Relationship between infection with Campylobacteriosis and genital disorder in ewes

Relationship between infection with Campylobacteriosis and genital disorder in ewes

Nawaf N. Dhaher¹, Bashar S. Noomi², Hiba Y. Khalaf², Sanaa S. Ahmed²

1Department of Internal Medicine, Surgery and Obstetrics, College of Vet. Med., Tikrit University, Iraq.

2Department of Microbiology, College of Vet. Med., Tikrit University,  Iraq.

Corresponding Author Email Address: Vetbashar@tu.edu.iq

ORCID: https://orcid.org/0000-0002-7890-0629

DOI:

Received:               ; Accepted

Abstract

Key words: Campylobacteriosis, genital disorder, abortion

Introduction

Many bacterial causes abortion in sheep. Campylobacter spp. one of them and may occur asymptomatic or cause abortions and infertility. Campylobacter spp. may also cause enteritis and ileitis (1). The main pathogenic species of Campylobacter are Campylobacter fetus subsp. fetus and Campylobacter jejuni which cause abortion in sheep. Thermophilic campylobacters (C. coli) may also cause abortions in sheep (2).

The first isolation of Campylobacter from aborted sheep fetuses was in 1909. While, it was named in 1963. Campylobacter is gram negative bacteria, curve, helical or gull-shape. It shedding throw feces, aborted fetuses’ placentas and uterine discharges, these sources may be contamination environment (3). Campylobacter transmission by ingestion of contaminated feed, water aborted material and via sexual transmission (4,5). The gross lesion appear on aborted or stillborn fetuses are subcutaneous edema, necrotic liver lesions or bronchopneumonia. On enlargement placental, cotyledons are covered with blood-tinged exudate revise. Microscopically main lesion appear on placenta are predominant in hilar zones, arterioles, leukocyte infiltration and accumulation of high numbers of bacteria within leukocyte (2,4).

Material and methods

Sampling: The current study conducted on Salahaldeen province in period from January/2021 to December/2021. Fifty abortion ewe’s cases were subjected in age 3-5 years, case history was taken and clinical examination preformed.

Twenty aborted fetus (abomasum content) and 30 vaginal swabs were collected in a period not more than 7 days. The samples were directed culturing in Brain-hart infusion broth (Himedia-India) and cultivated at 37°C for 48h.

DNA extraction: By use of ABIOpureTM Total DNA (ABIOpure, USA) according to Manufacturer's instructions. Primer used: three primers were used in the current study

a. Campylobacter fetus (give DNA product in size 143bp)

 F-5`-GCCAAGCTCATTTGCAGAGT-3`

R-5`-ACGCCAAAGACGGTGTGAG-3`

b. Campylobacter jejuni (give DNA product in size 364bp)

F- 5`-TGGAAGTGCTCCAGAAAAGG-3`

R- 5`-GCCAAATAATTCTTTTGCAAC-3`

c. Campylobacter coli (give DNA product in size 502bp)

F- 5-GGT ATG ATT TCT ACA AAG CGA G-3

R- 5-ATA AAA GAC TAT CGT CGC GTG-3

DNA reaction mixture: 25µM consist from:            3µM of DNA template, 12µM Master Mix, 1µM from each forward and reverse primers, and 4µM Nuclease Free Water.

Thermocycler programs: consist of: Initial Denaturation at 95°C for 5 min, then 30 cycles, each cycle consists of Denaturation at 95°C for 30 sec, Annealing: at 60°C for 60 sec and Extension: at 72°C for 60 sec. The final Extension: at 72°C for 7 min. Then hold at 10°C for 10 min.

Agarose Gel Electrophoresis: the electrophoresis done on agarose gel (2%) and Electrical power was turned on at 100v/mAmp for 75 min.

Results

The present study showed that out of 50 abortion cases, Campylobacter was detected in 13 cases with rate of 26% (Table 1). The current study demonstrated that 61.5% of positive cases of campylobacter spp. is accompany with placenta retention (Table 2). On the other hand, 75% of positive cases of Campylobacter spp. is a accompany with recurrent abortion that’s mean either the Campylobacter infection not produce effective immunity level or may other infection due to previous abortion (Table 3) The current study revealed that all the 13 detected cases were belong to species C. fetus, 2 cases were diagnosed as C. jejuni, and these two isolates detected as mix infection with Campylobacter fetus (Figure1 and 2). While, not detected any isolate belongs to C. Coli (Table 4).

Discussion

About 80% of abortions in ewes caused by Brucella, Toxoplasma, Campylobacter, and listeria. C. fetus and C. jejuni is the main Campylobacter species associated with sheep abortion.  All the aborted ewes and fetus should disengage in the field because they are a source of the spreading the infection (6). In the current study, Campylobacter caused abortion in a rate of 26%. This finding is in agreement with previous study in Suleimani province which is 26.8% (29 out of 108) (7). While, this rate is high than the rate recorded in Sistan region which is 7% (8). in Uruguay, Campylobacter recorded as abortion cause in a rate of 6% (9). The detected rate is less than the rate that recorded in Iran which is 50.6% (10).In the current study, the difference between infection recoded in compare with other study may due to geographic differences in the study area, seasonal of the study, techniques used in diagnosis, clinical signs management and control programs (11).The current study showed that high relationship between Campylobacter infection  and placenta retention, that’s due to the ability of Campylobacter to cause placentitis and the cotyledons separate easily from the caruncles. In addition, the placentitis lead to fibroblast activation which produce fibrin that leads to the attachment of placenta on uterus (12,13).The present study showed that 75% of abortion cases diagnosis as campylobacter infection were suffering from recurrent abortion, that’s may due to other causes of previous abortion such as Brucella, Salmonella, Toxoplasma and non-infection causes or due to expos to high dosage of bacteria by contact with aborted fetus, placenta, and uterine discharges (14, 15).Campylobacter spp. are common abortion causes in ewes, most abortion cases occur at the twelfth week of pregnancy and after 7-25 days after infection (17).

In the current study, C. coli was not detected. This finding is in agreement with previous studies (7,8,9,10). C. Coli is related to digestive system infection and is not highly related to abortion (18). In a study conducted in Baghdad, four Campylobacter spp. were isolated which were C. jejuni, C. coli, C. lari and C. fetus which from diarrheic and non-diarrheic cows (19).In the current study C. fetus is the main species of Campylobacter related to abortion in ewes. This finding is in agreement with the results of previous studies (7,8,9, and 10). C. fetus is the main cause of abortion in sheep. C. fetus expresses have a Para crystalline surface layer (S-layer) which makes bacteria resistant to serum killing by impairing C3b binding (20).Pathogenesis of C. fetus is include bacteremia, placentitis, uterine, and fetal infection, the fetus suffering from 1–2 cm orange/yellow necrotic foci in the liver. Placentitis with hemorrhagic, necrotic cotyledons and edematous or leathery areas between the cotyledons (21, 22).

Conclusion: highly relationship between Campylobacterosis with abortion, recurrent abortion, and placenta retention. C. fetus is the main species related to genital disorder in ewes.

Table 1: Number and Rate of positive cases of Campylobacter spp.

Table 2: The relationship between placenta retention and Campylobacter infection.

Table 3. The relationship between recurrent abortion and Campylobacter infection.

Table 4: Campylobacter species detected in the current study.

Figure 1. Agarose gel electrophoresis of MPCR for detection of Campylobacter spp. M: 100bp DNA marker, lens 1,4,5,6,8,10 positive to Campylobacter fetus which give band in size 143bp.

Figure 2. Agarose gel electrophoresis of MPCR for detection of Campylobacter spp. M: 100bp DNA marker, 18 positives to Campylobacter fetus which give band in size 143bp. Lens 11&14 mix infection with Campylobacter fetus which give band in size 143bp and Campylobacter jejuni give band in size 364bp.

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