Confirmation of a Campylobacteriosis Outbreak Associated with Chicken Liver Pâté Using PFGE and WGS.

In May 2012, an outbreak of campylobacteriosis occurred in southern Sweden at a wedding reception affecting 44 persons. A total of 17 cases were notified (13 were culture positive for Campylobacter spp.). Epidemiological investigation suspected chicken liver pâté as the source of infection. The liver pâté had been deliberately undercooked, lightly fried to keep the right texture and mixed with spices. Campylobacter isolates from six cases as well as three Campylobacter isolates from chicken flocks previously raised by the producer delivering the liver were subtyped using pulsed-field gel electrophoresis and whole-genome sequencing. Indistinguishable PFGE profiles were identified among five human and one chicken C. jejuni isolates as well among the two C. coli isolates, one from a human case and one from a chicken. WGS supported the PFGE findings; the six C. jejuni isolates belonged to one cluster. All these six isolates were of MLST type ST 50 (ST-CC 21). This study highlights the importance of a combination of strict biosecurity at the flock-level as well as adequate cooking of chicken liver to prevent transmission of Campylobacter to humans.

Associations between Campylobacter levels on chicken skin, underlying muscle, caecum and packaged fillets.

A study was performed with the aim to investigate associations between Campylobacter in chicken caecum, carcass skin, underlying breast muscle and packaged breast fillets. Samples were taken from 285 chickens from 57 flocks and analysed according to ISO 10272. Campylobacter spp. were isolated from caecal samples from 41 flocks. From birds of the same 41 flocks Campylobacter could be detected and quantified in 194 (68%) skin samples. Moreover, Campylobacter spp. were enumerated in 13 (5%) underlying muscle samples originating from 9 of the 41 flocks. The mean number of Campylobacter spp. in the 194 skin samples which could be counted was 2.3 log cfu/g and for the 13 underlying muscle samples 1.3 log cfu/g. Campylobacter could only be quantified in those breast muscle samples with a finding in corresponding skin sample. Five packaged chicken fillets were taken from each 25 of the 57 flocks and analysed both quantitatively and qualitatively. In qualitative analysis Campylobacter was detected in 79 (63%) fillets from 16 flocks and quantified in 24 (19%) samples from 11 flocks. The results showed a significant association (P < 0.05) between findings of Campylobacter on carcass skin (log cfu/g) and the proportion of Campylobacter positive breast muscle samples.

Viruses and bacteria in sputum samples of children with community-acquired pneumonia.

Few comprehensive studies have searched for viruses and bacteria in children with community-acquired pneumonia (CAP). We identified 76 children hospitalized for pneumonia. Induced sputum samples were analysed for 18 viruses by antigen detection and PCR, and for six bacteria by culture and PCR. Viruses were found in 72% of samples, bacteria in 91%, and both in 66%. Rhinovirus (30%), human bocavirus (18%) and human metapneumovirus (14%) were the most commonly detected viruses. Two viruses were found in 22% of samples and three in 8%. The most common bacteria found were Streptococcus pneumoniae (50%), Haemophilus influenzae (38%), and Moraxella catarrhalis (28%). Rhinovirus-S. pneumoniae was the most commonly found combination of virus and bacterium (16%). All six children with treatment failure had both viruses and bacteria detected in the sputum. Otherwise, we found no special clinical characteristics in those with mixed viral-bacterial detections. With modern molecular diagnostic techniques, there are high rates of both viral and bacterial identification in childhood CAP. The clinical significance of mixed viral-bacterial infections remains unclear, although we found a potential association between them and treatment failure.

Induced sputum in the diagnosis of childhood community-acquired pneumonia.

BACKGROUND: The usefulness of induced sputum in searching for causative agents of pneumonia in children has not been studied. METHODS: The study involved 101 children, aged 6 months to 15 years, treated for community-acquired pneumonia at Turku University Hospital (Turku, Finland) from January 2006 to April 2007. Nasopharyngeal aspirate samples were first collected through both nostrils. Sputum production was then induced by inhalation of 5.0% hypertonic saline for 5-10 min and a sputum sample was either aspirated or expectorated. The presence and amount of bacteria and viruses in paired nasopharyngeal aspirate and sputum specimens was analysed and compared using semiquantitative bacterial culture and quantitative PCR techniques. RESULTS: A good quality sputum specimen was obtained from 76 children. The possible causative agent was found in 90% of cases. Streptococcus pneumoniae (46%) and rhinovirus (29%) were the most common microbes detected. Newly discovered viruses human bocavirus and human metapneumovirus were detected in 18% and 13% of the children, respectively. One-quarter of all bacterial findings were only detected in sputum, and the amount of bacteria in the remainder of the sputum specimens compared with nasopharyngeal aspirate was higher in 14% and equal in 70%. The amount of rhinovirus in sputum was higher than in nasopharyngeal aspirate in 82%. CONCLUSIONS: Sputum induction provides good quality sputum specimens with high microbiological yield in children with community-acquired pneumonia. Induced sputum analysis can be useful in the microbiological diagnosis of childhood community-acquired pneumonia.

Pneumolysin polymerase chain reaction for diagnosis of pneumococcal pneumonia and empyema in children.

Streptococcus pneumoniae is the most important cause of childhood pneumonia and empyema, yet the diagnosis of pneumococcal infections by conventional methods is challenging. In this study, the clinical value of the pneumolysin-targeted real-time polymerase chain reaction (PCR) method for the diagnosis of pneumococcal pneumonia and empyema was evaluated with 33 whole blood samples and 12 pleural fluid samples. The analytical sensitivity of the PCR assay was 4 fg of pneumococcal DNA, corresponding to two genome equivalents of pneumococcal DNA per reaction. The PCR assay correctly detected all clinical isolates of S. pneumoniae tested, whereas all nonpneumococcal bacterial organisms tested were negative by PCR. In a clinical trial, S. pneumoniae was detected by PCR in the pleural fluid of 75% of children with empyema, increasing the detection rate of pneumococcus almost tenfold that of pleural fluid culture. However, in whole blood samples, PCR detected S. pneumoniae in only one child with pneumonia and one child with pneumococcal empyema and failed to detect S. pneumoniae in three children with blood cultures positive for S. pneumoniae. The present data indicate that pneumolysin-targeted real-time PCR of pleural fluid is a valuable method for the etiologic diagnosis of pneumococcal empyema in children. The ease and rapidity of the LightCycler technology (Roche Diagnostics, Mannheim, Germany) make real-time PCR an applicable tool for routine diagnostics. In the evaluation of blood samples, blood culture remains the superior method for the diagnosis of bacteremic pneumococcal disease.

Antimicrobial resistance in Staphylococcus spp., Escherichia coli and Enterococcus spp. in dogs given antibiotics for chronic dermatological disorders, compared with non-treated control dogs.

The aim of this study was to evaluate antimicrobial resistance in canine staphylococci, Escherichia coli and enterococci, which were isolated from 22 dogs with pyoderma and a history of previous antibiotic treatment, compared to bacterial isolates from 56 non-treated control dogs. Two isolates of each bacterial species per dog were investigated, if detected. Staphylococcal isolates from dogs with pyoderma (35 isolates) were more resistant to sulphatrimethoprim than the isolates from controls (56 isolates) (57% vs. 25%, p < 0.004). Multiresistance in staphylococci was also more common in dogs with pyoderma (29% vs. 9%, p = 0.02). A similar trend among isolates of E. coli was detected (24 and 74 isolates from treated and control dogs, respectively), but the differences were not significant. Resistance for macrolide-lincosamides was approximately 20% among staphylococci in both groups. Resistance to ampicillin among enterococci was 4%-7%. The age of the dogs might have an impact on resistance: multiresistance among staphylococcal isolates from younger dogs (< or = 5 years) was more common than in older dogs (26 years) (24%, vs. 0%, 63 and 27 isolates, respectively, p = 0.02). Staphylococci in younger dogs were more resistant to tetracycline (48% vs. 11%, p < 0.001) and sulphatrimethoprim (48% vs. 15%, p < 0.01) than those in older dogs. In contrast, the isolates of E. coli from older dogs tended to be more resistant, although a significant difference was detected only in resistance to tetracycline (13% vs. 2% of 40 and 50 isolates respecthely, p = 0.04)). The results of this small study indicate that resistance in canine staphylococci in the capital area of Finland is comparable with many other countries in Europe. Resistance in indicator bacteria, E. coli and enterococci, was low.

Use of phenotyping and genotyping to verify transmission of Escherichia coli O157:H7 from dairy farms.

A total of 80 human infections by Escherichia coli O157:H7 were documented in Finland in 1997 and 1998. Most were sporadic and their sources undetermined. Five cases not associated with one another, one of which led to secondary transmission within a family, could be traced to five different dairy farms. These five case patients (age range 2-17 years, median age 3 years) were hospitalised with bloody diarrhoea; two of them developed haemolytic uraemic syndrome. All nine human isolates obtained were sorbitol negative, carried the verocytotoxin 2 and eae genes, and produced verocytotoxin and enterohaemolysin. The phage and pulsed-field gel electrophoresis types of the human and bovine isolates from the corresponding farms were indistinguishable. The cattle (20-70 animals per farm) were monitored for up to 2 years after the human cases. The proportion of cattle excreting the type that caused the human infections varied from 3.2 to 66.7% when sampled soon after the human cases, and from 0.0 to 5.3% about a year or so later. On most of the farms, the animals excreted the pathogen intermittently. On one farm, Escherichia coli O157 isolates with other characteristics were also occasionally isolated. Although the infections were traced back to the farms, it could not be established whether the source was unpasteurised milk or direct or indirect contact with cattle. The results of this study emphasise the need for special recommendations for children visiting or living on a farm to prevent these infections.

Occurrence of Escherichia coli O157 in Finnish cattle.

Bovine faecal samples were collected during June-December 1997 at 14 major abattoirs slaughtering cattle in Finland. Escherichia coli O157 was isolated from 19 of the 1448 samples (1.31%) after enrichment and immunomagnetic separation (IMS). The positive faecal isolates originated from 16 farms and eight abattoirs. The occurrence of E. coli O157 was highest in July (8/204; 3.92%) and September (6/244; 2.46%). No E. coli O157 was detected in November and December, nor from the faecal samples from the northernmost region where cattle density is low. All of the isolates carried the eae gene and showed the enterohaemolytic phenotype. All except one were motile and had the flagella antigen H7. Seventeen of the isolates were positive for stx(2) gene and one carried both the stx(1) and stx(2) genes. Of the 17 isolates with stx genes, 16 were verocytotoxin-positive in a reversed passive latex agglutination test after polymyxin extraction but only eight without extraction. The isolates belonged to 10 different pulsed-field gel electrophoresis (PFGE) patterns. The most common PFGE pattern (1.42) was detected in eight isolates (42.1%). Four PFGE patterns (1.1; 1.6; 1.12; 1.14) were identical with those isolated from humans in Finland, suggesting that at least some human E. coli O157 infections may be of bovine origin.

Swimming-associated outbreak of Escherichia coli O157:H7.

In 1997 the first outbreak of Escherichia coli O157:H7 infections involving 14 cases occurred in Finland. A case was defined as a resident of Alavus with an episode of diarrhoea between 5 and 17 July 1997, and from whom E. coli O157:H7 was isolated from stool. The investigation included case searching and a population-based case control study. Five primary and eight symptomatic secondary cases of E. coli O157:H7 illness were detected. In the 10 days before the outbreak, all 5 primary patients (aged 3-8 years), but only 6 of 32 population controls from the same age range (Fisher's test, P < 0.001) and 4 of 10 sibling controls (P < 0.05) had visited (but had not necessarily bathed in) a shallow beach popular among young children. Four out of 5 primary cases had remained within 5 m of the beach while swimming and had swallowed lake water compared to 1 of 5 population controls. These analytical epidemiologic findings incriminated fresh lake water as the vehicle of E. coli O157:H7 transmission.

Transdermal estrogen with a levonorgestrel-releasing intrauterine device for climacteric complaints: clinical and endometrial responses.

OBJECTIVE: Our purpose was to study the effects of intrauterine release of a daily dose of 20 micrograms levonorgestrel by an intrauterine device on climacteric symptoms, bleeding pattern, and endometrial histologic features in postmenopausal women receiving transdermal estrogen replacement therapy. STUDY DESIGN: Forty parous postmenopausal women were randomly allocated into two groups for 1 year: 20 women receiving a continuous transdermal daily dose of 50 micrograms of estradiol had a levonorgestrel-releasing intrauterine contraceptive device inserted, and the control group of 20 women received a continuous oral dose of 2 mg of estradiol valerate and 1 mg of norethisterone acetate daily. The climacteric symptoms, bleeding patterns, endometrial thickness, and endometrial changes in biopsy samples were analyzed. Serum levels of estradiol in both groups and levonorgestrel levels in the intrauterine device group were also determined. RESULTS: Both treatment regimens effectively relieved climacteric symptoms. Spotting was more common in the intrauterine contraceptive device group than in the oral therapy group for the first 3 months. After that, the proportion of women without any bleeding was similar in both groups. Two patients in each group dropped out because of bleeding. CONCLUSION: These preliminary findings suggest that the levonorgestrel-releasing intrauterine contraceptive device is a useful alternative mode of progestin administration for certain selected women receiving estrogen replacement therapy.

PIP: The purpose was to study the effects of intrauterine release of a daily dose of 20 mcg levonorgestrel by an IUD on climacteric symptoms, bleeding pattern, and endometrial histologic features in postmenopausal women receiving transdermal estrogen replacement therapy. 40 parous postmenopausal women were randomly allocated into 2 groups for 1 year. They were required to be parous, to have an intact uterus, and to have had amenorrhea for at least 6 months but less than 5 years. 20 women received a combination of 50 mcg of estradiol per 24 hours delivered transdermally from a patch, and received estrogen pretreatment for 1 month to make insertion of a levonorgestrel-releasing IUD (Levonova), which was installed a month later, easier. This combination was continued for 1 year. The control group of 20 women received an established form of continuous oral estrogen and progestin with a daily dose of 2 mg of estradiol, and 1 mg of norethindrone acetate also administered for 1 year. Checkup visits were scheduled at 3, 6, and 12 months. The climacteric symptoms, bleeding patterns, endometrial thickness, and endometrial changes in biopsy samples were analyzed. The increase in estradiol concentration was similar in both groups, and the mean concentrations of levonorgestrel in the IUD group were 216 +or- 25 pg/ml at 3 months, 209 +or- 11 pg/ml at 6 months, and 212 + 10.5 pg/ml at 12 months. Both treatment regimens effectively relieved climacteric symptoms. The IUD group experienced more days of bleeding, primarily spotting, during the first 3 months than did the oral therapy group but the differences between the groups had disappeared by 6 months. Both treatments resulted in an atrophic endometrium developing from a proliferative one. Two patients in each group dropped out because of bleeding. The levonorgestrel-releasing IUD is a useful alternative mode of progestin administration for certain selected women receiving estrogen replacement therapy.

Maturation of vaginal and endometrial epithelium in postmenopausal breast cancer patients receiving long-term tamoxifen.

To assess the estrogenic effects of tamoxifen on vaginal and endometrial epithelium and to investigate whether these changes are associated with any pathological findings in the endometrium, 53 postmenopausal breast cancer patients receiving long-term tamoxifen and 52 control breast cancer patients without any hormonal treatment were examined. Pathological findings in the endometrium were evaluated by hysteroscopy and curettage. The main outcome measures were the maturation index in Papanicolaou (Pap) smears, estrogen-like epithelial changes in the endometrium, serum concentrations of gonadotropins, sex hormone-binding globulin (SHBG), estradiol (E2), and testosterone (T). Informative Pap smears showed estrogenic effects in 89% (41/46) of the tamoxifen group and in 49% (22/45) of the control group, and in endometrial aspiration samples in 71% (32/45) and in 41% (19/46), respectively. These changes were associated with increased concentrations of serum E2 in the control group but not in the tamoxifen group. All five patients (11%) with endometrial polyps in the control group showed estrogenic endometrial changes, whereas among 14 women with polyps in the tamoxifen group, 9 showed estrogenic changes and 5 endometrial atrophy. Endometrial adenocarcinoma was found in 3 patients in the tamoxifen group and in 2 in the control group. Pap smears showed atrophy in 2 patients in the former and in one in the latter group. These findings confirmed estrogen-like effects of tamoxifen on the vaginal and endometrial epithelium in postmenopausal breast cancer patients, but these were not closely associated with benign or malignant endometrial lesions.

Plasma insulin-like growth factor I and its binding proteins 1 and 3 in postmenopausal patients with breast cancer receiving long term tamoxifen.

BACKGROUND: Insulin-like growth factor I (IGF-I) is a potent mitogen for breast cancer cells. The majority of IGF-I in plasma is bound to IGF binding proteins (IGFBPs), which modulate the biologic effects of IGF-I. METHODS: Plasma concentrations of IGF-I, IGFBP-I, and IGFBP-3 were compared between 40 postmenopausal breast cancer patients receiving long term tamoxifen therapy and 39 breast cancer patients receiving no hormonal treatment. In an additional group of seven patients, serum levels of IGF-I and IGFBP-1 were determined before and during treatment at 6 and 12 months. RESULTS: The tamoxifen and the control groups did not differ with respect to age, parity, age at menopause, or body mass index. There were no significant differences in the mean concentrations (+/- standard error of the mean) of IGF-I (10.0 +/- 0.4 nmol/l and 11.2 +/- 0.5 nmol/l, respectively) and IGFBP-3 (3.2 +/- 0.1 mg/l and 3.1 +/- 0.1 mg/l, respectively), whereas the mean value of IGFBP-1 was significantly higher in the tamoxifen group (6.0 +/- 0.6 micrograms/l versus 2.8 +/- 0.3 micrograms/L, P = 0.0001). No significant differences were found in the insulin levels. During the treatment, concentrations of IGF-I decreased at 6 months and began increasing at 12 months. IGFBP-1 levels increased at 6 months and remained elevated at 12 months. CONCLUSIONS: The tamoxifen-induced increase in IGFBP-1 plasma levels may be an important mechanism modulating IGF-I action at the tissue level.

Endometrial changes in postmenopausal breast cancer patients receiving tamoxifen.

OBJECTIVE: To assess the estrogenic effect of tamoxifen and associated pathologic changes of the endometrium in postmenopausal breast cancer patients. METHODS: The endometrium of 103 gynecologically asymptomatic postmenopausal breast cancer patients was examined. Fifty-one had been treated with tamoxifen and 52 had not received any hormonal treatment. The two groups were similar in age, parity, age at menopause, and body mass index. RESULTS: Compared with the control subjects, the tamoxifen patients had a thicker endometrium (mean +/- standard deviation 10.4 +/- 5.0 versus 4.2 +/- 2.7 mm; P = .0001) and larger uterine volume (45 +/- 27 versus 25 +/- 11 cm3; P = .001), as determined by transvaginal sonography. Hysteroscopy showed an atrophic endometrium in 28% of the patients in the tamoxifen group, as compared with 87% of the control patients (P = .0001). Endometrial polyps were more frequent in the tamoxifen group (36 versus 10%; P = .004), which included one patient with atypical hyperplasia, one with adenomatous hyperplasia, and one with endometrial adenocarcinoma; two controls had endometrial adenocarcinoma. CONCLUSION: The results provide evidence for an estrogenic effect of long-term tamoxifen treatment on the postmenopausal uterus and show it to be associated with an increased occurrence of polyps.

Uptake and distribution of radioiodine, and the effect of ambient nitrate, in some fish species.

125I uptake by the thyroid was most pronounced in the smelt (Osmerus eperlanus), perch (Perca fluviatilis), rainbow trout (Salmo gairdneri), roach (Rutilus rutilus) and bleak (Alburnus alburnus). Unlike other tissues, only the muscle in the species studied no relative accumulation of 125I as compared to the ambient water. The Crusian carp (Cyprinus carassius) indicated the lowest levels of accumulation of 125I among the species tested. Thyroid radioiodine concentration was independent of the length of the fish in the rainbow trout and perch. In the rainbow trout, however, the liver and muscle radioiodine concentrations decreased significantly with increasing length of the fish. Exposure of rainbow trout to water containing supranormal concentration of nitrate (1500 micrograms/l) resulted in lowered 125I uptake. The same phenomenon was observed for the perch and Crusian carp, with low nitrate concentrations, while with higher nitrate concentrations, the uptake levels were again increased. Thyroid function, as judged from the conversion ratio and T/S ratio, was not affected by nitrate.