Campylobacter pdf

Antibiotic resistance in Campylobacter is emerging globally and has already been described by several authors and recognized by the WHO, as a problem of public health importance Greig, ; Takkinen et al. Most patients infected with Campylobacter spp. Antibiotics, generally macrolides, tetracycline and fluoro quinolones, are reserved for more severe cases. However, the increasing resistance to fluoro quinolones, tetracycline and erythromycin of C. Gentamycin is the only alternative to fluoroquinolones and macrolides, for systemic infections caused by Campylobacter spp.

Aarestrup and Engberg, Although comparable standardized procedures for susceptibility testing are available for a wide range of organisms, based on the guidelines defined by the National Committee for Clinical Laboratory Standards NCCLS , no internationally accepted criteria are available for susceptibility testing of Campylobacter spp.

As a result, there is a debate in the literature regarding the interpretation of the results of antimicrobial resistance Ge et al. Nevertheless, those studies comparing the evolution of antibiotic resistance patterns with time, observed rapid developments for both clinical Endtz, ; Rautelin et al. There is strong evidence that supports the hypothesis that the unregulated use of antimicrobial agents in food animal production has led to the emergence and spread of antibiotic resistance among Campylobacter spp.

The approval and use of fluoroquinolones in poultry in Europe and the USA were followed by increases in fluoroquinolone resistance in Campylobacter spp. A higher prevalence of multi resistant strains has been reported for animal and meat isolates than for human isolates EFSA, Campylobacter strains isolated from broiler chickens raised in conventional farms are significantly more resistant to antibiotics than those isolated from animals grown in organic regimes, since the use of antibiotics in the rearing of organic chickens is prohibited Luangtongkum et al.

For reasons that remain unclear, C. Studies by Gallay et al. However, other studies have suggested that resistance may persist for long time periods Nelson et al.

The avian species are the most common hosts for Campylobacter spp. Although all commercial poultry species can carry Campylobacter spp. Chicken meat comprises a substantial source of a high quality protein in most countries. Chicken meat is rich in essential amino acids along with vitamins and minerals. Lean chicken contains more protein than the same amount of lean roasted beef and the prices of chicken meat are lower than those of beef or pork.

Additionally, chicken by-products are eaten widely due to their low price, special taste, and the short time required for preparation. The consumption of chicken and chicken products, however, has been implicated over the recent years in a large number of outbreaks of acute campylobacteriosis in human populations worldwide, in both industrialized and developing countries, and especially in children, the elderly and immuno-suppressed patients Tauxe, ; Skirrow, ; Corry and Atabay, Most cases are associated with handling raw poultry, eating raw or undercooked poultry meat or cross-contamination of raw to cooked foods Butzler and Oosterom, ; Tauxe et al.

El-Shibiny et al. In a recent surveillance study in England and Wales, C. However, C. The intestinal tract of chicken, especially the cecum and colon, can harbor a large number of Campylobacter spp. Persistence and survival of Campylobacter spp. Previous studies reported that growth on skin stored at room temperature in a controlled atmosphere package is possible, increasing the risk for consumers if contaminated chicken is not adequately stored or handled Lee et al.

A study was carried out to investigate the effect of environmental temperatures over different seasons on the survival of C. Wills and Murray demonstrated that Campylobacter spp.

It was also reported that there was substantial variability in the intestinal colonization of C. Some published comparative information on the incidence of Campylobacter spp. The prevalence of Campylobacter spp. The high prevalence of Campylobacter spp. This is most likely attributable to the fact that pig carcasses undergo a communal scalding stage early in the slaughter process combined with the fact that the skin remains on the carcass following all of the dressing procedures Moore et al.

Contaminated shellfish have also been implicated as a vehicle in the dissemination of campylobacteriosis. Harvesting shellfish from Campylobacter -contaminated waters would appear to be the most likely cause of infection Wilson and Moore, Consumption of untreated water Schorr et al. In an ecological study in Sweden, positive associations were found between the incidence of Campylobacter spp.

There were similar associations with ruminant density. These observations suggest that drinking water and contamination from livestock might also be important factors in explaining at least a proportion of human sporadic campylobacteriosis cases Nygard et al. Raw milk has also been identified as a vehicle of human gastroenteritis caused by Campylobacter spp. Blaser et al. In addition to risks from food, especially poultry, and water consumption, contact with animals, either domestic pets or farm animals, presents another exposure pathway for human infection Kapperud et al.

Contamination of the environment by domestic and wild animal feces constitutes an additional risk for human infection via drinking Duke et al. Most infections are believed to result from the ingestion of contaminated food, although the role of other, non-food exposures in the epidemiology of sporadic campylobacteriosis is still unknown Brown et al.

Point source outbreaks are thought to be relatively uncommon compared to those by other major enteric pathogens, although there is increasing evidence for localized transmission Charlett et al. The primary source of contamination is believed to be animal feces. This is consistent with high carriage rates observed in poultry, pigs, and cattle Kramer et al.

Flock positivity for Campylobacter spp. Positive flocks are generally more frequent among organic and free-range chickens than among intensively reared birds, probably due to increased environmental exposure Hendrixson and DiRita, Consistent with exposure of the chickens to different environmental sources is the finding that organic and free-range chickens can be colonized with multiple genotypes of Campylobacter spp.

Newell and Wagenaar, Reducing levels of Campylobacter spp. Before control can be properly applied it is important to identify the sources and routes of infection in housed flocks. Horizontal transmission from the environment is considered to be the most likely source of Campylobacter spp. It is widely accepted that horizontal transmission within a flock occurs rapidly once individual birds are colonized by Campylobacter Carrillo et al.

Once established, it is very difficult to eliminate. High flock size, environmental water supplies, litter, insects, wild birds, rodents, fecal contact, personnel and other animals, may increase the risk of colonization and dissemination Aarts et al. Feed has not been implicated in the spread of Campylobacter spp.

White et al. The ubiquity of Campylobacter spp. However, Campylobacter spp. However, it is important to remember that, as with water, feed can act as a vehicle for horizontal transmission in a broiler house once Campylobacter spp.

There is continuing debate about the relative contribution of vertical transmission of Campylobacter spp. Clark and Bueschkens inoculated fertile chicken eggs with C.

Lindblom et al. The carrier rate of C. Kazwala et al. No differences between the types of Campylobacter spp. Cox et al. Moreover, the presence of Campylobacter in eggs and hatchery fluff may indicate the possibility of vertical transmission.

However, Callicott et al. Although this latter possibility was not excluded, it was considered as of little relevance. Although Campylobacter spp. Also, it is now recognized that campylobacters can attain the state of VBNC, that can lead to under-estimation or non-detection of the organism by culture-based techniques, yet cells in this state can still infect susceptible hosts poultry or humans, e. However, campylobacters are sensitive to drying or even low humidities, freezing and freeze—thaw stress, oxygen, etc.

Since poultry, especially of chicken as a widely consumed and relatively cheap source of meat, is the main source of human campylobacteriosis, this is the main focus of efforts to reduce human disease. As campylobacters are common in wild and domestic animals, and therefore in the environment, it is important to minimize contamination of chicken rearing houses from such sources.

Installing hygienic barriers between the external and internal environments, such as controlling the entry of farm personnel, strict hygienic routines such as washing and sanitizing of hands, changing boots and coveralls before entering, have been shown to be effective, but these barriers have often been found to be breached.

Rearing chickens in a free-range system has a much greater risk of infection compared to conventional production and therefore increased difficulties in control Humphrey et al.

Minimizing the amounts of or eliminating animal protein in feed, and sanitizing the water supply, have also been effective procedures. Another factor shown to result in spread of infection, is the practice of only partly emptying a rearing house, leaving some birds to grow further. The remaining birds show an increased level of infection Hald et al.

The use of antibiotics in food animal rearing, is no longer an acceptable approach since this has given rise to antibiotic-resistant strains, severely limiting the efficacy of antibiotics in treating human disease. However, the use of pre- and pro-biotics, i. Competitive exclusion Nurmi principle , generally successful for control of salmonellae, has not always been successful for controlling campylobacters Mead, Application of bacteriocin-producing bacteria e.

Similarly, application of bacteriophages lytic for C. Carvalho et al. Although there were phage-resistant strains detected, in one trial Carrillo et al. However, in the trials by Carvalho et al. Developing an effective phage treatment, by careful selection of lytic phages and use of a cocktail to minimize the appearance of phage-resistant strains, seems a viable means of reducing the level of infection in flocks and individual birds, although it is unlikely to eliminate the organisms.

However, reduction in numbers of campylobacters on carcasses, can lead to a corresponding reduction in human infections. Segregation of Campylobacter -positive flocks from negative flocks at the slaughter house, and slaughtering of the positive flocks, has proved to be an effective method of reducing spread of contamination Wagenaar et al.

The segregation of flocks was achieved by using a rapid testing protocol a 4-h gel-based PCR technique to identify positive from negative flocks. Strict cleaning practices after processing positive poultry were essential to the operation and certification Krause et al.

Along the processing line, there is a gradual reduction in the levels of campylobacters on the meat as a result of washing, de-feathering, submersion chilling, etc. If there is spread of fecal material from live birds or carcasses by rupture of the gut during evisceration, then there will be a local spread and subsequent contamination of later carcasses. Although there are no strict Critical Control Points i. Since Campylobacter spp. Since campylobacters can readily transfer and appear to attach to surfaces, cross-contamination needs to be avoided, and the current recommendation of transferring poultry from wrapping directly to the oven, rather than washing under running water, is a result of this need.

Routinely, hot water is used to wash working surfaces and utensils in order to control the presence of Campylobacter spp. Cogan et al. Dipping or spraying of carcasses or parts of poultry, with lactic acid, citric acid or hypochlorite, can achieve only maximal reductions of 1.

Humphrey et al. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. National Center for Biotechnology Information , U.

Front Microbiol. Published online Sep Author information Article notes Copyright and License information Disclaimer. This article was submitted to Frontiers in Food Microbiology, a specialty of Frontiers in Microbiology. Received Jun 8; Accepted Sep 7. This is an open-access article subject to a non-exclusive license between the authors and Frontiers Media SA, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and other Frontiers conditions are complied with.

This article has been cited by other articles in PMC. Abstract Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide. Keywords: Campylobacter spp. The Genus Campylobacter It is believed that the first report concerning Campylobacter was back in by Theodore Escherich who observed and described non-culturable spiral-shaped bacteria Vandamme, ; King and Adams, ; Vandamme et al.

Table 1 Reported D -values for Campylobacter jejun i and C. Open in a separate window. Detection, Isolation and Confirmation The sensitivity of Campylobacter spp. Table 2 Food products implicated in campylobacteriosis outbreaks occurring worldwide. Year Location Total no. Pathogenesis of Campylobacter Virulence factors Specific virulence mechanisms have not yet been clearly elucidated for Campylobacter spp. Flagella Motility, which increases under highly viscous conditions, is essential for colonization of the small intestine Jagannathan and Penn, ; Guerry, Cytolethal distending toxin Cytolethal distending toxin CDT is widely distributed among Gram-negative bacteria Ceelen et al.

Antimicrobial susceptibility of Campylobacter Antibiotic resistance in Campylobacter is emerging globally and has already been described by several authors and recognized by the WHO, as a problem of public health importance Greig, ; Takkinen et al.

Prevalence of Campylobacter spp. Control Strategies Although Campylobacter spp. On-farm As campylobacters are common in wild and domestic animals, and therefore in the environment, it is important to minimize contamination of chicken rearing houses from such sources. In processing plants Segregation of Campylobacter -positive flocks from negative flocks at the slaughter house, and slaughtering of the positive flocks, has proved to be an effective method of reducing spread of contamination Wagenaar et al.

In domestic and catering kitchens Since Campylobacter spp. Conflict of Interest Statement The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. References Aarestrup F. Antimicrobial resistance of thermophilic Campylobacter. Discrepancy between Penner serotyping and polymerase chain reaction fingerprinting of Campylobacter isolated from poultry and other animal sources.

The use of bacterial membrane fractions for the detection of Campylobacter species in shellfish. Rapid Methods Autom.

Second Report on Campylobacter. Advisory Committee on the Microbiological Safety of Food. Available at: www. Adak G. The Public Health Laboratory Service national case-control study of primary indigenous sporadic cases of Campylobacter infection. Use of systematic review to assist the development of Campylobacter control strategies in broilers.

Antibiotic resistance and resistance mechanisms in Campylobacter jejuni and Campylobacter coli. FEMS Microbiol. Campylobacter jejuni infections: update on emerging issues and trends. An outbreak of Campylobacter jejuni enteritis in a school of Madrid, Spain. Handling infected animals, without carefully washing hands afterwards, can also lead to illness.

Campylobacteriosis can cause mild to severe diarrhea, often with traces of blood in the stool. Most people also have a fever and stomach cramping, and some might have nausea and vomiting.

The illness usually lasts about one week. Some people infected with Campylobacter do not have any symptoms. In people with weakened immune systems, the bacteria can spread occasionally to the bloodstream and cause a life-threatening infection.

Campylobacteriosis is diagnosed when a laboratory test detects Campylobacter bacteria in stools, body tissue, or fluids. Most people with a Campylobacter infection will recover without treatment. If enrichments are bubbled during incubation, leave them in bags or ml flasks. The gas does not penetrate into a larger volume sufficiently to provide proper growth of campylobacters.

Request investigators collect liters for analysis. When collected, 5 ml of 1 M sodium thiosulfate should be added per liter of chlorinated water sample. These filters have a positive charge. The negatively charged Gram-negative organisms are more effectively retained in the filter. Filter larger volumes, especially those that are turbid, through 90 mm or larger diameter filters. Place filter unit into autoclavable pan. If filter clogs, wear sterile gloves and open filter holder unit to aseptically remove filter with sterile forceps.

Place filter into enrichment broth see below. Place another sterile filter in unit, reassemble, and continue filtering. Use as many filters as needed per subsample. When analyzing sea or other salt water, flush excess salt off filter by running , ml depending on filter size sterile phosphate buffer through the filter as the last of the sample is going through the filter.

Do this with every filter used for salt water analyses. Do not let filter become completely dry. Immediately transfer finished filter to broth. Campylobacters are very sensitive to drying and high salt concentrations. Place filter s in broth in the enrichment container. When using large filters, fragment with a sterile pipet. Be sure the broth covers the filter s. Enrichments incubated in Campy gas in anaerobe jars should be ml or less. Larger volumes should be divided into smaller amounts, aseptically dividing the filters.

Pipet 10 ml enrichment broth into sterile 50 ml Erlenmeyer flasks with foil tops. Place one swab into each flask, aseptically breaking off the sticks below the top of the flask. Replace covers loosely. Place flasks in anaerobe jar. To fit two layers of flasks in jar, place a cardboard circle over bottom layer, leaving space around the cardboard's edge for gas circulation.

Other milk types and ice cream. Adjust pH as in raw milk. Centrifuge a 50 g portion at 20, x g for 40 minutes. Discard supernatant and dissolve pellet not fat layer in 10 ml enrichment broth.

Transfer pellet to 90 ml enrichment broth. Ice cream and other frozen dairy products: melt and aseptically remove any candy or other solids before weighing out.

All water or shellfish samples are pre-enriched by the 5 h method. This method yields greater recovery for severely stressed organisms.

If analyzing for C. Incubate shaking enrichments h, except shellfish samples, which are incubated an extra 4 h. Dairy samples are incubated 48 hrs total. Incubate non-shaking enrichments h, except shellfish, which are incubated 48 h. Incubate samples for C. Analysts may choose from three methods for generating microaerobic conditions in enrichment broth. These are: bubbling the gas mixture through broth, shaking enrichments to incorporate the gas, or incubating in anaerobe jars with a modified atmosphere.

The first method uses the bubbler system that also can incorporate shaking the enrichments during incubation. The second uses heat-sealed, gassed, metalized poly pouches or evacuated and gassed flasks. The third method is the evacuated and gassed anaerobe jar or a jar that uses a Campy gas envelope. Choose this when other systems are not available. Exception: incubation of enrichments in 50 ml Erlenmeyer flasks i.

Double-bag enrichments to prevent bags from leaking bags can tear during shaking. Add about10 ml water to the outer bag for optimum heat trasfer to the broth. Fill excess space in basket with water-filled dilution bottles. Place 1 ml plastic pipet tip end into each bag and fasten tightly with a twist-tie. Insert the plugged end of each pipet into the tubing connected to the bubbler valves.

Open the main gas tank valve and set the pressure to lb with the regulator adjusting screw. This will give a flow rate of bubbles per sec into each bag figs. Tie bubbler tubing for each enrichment loosely together above the baskets to keep the bags standing upright. Bring the water level of the bath up slightly higher than the level of the broths in the bags. Replenish water as needed during the incubation period. Refer to the BAM, 7th ed.

Shaker bag system. Loosen the ring clamp holding the Schrader chuck and clip valve on the hose at the gas tank, and remove the valve. Open the main gas tank valve and set the regulator to 2 lb with the regulator adjusting screw. Use a new pipet for each bag. Heat seal each filled bag metalized poly pouch with a bag sealer.

Cut a very small corner from the top of the sealed end. Squeeze air from bag by pressing area above liquid until the area is flat. Insert pipet into open corner of bag and open on-off valve on the gas hose.

Fill area above the broth with gas. Repeat squeezing and gassing each bag 2 more times, ending with a gassing step. Quickly heat seal or tape the corner of the bag shut. Place gassed bags into baskets lined with plastic bags. Set the basket s onto a shaker incubator platform. Set shaker speed to rpm.

Guidelines for storing and maintaining jars. If a jar lid with gauges is knocked against a hard surface, a gauge can become misaligned. Mark new "0" place on gauge and adjust vacuum and gas readings accordingly. Store jars with screw clamps placed in jars so that one end is lying over lip of jar bottom. Prop lid against clamp to allow free flow of air and prevent mold build-up from damp jar.

If a jar will not hold vacuum or gas pressure, check for the following: cracks in the jar bottom, cracked or missing rubber rings or seals in the lids or a faulty valve stem. Replacement valve stems and a Schrader extractor tool are available from the jars' distributors or bicycle shops.

To replace stems, place prongs of extractor over valve stem and turn counter-clockwise until stem is removed. Drop new valve stem pin-head side up into valve and turn clockwise until meeting resistance. If cultures are used often, they can be kept at room temperature in semi-solid storage media G Control cultures can be ordered from ATCC. Labs should stock C. Inoculate broth or agar positive controls from a frozen culture by rubbing a moistened sterile swab against the culture and breaking off the swab end into broth or swabbing agar plate.

Incubate microaerobically. Incubate C. Wearing gloves, pipet 1. Use a sterile hockey stick to wash the growth to one end of each plate.

Transfer washings to a sterile test tube.