Study of spread the Campylobacter jejuni among children with Diarrhea
Nagham Shahin*, Nazih Daood
Department of Microbiology, Faculty of Pharmacy, Tishreen University, Lattakia, Syria
Campylobacter infection is recognized as a major cause of acute diarrhea in Humans especially children worldwide. This study was designed to determine the distribution of Campylobacter jejuni (C. jejuni) among children suffering from diarrhea in Syria. It is the first study dealing with campylobacteriosis caused by C. jejuni in Syria. The study was carried out on 73 stool samples of children with ages up to fourteen years admitted with diarrhea in two local hospitals during the period from September 2017 to December 2017. The samples were collected and analyzed to detect for the presence of Campylobacter jejuni by using Skirrow medium, and the incubation was at 42oC under microaerophilic conditions. The growth after 48 hours was provisionally identified by colonial morphology, oxidase test, catalase test, Gram staining and motility. The organisms were identified to species level by hippurate hydrolysis, resistance to Cephalothin and sensitivity to Nalidixic acid. This study showed that 5 of 73 stool samples were positive for campylobacter spp. (6.84%). three of which were positive for C. jejuni (4.10%) and two for C. coli (2.73%). In conclusion this study showed that Campylobacter spp. especially Campylobacter jejuni is frequently associated with childhood diarrhea in Syria. Furthermore Campylobacteriosis may be underdiagnosed because the detection of Campylobacter is not part of the routine stool culture.
Campylobacter organisms are Gram-negative spiral rods, non-spore forming1,2,3 measuring 0.2 to 0.9 microns wide and 0.5 to 5 microns long, motile by a polar unsheathed flagellum at one or both ends. They are microaerophilic, although there are some can grow aerobically or anaerobically3.
Campylobacter spp. especially Campylobacter jejun (C. jejuni) and Campylobacter coli (C. coli) are the most common and important causes of gastroenteritis in humans especially among infants and children in both developed and developing countries3,4 The infectious caused by the pervious bacteria is called campylobacteriosis, and it has been increasing worldwide, exceeding shigellosis and shigellosis5,6
Campylobacteriosis is food-borne zoonotic infection, which often acquired from consuming contaminated animal food products, milk and water3,7,8,9
Clinical manifestations of campylobacteriosis are largely undistinguishable from other bacterial gut infections and include fever, abdominal cramping, and diarrhea, with or without blood in stool. These symptoms can last from three days to one week, In some cases of C. jejuni infection, individuals may develop post infection complications associated with Guillain- Barré Syndrome or Miller Fischer Syndrome3,8,10,11
Campylobacter spp. are detected by various methods but culture using selective medias consider as the typical way for the laboratory detection. Recently, several antigen-based tests for the detection of Campylobacter have been developed, and have excellent sensitivity and specificity compared with the results of culture (> 89%) 12.
The aim of the current study is to determine the distribution of Campylobacter jejuni among children suffering from diarrhea in two hospitals in Syria, by culture stool samples Skirrow medium and incubate it at 42oC under microaerophilic conditions.
MATERIALS AND METHODS:
A total of 73 stool samples (41: males, 32: famales) were collected from children with ages up to fourteen years admitted with diarrhea in two local hospitals during the period from September 2017 to December 2017. Diarrhea was defined as per guidelines:
(three episodes of passing loose stools per day, for at least 2 days). Patients on antibiotic therapy three days before sample collection were excluded from this study. A detailed history of patients were recorded as following:
frequency of stools, its content, contact with animals and the nature of food eaten.
The samples were collected in clean plastic containers and transported to the microbiology laboratory immediately.
Cultivation of samples:
The samples were prepared by emulsify approximately 0.5 g of the sample in 5 ml of sterile NaCl (0.9%) to form an 1:10 dilution and Inoculate it onto the selective medium with a sterile disposable plastic spreader or cotton tipped swab.
The selective used media was Skirrow medium composes of: campylobacter agar base (Liofilchem, Italy) + Blood 5 % and supplement S.
After inoculate the sample, two antibiotics disks (Cephalothin and Nalidixic acid) were applied onto the surface of the medium to confirm the diagnosis, then the plates incubated at 42oC under microaerophilic conditions provided by a candle jar, for 48 hours.
Identification of Campylobacter jejuni:
Typically, the growth was provisionally identified by colonial morphology: size, color, edges, side views and surface of the colonies. the characters of Campylobacter family colonies are flat and gray with an irregular edge or raise and round with mucoid appearance although, some strains could appear tan or slightly pink, and it is not haemolytic (Fig. 1).
For confirmation the identification, the following tests were carried: oxidase test, catalase test, Gram staining and motility study. The organisms were identified to species level by hippurate hydrolysis, and susceptibility to the disks containing Cephalothin and Nalidixic acid (Fig. 2).
Fig. 1: Colonial morphology of Campylobacter on Skirrow medium
Culturing onto Skirrow medium:
This medium should grew only Campylobacter spp. because of the precence of the antibiotics, and also the role of incubation degree (42o), although information tests such as biochemical tests and gram stain should be done (Fig. 3).
Fig. 2: Campylobacter spp. on Skirrow medium with antibiotics disks (up: Nalidixic acid, down: Cephalothin)
Fig. 3: Campylobacter spp. under microscope
RESULTS AND DISSICUISON:
Out of the total of 73 stool samples, 5 were positive for Campylobacter spp. (6.84%) (Fig. 4). All of these positive samples distributed in Hospital 1, three of them were positive for C. jejuni (60%), and two were positive for C. coli (40%) (Fig. 5).
The three C. jejuni samples were from males children, their ages was 10, 7, 5 years, and two of them were brothers, the two C. coli samples were from male and female, their ages were 7 and 8 respectively.
All of the patients affected by Campylobacter spp. had abdominal pain and fever with or without other complaints. Of the three patients with C. jejuni isolates, two (66.6%) had symptoms of bloody diarrhea stool samples and vomiting. and one (33.3%) had watery diarrheal stool samples. A history of chicken meat consumption was noted in the all cases (100%).
As conclusion result of the study, Campylobacter spp. especially Campylobacter jejuni is frequently associated with childhood diarrhea in Syria. And there is a strong relation between consuming poultry meat and the infection with Campylobacter spp.
our study results matched with the results of the study carried by F. Dabboussi, et al. (2010) were they found that Campylobacter species is frequently associated with childhood diarrhoea in north Lebanon, Out of the total of 90 stool samples from children (aged 1 month to 10 years) presenting with diarrhoea were collected from 5 hospitals.10 were positive for Campylobacter species (11.1%): 1 for C .coli, 1 for C. jejuni, 2 for both C. jejuni and C. coli, which agree with the findings of our present study13.
In the study carried out by Shu-Chien Wang, et al., (2007) they found that Campylobacter is a major enteric pathogen in northern Taiwan, especially in children younger than 5 years (60.6%).
C. jejuni was found in 80 patients out of 104 patients with campylobacteriosis (76.9%), while Campylobacter coli was found in 24 patients (23.1%). which agree with the results of our study.
Moreover by A. M. Ali, et al. (2002) they found that Campylobacter jejuni is a frequent cause of diarrhoea/ dysentery in children, it was positive out of 80% of samples.
Furthermore, Parvin Hassanzadeh and Mohammad Motamedifar (2006) confirmed the presence of C. jejuni as a significant cause of gastroenteritis in Shiraz, Southwest Iran. Of the 114 samples, 40 proved to be positive for C. jejuni 11 (9.6%), and Maximum prevalence of C. jejuni was observed in patients aged 11–15 years.
Fig. 4: Diarrhea causes in our study
Fig. 5: Campylobacter spp. distribution in our study
In conclusion, This study confirmed that Campylobacter spp. especially Campylobacter jejuni is frequently associated with childhood diarrhea in Syria. Furthermore Campylobacteriosis may be underdiagnosed because the detect of Campylobacter spp. is not part of the routine stool culture.
1- Shu-Chien Wang, et al. Campylobacter enteritis in children in northern Taiwan a 7-year experience. Journal of Microbiology, Immunology and Infection. 2008; 41:408-413.
2- Yu-Chia Hsieh, et al. The Genus Campylobacter: A decade of progress. Clin. Microbiol. Rev. 1988,1 0172- 157
3- Sharon V. R. Epps, et al. Foodborne Campylobacter: Infections, Metabolism, Pathogenesis and Reservoirs. Int. J. Environ. Res. Public Health 2013; 10:6292-6304.
4- E. Oh, et al. Differential Survival of Hyper-Aerotolerant Campylobacter jejuni under Different Gas Conditions Original research. Available from: URL: http:// www.frontiersin.org/ vol-8-issue-954/2017/.
5- Altekruse, S.F, et al. Campylobacter jejuni and emerging foodborne pathogen. Emerging Infectious Diseases. 1999; 5(1): 28–35.
6- Akitoye O. Coker, et al. Human Campylobacteriosis in Developing Countries. Emerging Infectious Diseases. 2002; Vol. 8, No. 3: 237-43.
7- Joana Silva, et al. Campylobacter spp. as a foodborne pathogen: a review. 2011; Frontiers in Microbiology. Vol. 2, Article 200. URL: http://www.frontiersin.org/.
8- Julian M. Ketley. Pathogenesis of enteric infection by Campylobacter. Microbiology 1997; 143:5-21.
9- Humphrey T, O'Brien S, Madsen M. Campylobacters as zoonotic pathogens: a food production perspective. Int J Food Microbiol. 2007 Jul 15;117(3):237-57.
10- Iain A. Gillespie, et al. Investigating vomiting and/or bloody diarrhoea in Campylobacter jejuni infection. Journal of Medical Microbiology 2006; 55:741–746.
11- Martin J. Blaser. Epidemiologic and Clinical Features of Campylobacter jejuni Infections. The Journal of Infectious Diseases. 1997;176(2): 103–5.
12- A. James, et al. Detection of Campylobacter in Stool and Determination of Significance by Culture, Enzyme Immunoassay, and PCR in Developing Countries. J Clin Microbiol. 2014;52(4): 1074–1080.
Received on 18.05.2018 Modified on 10.07.2018
Accepted on 16.07.2018 © RJPT All right reserved