STUDIES ON SOME BACTERIA ASSOCIATED WITH ABORTION IN RABBITS

Assiut Vet. Med. J. Vol. 52 No. 109 April 2006

Dept. of Bacteriology, Animal Health Research Institute, Mansoura Provincial Laboratory.

STUDIES ON SOME BACTERIA ASSOCIATED WITH

ABORTION IN RABBITS

(With 4 Tables)

By M. EL-SAYED HATAB and M.M. ABD EL-LATIF

(Received at 6/3/2006)

دراسات على بعض البکتريا المصاحبة للإجهاض في الأرانب محمد السيد محمد حطب،محمود محمد محمود عبد اللطيف

الاستبيان المسببات البکتيرية المصاحبة للإجهاض في الأرانب تم جمع عدد 60 عينة من (۲۰ أرنبة مجهضة ونافقة حديثا و ۳۰ أرنبة مجهضة وحية و10 أرانب غير مجهضة مخالطة) من المزارع الخاصة بمحافظة الدقهلية وذلک بهدف فحصها بکتريولوجيا. وأظهر الفحص البکتريولوجی وجود 44 (۷۳٫۲%) حالة إيجابية للعزل البکتيري حيث تبين إصابة بعض الحالات ۲۶ (۰۹٫۱%) بنوع واحد من البکتيريا (عدوی فردية بينما تم عزل أجناس میکروب الليستيريا مونوسيتوجينس وکذلک ميکروب البأستيريلا مالتوسيدا في ۲۰% لکل منها أما العدوى البکتيرية المختلطة فتمثلت في عزل الميکروب القولوني (

E. Coli ) مع کل من الميکروب العنقودى الذهبي (Stapha aureus) وميکروب الباستیریلا مالتوسيدا في ۱۱٫۳۶ % ،

%۹,۰۹

على التوالى کذلک تم عزل ميکروب الباستیریلا مالتوسيدا مع ميکروب السالمونيلا في ۱۱٫36% أيضا ثم عزل الميکروب المکور العنقودي الذهبي مع ميکروب الإستربتوکوکس بيوجين في ۹,۰۹ % وقد تم تصنيف معزولات السالمونيلا إلى ۸ عترات (۸۸٫۹%) سالمونيلا تيفي ميوريم. کما تم عمل اختبار حساسية للميکروبات المعزولة حيث کانت معظم المعزولات حساسة لکل من الإنروفلدکاسين والجنتاميسين بينما کانت معزولات الليستيريا مونوسيتوجينس حساسة لکل من الأمبسلين والأموکسيسيللين والبنسلين. هذا وقد تمت مناقشة النتائج والتوصيات الواجب إتباعها للمحافظة على مزارع الأرانب وکذلک الثروة القومية.

SUMMARY

A total of 60 does (20 freshly & aborted dead, 30 live & aborted and 10

private farms at El-Dakahlia aborted) were collected from non Governorate for bacteriological examination. 44 cases (73.3%) of examined samples were positive for bacterial isolates, which indicated that Listeria monocytogenes and Pasteurella multocida were the

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common bacteria as a single infection in 59.1% of examined samples, however, E. coli with Staph. aureus and P. multocida; P. multocida with Salmonella and Staph. aureus with Strept. Pyogene were isolated as mixed infection in 40.9% of examined samples. The majority of Salmonella strains were identified as Salmonella typhimurium 88.9%. In - vitro sensitivity pattern of isolated strains proved that enrofloxacin and gentamycin were the most effective drugs for most isolates, except Listeria monocytogenes was sensitive to ampicillin, amoxicillin and penicillin.

Key words: Rabbits, abortion, Salmonella, P. multocida, antibiotic sensitivity.

INTRODUCTION

In Egypt rabbit breeding is considered as an important source of good quality and economic animal protein because it needs simple requirements to start as well as its short production cycle and large number of off springs. Rabbits are more sensitive to diseases of the reproductive tract. Interest has been focused on abortion in rabbits, since it is responsible for high economic losses.

Many microorganisms are commonly associated with infectious abortion in rabbits, Salmonella species, Pasteruella multocida, Listeria monocytogenes, E. coli and Staphylococcus aureus (Harwood, 1989; Boucher and Nouaille, 1996; Peters and Scheele, 1996 and Rosell, 2000).

Several outbreaks and sporadic cases of rabbit abortion were occure in our area at El-Dakahlia, therefore the present work was aimed to study the role of bacteria as a cusative agent of abortion in rabbits, also to determine in vitro the antibiotic sensitivity of the isolated microorganisms.

MATERIALS and METHODS

I-Samples:

A total of 60 does (50 aborted does with aborted foeti and aborted materials in addition to 10 non aborted does), were collected from different private farms at El-Dakahlia Governorate. a) Aborted and freshly dead does. Samples from uterus and its content, brain, spleen, liver, heart blood, aborted foeti and aborted materials.

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Assiut Vet. Med. J. Vol. 52 No. 109 April 2006 b) Aborted living does: Samples were vaginal swabs, aborted foeti and aborted materials. c) Non aborted does: Samples were vaginal swabs. All samples were collected and subjected to bacteriological examination. II-Media: a) Liquid media: Tryptose broth, peptone water, Selenit F-broth and Listeria enrichment broth. b) Solid media: Blood agar, Tryptose agar, MacConkey's agar, Xylose Lysine deoxycholate agar and Listeria enrichment agar (Oxoid). III- Isolation and identification:

The collected samples were transferred to sterile test tubes containing tryptose broth, selenite. F. broth and listeria enrichment broth, and incubated at 37°C for 18 - 24 hours, followed by subculturing on blood agar, MacConky's agar, Xylose Lysine deoxycholate agar and Listeria enrichment agar plates and incubated aerobically at 37°C for 24 48 hours. For isolation of Pasteurela multocida the blood agar plates were incubated in candle Jar at 37°C / 48 hours.

The growing colonies on various plates were examined morphologically, culturally and biochemically according to Edward and Ewing, (1972); Cruickshank et al., (1982); Finegold and Baron, (1986) and Carter and Cole (1991). IV -Serological identification of Salmonella:

The biochemically identified salmonella strains were subjected for serological identification as described by Edward and Ewing (1972), Kauffmann (1973) and the instruction of the manufacturer (Denka Selken Co. LTD, Tokyo, Japan). V-In vitro antibiotic sensitivity test:

The disc diffusion technique was performed on isolated bacteria using Muller-Hinton medium (Oxoid). Ten chemotherapeutic disks kindly supplied by Oxoid and namely ampicillin, amoxicillin, enrofloxacin, chloramphincol, gentamycin, streptomycin, pencillin, erythromycin, oxyteteracyline and trimethoprim – sulphamethoxazole.

The degree of sensitivity was determined and interpretated according to Koneman et al., (1994); Quinn et al., (1994) and Oxoid Manual (1998).

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RESULTS

Table 1: Incidence of single and mixed infection cases with bacteria in

examined does.

Single infection Mixed

Total No. of Condition of does

cases

infection cases

positive cases No.1 % No. % Non, aborted does 3 6 .82 2 4.55 Aborted & dead does 8 18.18 10 22.73

18 Aborted & live does 15 34.09 6 13.63

21 Total

26 59.0918 40.91

44 The percentage was calculated in relation to the total number of positive cases (44).

Table 2: Incidence of different mixed bacterial isolated recovered from

examined does.

Types of microorganisms

Condition of examined does

Aborted & Aborted &

Total Non-aborted

dead

five No. % No. % No. % No. % 1 2.272 4.55 2 4.555 11.36

6.82

2.27

9.06 6.82

4.55

11.36

E.Coli + Staph. Aureus E. coli + P. multocida P. muitocida + Salmonella Staph. aureus + Strept. Pyogenes

1

2.27 | 2

4.55

i

2.27 | 4 | 9.09

Table 3: Incidence of bacteria isolates from examined does.

Bacteria isolates

E.Coli Listeria monocytogenes Pasturella multocida Salmonella species ** Staphylococcus aureus Streptococcus pyogenes Total

Condition of does Non-aborted | Aborted & Aborted &

Total (10)* dead (20)* | live (30)* No. % No. % No. % No. %

2 20.00 5 25.00 5 16.66 | 12 | 20.00 0 | 0.00 6 30.00 6 20.00 12 20.00 0 | 0.00 6 30.00 6 1 20.00 1220.00 1*** 10.00 4 20.00 4 13.33 9 15.00 2 20.00 4 20.00 4 13.33 10 16.66 2 20.00 3 15.00 2 1 6.66 7 1 11.66

28

27

6 2

* The number of examined does. ** salmonella typhymurium (8), untypable (1) ***

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Table 4: In-vitro susceptability of important isolated microorganisms.

E coli (12)

L. monocytogenes P. multocida (12)* Salmonella (9)* Staph. aureus (10)* Antibiotic disc

Sensitive Activity Sensitive Activity Sensitive Activity Sensitive Activity Sensitive Activity

isolates

isolates

isolates

isolates

isolates Ampicillin 10 ug

- 1 0 12 100.00

TOT TO Amoxycilin 25 ug

1 -

0

12 100.00 -

0

-

0 Enrofloxacin 5 ug

12

100.00

66.66 12

100.00

100.00

90.00 Gentamycin 10 ug

12 100.00 10 33,33 12 100.00 9 100.00

90.00 Streptomycin 10 ug

50.00

66.66 Chloramephincol 30 ug 10 83.33 5 41.66

66.66

88.88

40.00 Pencillin 10 ug

- 01 1083.33 - 0

40.00 Erythromycin 15 ug

0

30.00 Oxyteteracyclin 30ug

50.00

41.66

50.00

66.66

40.00 Trimethoprim-sulpha

10 83.33 7 58.33

50,00

44.44 methoxazol 1.25-23.75 ug

S000

O

O

* The number of tested strains.

DISCUSSION

Nowdays, a great attention was payed toward the diseases caused by bacteria which affected the reproductive tract of rabbits because this group of diseases has been responsible for considerable economic loss to rabbit breeder. Peters and Scheele (1996) concluded that previous infection of the uterus by Listeria monocytogenes should be considered as a cause of infertility.

It is clear from the results presented in Table (1) that out of 44 positive cases from which pathogenic bacteria were isolated, 26 (59.09%) revealed single infection, while the remaining cases 18 (40.91%) were due to mixed infection. The isolated bacteria as mixed infections included E. coli with Staphylococcus aureus in 5 cases (11.36%); E. coli with Pasteurella multocida in 4 cases (9.09%); Pasteurella multocida with Salmonella in 5 cases (11.36%) and Staphylococcus aureus with Streptococcus pyogenes in 4 cases (9.09%). Table (2).

Bacteriological examination of the samples revealed that the isolated bacterial pathogens were, E. coli, Listeria monocytogenes, Pasteurella multocida, Salmonella spp., Staphyloccus aureus and Streptococcus pyogenes (Table 3). Nearly similar pathogens were isolated by (Harwood 1989; Peter and Scheele 1996; Rosel 2000 and Boucher et al. 2001). The results achieved revealed the prevalence of Listeria monocytogenes and Pasteurella multocida in this work whereas the frequency of isolation from aborted dead and live does were 12 isolates from each as a single cause of abortion. The role of the content

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of the pregnant uterus may Simply offer a more favorable environment for the organism to grow and thereby induce a more sever disease (Gray and Killinger, 1966).

Listeriosis is a septicemic diseases of rabbits characterized by sudden death or abortion or both (Flatt, 1974), the disease is caused by Listeria monocytogenes which is a facultative intracellular gram - positive, small rod, aerobic and facultatively anaerobic, found in soil, vegetation and faeces (Rebhun, 1987 and Jubb and Huxtable, 1993). Results given in Table (3) point out that Listeria monocytogenes could be isolated from 6 of 20 aborted and dead does with an incidence percentage 30%, also it could be isolated from 6 of 30 aborted and live does with an incidence percentage 20% and could not be detected in non aborted does.

The same organism was isolated previously from rabbits (Abd El-Waneas 1985; Abd El-Motelib et al., 1990; Peters and Scheele 1996 and Abd El-Ghaffar and Abd El-Gwad 1997).

Pasteurella multocida can be a virulent pathogen of rabbits, producing fatal septicemia, pneumonia, chronic rhinitis, otitis media as well as multiple absces (Lu et al., 1978) and is considered allover the world as one of the most dangerous diseases which affect rabbit industry (Deeb et al. 1990). The results achieved from Table (3) revealed that the pasteurella multocid could be isolated from aborted dead and live does with an incidence percentage 30, 20% respectively and could not be detected in non aborted does. Nearly similar results were reported by Boucher and Nouaille 1996, Rosell 2000 and Boucher et al. 2001).

Salmonellosis is a very important disease not only from the economic point of view but also from the public health aspect as it is zoonotic disease, it occurs world wide and its incidence is on the increase (Engler, 1988). Salmonellosis in rabbits is characterized by septicemia, acute enteritis and rapid death, while pregnant does commonly aborted (Sadek and Mostufa, 1970; Ghoniem et al; 1971 and Casaro et al., 1979).

The results given in Tables (2, 3) revealed that Salmonella could be isolated from one of non aborted does and from 4 each of aborted dead and live does with an incidence percentage 10, 20 and 13.33% respectively. On serotyping of the 9 recovered Salmonella organisms from examined samples, 8 (88.9% of which were recognized as Salmonella typhymurium and 1 (11.1%) was untyped. Some authors recorded Salmonella typhimurium from aborted rabbits (Harwood, 1989; Boucher and Nouaille, 1996 and Rosell, 2000).

eco

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Esherchia coli involved in many health problems in rabbits, they may be direct cause of disease (colibacillosis or colisepticaemia) or associated with other microorganisms. E. coli were isolated from 2 (20%), 5 (25%), 5(16.66%) of non aborted does, aborted dead does and aborted live does respectively Table (3). Similar finding were also reported by Boucher et al., (2001).

Staphylococcosis is a commonly occurring disease in both domestic and wild rabbit, this disease is caused by staphylococcus aureus and characterized by fatal septicemia or suppurative inflammation in nearly any organ or site. Through the present study, staphylococcus aureus was isolated from 2(20%); 4(20%) and 4(13.33%) of examined non-aborted does, aborted dead does and aborted live does respectively Table (3).

Streptococcus has been associated with acute septicemia in rabbits. The results recorded in Table (3) revealed that Streptococcus pyogenes was isolated from 2 (20%); 3 (15%) and 2 (6.66%) of examined non-aborted does, aborted dead does and aborted live does respectively.

In vitro sensitivity testing of isolates revealed that most isolates were highly sensitive to enrofloxacin and gentamycin, except Listeria monocytogenes was sensitive to ampicillin, amoxicillin and penicillin, nearly similar results were reported by Harwood, 1989; Boisivon et al., 1990, Diker et al., 1994; Abd El-Ghaffar and Abd El-Gwad 1997 and Abd El-Rahman et al., 2005.

Finally, great attention and efforts should be paid to overcome that problem. Such efforts include, medical care of does; eliminating potential sources of infection; special precautions can be instituted to detect and eliminate carrier animals and eliminate vehicle of transmission; complete hygienic measures must be followed in the farms and avoid the misuse of antibiotics.

op

Ivory.

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