Evaluation of Two Multiplex PCR-High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies.

Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)-high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene. However, higher intraspecies variation within C. coli and C. jejuni isolates was detected in mPCR1-HRM when compared with mPCR2-HRM. Both assays were rapid and required minimum interpretation skills for discrimination between and within Campylobacter species when using HRM curve analysis software.

Tricomonose e campilobacteriose em bovinos: revisão de literatura / Trichomoniasis and campylobacteriosis in cattle: a literature review / Tricomoniasis y campilobacteriosis en ganado: revisión de literatura

O aumento da produção de leite no Brasil tem sido essencial para o crescimento do país no mercado internacional, com destaque para a região Sul do país. Com o avanço tecnológico na atividade leiteira, avançou também o cuidado com a sanidade dos rebanhos, que engloba o manejo sanitário da reprodução desses rebanhos. A Tricomonose e a Campilobacteriose são doenças sexualmente transmissíveis que afetam bovinos em várias idades. Tritrichomonas foetus é o agente causador da Tricomonose e C. fetus subsp. venerealis o da Campilobacteriose, sendo, respectivamente, um protozoário e uma bactéria Gram-negativa. O diagnóstico dessas doenças se dá via coleta de lavado prepucial ou cervicovaginal, para a pesquisa de Tritrichomonas spp, e pelo swab prepucial ou cervicovaginal para a pesquisa de Campylobacter spp. Não há tratamento específico para essas doenças, visto que o controle e profilaxia baseiam-se na retirada dos machos portadores do rebanho, ou realização de descanso reprodutivo de quatro ou cinco estros nas fêmeas, já que as mesmas eliminam os agentes etiológicos. Diante do exposto é necessário reconhecer a necessidade de estudos relacionados a Tricomonose e Campilobacteriose em rebanhos leiteiros, principalmente aqueles inseridos em sistema de produção familiar, subsidiando essa revisão de literatura.(AU)

The increase in dairy production in Brazil has been essential to the country's growth in the international market, especially in the southern region of the country. With the technological advances in dairy farming, the caring for the health of herds has also increased, as well as the health management of the reproduction of these animals. Trichomoniasis and campylobacteriosis are sexually transmitted diseases that affect cattle at different ages. Tritrichomonas fetus is a protozoan that promotes Trichomoniasis and C. fetus subsp. venerealis is a gram-negative bacterium that promotes campylobacteriosis. In order to diagnose these diseases, the prepucial or cervicovaginal wash has been collected for Tritrichomonas spp. and the preputial or cervicovaginal swab has been collected for Campylobacter spp. There is no specific treatment for these diseases, since the control and prophylaxis are based on the removal of infected males, or in carrying out reproductive rest between four or five estrous cycles in females, since these actions can eliminate the etiologic agent. Thus, it is important to study trichomoniasis and campylobacteriosis in dairy herds, especially those placed in family farming, supporting this literature review.(AU)

El aumento de la producción de leche en Brasil ha sido esencial para el crecimiento del país en el mercado internacional, especialmente en la región sur del país. Con los avances tecnológicos en la industria lechera también se avanzó el cuidado con la salud de los rebaños, que incluye el manejo sanitario de la reproducción de esos rebaños. La tricomoniasis y la campilobacteriosis son enfermedades sexualmente transmisibles que afectan al ganado de diferentes edades. Tritrichomonas fetus es el agente causante de la tricomoniasis y C. fetus subsp. venerealis o de la campilobacteriosis, siendo, respectivamente, un protozoos y una bacteria Gram- negativa. El diagnóstico de esas enfermedades se produce a través de colección de lavado de prepucio o cervicovaginal, para la investigación de Tritrichomonas spp, y por el swab prepucio o cervicovaginal para la investigación de Campylobacter spp. No existe tratamiento específico para esas enfermedades, ya que el control y profilaxis se basan en la retirada de los machos portadores del rebaño, o realización de descanso reproductivo de cuatro o cinco celos en las hembras, pues las mismas eliminan los agentes etiológicos. Delante el expuesto, es necesario reconocer la necesidad de realizar estudios relacionados con la tricomoniasis y campilobacteriosis en rebaños lecheros, especialmente los situados en el sistema de producción familiar, subsidiando esa revisión de literatura.(AU)

Multilocus Sequence Typing analysis of human Campylobacter coli in Granada (Spain) / Tipificación mediante secuencias multilocus de cepas humanas de Campylobacter coli en Granada (España)

Introduction. Different subtypes of Campylobacter spp. have been associated with diarrhoea and a Multilocus Sequence Typing (MLST) method has been performed for subtyping. In the present work, MLST was used to analyse the genetic diversity of eight strains of Campylobacter coli. Material and methods. Nineteen genetic markers were amplified for MLST analysis: AnsB, DmsA, ggt, Cj1585c, CJJ81176-1367/1371, Tlp7, cj1321-cj1326, fucP, cj0178, cj0755/cfrA, ceuE, pldA, cstII, cstIII. After comparing the obtained sequences with the Campylobacter MLST database, the allele numbers, sequence types (STs) and clonal complexes (CCs) were assigned. Results. The 8 C. coli isolates yielded 4 different STs belonging to 2 CCs. Seven isolates belong to ST-828 clonal complex and only one isolate belong to ST-21. Two samples came from the same patient, but were isolated in two different periods of time. Conclusions. MLST can be useful for taxonomic characterization of C. coli isolates (AU)

Introducción. Diferentes subtipos de Campylobacter spp. se han asociado con diarrea y la técnica de tipado mediante análisis de secuencias de múltiples locus (MLST) se ha empleado para la tipificación genética. En el presente trabajo, la técnica MLST se utilizó para analizar la diversidad genética de ocho cepas de Campylobacter coli. Material y métodos. 19 marcadores genéticos fueron amplificados mediante el análisis MLST: AnsB, DmsA, ggt, Cj1585c, CJJ81176-1367/1371, Tlp7, cj1321-cj1326, fucP, cj0178, cj0755/cfrA, ceuE, pldA, cstII, cstIII. Después de comparar las secuencias obtenidas con la base de datos MLST para Campylobacter, se asignaron el número de los alelos, los secuenciotipos (STs) y los complejos clonales (CCs). Resultados. Las 8 cepas de C. coli aisladas mostraron 4 STs diferentes pertenecientes a 2 CCs. Siete aislamientos pertenecieron al complejo clonal ST-828 y sólo un aislado perteneció al ST-21. Dos aislados pertenecieron al mismo paciente, pero fueron obtenidos en diferentes periodos de tiempo. Conclusiones. La técnica MLST puede ser útil para la caracterización taxonómica de aislados de C. coli (AU)

Correlations between Campylobacter spp. prevalence in the environment and broiler flocks.

AIMS: To investigate (i) possible correlations between the presence of Campylobacter spp. in the surroundings of broiler farms and their incidence in flocks, and (ii) possible associations between weather conditions and the occurrence of Campylobacter spp. METHODS AND RESULTS: Farms were selected according to previous results from the Swedish Campylobacter programme. Samples were collected in and around broiler houses during the rearing period from 131 flocks on 31 farms, including sock samples from the ground outside, from the floor in the broiler houses and anterooms, and samples from insects, water, feed and ventilation shafts. CONCLUSIONS: As expected, there was a difference in Campylobacter isolation rates for different categories of farms regarding samples taken in the houses. However, there were no differences regarding the presence of Campylobacter spp. in the environment between producers that often deliver Campylobacter-positive slaughter batches and those that rarely deliver positive batches. Campylobacter spp. were more frequently found in the surroundings on rainy days when compared with sunny days. SIGNIFICANCE AND IMPACT OF THE STUDY: Physical barriers between outside and inside the houses appeared to be important for preventing Campylobacter spp. in the environment to be transferred into the broiler houses.

European survey on Campylobacter surveillance and diagnosis 2001.

Two surveys have been conducted on behalf of the Campylobacter Working Group with the aim of assessing the feasibility of a European network on human Campylobacter infections. The first survey, conducted in 18 countries, collected information about diagnostic methods used for surveillance purpose while the second one, conducted among 10 European countries, gathered data on diagnostic methods and procedures in primary laboratories. Seventeen of the 18 countries had a surveillance system for Campylobacter infections and 13 a national reference laboratory. The case definition used for the surveillance in all these countries included laboratory confirmation. No commonly applicable subtyping methods were applied. Concerning primary laboratories dealing with Campylobacter infections, only a few of them reported directly to the national level, and antimicrobial susceptibility testing was performed by about half of the laboratories. These surveys indicated that a basic infrastructure for a wide European Campylobacter surveillance exists.

Campylobacter species in health, gingivitis, and periodontitis.

At least seven Campylobacter species have been identified from subgingival sites. Campylobacter rectus has been implicated as a periodontal pathogen; however, association with periodontal infections of other Campylobacter species, especially the newly described Campylobacter showae, is unclear. This study examined which Campylobacter species were associated with periodontal health and disease. Subgingival Campylobacter species from initial and established periodontitis were compared with species from periodontally healthy subjects, including subjects with gingivitis. Campylobacter species were isolated on selective media and identified by whole-cell protein profiles (SDS-PAGE). Except for C. rectus, Campylobacter levels were frequently below the detection limit (2-5% of the microbiota) of non-selective culture methods. C. rectus and C. showae, including Campylobacter X, were found more frequently and in higher levels from diseased than from healthy periodontal sites. C. gracilis was the dominant Campylobacter species found in relatively shallow pockets; however, its presence was unrelated to periodontal health or disease. C. concisus was isolated in higher proportions from relatively shallow and healthy sites, compared with deeper pockets. C. curvus was unrelated to periodontal health or disease. Analysis of the study data confirmed the relationship of C. rectus with diseased subgingival sites and indicated that C. showae may also be associated with periodontal disease.

Repetitive and arbitrary primer DNA sequences in PCR-mediated fingerprinting of outbreak and sporadic isolates of Campylobacter jejuni.

PCR-mediated fingerprinting with combined repetitive and arbitrary DNA primers (ERIC-2 and 1026) was used to type Campylobacter jejuni from a milk-associated outbreak, and from sporadic cases of the same and allied HS serotypes. The 14 outbreak strains had identical or similar DNA band profiles whereas the 25 strains from sporadic infections were more heterogeneous with 18 different DNA profiles. Although PCR-based DNA fingerprints lacked serotype specificity, the method was fast, simple to perform and reproducible, provided defined technical protocols were adhered to precisely. Profiles were highly discriminatory but did not consistently match types based on other molecular methods. We conclude that AP-PCR has demonstrable potential for initial rapid investigation of outbreaks, and when used in conjunction with PFGE analysis of DNA restriction profiles, provides a high resolution strategy for accurately defining subtypes of C. jejuni.

Aneurisma de aorta abdominal infectado por Campylobacter fetus spp fetus: relato de cas e revisao de literatura / Abdominal aortic aneurysm infected with campylobacter fetus spp fetus

A infeccao de aneurisma de aorta por Campylobacter fetus spp fetus e rara, sendo o primeiro caso relato em 1971. Apresentamos um caso de aneurisma de aorta abdominal, com historia de dor abdominal, febre e calafrios, em que houve identificacao deste bacilo Gram negativo na cultura de fragmento de aorta colhido na operacao e visualizacao de microrganismo em estudo anatomo-patologico...

[Detection of Campylobacter pylori in gastric mucosa--relation to gastroscopy findings and degree of gastritis]. / Nachweis von Campylobacter pylori in der Magenschleimhaut-Beziehungen zu gastroskopischen Befunden und zum Gastritisgrad.

In 862 unselected gastroscopies we could confirm the high percentage of campylobacter pylori infection in the antral gastric mucosa strongly associated with chronic superficial gastritis and active inflammatory signs. Significant correlation with age concerned the normal findings only. In patients with duodenal ulcers we found in 91% gastritis and in 88% campylobacter pylori in antral mucosa. A small group with duodenal ulcers was campylobacter pylori-negative as well as without gastritis signs. Comparing the antrum and corpus gastric mucosa (250 pat.) the antral mucosa was representative for the whole stomach in view of gastritis and campylobacter pylori test, sufficiently for practical use. Between the test methods (culture, urease, microscopy) we favoured the microscopic test.

Campylobacter enteritis in Nottingham.

The incidence of campylobacter gastroenteritis in the population of Nottingham over a period of 3 years was studied. There was a seasonal variation with the highest number of cases occurring in the summer months. Campylobacter sp. were isolated from the stools of a total of 780 patients over this period. Of these 160 patients with gastroenteritis required admission to hospital. These patients' illness had an acute onset, and the predominant features were diarrhoea, severe abdominal pain, nausea and bright red blood with the stool. However not all the patients had diarrhoea. More than a third of the patients studied were less than 10 years old. The mean duration of symptoms was 4 days and the average stay in hospital was 5 days; some patients required prolonged admission (14 days). In a few cases campylobacter enteritis mimicked other clinical conditions including acute appendicitis. This study emphasises the importance of campylobacter enteritis as a cause of gastroenteritis in the community and the degree of morbidity associated with this illness.

[Campylobacteriosis in humans caused by subspecies intestinalis and fetus. Six new diseases (author's transl)]. / Campylobacteriosis des Menschen durch die Subspezies intestinalis und fetus unter Berücksichtigung sechs neuer Erkrankungen.

The subspecies intestinalis and fetus of Campylobacter fetus are opportunistic pathogens in humans. So far, some 200 types of disease due to these pathogens have been reported from all over the world, and six new diseases are briefly described in this paper. The most frequent disease is sepsis, followed by meningitis. While ticarcillin, erythromycin, clindamycin and tetracycline are the antibiotics which show activity against these species, a large degree of resistance is seen against cephalothin, cefazolin, cefuroxime, cefoxitin and cefotaxime. There is a moderate degree of sensitivity to gentamicin. The epidemiology of the disease is not clear, and about twice as many men are affected as women. Subspecies intestinalis was isolated in 95% of all diseases. Antibodies to Campylobacter fetus were detected in 3.9% of the population.

Campylobacter fetus infections in children.

There has been a gradual accumulation of reported Campylobacter fetus human infections since the first description in 1947. The taxonomy of these fastidious and morphologically confusing organisms has been recently revised and, in the past few years, through the use of selective culture medium for stool isolation C fetus has been implicated as a frequent paghogen in children. There are three relatively distinct patterns of human C fetus infection. The first, most frequent, pattern of disease is enteritis which is usually uncomplicated and due to C fetus subsp jejuni. A second form of disease consists of focal infections, often associated with vasculitis and/or chronic bacteremia. These infections, due to C fetus subsp intestinalis, are seen most often in older, debilitated, or chronically ill men. The third pattern, perinatal infections causing abortion, prematurity, and neonatal meningitis, is the least frequent, but these infections are usually fetal to the fetus or infant and are also due to C fetus subsp intestinalis.