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Article

Campylobacter Colonization and Diversity in Young Turkeys in the Context of Gastrointestinal Distress and Antimicrobial Treatment

by
Margaret Kirchner
1,
William G. Miller
2,
Jason A. Osborne
3,
Brian Badgley
4,
Jeffrey Neidermeyer
1 and
Sophia Kathariou
1,*
1
Department of Food, Bioprocessing, and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA
2
Produce Safety and Microbiology Research Unit, Agricultural Research Service, US Department of Agriculture, Albany, CA 94710, USA
3
Department of Statistics, North Carolina State University, Raleigh, NC 27695, USA
4
School of Plant and Environmental Sciences, Virginia Tech, Blacksburg, VA 24061, USA
*
Author to whom correspondence should be addressed.
Microorganisms 2023, 11(2), 252; https://doi.org/10.3390/microorganisms11020252
Submission received: 16 December 2022 / Revised: 10 January 2023 / Accepted: 11 January 2023 / Published: 19 January 2023
(This article belongs to the Special Issue Advances in Campylobacter: Molecular Epidemiology, Virulence Factors, Immune Response and Drug Resistance)
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Abstract

:
Young turkeys are vulnerable to undifferentiated gastrointestinal distress, including “irritable and crabby syndrome” (ICS), which compromises flock performance and is typically treated with a combination of penicillin and gentamicin (P/G). However, the effects of ICS and P/G treatment on Campylobacter remain poorly understood. We investigated the impact of ICS and P/G treatment on Campylobacter levels and diversity in four flocks from three turkey farms. Cecum and jejunum samples were analyzed weekly from day of hatch to week 4–5. All four flocks became colonized with multidrug resistant (MDR) Campylobacter jejuni and C. coli by week 2–3, and two developed ICS. ICS and P/G treatment did not significantly impact total Campylobacter levels or strain genotypes but impacted species and antimicrobial resistance (AMR) profiles. One flock was raised under antibiotic-free (ABF) conditions while another flock at the same farm was raised conventionally. The ABF flock did not develop ICS while its counterpart did. However, Campylobacter strains, AMR profiles and sequence types were generally shared between these two flocks. Our findings suggest that ICS and P/G treatment impacted Campylobacter population dynamics in commercial young turkey flocks, and that ABF flocks may become readily colonized by MDR strains from non-ABF flocks at the same farm.

1. Introduction

Campylobacter is a leading cause of human bacterial gastroenteritis worldwide [1] and causes approximately 800,000 cases of disease (campylobacteriosis) annually in the United States alone [2]. Campylobacteriosis is also the most prevalent bacterial antecedent to Guillain-Barré Syndrome which can have long-term debilitating effects and contributes to reactive arthritis and other sequelae [1,3]. The majority of campylobacteriosis cases are caused by C. jejuni (90%) followed by C. coli [2]. The consumption and/or handling of raw or under-cooked poultry is a major risk factor for developing campylobacteriosis [4,5,6]. FoodNet, a foodborne pathogen monitoring system, found that Campylobacter is a leading cause of foodborne infections, with higher incidence in 2019 compared to 2016–2018 [7]. Campylobacter colonization has been extensively investigated in broilers but has not been as well characterized in turkeys [1,8,9].
C. coli and C. jejuni isolated from poultry often exhibit acquired a host of antimicrobial resistance (AMR) traits, notably resistance against fluoroquinolones and macrolides [10,11,12]. Such antimicrobial-resistant Campylobacter spp. have also been reported from young turkeys on brooder farms [10,13,14,15].
Young turkeys (brooders), from day of hatch to week 5, can be vulnerable to infection and gastrointestinal distress syndromes which are often attributed to an abnormal microbiota or “dysbiosis” in the birds [16,17,18]. The consequences of such gastrointestinal distress syndromes can be disastrous for bird health, final bird weight, and profitability of the farm operation [19,20]. One such syndrome frequently experienced in young turkey flocks in eastern North Carolina causes birds to appear restless and distressed, go off of feed and fail to gain weight and is herein referred to as “irritable crabby syndrome” (ICS). In the poultry industry these gastrointestinal syndromes are often treated with antimicrobials [21]. However, with heightened concerns about AMR among pathogenic microorganisms and antimicrobial use in animal agriculture [22,23,24], there is increasing interest in antibiotic-free (ABF) methods of rearing poultry [22,23,25,26].
To develop potential strategies for ICS mitigation, it would be critical to elucidate the potential microbiological underpinnings and implications of ICS and the accompanying treatment on the bacteria in the turkey gastrointestinal tract, including those that are of food safety concern, such as Campylobacter. This prompted the development of a partnership between our laboratory and veterinarians in the turkey industry which aimed to elucidate Campylobacter levels as well as gastrointestinal community composition and possible microbial community shifts associated with ICS and the corresponding treatment of young turkey flocks. The microbiome investigations will be described in a separate presentation. The objective of the present study was to characterize the prevalence and diversity of Campylobacter in brooder turkeys with or without ICS and administration of the corresponding antimicrobial treatment. To address these objectives, we investigated Campylobacter populations, AMR profiles, and genotypes at weekly intervals in four commercial turkey brooder flocks, including one produced under ABF standards.

2. Materials and Methods

2.1. Turkey Flocks

Due to the severe disease burden imposed by ICS in young turkeys discussed above, the partnering turkey company veterinarians upon consultation with the corresponding growers identified four commercial flocks (Table 1) for participation in the study. Three of these flocks were grown under conventional (CONV) industry standards (flocks 1, 2 and 3), while one of the flocks (flock 4) was grown under antibiotic-free (ABF) conditions (Table 1). All four flocks were obtained from the same breeder and placed at the farms as day-old birds. The design of the farms, size of turkey houses, general management and biosecurity practices were the same for all four flocks and followed industry standards. Flocks 1 and 2 were raised on separate farms (farms A and B, respectively), while flocks 3 and 4 were raised in different houses of the same farm (farm C). Farms A, B and C were operated by different growers under control of the same vertical integrator and were located in the same region (eastern North Carolina, USA). All flocks were fed a high-protein and high-starch starter feed composed of corn, wheat, and soymeal from day of hatch to 3.5 weeks old, at which time they received a lower-protein, lower-starch, and increased-fat diet for the remainder of the brooder period. The feed of CONV flocks (flocks 1, 2 and 3) included animal byproducts, ionophores and the coccidiostats monensin or lasalocid. Feed for flock 4 (ABF) contained the coccidiostat diclazuril and lacked animal byproducts, which were replaced with vegetable oil. Bird density was similar for the three CONV flocks (16,000–17,200/turkey house) while the ABF flock had lower bird density, with 9000 birds in the turkey house (Table 1). Flocks 2 and 3 developed ICS between weeks 3 and 4 and were treated with antibiotics (combination of penicillin and gentamicin, P/G) administered at therapeutic levels upon detection of ICS (Table 1). Flock 1 remained free of ICS but was treated with copper sulfate between weeks 4 and 5, possibly reflecting routine practices at farm A.

2.2. Sample Collection, Campylobacter Isolation and Enumeration

As indicated above, the four commercial flocks (Table 1) were included in the study by turkey company veterinarians upon consultation with the growers for farms A, B and C. At placement (day-old birds) and once each week, 10 birds from each flock were randomly chosen by the turkey company veterinarians and euthanized at the company’s facility following the company’s animal welfare guidelines, as described before [14]. A segment (approx. 3–4 cm) of the jejunum and one cecum from each bird were placed in separate bags and labeled to indicate bird, date, flock number and flock age. Samples were shipped by the company veterinarians to our laboratory at North Carolina State University overnight on ice and kept at 4 °C until processing, typically within 4 h. The cecum and jejunum samples of each bird were processed individually from weeks 2–5. The day of hatch and week 1 samples were pooled for processing because it was unlikely that they would yield Campylobacter [10,15,27,28]. For early time points (day of hatch to week 3), samples were also enriched for Campylobacter as previously described [29]. To detect and enumerate Campylobacter, 0.1 g of sample was suspended in 1.0 mL of Mueller-Hinton broth (MHB) (Becton Dickinson, Sparks, MD, USA), diluted serially in MHB, and 10 μL of the serial dilutions were spotted onto blood-free modified charcoal cefoperazone desoxycholate agar (mCCDA; Oxoid, Hampshire, UK). Selected dilutions were also plated (100 μL) onto mCCDA. Plates were incubated under microaerobic conditions generated by a GasPak EZ Campy sachet (Becton, Dickinson and Co., Sparks, MD, USA) for 48 h at 42 °C. A minimum of one isolate from each positive bird was purified from mCCDA at each time point by streaking onto MHA (Mueller-Hinton broth with 1.2% agar). All isolates were stored at −80 °C as previously described [15,30].

2.3. Determination of Campylobacter Species, AMR profiles and Genotypes

The Campylobacter species of each isolate was determined by multiplex PCR using hip (5′-ATG ATG GCT TCT TCG GAT AG-3′ and 5′-GCT CCT ATG CTT ACA ACT GC-3′) and ceu (5′-GAT TTT ATT ATT TGT AGC AGC G-3′ and 5′-TCC ATG CCC TAA GAC TTA ACG-3′) primers to identify C. jejuni and C. coli, respectively, as previously described [15,30]. Briefly, reactions were performed using X-Taq DNA polymerase (Fisher, Fair Lawn, NJ, USA) in 25 mL with 0.5 μL of genomic DNA as a template. The reaction conditions included an initial denaturation at 95 °C for 5 min, followed by 30 cycles of 95 °C for 1 min, 50 °C for 1 min, and 72 °C for 2 min, with a final extension at 72 °C for 5 min. AMR profiles were determined as previously described on antibiotic-amended MHA [30]. Briefly, isolates were spotted (3.5 μL) in duplicate on MHA amended with tetracycline (16 μg/mL), streptomycin (64 μg/mL), erythromycin (8 μg/mL), kanamycin (64 μg/mL), nalidixic acid (32 μg/mL), ciprofloxacin (4 μg/mL), or gentamicin (50 μg/mL). MHA without added antibiotics and the pan-sensitive C. jejuni ATCC 33560 were used each time for quality assurance. Plates were incubated for 48 h at 42 °C under microaerobic conditions, and resistance was determined based on the visual assessment for confluent growth on both spots. For genotyping, multilocus sequence typing (MLST) was done using the primers tktFN/tktRN [31] together with primers aspAF1/aspAR1, atpAF/atpAR, glnAF/glnAR, gltAF/gltAR, glyAF/glyAR and pgmF1/pgmR1 [32]. Each amplification used 50 ng genomic DNA and 50 pmol each primer under the following conditions: 30 s at 94 °C, 30 s at 53 °C and 2 min at 72 °C (30 cycles) [32]. Sequencing reactions were performed using the same amplification primers, as described [32]. Alleles and sequence types were assigned using MLSTparser [32] and a database of C. jejuni and C. coli MLST alleles and STs. Novel allelic profiles were submitted to PubMLST (https://pubmlst.org/organisms/campylobacter-jejunicoli; accessed on 5 December 2022) for ST assignment. Minimum spanning trees (MSTs) were prepared using BioNumerics, as previously described [31].

2.4. Statistical Analysis

A t-test was used to compare species/AMR combinations before and after antibiotics within each flock and to compare the prevalence of species in the cecum and jejunum. A two-way ANOVA was used to investigate the prevalence and enumeration of Campylobacter within and between flocks.

3. Results and Discussion

3.1. Campylobacter Colonization Varies among Individuals and Intestinal Site

All four turkey brooder flocks became positive for Campylobacter by week 2 (flocks 2 and 3) or 3 (flocks 1 and 4) (Table 1). Previous samples were negative by direct plating as well as by enrichment. Interestingly, only the flocks that developed ICS had detectable Campylobacter in week 2 (Table 1). This opens up the possibility that the flocks colonized earlier were pre-disposed to ICS or vulnerable due to stress, or that Campylobacter could be a contributing factor for development of ICS. No noticeable changes in the levels of Campylobacter in the cecum or the jejunum were noted in flocks 2 and 3 after ICS and P/G treatment, while in flock 1 some decreases were noted after copper sulfate treatment, especially in the cecum (Figure 1 and Supplementary Table S1).
For flocks 2 and 3 only some of the birds were positive in the first week of Campylobacter detection (week 2) and the Campylobacter levels in the few positive birds were generally below 106 CFU/g in the cecum (Supplementary Table S1). In contrast, for flocks 1 and 4 that did not develop ICS the birds were all positive in the very week that Campylobacter was detected, i.e., week 3, and the Campylobacter CFU/g cecum exceeded 106 for most birds (Supplementary Table S1). These findings indicate a surprisingly sudden onset of high-level Campylobacter colonization in ICS-free flocks, while in those that developed ICS the onset was more gradual, similarly to what was described previously for turkey colonization with Campylobacter [28]. It is possible that the ICS-free flocks were also colonized in week 2 but the numbers were too low for the detection methods that we employed, even with the inclusion of enrichments. In the weeks that followed, the birds from all four flocks were all positive with the Campylobacter levels in the cecum ranging between 1 × 107 and 1 × 109 CFU/g (with only few occasional exceptions (Supplementary Table S1), as reported previously [28].
Even though most birds tested positive for Campylobacter in the cecum and jejunum after the initial detection, noticeable bird-to-bird variation in CFU Campylobacter/g cecal or jejunal content was observed (Figure 1 and Supplementary Table S1). A low level of Campylobacter in the cecum was not always accompanied with a low level in the jejunum (Supplementary Table S1). Campylobacter CFU/g values in the cecum (maximum 109 CFU/g) were at least 3-log higher than in the jejunum (maximum 106 CFU/g) (Figure 1). This is in agreement with a previous study that documented lower Campylobacter colonization in the jejunum versus the cecum [28]. The difference in colonization between intestinal sites in this study ranged between 2–3 logs (Supplementary Table S1).
As indicated above all birds in the ABF flock were positive starting with week 3, and the average CFU Campylobacter/g of cecal content was similar to the levels noted with the other three flocks (Figure 1). Thus, the level of colonization was overall similar and comparable across all flocks regardless of management (ABF vs. conventional) or treatment. It appears that the ABF production practices did not affect Campylobacter levels in the cecum and jejunum, as previously demonstrated in European organic flocks [33,34]. However, it is worthy of note that flock 4 (ABF flock) yielded Campylobacter-positive samples one week later than flock 3, which was housed at the same farm and grown under conventional industry practices (Table 1). Later onset of Campylobacter colonization in this flock could be associated in part with the slower weight gain generally observed with ABF turkeys, similarly to previous findings in birds grown without antibiotics [19,21,35].
Campylobacter isolates (n = 372) included 90, 90, 96 and 96 isolates from flocks 1, 2, 3 and 4, respectively (Table 2). All isolates were identified as either C. coli or C. jejuni and C. coli was most frequently isolated overall (57.8%), except for flock 3 where C. coli and C. jejuni were present at about equal proportions (Table 2). Interestingly, C. coli was overall significantly more common (p < 0.05) in the cecum (86.3%) than in the jejunum (13.2%) while C. jejuni was significantly more common (p < 0.05) in the jejunum (74.4%) than in the cecum (25.6%) (Table 2). Flock 2 was the only flock that had more C. coli (62.5%) than C. jejuni (37.5%) in the jejunum (Table 2). The mechanisms underlying the apparent predilection of C. jejuni for the jejunum, and C. coli for the cecum, remain to be elucidated. Strong preferential association of C. coli and C. jejuni with the cecum and the jejunum, respectively, has been repeatedly noted before in our laboratory in assessments of gastrointestinal samples from turkeys and other birds (chickens, guineafowl) grown commercially (R. M. Siletzky and S. Kathariou, unpublished findings).

3.2. Diversity of Campylobacter AMR Profiles and MLST Sequence Types Vary by Flock

By combining both species and AMR profile, 12 (six each in C. coli and C. jejuni) total distinct species/AMR combinations were identified, including eight in flock 1 and six each in flocks 2–4 (Table 2). MLST was employed to genotype isolates from week 2 (n = 3), 3 (n = 15), 4 (n = 22), and 5 (n = 13) from all four flocks. One isolate for each individual species/AMR combination within each flock was chosen for MLST, typically from the cecum. When specific species/AMR combinations were detected in multiple weeks, isolates from multiple time points were included (Supplementary Table S2).
Multidrug resistance (MDR), i.e., resistance to three or more antimicrobial classes, was seen in 90% of the isolates, and all isolates were resistant to tetracycline. Erythromycin resistance was only seen among C. coli isolates, which is consistent with previous studies of Campylobacter from turkeys from this region [14,30,36]. AMR profile designations were created for each isolate by using the first letter of each antibiotic the isolate was resistant to, except for the (fluoro)quinolones nalidixic acid and ciprofloxacin, which were represented by “Q”. The two most predominant species/AMR profiles were C. coli resistant to all tested antimicrobials i.e., tetracycline (T), streptomycin (S), erythromycin (E), kanamycin (K), gentamicin (G), nalidixic acid and ciprofloxacin (Q), i.e., ccTSEKQG, and C. jejuni resistant to all tested antimicrobials except for erythromycin, i.e., cjTSKQG, accounting for 37.6% and 34.4%, respectively of the 372 isolates (Table 2). The AMR profiles TSKQG, TSKQ, TK, and TKQ were only found in C. jejuni and none of the C. jejuni isolates exhibited resistance to erythromycin while erythromycin resistance was commonly encountered in C. coli, similarly to previous studies of Campylobacter from turkeys in eastern North Carolina [14,15,30,36].
MLST analysis indicated that overall C. coli and C. jejuni exhibited similar diversity and 24 STs were totally identified, 11 and 13 in C. jejuni and C. coli, respectively (Table 3 and Figure 2A). Several of these STs have been repeatedly encountered in other studies of turkeys from eastern North Carolina [14,30,31,36]. However, 15 of the 24 STs were novel, including six in C. coli and nine in C. jejuni (Table 3). Of the 15 novel STs, eight were closely related (maximum of 2 allelic differences) to known STs identified in the four flocks and six C. jejuni STs (8522, 8524, 8525, 8526, 8527 and 8542) were closely related to each other but not to a known ST within the flocks (Figure 2A). The two most common STs in the C. coli isolates were STs 1604 (n = 10) and 8531 (n = 7), a novel ST closely related to ST-1161, 1192, 8534, and 8533, and the two most common C. jejuni STs were 8227 (n = 9) and 1839 (n = 11). These dominant C. coli and C. jejuni STs were shared among different flocks and encountered in birds of diverse ages (Figure 2A,B). Novel STs were found in all flocks with the highest number found in flock 4 (n = 6) and the lowest in flock 3 (n = 2) (Table 3).
Farms A (flock 1), B (flock 2) and C (flocks 3 and 4) were within 37 miles of each other and managed by the same company. This overlap and relative proximity may account for the similarity of STs across the different flocks and suggests that certain C. jejuni and C. coli STs may persist in commercial turkey production in eastern North Carolina (Figure 2A, Table 3).
The apparent similar levels of diversity within C. jejuni and C. coli is in contrast with findings with isolates from three turkey farms in Ohio, where higher diversity was noted in C. coli [10]. The gastrointestinal site (e.g., cecum vs. jejunum) from which the isolates originated was not reported in this previous study [10]. The fact that isolates both from jejunum and cecum were included in the current study, and C. jejuni was found to be noticeably more common in the jejunum, may have enhanced the opportunity to survey diverse strains of both species colonizing the birds. It is also noteworthy that none of the 11 C. jejuni STs in our study, and only two of the 13 C. coli STs (STs 889 and 1119, encountered in just three of our C. coli strains) overlapped with those from the turkeys from the farms in Ohio [10], suggesting noticeable regional diversity in the genotypes of C. jejuni and C. coli colonizing commercial turkeys. The underlying reasons remain to be elucidated but may be related to management practices.
MLST identified two main clusters in C. coli as well as two main clusters in C. jejuni (Figure 2A). These clusters in C. jejuni tended to form around AMR profiles, with cjTSKQG and cjTKG belonging to the same or closely related STs (Figure 2C). Most species/AMR combinations were of the same or closely related STs (Figure 2C). Two notable exceptions were the C. jejuni profiles cjTKQG and cjTSKQ, which were identified in two clearly-distinct clusters, each consisting of highly related STs (Figure 2C).

3.3. Penicillin and Gentamicin Treatment Are Associated with a Shift in Certain Campylobacter AMR Profiles

As indicated earlier, flocks 2 and 3 developed ICS between weeks 3 and 4 and were P/G-treated (Table 1), allowing the opportunity to assess potential impacts of ICS and P/G treatment on Campylobacter species and strain distributions (Figure 3). In flock 2, ccTKQG was a species/AMR profile unique to this flock. This profile was dominant in the cecum and jejunum of flock 2 in week 2 and also encountered in week 3 (Table 2). Another species/AMR profile, ccTEKQG, first appeared in flock 2 in week 3, composing a majority of the isolates in the cecum (11/20) and jejunum (6/8). Interestingly, however, neither ccTKQG nor ccTEKQG were detected in flock 2 after P/G treatment (Table 2, Figure 3). Instead, after P/G treatment the dominant species/AMR profiles in flock 2 shifted completely to ccTESKQG and cjTSKQG in the cecum and jejunum, respectively (Table 2, Figure 3). Treatment with P/G was followed by a significant shift in species/AMR profiles in flock 2 with a decrease in ccTKQG and ccTEKQG (p < 0.0001) and a concomitant increase in cjTSKQG (p < 0.0001).
In flock 3, ccTEKQG was detected in flock 3 only after P/G treatment (Table 2, Figure 3). The ccTEKQG isolates identified in this flock as well as those in the ABF flock 4 housed at the same farm were ST-8531, while ccTEKQG from flock 2 had the markedly distinct STs 8523 and 1149. which actually belonged to a different C. coli cluster (Figure 2D). These findings suggest that ccTEKQG was independently introduced into the farm that housed flock 2 and the one that housed flocks 3 and 4, and might not be affected by the antibiotics administered to flock 3. Similarly to flock 2, a significant overall increase in cjTSKQG (p = 0.0005) after antimicrobial treatment was seen in flock 3. The jejunum of flock 3 birds at week 3 also appeared to have greater strain diversity than in weeks 4 or 5, after P/G had been administered. Even though cjTSKQG and two other profiles (cjTKQG, cjTKG) were detected in similar proportions in week 3 prior to treatment, all but one post-treatment isolates from week 4, and all those from week 5, were cjTSKQG (Table 2, Figure 3). The post-treatment increases in cjTSKQG in both flocks 2 and 3 raise the possibility that frequent P/G treatment of young turkeys for ICS and similar gastrointestinal disturbances may contribute to the apparent dissemination of multidrug resistant cjTSKQG strains in eastern North Carolina observed here and in other studies [14,36].

3.4. Impacts of Copper Sulfate Treatment on Campylobacter Diversity

Flock 1 was reported to be healthy throughout the study period but was treated with copper sulfate, an antimicrobial, between weeks 4 and 5. The most common species/AMR combinations in this flock were ccTSEKQG and cjTSKQG (Table 2). ST-8086, only found in flock 1, and ST-889 comprised the ccTSEKQG isolates from flock 1 while STs 8227 and 8528 comprised the cjTSKQG isolates (Figure 2D). Four other C. jejuni and C. coli STs were only seen in flock 1 and were closely related to at least one other ST (Figure 2A). Other species/AMR combinations were transient (i.e., detected only during one week), including ccTEKG and cjTK, which were only detected in flock 1 (Table 2). Both of these unique species/AMR combinations had novel STs.
Before copper treatment of flock 1, ccTSEKQG was dominant in the cecum and cjTSKQG was dominant in the jejunum with four other species/AMR combinations also encountered both in the cecum and the jejunum (Table 2, Figure 3). After copper treatment, ccTSEKQG and cjTSKQG remained dominant in the cecum and jejunum but the number of additional species/AMR combinations decreased from five to two in the cecum and from five to three in the jejunum (Table 2, Figure 3). Even though the small numbers of STs did not allow statistical assessments of significance, the findings suggest the possibility that copper sulfate treatment may be accompanied with decreased diversity in species/AMR profile combinations.

3.5. Campylobacter Strains Are Largely Similar between ABF and Non-ABF Flocks on the Same Farm

Flocks 3 (ABF) and 4 (non-ABF) were raised on the same farm (Table 1). This allowed us to compare Campylobacter diversity, colonization, and genotypes between a non-ABF flock that contracted ICS and was treated and a flock that was raised under ABF conditions and did not contract ICS. The flocks were highly comparable in respect to Campylobacter diversity, sharing the same dominant species/AMR combinations, ccTSEKQG and cjTSKQG, throughout the study and many of the same transient species/AMR combinations (Table 2). The exceptions were ccTSEQG and ccTKG, detected only in flock 3 and 4, respectively. The presence of MDR observed here was markedly higher than observed in previous studies of poultry flocks raised without antibiotics [25,37,38]. This could be attributed to the ABF flock being raised on the same farm as flocks that were colonized with MDR Campylobacter strains. Flocks 3 and 4 shared two C. coli STs, 1604 and 8531, and two C. jejuni STs, 8227 and 1839 (Figure 2D). There were several STs only found in flock 4, including STs 8542, 8525 and 8526 in C. jejuni and STs 8521, 8533 and 1192 in C. coli (Table 3). These C. jejuni STs were all closely related to each other and encompassed cjTKQG, cjTSKQ, and cjTKG (Figure 2D); however, among C. coli, ccTSEKQG with ST-8521 was only found in flock 4, and the two ccTKG isolates in flock 4 had distinct STs (1192 and 8533) not found in flock 3 (Figure 2D).
Three of the transient species/AMR combinations, cjTKQG, cjTKG, and ccTEKQG, which made up 30.8%, 19.2% and 38.5%, respectively, of all non-major species/AMR combinations, appeared in flock 3 either before or concurrently with flock 4 (Table 2). This suggests that they were introduced to farm C where flocks 3 and 4 were housed and were not consistently present in young turkeys at other farms. Since non-ABF flocks were concurrently raised in the farm (farm C) that housed flock 4, the environment would be expected to be similar to commercial farms operating under standard industry practices. Commercial turkey farms in this region were previously found to be colonized with MDR C. jejuni and C. coli. [13,14,15,36]. This established presence of MDR strains in the region could also explain why the species/AMR combinations found in flocks 3 and 4 were so comparable (Table 2).

4. Conclusions

Our findings were based on the analysis of four brooder turkey flocks in eastern North Carolina, a major turkey-producing region in the United States. Clearly, additional studies are needed with larger numbers of turkey flocks, and in diverse regions. It would be also desirable to monitor the birds subsequent to the brooder period, which was not feasible in our study due to the logistics of turkey production where the brooders would be transported to different and often distantly located grow-out farms. Nonetheless, our findings indicate that ICS and P/G treatment noticeably impacted Campylobacter population dynamics and diversity in commercial young turkey flocks. The prevalence of certain C. jejuni and C. coli strains with multidrug resistance profiles increased significantly the week after penicillin/gentamicin treatment in both ICS-afflicted flocks, raising the possibility that gastrointestinal distress episodes in young turkeys and the accompanying antimicrobial treatment may contribute the high prevalence of such multidrug-resistant strains in commercial turkey production. Thus, flock management strategies to mitigate gastrointestinal distress in young turkeys may also contribute to reductions in the prevalence of multidrug-resistant Campylobacter in the flocks. Another novel finding was the significant association of C. coli and C. jejuni with the turkey cecum and jejunum, respectively. Thus, accurate surveillance of Campylobacter in turkey flocks may benefit from inclusion of both of these gastrointestinal tract sites. Lastly, the similarity of species/AMR profiles across flocks, especially flocks 3 (conventional) and 4 (ABF) which were housed on the same farm, serves to underscore that in-flock management practices are not always sufficient to mitigate the high prevalence of antimicrobial resistance in Campylobacter from young turkeys. The environment in which the young turkeys are raised, including proximity to other flocks, along with management practices and the persistence of AMR genes, can all contribute to the AMR profiles of C. jejuni and C. coli that colonize the flocks.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/microorganisms11020252/s1, Supplementary Table S1: Campylobacter content in the cecum and jejunum of individual birds; Supplementary Table S2: Campylobacter isolates typed by MLST.

Author Contributions

Conceptualization, M.K., B.B. and S.K.; methodology, M.K., W.G.M. and S.K.; formal analysis, M.K., J.A.O., W.G.M. and S.K.; investigation, M.K. and J.N.; resources, S.K. and W.G.M.; writing—original draft preparation, M.K.; writing—review and editing, M.K., S.K., W.G.M. and J.A.O.; supervision, S.K.; project administration, S.K. and B.B.; funding acquisition, S.K., B.B. and W.G.M. All authors have read and agreed to the published version of the manuscript.

Funding

This work was partially funded by the Dean’s Enrichment Fund at the College of Agriculture and Life Sciences at North Carolina State University, the USDA National Institute of Food and Agriculture, award 2018-67017-27927, and the Virginia Tech College of Agriculture and Life Sciences: Regional Collaboration Program. This work was also supported by CRIS project 2030-42000-055-00D from the United States Department of Agriculture, Agricultural Research Service (https://www.ars.usda.gov/research/project/?accnNo=440168, accessed on 17 May 2022).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data for this manuscript can be found in the Supplementary Materials. Additional information regarding novel or existing MLST alleles and sequence types can be obtained from PubMLST (https://pubmlst.org/, accessed on 5 December 2022).

Acknowledgments

We are grateful to Donna Carver (Prestage Department of Poultry Science, North Carolina State University) for her insights and support regarding numerous aspects of the study. We thank the turkey company veterinarians and service staff for availability of the turkey samples utilized in this study and information about the four flocks that participated in the study. We also thank Emma Yee for MLST sequencing.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Kaakoush, N.O.; Castano-Rodriguez, N.; Mitchell, H.M.; Man, S.M. Global epidemiology of Campylobacter infection. Clin. Microbiol. Rev. 2015, 28, 687–720. [Google Scholar] [CrossRef] [Green Version]
  2. Scallan, E.; Hoekstra, R.M.; Angulo, F.J.; Tauxe, R.V.; Widdowson, M.A.; Roy, S.L.; Jones, J.L.; Griffin, P.M. Foodborne illness acquired in the United States—Major pathogens. Emerg. Infect. Dis. 2011, 17, 7–15. [Google Scholar] [CrossRef]
  3. Nyati, K.K.; Nyati, R. Role of Campylobacter jejuni infection in the pathogenesis of Guillain-Barre syndrome: An update. Biomed. Res. Int. 2013, 2013, 852195. [Google Scholar] [CrossRef] [Green Version]
  4. Buettner, S.; Wieland, B.; Staerk, K.D.; Regula, G. Risk attribution of Campylobacter infection by age group using exposure modelling. Epidemiol. Infect. 2010, 138, 1748–1761. [Google Scholar] [CrossRef] [Green Version]
  5. Kittl, S.; Korczak, B.M.; Niederer, L.; Baumgartner, A.; Buettner, S.; Overesch, G.; Kuhnert, P. Comparison of genotypes and antibiotic resistances of Campylobacter jejuni and Campylobacter coli on chicken retail meat and at slaughter. Appl. Environ. Microbiol. 2013, 79, 3875–3878. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  6. Mughini Gras, L.; Smid, J.H.; Wagenaar, J.A.; de Boer, A.G.; Havelaar, A.H.; Friesema, I.H.; French, N.P.; Busani, L.; van Pelt, W. Risk factors for campylobacteriosis of chicken, ruminant, and environmental origin: A combined case-control and source attribution analysis. PLoS ONE 2012, 7, e42599. [Google Scholar] [CrossRef] [Green Version]
  7. Tack, D.M.; Ray, L.; Griffin, P.M.; Cieslak, P.R.; Dunn, J.; Rissman, T.; Jervis, R.; Lathrop, S.; Muse, A.; Duwell, M.; et al. Preliminary incidence and trends of infections with pathogens transmitted commonly through food—Foodborne Diseases Active Surveillance Network, 10 U.S. Sites, 2016–2019. MMWR Morb. Mortal. Wkly. Rep. 2020, 69, 509–514. [Google Scholar] [CrossRef]
  8. Kalupahana, R.S.; Kottawatta, K.S.; Kanankege, K.S.; van Bergen, M.A.; Abeynayake, P.; Wagenaar, J.A. Colonization of Campylobacter spp. in broiler chickens and laying hens reared in tropical climates with low-biosecurity housing. Appl. Environ. Microbiol. 2013, 79, 393–395. [Google Scholar] [CrossRef] [Green Version]
  9. Newell, D.G.; Fearnley, C. Sources of Campylobacter colonization in broiler chickens. Appl. Environ. Microbiol. 2003, 69, 4343–4351. [Google Scholar] [CrossRef] [Green Version]
  10. Kashoma, I.P.; Kumar, A.; Sanad, Y.M.; Gebreyes, W.; Kazwala, R.R.; Garabed, R.; Rajashekara, G. Phenotypic and genotypic diversity of thermophilic Campylobacter spp. in commercial turkey flocks: A longitudinal study. Foodborne Pathog. Dis. 2014, 11, 850–860. [Google Scholar] [CrossRef]
  11. Luangtongkum, T.; Jeon, B.; Han, J.; Plummer, P.; Logue, C.M.; Zhang, Q. Antibiotic resistance in Campylobacter: Emergence, transmission and persistence. Future Microbiol. 2009, 4, 189–200. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  12. Lutgen, E.M.; McEvoy, J.M.; Sherwood, J.S.; Logue, C.M. Antimicrobial resistance profiling and molecular subtyping of Campylobacter spp. from processed turkey. BMC Microbiol. 2009, 9, 203. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  13. Lee, B.C.; Reimers, N.; Barnes, H.J.; D’Lima, C.; Carver, D.; Kathariou, S. Strain persistence and fluctuation of multiple-antibiotic resistant Campylobacter coli colonizing turkeys over successive production cycles. Foodborne Pathog. Dis. 2005, 2, 103–110. [Google Scholar] [CrossRef]
  14. Niedermeyer, J.A.; Ring, L.; Miller, W.G.; Genger, S.; Lindsey, C.P.; Osborne, J.; Kathariou, S. Proximity to other commercial turkey farms affects colonization onset, genotypes, and antimicrobial resistance profiles of Campylobacter spp. in turkeys: Suggestive evidence from a paired-farm model. Appl. Environ. Microbiol. 2018, 84, e01212-18. [Google Scholar] [CrossRef] [Green Version]
  15. Smith, K.; Reimers, N.; Barnes, H.J.; Lee, B.C.; Siletzky, R.; Kathariou, S. Campylobacter colonization of sibling turkey flocks reared under different management conditions. J. Food Prot. 2004, 67, 1463–1468. [Google Scholar] [CrossRef]
  16. Barnes, H.J.; Guy, J.S.; Vaillancourt, J.P. Poult enteritis complex. Rev. Sci. Tech. 2000, 19, 565–588. [Google Scholar] [CrossRef]
  17. Oakley, B.B.; Lillehoj, H.S.; Kogut, M.H.; Kim, W.K.; Maurer, J.J.; Pedroso, A.; Lee, M.D.; Collett, S.R.; Johnson, T.J.; Cox, N.A. The chicken gastrointestinal microbiome. FEMS Microbiol. Lett. 2014, 360, 100–112. [Google Scholar] [CrossRef]
  18. Teirlynck, E.; Gussem, M.D.; Dewulf, J.; Haesebrouck, F.; Ducatelle, R.; Van Immerseel, F. Morphometric evaluation of “dysbacteriosis” in broilers. Avian Pathol. 2011, 40, 139–144. [Google Scholar] [CrossRef] [Green Version]
  19. Danzeisen, J.L.; Calvert, A.J.; Noll, S.L.; McComb, B.; Sherwood, J.S.; Logue, C.M.; Johnson, T.J. Succession of the turkey gastrointestinal bacterial microbiome related to weight gain. PeerJ 2013, 1, e237. [Google Scholar] [CrossRef] [Green Version]
  20. Day, J.M.; Zsak, L. Investigating turkey enteric picornavirus and its association with enteric disease in poults. Avian Dis. 2015, 59, 138–142. [Google Scholar] [CrossRef]
  21. Feighner, S.D.; Dashkevicz, M.P. Subtherapeutic levels of antibiotics in poultry feeds and their effects on weight gain, feed efficiency, and bacterial cholyltaurine hydrolase activity. Appl. Environ. Microbiol. 1987, 53, 331–336. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  22. Allen, H.K.; Stanton, T.B. Altered egos: Antibiotic effects on food animal microbiomes. Annu. Rev. Microbiol. 2014, 68, 297–315. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Diarra, M.S.; Malouin, F. Antibiotics in Canadian poultry productions and anticipated alternatives. Front. Microbiol. 2014, 5, 282. [Google Scholar] [CrossRef] [Green Version]
  24. Lammie, S.L.; Hughes, J.M. Antimicrobial resistance, food safety, and One Health: The need for convergence. Annu. Rev. Food Sci. Technol. 2016, 7, 287–312. [Google Scholar] [CrossRef]
  25. Cui, S.; Ge, B.; Zheng, J.; Meng, J. Prevalence and antimicrobial resistance of Campylobacter spp. and Salmonella serovars in organic chickens from Maryland retail stores. Appl. Environ. Microbiol. 2005, 71, 4108–4111. [Google Scholar] [CrossRef] [Green Version]
  26. El-Adawy, H.; Ahmed, M.F.; Hotzel, H.; Tomaso, H.; Tenhagen, B.A.; Hartung, J.; Neubauer, H.; Hafez, H.M. Antimicrobial susceptibilities of Campylobacter jejuni and Campylobacter coli recovered from organic turkey farms in Germany. Poult. Sci. 2015, 94, 2831–2837. [Google Scholar] [CrossRef] [PubMed]
  27. El-Adawy, H.; Hotzel, H.; Tomaso, H.; Neubauer, H.; Hafez, H.M. Elucidation of colonization time and prevalence of thermophilic Campylobacter species during turkey rearing using multiplex polymerase chain reaction. Poult. Sci. 2012, 91, 454–459. [Google Scholar] [CrossRef] [PubMed]
  28. Wallace, J.S.; Stanley, K.N.; Jones, K. The colonization of turkeys by thermophilic campylobacters. J. Appl. Microbiol. 1998, 85, 224–230. [Google Scholar] [CrossRef]
  29. Gharst, G.; Hanson, D.; Kathariou, S. Effect of direct culture versus selective enrichment on the isolation of thermophilic Campylobacter from feces of mature cattle at harvest. J. Food Prot. 2006, 69, 1024–1027. [Google Scholar] [CrossRef]
  30. Gu, W.; Siletzky, R.M.; Wright, S.; Islam, M.; Kathariou, S. Antimicrobial susceptibility profiles and strain type diversity of Campylobacter jejuni isolates from turkeys in eastern North Carolina. Appl. Environ. Microbiol. 2009, 75, 474–482. [Google Scholar] [CrossRef]
  31. Miller, W.G.; Englen, M.D.; Kathariou, S.; Wesley, I.V.; Wang, G.; Pittenger-Alley, L.; Siletz, R.M.; Muraoka, W.; Fedorka-Cray, P.J.; Mandrell, R.E. Identification of host-associated alleles by multilocus sequence typing of Campylobacter coli strains from food animals. Microbiology 2006, 152, 245–255. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  32. Miller, W.G.; On, S.L.W.; Wang, G.; Fontanoz, S.; Lastovica, A.J.; Mandrell, R.E. Extended multilocus sequence typing system (MLST) for Campylobacter coli, C. lari, C. upsaliensis and C. helveticus. J. Clin. Microbiol. 2005, 43, 2315–2329. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  33. Hoogenboom, L.A.; Bokhorst, J.G.; Northolt, M.D.; van de Vijver, L.P.; Broex, N.J.; Mevius, D.J.; Meijs, J.A.; Van der Roest, J. Contaminants and microorganisms in Dutch organic food products: A comparison with conventional products. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. 2008, 25, 1195–1207. [Google Scholar] [CrossRef] [PubMed]
  34. Van Overbeke, I.; Duchateau, L.; De Zutter, L.; Albers, G.; Ducatelle, R. A comparison survey of organic and conventional broiler chickens for infectious agents affecting health and food safety. Avian Dis. 2006, 50, 196–200. [Google Scholar] [CrossRef]
  35. McCrackin, M.A.; Helke, K.L.; Galloway, A.M.; Poole, A.Z.; Salgado, C.D.; Marriott, B.P. Effect of antimicrobial use in agricultural animals on drug-resistant foodborne Campylobacteriosis in humans: A systematic literature review. Crit. Rev. Food Sci. Nutr. 2016, 56, 2115–2132. [Google Scholar] [CrossRef]
  36. Good, L.; Miller, W.G.; Niedermeyer, J.; Osborne, J.; Siletzky, R.M.; Carver, D.; Kathariou, S. Strain-specific differences in survival of Campylobacter spp. in naturally contaminated turkey feces and water. Appl. Environ. Microbiol. 2019, 85, e01579-19. [Google Scholar] [CrossRef]
  37. Luangtongkum, T.; Morishita, T.Y.; Ison, A.J.; Huang, S.; McDermott, P.F.; Zhang, Q. Effect of conventional and organic production practices on the prevalence and antimicrobial resistance of Campylobacter spp. in poultry. Appl. Environ. Microbiol. 2006, 72, 3600–3607. [Google Scholar] [CrossRef] [Green Version]
  38. Noormohamed, A.; Fakhr, M.K. Prevalence and antimicrobial susceptibility of Campylobacter spp. in Oklahoma conventional and organic retail poultry. Open Microbiol. J. 2014, 8, 130–137. [Google Scholar] [CrossRef]
Microorganisms 11 00252 g001 550
Figure 1. Campylobacter load within the cecum and jejunum from the four turkey flocks. The average Campylobacter load of the ten sampled birds from (A) flock 1, (B) 2, (C) 3, and (D) 4 are shown for each time point starting with week 1. Red and black dots represent CFU/g counts from the jejunum and cecum, respectively. A thick black line and a thick red line along the x axis indicate the time of administration of copper sulfate and penicillin/gentamicin, respectively. The limit of detection (horizontal black dotted lines) was 1.00 × 103 CFU/g in the cecum and 1.00 × 101 CFU/g in the jejunum.
Figure 1. Campylobacter load within the cecum and jejunum from the four turkey flocks. The average Campylobacter load of the ten sampled birds from (A) flock 1, (B) 2, (C) 3, and (D) 4 are shown for each time point starting with week 1. Red and black dots represent CFU/g counts from the jejunum and cecum, respectively. A thick black line and a thick red line along the x axis indicate the time of administration of copper sulfate and penicillin/gentamicin, respectively. The limit of detection (horizontal black dotted lines) was 1.00 × 103 CFU/g in the cecum and 1.00 × 101 CFU/g in the jejunum.
Microorganisms 11 00252 g001
Microorganisms 11 00252 g002 550
Figure 2. Minimum spanning tree of the Campylobacter MLST sequence types. Minimum spanning tree showing the clustering of the MLST-based STs of 69 C. coli and C. jejuni isolates. Each tree is colored based on (A) flock, (B) week, (C) AMR profile, or (D) AMR profile by color and flock by number. The tree was created in BioNumerics (v. 7.6). Each ST is represented by a circle with each segment in the circle corresponding to an individual isolate. Sequence types in blue font are C. jejuni while those in black font are C. coli. Thick short lines connecting STs indicate single-locus differences; thin lines indicate two-locus differences; a longer, thinner line indicates a three-locus difference; and black dotted lines represent > four-allele differences. The gray dotted line between STs 8528 and 8534 separates C. jejuni STs in the bottom (blue font STs) from C. coli STs in the top (black font STs).
Figure 2. Minimum spanning tree of the Campylobacter MLST sequence types. Minimum spanning tree showing the clustering of the MLST-based STs of 69 C. coli and C. jejuni isolates. Each tree is colored based on (A) flock, (B) week, (C) AMR profile, or (D) AMR profile by color and flock by number. The tree was created in BioNumerics (v. 7.6). Each ST is represented by a circle with each segment in the circle corresponding to an individual isolate. Sequence types in blue font are C. jejuni while those in black font are C. coli. Thick short lines connecting STs indicate single-locus differences; thin lines indicate two-locus differences; a longer, thinner line indicates a three-locus difference; and black dotted lines represent > four-allele differences. The gray dotted line between STs 8528 and 8534 separates C. jejuni STs in the bottom (blue font STs) from C. coli STs in the top (black font STs).
Microorganisms 11 00252 g002
Microorganisms 11 00252 g003 550
Figure 3. Campylobacter species and AMR profile pre- and post-treatment in isolates from the cecum and the jejunum of the three turkey flocks. Distribution of Campylobacter species/AMR profiles among isolates from the cecum and jejunum before (A) and after (B) treatment of the flocks which underwent treatment with copper sulfate (flock 1) or penicillin/gentamicin (flocks 2 and 3).
Figure 3. Campylobacter species and AMR profile pre- and post-treatment in isolates from the cecum and the jejunum of the three turkey flocks. Distribution of Campylobacter species/AMR profiles among isolates from the cecum and jejunum before (A) and after (B) treatment of the flocks which underwent treatment with copper sulfate (flock 1) or penicillin/gentamicin (flocks 2 and 3).
Microorganisms 11 00252 g003
Table
Table 1. Turkey flocks investigated in this study.
Table 1. Turkey flocks investigated in this study.
Dates 1FlockFarmRearing Method 2Birds per HouseDuration of Life (Weeks)Initial Campylobacter ColonizationICS Treatment 3Time of Treatment
19 April 2016–25 May 20161ACONV16,0005Week 3NoCuWeeks 4–5
19 April 2016–18 May 20162BCONV17,2004Week 2YesP/GWeeks 3–4
26 April 2016–1 June 20163CCONV17,0005Week 2YesP/GWeeks 3–4
26 April 2016–1 June 20164CABF90005Week 3NoN/AN/A
1 Date format is Month / Day/ Year. 2 “CONV” designates conventional flock rearing practices and “ABF” designates antibiotic-free rearing practices, as detailed in the Materials and Methods. 3 “Cu”, flock treated with copper sulfate, “P/G”, flock treated with penicillin and gentamicin. “N/A” (non-applicable) indicates a flock for which no known antimicrobial treatment was administered.
Table
Table 2. Distribution of Campylobacter species and AMR profiles in the cecum and jejunum from the four turkey flocks. Campylobacter Species/AMR profiles 1.
Table 2. Distribution of Campylobacter species and AMR profiles in the cecum and jejunum from the four turkey flocks. Campylobacter Species/AMR profiles 1.
FlockWeek 2Intestinal SiteccTSEKQGccTKGccTEKGccTKQGccTSEQGccTEKQGTotal CccjTSKQGcjTSKQcjTKQGcjTKGcjTKcjTKQTotal Cj
11Cecum00000000000000
Jejunum00000000000000
2Cecum00000000000000
Jejunum00000000000000
3Cecum1300000137001008
Jejunum1000001223001026
4Cecum910000100011002
Jejunum10100028000008
5 *Cecum1010000110000000
Jejunum10000017100008
21Cecum00000000000000
Jejunum00000000000000
2Cecum0001900190000000
Jejunum0001700170000000
3Cecum5004011200000000
Jejunum00020681020003
4 *Cecum70000072010014
Jejunum000000090300012
31Cecum00000000000000
Jejunum00000000000000
2Cecum50000051000001
Jejunum30000032000002
3Cecum1000010110000000
Jejunum30000032033008
4 *Cecum800006140001001
Jejunum00000117020009
5 *Cecum10000001090100010
Jejunum1000001170000017
41Cecum00000000000000
Jejunum00000000000000
2Cecum00000000000000
Jejunum00000000000000
3Cecum2500000250000000
Jejunum2000002100000010
4Cecum1210000130000000
Jejunum100002390200011
5Cecum1110001130100001
Jejunum2000002150010016
Total Cecum1254023118171191330127
Jejunum1501190944109412410130
Overall1404142127215128515711157
1 The isolate identifiers consist of the species designation (cc and cj indicating C. coli and C. jejuni, respectively) followed by the AMR profile determinized by assessing resistance to tetracycline (T), streptomycin (S), erythromycin (E), gentamicin (G), kanamycin (K), and the (fluoro)quinolones nalidixic acid and ciprofloxacin (Q), with Q indicating resistance to both nalidixic acid and ciprofloxacin. For instance, C. coli resistant to all tested antibiotics would be denoted as a ccTSEKQG, C. jejuni resistant to all tested antibiotics except erythromycin would be designated cjTSKQG, and C. coli resistant to tetracycline, kanamycin and gentamicin but none of the other tested antimicrobials would be designated ccTKG. Gray shadowing indicates detection of the indicated species/AMR profiles in the number of isolates shown. 2 * indicates post-treatment.
Table
Table 3. Campylobacter jejuni and C. coli sequence types identified in the four turkey flocks.
Table 3. Campylobacter jejuni and C. coli sequence types identified in the four turkey flocks.
ST 1Clonal Complex 1SpeciesAMR Profiles 2Number of Isolates Flock 1Flock 2Flock 3Flock 4
889828C. coliTSEKQG 21100
1119828C. coliTSEKQG 10100
1149828C. coliTEKQG20200
11611150C. coliTKQG40400
11921150C. coliTKG10001
1604828C. coliTSEKQG (n = 9), TSEQG (n = 1)100046
8086828C. coliTSEKQG11000
8521828C. coliTSEKQG10001
8523828C. coliTEKQG10100
85311150C. coliTEKQG (n = 6), TKQG (n = 1)70133
85321150C. coliTEKQ (n = 1), TKG (n = 1)22000
85331150C. coliTKG10001
85341150C. coliTKQG10100
1839UnknownC. jejuniTSKQG (n = 9), TKQG (n = 1), TSKQ (n = 1)111172
8227UnknownC. jejuniTSKQG (n = 6), TKQG (n = 2), TKQ (n = 1)91431
8522353C. jejuniTKG (n = 2), TKQG (n = 1), TK (n = 1)44000
8524UnknownC. jejuniTKG11000
8525353C. jejuniTKQG10001
8526UnknownC. jejuniTKG10001
8527353C. jejuniTKG10010
8528UnknownC. jejuniTSKQG (n = 1), TSKQ (n = 1),22000
8529UnknownC. jejuniTKQG10100
8530UnknownC. jejuniTKQG10100
8542353C. jejuniTKQG (n = 1), TSKQ (n = 1), TKG (n = 1)30003
1 Sequence types (STs) were determined by multilocus sequence typing (MLST) as described in Materials and Methods. Designations in bold indicate novel STs. “Unknown” CC indicates singleton STs not currently known to belong to a known CC. 2 AMR profile identifiers are as described for Table 2. When multiple AMR profiles were encountered among isolates of the same ST, the numbers of isolates with each profile are in parentheses.
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Kirchner, M.; Miller, W.G.; Osborne, J.A.; Badgley, B.; Neidermeyer, J.; Kathariou, S. Campylobacter Colonization and Diversity in Young Turkeys in the Context of Gastrointestinal Distress and Antimicrobial Treatment. Microorganisms 2023, 11, 252. https://doi.org/10.3390/microorganisms11020252

AMA Style

Kirchner M, Miller WG, Osborne JA, Badgley B, Neidermeyer J, Kathariou S. Campylobacter Colonization and Diversity in Young Turkeys in the Context of Gastrointestinal Distress and Antimicrobial Treatment. Microorganisms. 2023; 11(2):252. https://doi.org/10.3390/microorganisms11020252

Chicago/Turabian Style

Kirchner, Margaret, William G. Miller, Jason A. Osborne, Brian Badgley, Jeffrey Neidermeyer, and Sophia Kathariou. 2023. "Campylobacter Colonization and Diversity in Young Turkeys in the Context of Gastrointestinal Distress and Antimicrobial Treatment" Microorganisms 11, no. 2: 252. https://doi.org/10.3390/microorganisms11020252

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