 1969). Dingle and McNab (1985) used triangular sutures for the peritoneum and muscle layers. Belay et al. (1993) sutured the exteriorized edges of the colon to abdominal musculature and skin. Jirjis et al. (1997) everted the colon on itself over a distance of 0.5 cm. Most of the reports mentioned above have indicated consistent problems of recurring blockage, scabbing or closure of the opening, short-term survivability, colon protrusion, and colon withdrawal or retraction into abdomen.

To collect urine and feces, Ariyoshi and Morimoto (1956) sutured a plastic sleeve (5 x 5 cm) to the skin around an artificial anus and sutured a polyethylene rubber beaker to the cloacal opening to collect urine. A metal ring was sutured to the surrounding of the whole pelvic and posterior abdominal area (Paulson, 1969). Richardson et al. (1960) and Fussell (1969) used a Hessian harness to provide support when attaching a collection bag around the colostomy. Belay et al. (1993) used a plastic sleeve with a carbon ring to attach a collection bag around the cloaca. Jirjis et al. (1997) used an oval collection fitting attached to the skin around the cloaca and a plastic bag to collect the urine. All these excrement collection devices may be suitable for the short term and require frequent removal of urine from the collection fittings to reduce the discomfort of the birds, and may not be suitable for egg collection in egg-laying birds. It is important to have a proper excrement collection system, especially for breeder hens, for which both eggs and urine need to be trapped. To our knowledge, there are no reports of colostomy using broiler breeder hens. Our main objectives were to improve 1) the colostomy procedure in broilers and broiler breeder hens, 2) the collection device for urine collection in broilers, and 3) both urine and egg collection for broiler breeder hens.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Cobb-700 broiler breeder hens (25 to 30 wk old) and Cobb-500 broiler chicks (3 to 4 wk old) of uniform body weights were selected for colostomy. Feed was withdrawn for 24 h before anesthetizing the birds, and the birds were given procain penicillin and benzathin penicillin (CombiPen-48, 0.1 mL/kg i.m., Bimeda Inc., Irvindale, CA). Water was withdrawn 1 to 2 h prior to anesthetizing the birds. Ketamine HCl (10 to 30 mg/kg i.m., Ketaset, Fort Dodge Laboratories Inc., Fort Dodge, IA) in combination with Xylazine (2 to 6 mg/kg i.m., Sedazine, Fort Dodge Laboratories Inc.; Plumb, 2002) was used to anesthetize the birds. After placing the bird on its right side on the surgery table and retracting the right leg caudally and the left leg cranially, feathers were removed from an area of approximately 30 cm2 on the left (on the right if the right side is approached) lateral side cranial to the cloaca, and the surgical site was prepared with tincture of iodine. The left wing was abducted and retracted caudally away from the surgical site. A circular (1.5-cm) skin incision was made approximately 2.5 cm cranial to the cloaca on the left ventrolateral abdomen (if right side is approached, then the skin incision would be made on the right ventrolateral abdomen at a distance of 2.5 cm from the cloaca) at an angle of 30° with respect to the midline of the abdomen. The incision was extended through the subcutaneous tissue, abdominal musculature, air sac, and peritoneum. The peritoneal fascia was pierced at 3 points in a triangular shape and held temporarily (Figure 1Go) using 4-0 silk suture (Silk-black braided, Ethicon, Inc., Somerville, NJ) as a stay suture before entering the abdominal cavity. The distal colon to be exteriorized was gently directed to the surgical site by inserting a blunted sterile rod (5 mm in diameter) through the cloacal orifice and applying a gentle traction with a Babcock intestinal forceps (Miltex, Inc., York, PA). The distal part of the colon was isolated using a Babcock or Doyen intestinal forceps (Miltex, Inc.). The part of the colon segment to be colostomized was manually milked distal and proximal to the area to be clamped. The area beneath the colonic segment to be transected was packed with a sterile gauze pad soaked in sterile saline solution that contained 10% Gentamycin (American Pharmaceutical Partners, Inc., Schaumburg, IL). The colon was cross-clamped with the Doyen intestinal forceps (Miltex, Inc.) approximately 1.5 to 2 cm from the proximal cloaca and completely transected proximally to the forceps, and the segment to be colostomized was withdrawn through the surgical incision using saline-soaked gauge sponges. The distal colonic segment was ligated with 3-0 absorbable surgical sutures (9'-vicryl polygalactin 910, Ethicon, Inc.). The exposed edges of the distal colonic segment proximal to the ligature were first sponged using a Betadine-soaked swab and then cleaned with sterile gauze soaked in sterile saline solution that contained 10% Gentamycin (American Pharmaceutical Partners, Inc.). A drop of Genatmycin was placed onto the exposed edges of the distal colonic segment before returning it to the abdominal cavity. The packed gauze pad was carefully removed. The isolated proximal colonic segment was stabilized temporarily outside the abdomen using a Babcock intestinal forceps (Miltex, Inc.). The exposed colonic segment was kept moist with sterile physiological saline, and hemorrhages (if any) were controlled by ligating and cauterizing the blood vessels. The temporarily held peritoneal fascia was sutured to the colonic segment by passing suture through the seromuscular layer and not through the mucosa. The mesentery was ligated at the skin level to prevent continued bleeding of the colostomy stoma. About 0.5 to 1 cm of the colostomy stoma was kept as a final length of colostomy. Using a triangulation technique (Corman, 1989), 3 sutures were placed, consisting of the seromuscular aspect of the colon segment and the skin and subcuticular skin. Finally, all exteriorized edges of the transected proximal colonic segment were sutured after mucosal eversion to the skin with simple interrupted absorbable sutures (Figure 2Go). Immediately after completing the colostomy, a length of approximately 2 cm of saline-soaked gauze (Figure 3Go) was placed inside the exteriorized colon to prevent blockage of the opening caused by a clot from bleeding. Normally, a bird would expel this gauze while passing feces. The surgical area was coated with triple antibiotic cream.


Figure 1
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  		Figure 1. View of the operation showing peritoneal fascia before entering the abdominal cavity.

 

Figure 2
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  		Figure 2. General appearance of the colostomy area immediately after surgery.

 

Figure 3
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  		Figure 3. General appearance of the colostomy area, with saline-soaked gauze placed inside the artificial anus of an anesthetized broiler breeder hen.

 
Postoperative Care

The birds were placed in individual cages. The regular broiler diet for broilers and the breeder diet for breeder hens were continued at 50% of normal feeding quantity for 3 d, and the birds continued to be fed similar to noncolostomized birds. The colostomized birds were given a combination of procaine penicillin and benzathin penicillin (CombiPen-48 0.1 mL/kg per d i.m., Bimeda Inc.) or gentamycin (2.5 mg/kg twice a day, American Pharmaceutical Partners, Inc.) postoperatively for 3 to 5 d. The surgical site was observed daily and cleaned gently with warm water to remove feces and foreign matter, if any. The colostomized broiler breeder birds were selected for the experiment after watching them for 10 to 15 d of egg laying, compared with noncolostomized birds, before feeding the experimental diets. The colostomized broiler birds were selected for the experiment after watching them for a continued period of 8 to 10 d and comparing them with noncolostomized birds for water and feed consumption before feeding the experimental diets. After collecting feces and urine during the experimental days, all colostomized broiler breeder and broiler birds were maintained by feeding a standard breeder diet and a broiler diet, respectively, for their continued period of survivability.

Urine-Collection Bags

An individual colostomy bag of suitable size [~10 cm. flange diameter for broiler breeder birds (Figures 4Go and 5Go) and 3.2 to 3.8 flange diameter for broiler birds (Figure 6Go)] was sutured to the skin around the rectum with 3 to 4 sutures using 2-0 nonabsorbable suture (Ethilon-nylon black monofilament, Ethicon, Inc.) The excreta was collected on a tray. The general appearance of samples of feces from noncolostomized and colostomized birds is shown in Figures 7Go and 8Go.


Figure 4
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  		Figure 4. Colostomized broiler breeder hen showing feces coming out of the artificial opening and urine in the urine-collection bag.

 

Figure 5
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  		Figure 5. Colostomized broiler breeder hen showing the presence of an egg in the urine-collection bag immediately after egg-laying.

 

Figure 6
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  		Figure 6. Colostomized broiler bird showing feces coming out of the artificial opening and urine in the urine-collection bag.

 

Figure 7
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  		Figure 7. Samples of excreta collected from colostomized (left) and noncolostomized (right) broiler breeder hens.

 

Figure 8
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  		Figure 8. Excreta samples collected from colostomized (left) and noncolostomized (right) broiler birds.

 
Euthanasia

Daily monitoring of birds was done to detect infections and feed or water refusal. Birds that exhibited these symptoms were removed from the study group and euthanized using CO2 inhalation. The animal use protocol and the procedure for colostomy were approved by the University of Arkansas Institutional Animal Care and Use Committee.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
As anticipated, the colostomized broilers and broiler breeder hens recovered completely in 8 to 10 d. The behavior of the colostomized birds was observed to be normal compared with that of noncolostomized birds. Colostomized broilers consumed an average of 927 g during the 40- to 45-d period compared with the 816 g of feed consumption of noncolostomized broilers. The weight gain of colostomized broilers was from 1.96 kg (initial weight on d 40) to 2.42 kg (d 45), whereas the weight gain of noncolostomized broilers was from 2.28 kg (d 40) to 2.7 kg (d 45), with a feed:gain ratio of 2:1 for both colostomized and noncolostomized broilers. The increase in weight gain for colostomized birds could be attributed to compensatory growth that was stopped because of the stress of surgery, starvation for a day before surgery, and limited feeding for 3 d after surgery. Both colostomized and noncolostomized broiler breeder hens were observed to consume the same 146 g/d (precision fed) at the same pace. Of the 16 colostomized broilers, 4 exhibited symptoms of recurring blockage. Of the 10 breeder hens, 1 exhibited recurring blockage, 1 showed complete closure of the artificial opening (Figure 9Go), and 1 exhibited protrusion of the colon (Figure 10Go). The broilers and breeder hens were humanely euthanized after keeping them for up to 9 wk of age and up to 6 mo after lay, respectively. At the end of the sixth month, 5 breeder hens were showing normal behavior, consuming feed normally and laying eggs normally compared with noncolostomized breeder hens. Two breeder hens had a protrusion of the colon after approximately 3 mo of continued egg laying. The consistency of the feces in both broilers and breeder hens appeared to be unchanged during their survival period. Details on the successful use and application of the colostomized broilers and broiler breeder hens for nutritional studies have been mentioned elsewhere (Manangi and Coon, 2006a,b,c).


Figure 9
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  		Figure 9. Colostomy complication: closure of the artificial opening in a colostomized broiler breeder hen.

 

Figure 10
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  		Figure 10. Colostomy complication: protrusion of the colon in a colostomized broiler breeder hen.

 

   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
The combination of Ketamine and Xylazine used in the present study was more convenient and required less professional help to maintain the anesthesia for up to 1 h in both broilers and breeder hens. Some of the birds exhibited tetanic convulsions either during the surgical procedure or during recovery from anesthesia. The literature indicates the use of different anesthetics to anesthetize the birds. Ainsworth (1965) reported the use of pentobarbital sodium i.v. followed by 60 mg of phenobarbitone i.m. for maintenance, along with ether supplementation (if necessary). Studies by Rothchild (1947), Dixon (1958), Colvin et al., (1966), and Paulson (1969) reported using 6% sodium pentobaribital or pentobarbitone i.v., whereas Tao et al. (1969) used chloral hydrate, magnesium sulfate, and a pentobarbital mixture i.v. Okumura (1976) used diluted tribromethyl alcohol solution i.p. plus a procaine HCl local anesthetic. Anesthetic induction and maintenance was achieved with Ketamine HCl and halothane in oxygen in the study by Belay et al. (1993), whereas Jirjis et al. (1997) used sodium pentobarbital i.v.

The colostomy technique differs from those of Belay et al. (1993) and Jirjis et al. (1997) in that 1) the seromuscular coat of the colon was ligated to the peritoneal tissue at 3 points in a triangular shape using 4-0 silk suture, 2) the mesentery was ligated at the skin level to prevent continued bleeding of colostomy stoma, and 3) 3 sutures were placed using a triangulation technique that consisted of suturing the seromuscular aspect of the transected proximal colonic segment and the skin, and finally suturing all exteriorized edges of the transected proximal colonic segment after mucosal eversion to the skin with simple interrupted sutures using absorbable suture. The triangulation suture technique (Dingle and McNab, 1985) involved suturing both the peritoneum and the muscle layers to the serosa of the exteriorized colon, but to our knowledge, details on this procedure are not available in the literature. The triangulation suture technique has been indicated for ileostomy procedures in humans (Corman, 1989), with the satisfactory result of keeping the stoma intact for a longer period. In the present study, the use of colostomy bags of suitable size to collect urine from broilers and both urine and eggs from breeder hens offered a convenient means of excrement collection. A colostomy bag of appropriate size, which is placed around the cloaca, rests on the cage with less pressure on the bird and offers more comfort. This type of collection device is more useful especially for breeder hens, where the collection of both eggs and urine is involved.

Postoperatively, polyurea was observed for the first 2 to 3 d in a few of the colostomized broiler breeder hens, whereas the colostomized broilers showed no symptoms of polyurea. Polyurea was not a concern in the present study because the birds were used for metabolic studies after 10 d, in the case of broilers, and at least 2 wk after operation, in the case of broiler breeder hens. Polyurea or diuresis for 3 d after operation has been reported elsewhere (Hester et al., 1940; Colvin et al., 1966; Paulson, 1969). The variation in urine volume could be attributed to physiological conditions of the bird, type and intake of feed, and environmental conditions (Ariyoshi and Morimoto, 1956). Constipation was not observed in the present study in both the colostomized broilers and broiler breeder hens. Ariyoshi and Morimoto (1956) reported constipation problems in colostomized birds after 2 to 3 wk of operation. In the present study, postoperative complications such as closure of the wound, protrusion of the colon, and ballooning of the intestine caused by intestinal stasis, atony, or closure of the wound were observed in a few birds. Jirjis et al. (1997) reported the problems of narrowing of the artificial anus, difficulty in passing feces, and gradual encroachment of skin surrounding the artificial anus. Various methods have been tried to prevent obstruction in passing feces. Ariyoshi and Morimoto (1956), Richardson et al. (1960), Colvin et al. (1966), Polin et al. (1967), and Tao et al. (1969) have tried placing cannulas within the artificial anus, but they later concluded that the cannulas were rejected as foreign bodies and were not suitable for long-term colostomies. Polin et al. (1967) tried irrigating the colon with 2 mL of warm tap water, which was unsuccessful. Paulson (1969) and Jirjis et al. (1997) placed tension sutures in the skin around an artificial anus, and Belay et al. (1993) used rubber cannulas. One of our preliminary studies resulted in a futile attempt to place a rubber cannula (polyethylene tubing) to prevent closure of the wound and obstruction in passing feces in birds that had a colostomy with no mucosal eversion of the exteriorized colon. Retraction or withdrawal of the colon into the abdominal cavity is also known to be a problem (Dingle and McNab, 1985), in addition to protrusion of the colon. Postoperative infection has also been reported, but antibiotic therapy for 3 to 5 d after operation helped to minimize infection. No pecking behavior was observed in the present study. Rothchild (1947) reported pecking at the operation site and therefore constructed a wire guard around the colostomy area, whereas others (Fussell, 1969; Tao et al., 1969) indicated that birds normally did not peck at the operation site.

In conclusion, the colostomized broilers could be kept for several days and the colostomized broiler breeder hens for several months to collect urine, eggs (for breeder hens), and feces separately without the problem of cross-contamination.

Received for publication September 28, 2006. Accepted for publication December 27, 2006.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Ainsworth, L. 1965. Surgical procedure for exteriorization of the uretheral opening of the hen. Poult. Sci. 44:1561–1564.

Ariyoshi, S., and H. Morimoto. 1956. Studies on the nitrogen metabolism in the fowl. 1. Separation of urine for the nutritional balance studies. Bull. Nat. Inst. Agric. Sci. (Tokyo) G12:37–44.

Belay, T., K. E. Bartels, C. J. Wiernusz, and R. G. Teeter. 1993. A detailed colostomy procedure and its application to quantify water and nitrogen balance and urine contribution to thermo-balance in broilers exposed to thermoneutral and heat-distressed environments. Poult. Sci. 72:106–115.[Web of Science][Medline]

Buss, E. G., P. Merkur, and R. B. Guyer. 1980. Urinary excretion of calcium in the presence or absence of shell formation by chickens producing thick or thin shells. Poult. Sci. 59:885–887.[Web of Science][Medline]

Colvin, L. B., C. R. Creger, J. R. Couch, T. M. Ferguson, and M. N. A. Ansari. 1966. A simplified method for separation of urine and feces in the immature fowl. Proc. Soc. Exp. Biol. Med. 123:415–417.[Medline]

Corman, M. L. 1989. Colon and Rectal Surgery, 2nd ed. J. B. Lippincott, Philadelphia, PA.

Coulson, E. J., and J. S. Hughes. 1930. Collection and analysis of chicken urine. Poult. Sci. 10:53–58.

Plumb, D. C. 2002. Veterinary Drug Handbook, 4th ed. Pharma Vet Publishing, White Bear Lake, MN.

Davis, R. E. 1927. The nitrogenous constituents of hens’ urine. J. Biol. Chem. 74:509–515.[Free Full Text]

Dingle, J. G., and J. M. McNab. 1985. Colostomy of the fowl. World’s Poult. Sci. J. 41:114–123.

Dixon, J. M. 1958. Investigation of urinary water reabsorption in the cloaca and rectum of the hen. Poult. Sci. 37:410–414.

Dixon, J. M., and W. S. Wilkinson. 1957. Surgical technique for the exteriorization of the ureters of the chicken. Am. J. Vet. Res. 18:665–667.[Web of Science][Medline]

Fussell, M. H. 1969. A method for the separation and collection of urine and feces in the fowl Gallus domesticus. Res. Vet. Sci. 10:332–337.[Web of Science][Medline]

Hart, W. M., and H. E. Essex. 1942. Water metabolism of the chicken (Gallus domesticus) with special reference to the role of cloaca. Am. J. Physiol. 136:657–668.[Free Full Text]

Hester, H. R., H. E. Essex, and F. C. Mann. 1940. Secretion of urine in the chicken (Gallus domesticus). Am. J. Physiol. 128:592–600.[Free Full Text]

Jirjis, F. F., P. E. Waibel, G. E. Duke, and J. M. McNab. 1997. An improved colostomy technique in turkeys. 1997. Br. Poult. Sci. 38:603–606.[Web of Science][Medline]

Manangi, M. K., and C. N. Coon. 2006a. Calcium particle size effects on excreta, and urinary Ca and P changes in broiler breeder hens. Poult. Sci. 85(Suppl. 1):33. (Abstr.)

Manangi, M. K., and C. N. Coon. 2006b. Effects of varied levels of dietary nonphytate P and Ca on P excretion and relationship between plasma inorganic P and urinary excretion of P in broilers. Poult. Sci. 85(Suppl. 1):168. (Abstr.)

Manangi, M. K., and C. N. Coon. 2006c. Phosphorus utilization and environmental concerns. Pages 97–111 in Proc. 4th Mid-Atlantic Nutr. Conf., Timonium, MD. University of Maryland, College Park.

Okumura, J. 1976. Method of colostomy and cannulation of the chicken. Br. Poult. Sci. 17:547–551.[Web of Science][Medline]

Paulson, G. D. 1969. An improved method for separate collection of urine, feces, and expiratory gases from the mature chcken. Poult. Sci. 48:1331–1336.

Polin, D., E. R. Wynosky, M. Loukides, and C. C. Porter. 1967. A possible urinary backflow to ceca revealed by studies on chicks with artificial anus and fed amprolium C14 or thiamin C14. Poult. Sci. 46:88–93.[Web of Science][Medline]

Richardson, C. E., A. B. Watts, W. S. Wilkinson, and J. M. Dixon. 1960. Techniques used in metabolism studies with surgically modified hens. Poult. Sci. 39:432–440.

Rothchild, I. 1947. The artificial anus in the bird. Poult. Sci. 26:157–162.

Tao, R., R. J. Belzile, G. J. Brisson, and J. A. Awad. 1969. Technique of colostomy in chickens. Am. J. Vet. Res. 30:1067–1070.[Web of Science][Medline]





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