Campylobacter Fetus Research Articles

Whole-Genome Sequencing of Multidrug-Resistant Campylobacter fetus subsp. fetus NWU_ED24, Isolated from a Bovine Sheath Wash Sample.

Campylobacter fetus subsp. fetus is an opportunistic human pathogen that is frequently identified as a cause of intestinal infections as well as bloodstream infections. This bacterium is well known to cause spontaneous abortions in sheep and cows. The strain reported in this study was isolated from a preputial wash sample from a bull in South Africa.

Genome Sequence of Campylobacter fetus subsp. venerealis NW_ED23, Isolated from Bovine Sheath Wash.

Campylobacter fetus subsp. venerealis is the causative agent of bovine genital campylobacteriosis, which is mostly characterized by reproduction problems. The strain reported in this study was isolated from bull sheath wash in South Africa.

Prevalence, molecular typing and antimicrobial susceptibility of Campylobacter spp. isolates in northern Spain.

To analyse the prevalence, genetic diversity and antimicrobial susceptibility of Campylobacter spp. in northern Spain. Campylobacter was isolated from 139 samples of broiler meat and faecal dropping of broiler and swine with a prevalence of 35·4, 62 and 42·8%, respectively. Campylobacter jejuni (n=55) and Campylobacter coli (n=31) were identified by multiplex-PCR in meat, faeces and human clinical samples while Campylobacter fetus (n=3) was exclusively detected in the latter. Fingerprinting by flaA-RFLP and PFGE revealed 68 different genotypes from the 89 isolates with a Biodiversity Simpson's index of 0·98. The 86·5% of the isolates were resistant to ciprofloxacin, 85·4% to tetracycline and 49·4% to erythromycin; only three genotypes were susceptible to the three antimicrobial drugs. Multidrug resistance was detected in the 40·7% of the isolates. Campylobacter remains prevalent in northern Spain with a high biodiversity degree. About 93·3% of the isolates were resistant to one or more drugs. Although different measures are taken to control Campylobacter, the detection of isolates resistant to the drugs used in the treatment of campylobacteriosis is still high, including different species and genotypes. This evidences the need of additional strategies against this pathogen.

Genotyping and antibiotic resistance patterns of Campylobacter fetus subsp.venerealis from cattle farms in India.

Bovine genital campylobacteriosis caused by Campylobacter fetus subsp. venerealis (Cfv) is of considerable economic importance to the cattle industry worldwide. Cfv causes syndrome of temporary infertility in female cattle, early embryonic mortality, aberrant oestrus cycles, delayed conception, abortions and poor calving rates. In the present study, a total of 200 samples obtained from vaginal swabs, cervicovaginal mucous (CVM), preputial washes and semen straws were investigated that were obtained from organized cattle farm of MLRI, Manasbal and unorganized sectors. Out of a total of 200 samples, 49 (47·57%) vaginal swabs, 1 (3·33%) preputial wash and 8 (25%) carried out CVM samples were positive for Cfv, whereas none of the semen straws were positive for Cfv. A total of eleven isolates of Cfv were recovered. PFGE (Pulse field gel electrophoresis) analysis revealed four different pulsotypes (I-IV) circulating in the screened farms. A common pulsotype circulating among farms could not be established. Insertion element (ISCfe1), a 233bp amplicon of Cfv, was sequenced and the sequence was deposited in GenBank (accession no: MK475662).

Effects of Campylobacter fetus on bull sperm quality

Campylobacter fetus is a gram-negative, motile, spiral or S-shaped bacterium, which induces campylobacteriosis. This disease causes decrease productivity of cattle. Although considerable research has been done on the role of C. fetus on female fertility, little is known about the effect on bulls. The aim of this work was to evaluate the effect of C. fetus subsp. fetus (Cff) and C. fetus subsp. venerealis (Cfv) on bull sperm quality. Samples of frozen semen (n = 29 straws) were each distributed into three groups: two of them incubated with the microorganism (Cff, Cfv) and a control group. The proportions of live spermatozoa, with functional membrane and true acrosomal reaction in control group were significantly (P < 0.01) greater than those observed in Cff and Cfv groups. However, no significant differences (P > 0.05) were found in sperm chromatin structure among treatments. In adhesion assay, proportions of spermatozoa with adhered Campylobacter were similar for both subspecies. Results confirm that Cff and Cfv have the same ability to bind in an irreversible way to bull spermatozoa and to affect sperm quality. It is proposed that adherence could be considered as the main cause of sperm alterations, and also an important step of pathogenesis and venereal transmission.

Open Database Searching Enables the Identification and Comparison of Bacterial Glycoproteomes without Defining Glycan Compositions Prior to Searching

Mass spectrometry has become an indispensable tool for the characterization of glycosylation across biological systems. Our ability to generate rich fragmentation of glycopeptides has dramatically improved over the last decade yet our informatic approaches still lag behind. Although glycoproteomic informatics approaches using glycan databases have attracted considerable attention, database independent approaches have not. This has significantly limited high throughput studies of unusual or atypical glycosylation events such as those observed in bacteria. As such, computational approaches to examine bacterial glycosylation and identify chemically diverse glycans are desperately needed. Here we describe the use of wide-tolerance (up to 2000 Da) open searching as a means to rapidly examine bacterial glycoproteomes. We benchmarked this approach using N-linked glycopeptides of Campylobacter fetus subsp. fetus as well as O-linked glycopeptides of Acinetobacter baumannii and Burkholderia cenocepacia revealing glycopeptides modified with a range of glycans can be readily identified without defining the glycan masses before database searching. Using this approach, we demonstrate how wide tolerance searching can be used to compare glycan use across bacterial species by examining the glycoproteomes of eight Burkholderia species (B. pseudomallei; B. multivorans; B. dolosa; B. humptydooensis; B. ubonensis, B. anthina; B. diffusa; B. pseudomultivorans). Finally, we demonstrate how open searching enables the identification of low frequency glycoforms based on shared modified peptides sequences. Combined, these results show that open searching is a robust computational approach for the determination of glycan diversity within bacterial proteomes.

Pengembangan Media Transpor untuk Koleksi Sampel Preputium, untuk deteksi Bovine Genital Campylobacteriosis

Campylobacter fetus subsp. Venerealis (Cfv) is bacteria causing contagious genital diseases in cows called Bovine Genital Campylobacteriosis (BGC) or vibriosis. Isolation of Cfv is difficult, because the bacteria are fragile and need specific nutrients and oxygen (5-10%). The transport media is very important to maintain Cfv survival before culturing in laboratory. The aim of this study was to modify a new transport media as an alternative media for Cfv. Developed media capability was compared to Weybridge media, and Phosphate Buffered Saline (PBS). All transport media was contaminated by Cfv with concentrations of 105,104,103,102,101 (CFU/ml), and was stored for &lt;6, 24, 48, 72, and 96 hours in each transport medium before culturing on blood agar, all in triplicate. The quality of transport media was analyzed based on bacterial growth on blood agar. PCR test was used as a confirmatory test of growing bacteria cultured on blood agar. Based on culture results, Cfv stored in three transport mediums for &lt;6 hours, Cfv grew on blood agar from all concentration levels provided. Cfv stored for 24, 48, 72 and 96 hours on PBS did not grow, whereas on developed media and Weybridge media, the bacteria could grow and enrichment occurs at all concentration levels given. This study indicated the developed media can be used as an alternative transport medium for Cfv bacteria.

Influence of Protein Glycosylation on Campylobacter fetus Physiology.

Campylobacter fetus is commonly associated with venereal disease and abortions in cattle and sheep, and can also cause intestinal or systemic infections in humans that are immunocompromised, elderly, or exposed to infected livestock. It is also believed that C. fetus infection can result from the consumption or handling of contaminated food products, but C. fetus is rarely detected in food since isolation methods are not suited for its detection and the physiology of the organism makes culturing difficult. In the related species, Campylobacter jejuni, the ability to colonize the host has been linked to N-linked protein glycosylation with quantitative proteomics demonstrating that glycosylation is interconnected with cell physiology. Using label-free quantitative (LFQ) proteomics, we found more than 100 proteins significantly altered in expression in two C. fetus subsp. fetus protein glycosylation (pgl) mutants (pglX and pglJ) compared to the wild-type. Significant increases in the expression of the (NiFe)-hydrogenase HynABC, catalyzing H2-oxidation for energy harvesting, correlated with significantly increased levels of cellular nickel, improved growth in H2 and increased hydrogenase activity, suggesting that N-glycosylation in C. fetus is involved in regulating the HynABC hydrogenase and nickel homeostasis. To further elucidate the function of the C. fetus pgl pathway and its enzymes, heterologous expression in Escherichia coli followed by mutational and functional analyses revealed that PglX and PglY are novel glycosyltransferases involved in extending the C. fetus hexasaccharide beyond the conserved core, while PglJ and PglA have similar activities to their homologs in C. jejuni. In addition, the pgl mutants displayed decreased motility and ethidium bromide efflux and showed an increased sensitivity to antibiotics. This work not only provides insight into the unique protein N-glycosylation pathway of C. fetus, but also expands our knowledge on the influence of protein N-glycosylation on Campylobacter cell physiology.

Causes of abortion in dairy cows in Uruguay

ABSTRACT: A case series study was conducted to determine the frequency of causes of abortion in dairy cattle in Uruguay. The sample size of 102 cases was composed of 53 fetuses, 35 fetuses with placentas, and 14 placentas without an associated fetus. All cases underwent gross and microscopic pathologic examinations as well as microbiological and serological testing. The etiology was determined in 54 (53%) of cases, 51 of which were caused by infectious agents. Within the observed 102 cases, 30 (29%) were caused by Neospora caninum, six (6%) by Coxiella burnetii and two (2%) by Campylobacter fetus subsp. venerealis. Bovine Parainfluenza-3 virus and Salmonella enterica serovar Newport caused one abortion each. Opportunistic bacteria (Escherichia coli, Streptococcus sp., Staphylococcus sp., Mannheimia sp., Trueperella pyogenes, and Providencia stuartii) were associated with 11 abortions. In two cases the fetal death was attributed to dystocia, and in one case the fetus had a congenital mesothelioma. Bovine viral diarrhea virus (BVDV) infection was identified in three fetuses; two of which were co-infected with and had typical lesions of N. caninum. No lesions were observed in the other fetus infected by BVDV. Leptospira interrogans was identified in one fetus without lesions. Despite the relatively low overall success rate in establishing an etiological diagnosis in cases of abortion in cattle, a systemic workup of bovine abortion is necessary to establish prevention and control strategies. This also facilitates monitoring and surveillance of reproductive diseases in dairy cattle, some of which represent a risk to public health.

Co-occurrence of Campylobacter Species in Children From Eastern Ethiopia, and Their Association With Environmental Enteric Dysfunction, Diarrhea, and Host Microbiome.

High Campylobacter prevalence during early childhood has been associated with stunting and environmental enteric dysfunction (EED), especially in low resource settings. This study assessed the prevalence, diversity, abundance, and co-occurrence of Campylobacter spp. in stools from children in a rural area of eastern Ethiopia and their association with microbiome, diarrhea, and EED in children. Stool samples (n = 100) were collected from randomly selected children (age range: 360–498 days) in five kebeles in Haramaya District, Ethiopia. Diarrhea, compromised gut permeability, and gut inflammation were observed in 48, 45, and 57% of children, respectively. Campylobacter prevalence and species diversity were assessed using PCR and meta-total RNA sequencing (MeTRS). The prevalence of Campylobacter spp. in the children's stools was 50% (41–60%) by PCR and 88% (80–93.6%) by MeTRS (P < 0.01). Further, seven Campylobacter species (Campylobacter jejuni, Campylobacter upsaliensis, Campylobacter hyointestinalis, Campylobacter coli, Campylobacter sp. RM6137, uncultured Campylobacter sp., and Campylobacter sp. RM12175) were detected by MeTRS in at least 40% of children stools in high abundance (>1.76-log read per million per positive stool sample). Four clusters of Campylobacter species (5–12 species per cluster) co-occurred in the stool samples, suggesting that Campylobacter colonization of children may have occurred through multiple reservoirs or from a reservoir in which several Campylobacter species may co-inhabit. No associations between Campylobacter spp., EED, and diarrhea were detected in this cross-sectional study; however, characteristic microbiome profiles were identified based on the prevalence of Campylobacter spp., EED severity, and diarrhea. Forty-seven bacterial species were correlated with Campylobacter, and 13 of them also correlated with gut permeability, gut inflammation and/or EED severity. Forty-nine species not correlated with Campylobacter were correlated with gut permeability, gut inflammation, EED severity and/or diarrhea. This study demonstrated that (1) in addition to C. jejuni and C. coli, multiple non-thermophilic Campylobacter spp. (i.e., Campylobacter hyointestinalis, Campylobacter fetus, and Campylobacter concisus) were frequently detected in the children's stools and (2) the Campylobacter, gut permeability, gut inflammation, EED severity, and diarrhea were associated with characteristic microbiome composition. Additional spatial and longitudinal studies are needed to identify environmental reservoirs and sources of infection of children with disparate Campylobacter species and to better define their associations with EED in low-income countries.

Fluorescence in situ hybridization in species-specific diagnosis of ovine Campylobacter abortions

Campylobacter infection is a leading cause of ovine abortion worldwide. Campylobacter fetus and C. jejuni are the major species involved. We report herein on abortion storms in 4 Danish sheep flocks. Initially, no pathogenic bacteria were isolated from placental and fetal tissues on aerobic and selective media despite the presence of severe suppurative and necrotizing placentitis with numerous bacteria located intracellularly in trophoblasts. Fluorescence in situ hybridization (FISH) was then applied on abortion material from 13 cases; species-specific oligonucleotide probes directed against either C. fetus or C. jejuni were used in combination with a general bacterial probe. C. fetus was detected as the only lesion-associated bacterial species in 4 cases from 2 flocks, and C. jejuni in 6 cases from the other 2 flocks, thereby establishing the likely etiology of the abortion storms in all 4 flocks. FISH is a useful detection tool in culture-negative cases with tissue lesions suggestive of bacterial infection. Furthermore, FISH is a fast and economical method to detect and identify the zoonotic agent Campylobacter within ovine abortion material.

English

Campylobacter is a major agent of gastroenteritis worldwide. The incidence and prevalence of campylobacteriosis have been increasing in both developed and developing countries over the last decade. In this study, 197 strains of successive Campylobacter-like were identified in French laboratories in September 2018. Bacterial isolates from clinical samples were identified with a mass spectrometer (Ultraflex III TOF/TOF and the BIOTYPER database from Bruker Daltonics). Of the 197 isolates tested, 143 were identified as Campylobacter jejuni (72.59%), 28 as Campylobacter coli (14.21%), 2 as Arcobacter butzleri (1.02%), 1 as Campylobacter fetus and 1 as Campylobacter lari with (0.51% each) by the MALDI-TOF mass spectrometry. Isolation rate of Campylobacter was highest in the 0 - 9 age group (22%). The proportion of male and female patients was 59.4% (CI 95% = 52.2-66.3) and 40.6% (CI 95% = 33.7-42.8) respectively. Sixty strains (30.5%) were resistant to tetracycline and 52 (26.4%) resistant to ampicillin. This study showed that the MALDI-TOF mass spectrometry is a rapid and accurate identification method of Campylobater spp in patients treated in private French laboratories. Key words: Campylobacter, identification, MALDI-TOF, patients, France.

Virulence Genes, Antibiotic Susceptibility Profiles and Molecular Characterization of Campylobacter Species Isolated from Different Slaughterhouses

The aim of this study was to investigate the frequency of Campylobacter species, to detect the antibiotic resistance profiles and the virulence genes and to determine the clonal proximity of the isolates in the samples of cutting board, slaughterhouse waste water, wall, knife and carcass from three different slaughterhouses in Kayseri region. For this purpose, a total of 150 samples, 10 of each from knife, wall, cutting board, carcass smear sample and slaughterhouse wastewater were collected from each of the three types of slaughterhouses in 2018 in Kayseri. For the isolation of the Campylobacter species, following preenrichment, the suspensions were inoculated onto modified charcoal cefoperazone desoxycholate (CCD) agar and were incubated at 37°C under microaerophilic condition for 48-72 hours. Suspicious colonies with gray-white color were recovered and subjected to phenotypical (Gram staining, oxidase, catalase test, and motion test) tests. Multiplex polymerase chain reaction (mPCR) was used for the molecular identification of the Campylobacter species. Antimicrobial susceptibilities of the isolates identified at the species level were detected by using the disk diffusion test and antibiotic gradient test. Virulence genes (iam, cadF, cdtA, flaA, ceuE, cdtC, cdtB and virB11) among the isolates were evaluated by PCR. The molecular typing of the isolates determined at species level was performed by Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR). In the study, 17 (11.3%) of the 150 samples taken from the slaughterhouse were found to be suspicious in terms of Campylobacter spp. and as a result of phenotypic identification tests, all of the isolates were verified as Campylobacter spp.. As a result of mPCR; eight of the isolates were identified as Campylobacter jejuni, eight as Campylobacter fetus and one as Campylobacter coli. The isolation of the Campylobacter species from different sources was found to be higher in slaughterhouse wastewater than those of others (p<0.001) and the difference in the proportional distribution of the Campylobacter species obtained from various sources was statistically significant (p<0.05). As a result of the disk diffusion test, while, all C.jejuni isolates were resistant to ciprofloxacin, 87.5%, 25%, 25% and 12.5% of C.jejuni isolates were resistant to enrofloxacin, neomycin, amoxicillin/clavulanic acid, and erythromycin, respectively. In addition, 25%, 25% and 12.5% of C.fetus isolates were resistant to amoxicillin/clavulanic acid, neomycin and gentamicin, respectively. C.coli isolate was not resistant to any of the antibiotics tested. Antibiotic gradient test results were found to be compatible with the disc diffusion test results. One of the virulence genes examined, virB11, was not detected in any of the isolates. Moreover, iam gene was not present in C.fetus and C.coli isolates, but only in one C.jejuni isolate. The flaA gene was detected in six C.jejuni isolates. C.coli isolate and seven C.jejuni and seven C.fetus isolates were positive in terms of the cdtC gene. The cdtA, cdtB, ceuE and cadF genes were found to be positive in all C.jejuni isolates. All isolates analyzed in the study demonstrated different ERIC-PCR profiles. In conclusion, it was shown that Campylobacter strains isolated from slaughterhouses were resistant to the most of the current antibiotics. Moreover, the presence of highly virulent Campylobacters in the slaughterhouse environment threatens public health due to the risk of contamination of the humans via carcasses and foods. Therefore, it is recommended that strict hygiene rules should be followed to reduce Campylobacter species contamination in slaughterhouses.

Bovine genital campylobacteriosis: main features and perspectives for diagnosis and control

ABSTRACT: Bovine genital campylobacteriosis (BGC) is a venereal disease caused by Campylobacter fetus subsp. venerealis. In countries with large cattle herds, such as Brazil, where the use of natural breeding as a reproductive strategy is a common practice, BGC is considered an important cause of reproductive failure and economic losses. In these cases, the bull is the asymptomatic carrier of the bacterium and the infected females can have infertility and even abortions. The techniques for the diagnosis of C. fetus are isolation in culture medium and identification by biochemical tests, immunofluorescence, immunoenzymatic assays and molecular techniques. Disease control is based on vaccination with bacterins. This review described the epidemiology, etiology, pathogenesis, and advances in the diagnosis and control of BGC.

Ovine Abortion Associated with Campylobacter fetus subsp. fetus ST2 in Turkey

Ovine Abortion Associated with Campylobacter fetus subsp. fetus ST2 in Turkey

167 Improving the reproductive performance of beef cattle following multivalent viral-bacteria vaccination and mineral supplementation

In beef cattle production under pasture-based tropical conditions, the minerals most likely to be lacking are Ca, P, Na, Co, Cu, I, Se, and Zn, all of which are required for optimal growth and reproduction. Phosphorous deficiency, in particular, is a major problem for grazing cattle in many tropical regions. On the other hand, reproductive infectious diseases are the greatest threat to the production and profitability of beef cattle herds. To overcome reproductive failure, the present study aimed to evaluate the effects of injectable mineral supplementation and vaccination against reproductive diseases on synchronised Nelore females for fixed-time AI. A total of 1361 females were enrolled, and blood was collected from a random population (n=150) to measure serum antibodies against Leptospira sp., infectious bovine rhinotracheitis, and bovine viral diarrhea, mineral status, and creatinine. With Day 0 as the beginning of the synchronisation programme and Day 10 as the day of fixed-time AI, females were randomly assigned as follows. In the control treatment (n=365), no additional treatment was administered. In the Vaccine treatment (n=314), at Days −21 and 0 females received a 5-mL subcutaneous injection of BovigenRepro (Virbac; inactivated vaccine containing infectious bovine rhinotracheitis 1 and 5; bovine viral diarrhea 1 and 2; Campylobacter fetus ssp. fetus, venerealis, and venerealis biotype intermedius; and Leptospira pomona, wolffi, hardjo prajitno, icterohaemorrhagiae, canicola, copenhageni, bratislava, and hardjo bovis in an adjuvant of 10% Al(OH)3 with Se). In the Suppl treatment (n=314), at Day 0 females received a 15-mL intramuscular injection of Fosfosal (Virbac; 100mL containing Na glycerophosphate (14g), monosodium phosphate (20.1g), copper chloride (0.4g), potassium chloride (0.6g), magnesium chloride (2.5g), and sodium selenite (0.24g). In the SupplVacc treatment (n=363), females received a 5-mL subcutaneous injection of the vaccine at Days −21 and 0 as well as a 5-mL intramuscular injection of Fosfosal at Day 0. Pregnancy was verified by transrectal ultrasound at Days 40 and 100. Body condition score was recorded on Days −21 and 40. Data were analysed using the GLIMMIX procedure of SAS (SAS University edition, SAS Institute Inc.). When significant, Tukey and least squares means tests were used to compare means. Serum analysis showed that 89% of the animals had seroprevalence to at least two diseases. Average blood P concentration was 7.2mg dL−1. Pregnancy was affected by Ca levels (pregnant=2.55mg dL−1; non-pregnant=2.33mg dL−1; P&amp;lt;0.05) and creatinine levels (pregnant=5.85mg dL−1; non-pregnant=5.76mg dL−1; P&amp;lt;0.05). The average BCS was 3.2 (1=skinny; 5=fat), and there was no effect of time (Days −21 to 40). Pregnancy verified by transrectal ultrasound at Day 40 for the Vaccine, Suppl, SupplVacc, and control treatments was 54, 52, 57, and 49% (P=0.11), respectively. There was a tendency of higher pregnancy verified by transrectal ultrasound at Day 90 for the SupplVacc group (55%) compared with the control group (45%; P&amp;lt;0.1). There was no effect of pregnancy loss among groups that were vaccinated (control=6%, vaccine=5%, Suppl=4%, and SupplVacc=3%). Considering only pluriparous cows, there was an increase in pregnancy per AI in the SupplVacc group (59%) compared with the control group (51%; P&amp;lt;0.05). Once females presented a very good BCS, the supplement effect was only an immunostimulant. Starting a breeding season with supplementation and vaccination can increase pregnancy rate and profitability for farmers.

Abortion outbreak caused by Campylobacter fetus subspecies venerealis and Neospora caninum in a bovine dairy herd

In November 2015, an abortion outbreak occurred in a commercial dairy herd of 650 Holstein cows in Florida department, Uruguay. Forty-five cows aborted within three weeks. Five fetuses were subjected to gross and microscopic pathologic examination, and microbiological testing. One fetus had fibrinous epicarditis and peritonitis, and neutrophilic bronchopneumonia. Campylobacter fetus subsp. venerealis was detected by direct immunofluorescence, isolated and identified by PCR and sequencing of the 16S rDNA in the abomasal fluid and/or lung. Histologic examination of two other fetuses revealed non-suppurative necrotizing encephalitis, lymphohistiocytic myositis and myocarditis, and lymphocytic interstitial nephritis. In these fetuses, N. caninum antigen was detected intralesionally by immunohistochemistry, and N. caninum DNA was amplified by PCR on formalin-fixed paraffin-embedded brain. Antibodies against N. caninum were detected by indirect immunofluorescence in 10 of 27 cows, with titers ranging from 1/200 to 1/3200. The results indicate that two abortigenic microorganisms may coexist and cause contemporaneous abortion in a herd. We highlight the importance of performing multiple diagnostic tests in various aborted dams and fetuses from the same herd for the etiologic confirmation of bovine abortion syndrome.

Complete Genome Sequences of the Campylobacter fetus subsp. venerealis, Campylobacter lari subsp. concheus, Campylobacter sputorum bv. sputorum, and Campylobacter volucris Type Strains.

Campylobacter spp. are recovered from a wide variety of sources, including birds, livestock, shellfish, and human clinical samples. We present here the complete genomic data for the type strains of Campylobacter fetus subsp. venerealis, Campylobacter lari subsp. concheus, Campylobacter sputorum bv. sputorum, and Campylobacter volucris.

Histopathologic characterization of the reproductive organs of heifers experimentally infected with Campylobacter fetus venerealis

Background: Bovine campylobacteriosis is a venereal disease due to infection with Campylobacter fetus venerealis. It causes mainly reproductive failures that lead to considerable economic losses. Objective: To perform a histopathological description of the mucosa from reproductive organs of heifers experimentally infected with Campylobacter fetus venerealis. Methods: Twelve 15-18-months-old Aberdeen Angus heifers were treated for estrous synchronization and exposed to natural breeding. They were then randomly divided into two groups: group A (n=9) was inoculated with C. fetus venerealis; group B (n=3, control) was inoculated with a placebo. Ultrasonography was performed at days 29, 38, and 42 post-breeding, and plasmatic progesterone levels were quantified using ELISA to confirm pregnancies. Animals in group A with plasma progesterone levels below 1 ng/mL and/or diagnosed as non-pregnant were further divided into three subgroups: A1 (n=4), euthanized at day 30 post-breeding; A2 (n=3), euthanized at day 40 post-breeding and A3 (n=2), euthanized at day 55 postbreeding. Heifers from group B, all diagnosed as pregnant, were euthanized each at day 30, 40, and 55 days post-breeding as well. Histological sections from every group were taken from oviducts, uterus, and vagina. Results: Lymphocytic inflammation was the most common lesion in all infected heifers. Trophoblast cells were found in the non-pregnant heifers euthanized at days 40, and 55 post-breeding. The inflammatory process with the presence of lymphoid cells probably altered the balance in the activity of maternal lymphoid cells, as well as gene expression of the trophoblast, finally affecting the embryo survival. Conclusion: This work contributes to the understanding of the histopathological process involved in post-mating infection of Campylobacter fetus bovine.

L-cysteine transporter-PCR to detect hydrogen sulfide-producing Campylobacter fetus.

Phenotypic differences between Campylobacter fetus fetus and C. fetus venerealis subspecies allow the differential diagnosis of bovine genital campylobacteriosis. The hydrogen sulfide production, for example, is a trait exclusive to C. fetus fetus and C. fetus venerealis biovar intermedius. This gas that can be biochemically tested can be produced from L-cysteine (L-Cys). Herein, we report a novel multiplex-PCR to differentiate C. fetus based on the evaluation of a deletion of an ATP-binding cassette-type L-Cys transporter that could be involved in hydrogen sulfide production, as previously described. A wet lab approach combined with an in silico whole genome data analysis showed complete agreement between this L-Cys transporter-PCR and the hydrogen sulfide production biochemical test. This multiplex-PCR may complement the tests currently employed for the differential diagnosis of C. fetus.