Campylobacteriosis in poultry

Campylobacteriosis in poultry

Engy A. Hamed, May Fathy Abdelaty, and Heba Roshdy

Reference Laboratory for Veterinary Quality Control on Poultry production, Animal Health Research Institute, Agricultural Research Center (ARC). Nadi El-Seid Street, Dokki P.O. Box 246, Giza 12618, Egypt.

• Abstract

Campylobacter is one of important microorganisms which responsible for food poising in human. Poultry play an important role of transmission of thermoplastic Campylobacter because it considered commensally Bacteria in gastrointestinal tract of poultry due to the body temperature of poultry 42 °C which the optimal temperature for growing of Campylobacter. Campylobacter is Gram negative short curved bacilli; motile by single bipolar flagellum has special movement (corkscrew-like). In Egypt many researchers try to isolate Campylobacter from poultry and poultry by product with incidence ranged from 4:44% since 1986 till now. The main rout of transmission of Campylobacter inside the farm is horizontal rout through contamination of food and water by the infected fecal matter or through any other vehicles but the vertical rout transmission is not confirmed till now. Application of good biosafety and biosecurity measure help in decreasing transmission of Campylobacter and other microorganisms.

Key words: Campylobacter, poultry, public health, incidence, prvention

• Introduction

Campylobacter is a gram-negative bacteriumits name is derived from Greek word “Kampylos“which means curved(keeneret al.,2004). Campylobacter is one of the important bacteria that can transmit to human through consumption of poultry meat and poultry byproducts and causes food poisoning outbreaks in developing and developed countries, Campylobacter cause many outbreaks in recent years the severity of these outbreaks were depend on many reasons like anycase had been reported or not, the level of country and this country has good labs that can isolate Campylobacter(Kaakoush et al., 2015)

Thermophilic Campylobacter species especiallyC. jejuniand C. coliwereadapted to colonize in the intestinal tract of domestic poultry(chickens, turkeys, ducks, and geese) beside the internal body temperature of poultry which 42 ̊ C that considered the optimum temperature for Campylobacter growth, also poultry body give good bacteriological conditions for Campylobacter to grow and colonize. In the pastCampylobacter lesions were named as vibronic hepatitisonce this lesion become very rare thin now a day return to be a very important problem in free-rang laying hens which causes high mortality and drop in egg production and cause new lesion in liver(multi focal small white gray lesion in liver) calledspotty liver and this caused by a new Campylobacter species named by C. hepaticusalso C.  jejunireported that it responsible for intestinal inflammation and diarrhea (Zhang and Sahin 2020; Golz et al., 2014; Dasti et al., 2009)

The prevalence rate of Campylobacter species in the poultry farms in age of slaughter in some farms may reach to 100%, Campylobacter isolated from poultry without evidence of any disease but it turns to diseases form like diarrhea and intestinal inflammation under certain conditions like experimental challenges (Golz et al., 2014; Humphrey et al., 2014).Found there are negative relationships between Campylobacter colonization with intestinal tract function and growth performance (Awad et al., 2015; Williams et al., 2013)

• History of Campylobacter isolation from poultry in Egypt

There are many researchers tried to isolate Campylobacterfrom  live poultry and poultry carcasses in slaughter houses during preparation before sold in markets in Egyptsince 1986 till now(Nouman et al., 1986; Hefnawy et al., 1989), or trying to found methods which may help in decreasing the count of Campylobacteron poultry carcass like adding ascorbic acid on carcass before distributing in markets (Khaled et al., 1990)other researcher tried toisolate Campylobacter from life and dead of different poultry species and the prevalence was ranged from 4% to 44% from 1991 till now (Ezzat et al., 1991 ; El- Gedaway 2004; Eid et al., 2018 and Hamed et al., 2023).

• Morphology of Campylobacter

Campylobacter was firstly identified as C. fetus subsp. jejunibyLuechtefeld et al. (1980) asbased vibiro forms motile growth at 42⁰C but not 25⁰C.it drives from family Campylobacteraceae, The genus Campylobacter are about 30 species and sub species like C.jejuni, C. coli, C. Lari, C. hepaticus(Lastovice et al., 2014; On 2013). Campylobacter is Gram negative curved bacilli takes many shapes spiral, S shape, gull wing or comma shape(keener et al., 2004), its size ranged from 0.2-0.8µm wide and 0.5-6.0µm long and cell can turn to coccoid form due to any stress, nonspore-froming, motile by single bipolar flagellum and has characteristic movement (corkscrew-like), it can’t ferment carbohydrate, oxidase and catalase positive while indole negative (Evans 2001; Byrd et al., 2007; Lastovica et al., 2014).

• Transmission of Campylobacter

Most common rote of Campylobacter transmission inside the poultry farmsis the Horizontal rote of transmission, through contaminated food or water with fecal matter of infected birds, contaminated litter, rodent, insect, house flies, farm worker, domestic pets, and transmission of farm equipment through farm’s houses. Due to that Campylobacter is fastidious microorganisms and it is very sensitive to exposure to the Oxygen and the normal temperature due to that Campylobacter can’t grow in fresh dry litter, fresh feed before contaminated with poultry fecal matter so applying the good biosecurity measures inside the farm and maintenance and cleaning the litter and equipment lead to decrease farm contamination with Campylobacter and other microbes (Newell et al., 2003;  Line 2002; Kassem et al., 2017; Adel et al., 2023).There is an argument about that Campylobacter can transmit vertically or not, due to low number of research which in this point in spite of there is some researchers were isolate Campylobacter from one day old chick, reproductive tract, semen of healthy bird, and from inner and outer shell of egg there is no confirmation that Campylobacter can transmit vertically (Byrd et al., 2007, Cox et al., 2002;  Cox et al., 2012; Hamed et al., 2014;Newell et al., 2003).

• Incidence of Campylobacter

Campylobacteriosis occurrence has been developing globally in recent years (Luangtongkum et al., 2009), leading to a significant increase in public health illnesses. The total number of sick individuals and the rate of death have both risen (Euro Surveill., 2015). Campylobacter jejuniis the most frequent related agent. Livestock have an essential role in the transmission of Campylobacteriosis to individuals.

In the United States, the Foodborne Diseases Active Surveillance Network (FoodNet) informed an occurrence of culture-confirmed infections linked to Campylobacter in the FoodNet sites of 12.7 per 100 thousand individuals in 2006 (Ailes et al. 2008). These findings reflect a reduction of thirty percent from the 1996 level, nevertheless the frequency stays over the national health target. Most other parts of the world have higher illness occurrences with almost 400 episodes per 100,000 residents in New Zealand in 2003 (Baker et al. 2007). Campylobacteriosis was among the most commonly reported infectious disease in humans in the European Union starting in 2004. In 2009, the collective EU reporting rate was 45.6 incidents per 100,000 people (EFSA and ECDC 2011). Campylobacter enteritis in humans is principally caused by C. jejuni (EFSA and ECDC 2011). In 2009, C. jejuni was responsible for 90% of the cases classified as that species. Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis individually contributed 2.5%, 0.2%, and 0.01% of the isolates. The other speciate isolates included other (unknown) taxa. Geographical or country disparities may still exist however: in Bosnia and Herzegovina, for instance, a higher incidence of C. coli in sporadic infections in humans (30%) has been reported (Uzunovic'- Kamberovic' et al. 2007).

Campylobacter prevalence in poultry and non-poultry farm animals varies by season, age of the animal, flock size and kind, diet, animal husbandry practices, and geography as a whole C. jejuni is the most isolated species (Kuana et al. 2008, Messens et al. 2009, Na¨ther et al. 2009, EFSA 2010, EFSA and ECDC 2011). In pigs, however, C. coli contributed to 84% of all isolates in 2009. unexpectedly in certain nations, such as Spain, Luxembourg, Slovenia, and Bosnia and Herzegovina, a significantly greater amount of C. coli than C. jejuni was additionally isolated from poultry samples (Zorman et al. 2006; EFSA and ECDC 2011),This is undoubtedly dependable, at least in particular, for the higher incidence of C. coli in sporadic infections among people in the latter two nations.

The possibility of a poultry flock getting colonized enhances during rearing, with 60%-80% of examined broiler flocks globally testing positive for Campylobacter species at slaughtering age (Herman et al. 2003, Rasschaert et al. 2006, Kuana et al. 2008, Reich et al. 2008, EFSA 2010). There are significant differences in occurrence across the EU. In 2008, almost every broiler batches in Northern European nations such as Estonia, Norway, and Finland were clear of Campylobacter, but Luxembourg had an unexpectedly high rate of 100%. In overall, C. jejuni has been found to be the most common species. In the European Union, 41% of the examined flocks were colonized especially by C. jejuni, while in Brazil, the proportion reached to 55%. Invaded avian populations could contaminate the environment around them, permitting Campylobacter to multiply and spread across other farms or individuals (Jonsson et al., 2010).

Campylobacter was isolated from chicken cloacal swabs obtained form  pressing plant in Australia by 82% ,  from fecal samples from broiler  by 67% I Sicily,  from frozen and chilled chicken carcasses by 20.3 % in Yogoslavia ,  found in 64.7% of chicken wing samples in Northern Ireland and UK, and isolated from 213 broiler carcasses by 49.2% in Italia, also isolated from retail poultry in Thailand and South Africa (Shankar et al., 1982,  Noto and Aiello 1988; Kalember and Paunovic  1989; Flynn et al., 1994; Manfreda et al., 2006; Hodaka S., Shigeki Y. (2009)).

Both freezing and refrigeration can change the amount of Campylobacter, therefore the quantity of Campylobacter may be decreased during thawing and freezing, even though the bacteria remain alive for a minimum of twelve months at -18°C (Beuchat, 1986). When a diseased bird is slaughtered, Campylobacter organisms can be conveyed from the intestines to the meat. Campylobacter was detected in 47% of raw chicken carcasses sold at grocery shops. Campylobacter has been detected in multiple poultry- processing factories, and the two Campylobacter species were identified in 32% of the chicken products acquired from restaurants. (Buchat, 1986).

Infected broiler provides the most significant hazard to consumers (Sahin et al. 2015). In comparison, layers deserved scant consideration; layers may be the main reservoirs for antimicrobial-resistant Campylobacter. This is crucial since layer farming is a huge business in Egypt. Thus, these hens carrying antibiotic resistant Campylobacter potentially pose a risk of contamination of the environment and subsequently spread to farm animals, wildlife, and workers (Ahmed et al., 2013).

 Campylobacter, in instance, could persist for many days in layer feces and poultry litter, indicating that using these wastes as fertilizer. on land can lead to the transmission of disease (Kassem et al., 2010; Ahmed et al., 2013).

• Incubation period

Experimentally Campylobactercolonization happened after one day post-infection, in few cases we can found the clinical signs of Campylobacter (diarrhea, mucosal damages and spotty liver after about 3:5 day after infection, one the colonization was happened it can survive for several weeks(Knudsen et al., 2006; Sahin et al., 2003; Van et a., 2017).

• Pathogenesis

Campylobacter has special and complicated pathological mechanisms that it considered the only bacteria has ability to execute N-linked glycosylation of more than 30 proteins related to virulence genes responsible for colonization, adhesion,and invasion. Also presence of flagellum which not only used for its motility to reach to the cite of infection but also has ability for secretion of Campylobacter invasive antigens (Cai). The Cytolethal toxins which excreted from Campylobacter speciesCdt A, B, C, DnaJ (heat shock protein), PldA ( phospholipase A), Cadf(Campylobacteradhesion to fibronectin), CmeR (a pleiotropic regulator) CmeABC ( multidrug efflux pump) and other are responsible as a major pathogenicity-associated factor (Zang and Sahin 2020; Kumar et al., 2016); which, also it has other virulence factors help in its adaptation inside host  and has a biological mechanism of colonization resistance especially young poultry body without any generations of signs and has an ability to survive in very bad condition in the gastrointestinal tract of poultry (Dasti et al., 2009; Zhang and Sahin 2020). The cycle of Campylobacterinside the host begins from entrance of infectious dose of Campylobacter to the host body mainly though ingestion contaminated food or water with fecal matter contains a very small dose of Campylobacterabout 800 organisms(Wallis 1994).Many research’s failed to isolate Campylobacter in age less than two or three weeksin spiteof the active positive infection (Kalupahana et al., 2013), in spite of this we can isolate Campylobacter from gizzard, intestinal tract, liver, spleen, gall bladder, and blood (Zhang and Sahin 2020).

Steps of cycle of Campylobacter inside the poultry body firstly Campylobacter presents in mucous layer of the crypts and not adhere directly to the intestinal epithelial cell, then usually no gross or microscopic lesions are detected in bird, finally invasion of Campylobacter inside the epithelial cell of intestinal tract. Sometimes even the invasion happens without any clinical signs can observed (Van Deun et al., 2008). Once invasion was happened bird can excrete large number of Campylobacter in feces may reach to (10⁹cfu/g feces) for a long period till slaughter time, Campylobacter colonization and appearance of clinical sings depend on many factors like kind of feed, age, and infected strain (Sahin et al., 2015; Han et al., 2016; Chaloner et al., 2014).

• Diagnosis

8.1- Culturing method

It is most usable methods for detection of thermophilic Campylobacter, it is highly fastidious and slow growing microorganism need especial conditions and requirements for growth, it needs microaerophilic atmosphere (5% O2, 10% CO2, 85% N2), need high temperature for growth 42⁰C for optimal growth in laboratory conditions, it can’t growth in under 31°C. (Hazeleger et al., 1998; Sahin et al., 2003). Isolation if Campylobacter from fecal matter and environmental samples needs special requirements in the used media and atmospheric conditions to help In Campylobacter due to highly contaminated conditions first selective media used for isolation of Campylobacter is Skirrow media (Skirrow 1977). From this time there are more than 40 selective agar and broth media made for isolation of Campylobacter, some of most common media used for Campylobacter isolation like Preston, Karmali, modified charcoal cefoperazonedeoxycolate agar (mCCDA) and Campylobacter selective chromogenic media (CASA), nowadays culturing method use for isolation of Campylobacter from food and chicken meat (Lynch et al., 2011).

Media which used in culturing Campylobacter must contain some kind of antibiotic which stop growing other microorganism rather than Campylobacter like (polymyxin, vancomycin, trimethoprim, cefoperazone, and nystatin (Lin et al., 2003) also Campylobacter is sensitive to high oxygen level that must media also contain component neutralize the toxic effect of oxygen radical like charcoal in mCCDA media, hematin in Preston medial (Corry et al., 1995). Using broth medium for enrichment before agar plating is good and important step in case of samples contain small number while in case of highly contaminated fecal matter direct plating method is the best choice (Jacobs-Reitsma et al., 2008; Musgrove et al., 2001). Increase time of pre-enrichment step more than 24 hours in highly contaminated samples may lead to decrease isolation rate (Zhang and Sahin 2020).

8.2 Molecular techniques

Because of the culturing methods may takes long time molecular techniques may be using for direct detection of Campylobacter but it mostly use for confirmation and identification of Campylobacter after culturing methods. Also we can use multiple probs which can detect more than Campylobacter species in the same time (Elvers et al., 2011; Fontenot et al., 2014; On 2013).

• Prevention and control

The main rout of entrance infections of Campylobacter into the poultry houses is the herozontal transmission route  which happened through insect, rodent, contaminated equipment or contamination of water and feed with fecal mater of infected birds, so the most important route for prevention and control of entrance of Campylobacter to poultry farm is applying the biosecurity measures (Gibbens et al., 2001; Herman’s et al., 2011). There are other routes for controlling of bacterial infection like vaccination but there is no commercial vaccine for Campylobacter(). In recent studies found that there is a good effect of using bacteriophage of Campylobacter and using of probiotics in reducing the Campylobacter colonization in the intestinal tract of poultry (Wageneer et al., 2005; Fischer et al., 2013; Hakeem and Lu 2021)

• Conclusion

Campylobacter infect all poultry species ,Campylobacter infecting in poultry cause high intestinal colonization with high morbidity and cause drop in egg production and spotty liver but mostly colonization happened without any clinical signs. It is unique bacteria that it has mor than 30 type of protein related to virulence genes. The best methods for detection of Campylobacter is culturing method  it need especial condition (microeariphillic condition and high temperature 42⁰C), due to PCR methods not detect is the bacteria is life of dead. Poultry considered the main source of human infections with Campylobacter due to its  intestinal tract give a good condition for Campylobacter growth.