The Veterinary Record, June 7, 1997 605

MARIN, C., JIMENEZ DE BAGUES, M. P., BARBERAN, M. & BLASCO, J. M. PHILIPPON, A., RENOUX, G. & PLOMMET, M. (1971) Anntcales de Recherches
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SAWYER, M. (1974) Jouirnial of the Ainericon Veterincar! Medical Association RICHARDSON, M., CONNER, G. H., BECK, C. C. & CLARK, D. T. (1971)
164, 295 Immunology 21, 795

Short Communications
Enzyme-linked immunoelectro- TABLE 1: Sensitivity and specificity of the enzyme-linked immuno-
electrotransfer blot (EITB) assay for the diagnosis of sheep hydatidosis
transfer blot assay for diagnosis of Sensitivity (%) Specificity (%)
hydatidosis (Echinococcus Peruvian sheep Serum samples Serum samples
naturally infected from USA from cestode-free
granulosus) in sheep EITB assay band
(kDa)
with E granulosus
(n = 94)
sheep
(n = 36)
Peruvian sheep
(n = 43)
8 only 31 97-2 100
P. Moro, M. Verastegui, R. H. Gilman, N. Falcon, 16 only 0 100 100
T. Bernal, C. Gavidia, A. Gonzalez, V. Malqui, 21 only 0 100 100
8and 16 17 100 100
M. H. Moro, E. Dueger 8 and 21 1 100 100
All three bands 24 100 100
Any or all bands 73 100 100
Veterinary Record ( 1 997) 140, 605-606
THE diagnosis of Echinococcus granulosus in livestock is most sheep has not been evaluated. The present study describes the
commonly made at slaughter. Abattoirs which are strictly regulat- application of an enzyme-linked immunoelectrotransfer blot (EITB)
ed can often provide acceptable prevalence data. However, in assay for the diagnosis of hydatidosis in sheep.
many areas where the disease is endemic home slaughter is prac- To determine sensitivity, 94 serum samples were obtained from
tised and few abattoirs have adequate veterinary supervision. In sheep (Junin breed) naturally infected with hydatidosis at an abat-
addition, postmortem diagnosis of the disease is of little use in toir in an endemic region of Peru. Blood samples were collected
areas of low prevalence, where segregation of infected animals is just before slaughter and transferred at 4C to Lima, where they
essential for disease control. The development of a sensitive, spe- were centrifuged and sera aliquoted and stored at -20C before
cific and reproducible serological assay for livestock would pro- analysis. The number and location of cysts as well as presence of
vide a useful epidemiological tool for the antemortem study and other parasites were recorded postmortem.
control of hydatid disease. Two groups of sheep were used to determine the specificity of
Although efforts to develop sensitive and specific immunodiag- the assay. Blood samples from 43 sheep (Junin breed) raised in a
nostic tests in humans have been relatively successful, similar cestode-free environment in the city of Lima were collected in
tests developed for livestock have been hampered by low speci- Vacutainer tubes and after separation from the clot, the sera were
ficity and cross-reactivity with other cestode species (Sweatman aliquoted and stored at -20C. Sera from 36 Columbia and Border
and others 1963, Martinez Gomez and others 1980, Craig and Leicester sheep from the National Animal Disease Center in
Rickard 1981, Young and Heath 1984). An immunoblot assay Ames, Iowa (kindly provided by Dr R. Cutlip) were also tested.
based on the identification of three specific antigens of 8, 16 and Cross-reactivity was tested with sera from four sheep experi-
21 kDa is currently used in humans with a sensitivity of 65 per mentally infected with Taenia ovis (kindly provided by Dr M.
cent and a specificity of 100 per cent (Verastegui and others Lightowlers) and one serum sample from a lamb which had T
1992). These antigens are also present in hydatid-infected farm hydatigena present in the peritoneal cavity after being experimen-
animals; however, their use in the diagnosis of hydatidosis in tally infected as described by Craig and Rickard (1981).
The ELITB assay was performed as previously described (Tsang
and others 1989, Verastegui and others 1992) except that sheep
serum was tested at a 1/10 dilution and a conjugate of horseradish
P. Moro, V. Malqui, Department of Pathology, M. Verastegui, peroxidase-rabbit antibody to sheep immunoglobulin G was used.
Department of Pathology and Department of Microbiology, Universidad The overall sensitivity of the EITB assay in 94 sheep was 73 per
Peruana Cayetano Heredia, PO Box 5045, Lima, Peru cent (69/94) and the specificity was 98.7 per cent (1/79) (Table 1).
R. H. Gilman, Department of International Health, Johns Hopkins One serum sample from a sheep from the USA was positive to the 8
University School of Hygiene and Public Health, Baltimore, Maryland kDa band. Sensitivity was 46 per cent (13/28) for light infections
21205, USA (one to five cysts <2 cm in diameter) and 85 per cent (56/66) for
N. Falcon, T. Bernal, C. Gavidia, A. Gonzalez, Public Health Section, heavy infections (one to 10 cysts >2 cm in diameter). The 8 kDa
School of Veterinary Medicine, Universidad Nacional Mayor de San band was present in all seropositive animals (Table 1).
Marcos, Apartado 03-5113, Lima 03, Peru Sixty-six animals (70 per cent) had cysts in both the lungs and
M. H. Moro, Department of Immunology, Mayo Clinic, Rochester, liver, 25 (27 per cent) had cysts only in the lung and three (3 per
Minnesota 55905, USA cent) had cysts only in the liver. The sensitivity of the immunoblot
E. Dueger, University of California at Davis, School of Veterinary according to cyst location was 83, 52 and 33 per cent, respectively
Medicine, California, USA (Table 2). No cross-reactions were noted in the sera of experimen-
Correspondence to Professor Gilman tally infected animals.
606
TABLE 2: Sensitivity of the enzyme-linked immunoelectrotransfer blot
(EITB) assay for the diagnosis of sheep hydatidosis according to the
organ affected
Number of Number (%) seropositive
Sensitivity sheep in any or all bands
Lung 25 13(52)
Liver 3 1 (33)
Multiple* 66 55 (83)
Overall 94 69 (73)
* Cysts in lung and liver
The EITB assay for the serodiagnosis of hydatidosis in sheep
demonstrated a moderate sensitivity (73 per cent) and high speci-
ficity (98-6 per cent). No cross-reactions were observed in animals
infected with T ovis or T hydatigena. The sensitivity was higher
for animals with heavy infections. Major drawbacks for the diag-
nosis of hydatidosis in sheep in the past have been the relatively
low sensitivity of the assays employed (Lightowlers and others
1984) and cross-reactivity with related cestodes such as T ovis and
Thydatigena (Craig and Rickard 1981, Young and Heath 1984).
Although the sensitivity of this EITB assay is similar to that of
the indirect haemagglutination or enzyme-linked immunosorbent
assays (Martinez Gomez and others 1980, Young and Heath
1984), the EITB assay has the advantage of high specificity and no
observed cross-reactions with other taenid infections. Further
refinements in the immunoblot assay, such as the use of purified
antigens, might result in higher sensitivities.
Use of enzyme-linked immunosorbent assays for echinococcal
diagnosis in sheep have resulted in cross-reactions against T ovis
and T hydatigena, even when hydatid antigens purified by affinity
chromatography were used (Craig and Rickard 1981). The EITB
described here may be suitable as a field tool to determine the
prevalence of ovine hydatidosis, especially in areas where T ovis
and T hydatigena are highly endemic.
Acknowledgements. This study was funded in part by NIH grant
- hypogastrium (three cases) in which partial resection of both the
No. 1-UOI A135894-01 and Consejo Nacional de Ciencia y
Tecnologia (CONCYTEC), Lima, Peru. The authors thank J. Moro,
N. Perez-Palma, J. B. Phu, D. Sara, P. M. Schantz, G. Leguia and
G. Montes for editorial help.
References
CRAIG, P. S. & RICKARD, M. D. (1981) International Journalfor Parasitology 11,
441
LIGHTOWLERS, M. W., RICKARD, M. D., HONEY, D., OBENDORF, D. L. &
MITCHELL, G. F. (1984) Australian Veterinary Journal 61, 101
MARTINEZ GOMEZ, F., HERNANDEZ RODRIGUEZ, S., NAVARRETE
LOPEZ-COZAR, I. & CALERO CARRETERO, R. (1980) Veterinary
Parasitology 7, 33
SWEATMAN, G. K., WILLIAMS, R. J., MORIARTY, K. M. & HENSHALL, T. C.
(1963) Research in Veterinary Science 4, 187
TSANG, V. C. W., BRAND, J. A. & BOYER, A. E. (1989) Journal of Infectious
Diseases 159, 50
VERASTEGUI, M., MORO, P., GUEVARA, A., RODRIGUEZ, T., MIRANDA, E.
& GILMAN, R. H. (1992) Journal of Clinical Microbiology 30, 1557
YOUNG, W. K. & HEATH, D. D. (1984) Research in Veterinary Science 36,24
The Veterinary Formulary
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6 Melville Terrace, Edinburgh EH9 IND, telephone 0131 667 8295,
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The Veterinary Record, June 7, 1997
Commercial polyester fabric repair
of abdominal hernias and defects
M. Shoukry, M. El-Keiey, M. Hamouda, S. Gadallah
Veterinary Record (1997) 140, 606-607
AN extensive literature has accumulated on the use of various
synthetic materials for reinforcing hernia repair and for bridging
tissue defects with variable results. The indications for prosthetic
hemiorrhaphy (Wion 1957, Adler 1962, Gilsdrof and Shea 1975,
Karakousis and others 1975, Larson and Harrower 1978,
Tulleners and Fretz 1983), and the technique of implantation
(Johnson 1969, Scott 1979, Tulleners and Fretz 1983) have been
described by many authors.
The aim of the present work was to study the feasibility of
using commercial polyester fabric (CPF) as a prosthetic mesh for
the reconstruction of major abdominal hernias and defects in
experimental and clinically affected animals.
The subjects of the experimental work were 12 dogs, three
goats and three donkeys. The dogs were premedicated with
xylazine (2-0 mg/kg intramuscularly [Rompun; Bayer]) and
ketamine (5-0 mg/kg intravenously [Ketavet; Parke Davis]) before
thiopental sodium (10 mg/kg intravenously [Nesodnal; Specia
Paris]) anaesthesia. The calves and goats were premedicated with
xylazine (0-2 mg/kg) before epidural and local infiltration anaes-
thesia. The donkeys were premedicated with detomidine (20
pg/kg intravenously [Demosedan; Formos Group]) before
thiopental sodium (2-5 mg/kg intravenously) anaesthesia. An arti-
ficial abdominal defect was then produced by resection of a piece
of abdominal muscle in three different abdominal regions, such as
the epigastrium (three cases) in which partial resection of the rec-
tus abdominis muscle unilaterally, both the external and internal
abdominal oblique muscles and the transverse abdominis muscle
was made; the mesogastrium (12 cases) in which bilateral partial
resection of the rectus abdominis muscle was made; and the
external and internal abdominal oblique muscles and the trans-
verse abdominis muscle was made. These defects measured 5 x 12
cm in the dogs and goats and 12 x 20 cm in the donkeys.
An appropriate piece of sterilised CPF was implanted either in
two layers in small animals or in four layers (folded) in large ani-
mals in the abdominal defect.
The techniques of implantation used were retroperitoneal/sub-
fascial (six cases) in which the CPF was implanted between the
internal rectus sheath and peritoneum (Fig la); intraperitoneal
implantation either with (three cases) or without (three cases)
omentalisation (Fig lb); and double sandwich (three cases) in
which one layer of CPF was implanted between the internal rectus
sheath and peritoneum and the other layer was implanted superfi-
cial to the external rectus sheath (Fig ic). The two layers of CPF
were sutured in position using a staple stitch (Johnson 1969). The
subcutaneous tissues were apposed over the prosthetic material
using Vicryl 0 suture in a simple continuous pattern. The skin was
closed routinely. Preoperative prophylactic ampicillin (25 mg/kg
intravenously) was administered to all animals twice daily for
seven days and they were kept under observation to record any
complications.
Morphological and histological examinations were carried out
on full thickness sections after euthanasia at one, two, three, four,
five and six months postoperatively. The histological sections
were stained with haematoxylin and eosin and Masson's
trichrome.
The clinically affected animals comprised two bovine and two
buffalo calves (aged six to 18 months and weighing 100 to 150
M. Shoukry, M. El-Keiey, M. Hamouda, S. Gadallah, Departments of
Surgery and Pathology, Faculty of Veterinary Medicine, Cairo University,
Giza, Egypt 12211