Genomic characterisation, detection of genes encoding virulence factors and evaluation of antibiotic resistance of Trueperella pyogenes isolated from cattle with clinical metritis.

Trueperella pyogenes is one of the most important microorganisms causing metritis in post-partum cattle. Co-infection with other bacterial species such as Escherichia coli or Fusobacterium necrofurom increases the severity of the disease and the persistence of bacteria in utero. The aim of this study was to investigate the frequency of T. pyogenes strains, and their virulence and antimicrobial resistant profiles in metritis cases. The study was carried out on 200 samples obtained from metritis discharges of postpartum cattle on 18 farms around Tehran, Iran. Sixty-five T. pyogenes isolates (32.5%) were identified, of which 16 isolates were detected as pure cultures and the other 49 isolates from cultures most commonly mixed with E. coli or F. necrofurom. In terms of diversity in biochemical characteristic of T. pyogenes strains, 8 different biotypes were identified among the isolates. Single or multi antimicrobial resistance was observed in 48 isolates (73.9%), which was mostly against trimethoprim sulfamethoxazole, azithromycin, erythromycin and streptomycin. The tetracycline resistance gene tetW and macrolide resistance genes ermB and ermX were detected in 30, 18 and 25 isolates, respectively. In the screening of genes encoding virulence factors, fimA and plo genes were identified in all tested isolates. Genes encoding nanP, nanH, fimC, fimG, fimE and cbpA were detected in 50, 54, 45, 40, 50 and 37 of isolates, respectively. Thirteen different genotypes were observed in these T. pyogenes isolates. A significant association between clonal types and virulence factor genes, biochemical profile, CAMP test result, severity of the disease and sampling time was detected.

Prevention and treatment strategy in pregnant women with group B streptococcal infection.

Group B streptococcus (GBS; Streptococcus agalactiae) are encapsulated gram-positive cocci belonging to Lancefield group B, that frequently colonizes the human genital and gastrointestinal tracts. It is an important cause of illness in three categories of population: infants, pregnant women, and adults with underlying medical conditions. In pregnant women and postpartum women, GBS is a frequent cause of asymptomatic bacteriuria, urinary tract infection, upper genital tract infection (i.e. intraamniotic infection or chorioamnionitis), postpartum endometritis (8%), pneumonia (2%), puerperal sepsis (2%), and bacteremia without a focal site (31%). It also can cause focal infections such as pneumonia, meningitis, and endocarditis, albeit rarely. Invasive maternal infection with GBS is associated with pregnancy loss and preterm delivery. Prior to the widespread use of maternal intrapartum chemoprophylaxis, maternal colonization with GBS conferred an increased risk of chorioamnionitis, and early postpartum infection. The serotype distribution of invasive GBS infection in pregnant women is similar to that of early-onset neonatal disease. The most common GBS serotypes causing invasive disease in adults and neonates are Ia, Ib, III, and V. Vaccination of adolescent women is considered an ideal solution. However, recent reports (April 2015) have shown that serotype IV GBS is emerging in pregnant carriers and causing infections in neonates and adults. This emergence is of concern because GBS conjugate vaccines that are being developed to prevent invasive disease may protect only against serotypes Ia, Ib, II, III, and V, or combinations thereof. Though research for the development of such a vaccine is underway, a good candidate vaccine has yet to surface.

Strategy for the treatment of puerperal metritis and improvement of reproductive efficiency in cows with retained placenta.

The objective of this study was to improve the reproductive efficiency of dairy cows with puerperal metritis (PM) subsequent to retained placenta (RP) using a two-step treatment strategy. A total of 188 postpartum cows, aged from 2 to 8 years, were utilised for 2 experiments. In Experiment 1, cows affected with RP/PM were randomly assigned to two treatment groups. Cows in Group A (n = 17) were treated with 600 mg of ceftiofur intramuscularly for 3 days followed by intrauterine lavage with 0.1% chlorhexidine and infusion with 0.5% povidoneiodine, while cows in Group B (n = 16) received two intrauterine infusions, first with 5 g of oxytetracycline and then with 0.5% povidone-iodine. Cows with normal postpartum findings were regarded as the healthy control group (n = 26). Ultrasonographic examination revealed that the ovarian activities including the appearance of a dominant follicle and days to first ovulation of the cows in Group A during the early postpartum period differed from those of Group B (P < 0.05), which coincided with the results of uterine swabbing for bacteriology. In Experiment 2, cows with normal postpartum findings were allocated to Group D (n = 78), which received an ovulation protocol (GnRH - 7 d PGF2α - 48 h hCG - 24 h AI) on day 50 ± 2 postpartum. Cows affected with PM were randomly divided into two groups, Group E (n = 25) combined the treatments applied in Groups A and D, while Group F (n = 26) repeated the treatment administered in Group E except for uterine lavage. The results indicated that the pregnancy rate within 150 days postpartum and the mean days open in Group E (76.0% and 106.3 ± 4.6 days, respectively) were significantly different from those in Group F (38.5% and 137.9 ± 10.9 days, respectively) (P < 0.05). This study suggests that reproductive efficiency could be improved by using the two-step treatment to regulate uterine involution and an early resumption of ovarian function in cows with RP/PM.

Short-lived sensitization of cardiovascular outcomes of postpartum endotoxemia in preeclamptic rats: Role of medullary solitary tract neuroinflammation.

Preeclampsia (PE) is a pregnancy-related disorder with serious maternal complications. Considering the increased importance of postpartum infection in maternal morbidity and mortality, we investigated whether preeclamptic maternal programming alters cardiovascular consequences of endotoxemia in rats and the role of cardiac and brainstem neuroinflammation in this interaction. Preeclampsia was induced by oral administration of L-NAME (50 mg/kg/day) for 7 days starting from day 14 of conception. Changes in blood pressure, heart rate, and cardiac autonomic function caused by lipopolysaccharide (LPS, 5 mg/kg i.v.) were assessed in mothers at 3 weeks (weaning time) and 9 weeks postnatally. Compared with respective non-PE counterparts, LPS treatment of weaning PE mothers caused significantly greater (i) falls in blood pressure, (ii) rises in heart rate and left ventricular contractility (dP/dtmax), (iii) reductions in time and frequency domain indices of heart rate variability and shifts in cardiac sympathovagal balance (low-frequency/high-frequency ratio, LF/HF) towards parasympathetic dominance, and (iv) attenuation of reflex bradycardic responses measured by the vasoactive method. The intensified LPS effects in weaning PE rats subsided after 9 weeks of delivery. Immunohistochemical studies showed increased protein expression of nuclear factor kappa B (NF-κB) in brainstem neuronal pools of the nucleus of the solitary tract (NTS), but not rostral ventrolateral medulla (RVLM), in endotoxic PE weaning rats compared with non-PE rats. Cardiac NF-κB expression was increased by LPS but this was similarly noted in PE and non-PE rats. Together, preeclamptic maternal programming elicits short-term exacerbation of endotoxic cardiovascular and autonomic derangements due possibly to exaggerated NTS neuroinflammatory insult.

Heart rate variability in dairy cows with postpartum fever during night phase.

Autonomic nervous function evaluated by heart rate variability (HRV) and blood characteristics were compared between Holstein Friesian cows that developed postpartum fever (PF; n = 5) and clinically healthy (CH; n = 6) puerperal cows in this case-control study. A cow was defined as having PF when its rectal temperature rose to ≥39.5°C between 1 and 3 days postpartum. We recorded electrocardiograms during this period using a Holter-type electrocardiograph and applied power spectral analysis of HRV. Comparisons between the groups were analyzed by t test or Mann-Whitney U test, and the relationship between rectal temperature and each parameter was analyzed using multiple regression analysis. Heart rate was higher in PF cows than in CH cows (Mean ± SE, 103.3 ± 2.7 vs. 91.5 ± 1.7 bpm). This result suggested that PF cows had a relatively dominant sympathetic nervous function. Total (44,472 ± 2,301 vs. 55,373 ± 1,997 ms) and low frequency power (24.5 ± 3.8 vs. 39.9 ± 5.3 ms) were lower in PF cows than in CH cows. These findings were possibly caused by a reduction in autonomic nervous function. The total white blood cell count (54.3 ± 5.1 vs. 84.5 ± 6.4 ×102/µL) and the serum magnesium (2.1 ± 0.1 vs. 2.4 ± 0.1 mg/dL) and iron (81.5 ± 8.0 vs. 134.4 ± 9.1 µg/dL) concentrations were lower and the serum amyloid A concentration (277 ± 33 vs. 149 ± 21 µg/mL) was higher in PF cows than in CH cows. These results imply that more inflammation was present in PF cows than in CH cows. Multiple regression analysis showed that both of low frequency power and concentration of serum iron were associated with rectal temperature. We found differences in changes in hematologic results, biochemical findings, and HRV patterns between PF cows and CH cows.

Screening asymptomatic households for Streptococcus pyogenes pharyngeal carriage as a part of in-hospital investigation of puerperal sepsis.

BACKGROUND: Invasive group A streptococcal (iGAS) infection in the peripartum setting is a rare but devastating disease occasionally occurring as a health care-associated infection (HAI). Current guidelines suggest enhanced surveillance and streptococcal isolate storage after a single case of iGAS, as well as a full epidemiological investigation that includes screening health care workers (HCWs) from several sites after 2 cases. Current guidelines do not recommend routine screening of household members of a patient with iGAS. METHODS: We conducted studies of 3 patients with iGAS puerperal sepsis and related epidemiologic and molecular investigations. RESULTS: Identical GAS emm gene types were found in pharyngeal cultures of 3 asymptomatic spouses of patients with iGAS puerperal sepsis. HCWs screened negative for GAS, and emm typing indicated that other iGAS cases from this hospital were sporadic and not related to the puerperal cases. CONCLUSIONS: The concurrent presence of the same emm type in a household member practically excludes the option of an inadvertent HAI or facility outbreak. Hence, we suggest that screening close family members for asymptomatic GAS carriage should be performed early as a part of infection prevention measures, as doing so would have significant utility in saving time and resources related to a full epidemiological inquiry.

Does histologic chorioamnionitis correspond to clinical chorioamnionitis?

OBJECTIVE: To evaluate the degree to which histologic chorioamnionitis, a frequent finding in placentas submitted for histopathologic evaluation, correlates with clinical indicators of infection in the mother. STUDY DESIGN: A retrospective review was performed on 52 cases with a histologic diagnosis of acute chorioamnionitis from 2,051 deliveries at University Hospital, Newark, from January 2003 to July 2003. Third-trimester placentas without histologic chorioamnionitis (n = 52) served as controls. Cases and controls were selected sequentially. Maternal medical records were reviewed for indicators of maternal infection. RESULTS: Histologic chorioamnionitis was significantly associated with the usage of antibiotics (p = 0.0095) and a higher mean white blood cell count (p = 0.018). The presence of 1 or more clinical indicators was significantly associated with the presence of histologic chorioamnionitis (p = 0.019). CONCLUSION: Histologic chorioamnionitis is a reliable indicator of infection whether or not it is clinically apparent.

Defining postpartum uterine disease in cattle.

Uterine function is often compromised in cattle by bacterial contamination of the uterine lumen after parturition, and pathogenic bacteria often persist, causing uterine disease, a key cause of infertility in cattle. However, the definition or characterization of uterine disease frequently lacks precision or varies among research groups. The aim of the present paper was to provide clear clinical definitions of uterine disease that researchers could adopt. Puerperal metritis should be defined as an animal with an abnormally enlarged uterus and a fetid watery red-brown uterine discharge, associated with signs of systemic illness (decreased milk yield, dullness or other signs of toxemia) and fever > 39.5 degrees C, within 21 days after parturition. Animals that are not systemically ill, but have an abnormally enlarged uterus and a purulent uterine discharge detectable in the vagina, within 21 days post partum, may be classified as having clinical metritis. Clinical endometritis is characterised by the presence of purulent (> 50% pus) uterine discharge detectable in the vagina 21 days or more after parturition, or mucuopurulent (approximately 50% pus, 50% mucus) discharge detectable in the vagina after 26 days post partum. In the absence of clinical endometritis, a cow with subclinical endometritis is defined by > 18% neutrophils in uterine cytology samples collected 21-33 days post partum, or > 10% neutrophils at 34-47 days. Pyometra is defined as the accumulation of purulent material within the uterine lumen in the presence of a persistent corpus luteum and a closed cervix. In conclusion, we have suggested definitions for common postpartum uterine diseases, which can be readily adopted by researchers and veterinarians.

Clinical chorioamnionitis and histologic placental inflammation.

OBJECTIVE: To estimate the rate of histologic chorioamnionitis in the presence of diagnosed clinical chorioamnionitis and determine whether clinical markers of maternal and neonatal infection are associated with histologic chorioamnionitis. METHODS: We identified singleton pregnancies from 1996 in which discharge diagnoses included clinical chorioamnionitis and reviewed maternal and neonatal records for clinical evidence of chorioamnionitis and suspected or confirmed neonatal infections. Placentas were examined for acute histologic chorioamnionitis. RESULTS: One hundred thirty-nine pregnancies with the discharge diagnosis of maternal clinical chorioamnionitis were included. Eighty-six (61.9%) had the clinical diagnosis supported by histologic chorioamnionitis. Histologic chorioamnionitis was associated with an earlier gestational age at delivery (35.7+/-6.5 weeks versus 38.6+/-2.9 weeks, P = .002), lower epidural usage (72.1% versus 92.5%, P = .004), less internal monitoring (47.7% versus 75.5%, P = .001), and possible neonatal sepsis (60.5% versus 35.8%, P = .005). For 19 of 71 (26.8%) infants with possible neonatal sepsis, placentas did not show histologic chorioamnionitis. CONCLUSION: Clinical chorioamnionitis and possible neonatal infection were not supported by histologic evidence for infection in 38.1% and 26.8% of cases, respectively, suggesting other noninflammatory causes of signs and symptoms.

Fistula-in-ano after episiotomy.

BACKGROUND: In the past 2 years, we treated three women with fourth-degree lacerations or episiotomy infections presenting with persistent pain and drainage not responding to standard treatment. CASES: These women were referred for evaluation 5 weeks, 3.5 months, and 2 years postpartum. After diagnosing fistula-in-ano, we treated them with fistulotomy and curettage, which resolved the problem. CONCLUSION: When a patient presents with pain or drainage at her episiotomy site, fistula-in-ano should be considered.

An immunohistological study of the uterus of mares following experimental infection by equid herpesvirus 1.

Twelve Welsh Mountain pony mares in late gestation were infected intranasally with EHV-1 (AB4 isolate) at dose rates from 10(3) to 10(7.3) TCID50. This resulted in 3 cases of paresis, at Days 9, 10 and 12 after inoculation, and 5 abortions, at Days 6, 9, 18, 19 and 20. Euthanasia was performed between Days 6 and 21, with collection of uterine specimens for histopathology, virus isolation and immunoperoxidase staining from the pregnant horn, non-pregnant horn and body. EHV-1 replication in endometrial vessels was detected as early as Day 6 and was maximal at Days 9-11, when widespread thromboischaemic damage was present. By Days 15-19 in mares remaining pregnant, EHV-1 antigen expression in the endometrium was sparse, despite residual lesions but little associated thrombosis. Endometrial vascular pathology varied considerably in degree and extent, and no consistent predilection sites for replication within the uterus were apparent.

Endometrial biopsy in Holstein-Friesian dairy cows. II. Correlations between histological criteria.

Endometrial biopsies were taken for histological assessment from 97 cows which calved in a commercial dairy herd between April and August 1984. The main objectives of this study were to analyze the interrelationships among histological criteria and to identify a shortlist of histological parameters to be included in subsequent analysis of associations with results of bacteriological culture, clinical findings and reproductive performance. Epithelial height and segmented cell counts were highly correlated within biopsy, between horns and between days. Subjective assessment of inflammation in the epithelium and/or stratum compactum generally identified biopsies which had any inflammation present. Cows which had inflammation in a biopsy from day 26 were likely to show inflammatory changes at day 40. Quantitative and subjective assessments of gland number, dilation and fibrosis were highly correlated. There was a positive association between the number of cross sections and the diameter of glands, and both of these criteria were negatively correlated with fibrosis and inflammatory changes. There may be different functional significance of the same histological finding at a different number of days postpartum.

[Clinical pathological analysis of peripartum maternal mortality (author's transl)]. / Klinisch-pathologische Analyse von peripartalen mütterlichen Todesfällen.

All maternal deaths which occurred in relation to labour at the Department of Obstetrics and Gynaecology, University Hospital, Graz, between 1963 and 1978 were reviewed and analysed. 24 mothers died over this 15-year period. The maternal death rate was 0.34 per thousand. In all cases a post mortem examination was performed. The youngest woman was 19, the oldest 42 years old. Most of the decreased mothers were primiparae. The main cause of maternal death was post-partum haemorrhage, followed by infection of the uterus especially after Caesarean section. Three mothers died of liver distrophy. The remaining causes were eclampsia (2 cases); pulmonary artery embolism after Caesarean section (2 cases); irreversible shock from amniotic fluid and air embolism (one case each); uraemia due to glomerulo- or pyelonephritis (one case each); malignant melanoma (one case). It is demonstrated that special risk factors are advanced maternal age, low social status and lack of antenatal care.

Bacterial cure and somatic cell count response of dairy cows with a positive California Mastitis Test at calving to therapy with cephapirin sodium.

Cows without signs of clinical mastitis were evaluated by the California Mastitis Test at calving (Day 0). Milk samples from 117 of 184 quarters (64 cows) were positive by this test for mastitis and were submitted for bacterial culture and determination of somatic cell counts. Cows with infected quarters were randomly allocated to treatment with cephapirin sodium by intramammary infusion or to be untreated as controls. Two and 4 weeks following calving, milk was again sampled from the infected quarters and tested. By the 4-week evaluation, the quarters treated with cephapirin sodium had significantly (P < or = .05) fewer positive bacterial cultures and somatic cell counts were significantly (P < or =.05) reduced compared with untreated control quarters.

Hyperacute Escherichia coli mastitis of cattle in the immediate post-partum period.

The pathology of hyperacute coliform mastitis was studied in 5 post-parturient cows. In all infected quarters infiltration of neutrophils was negligible. In all except one case there was severe damage to the ductular and secretory system, involving most areas of the gland. Bacteria were dense in infected alveoli, and there was evidence of substantial phagocytosis of bacteria by the secretory epithelium. The exception showed a large lesion in the middle of the gland from which the spread was ductular; other infections were consistent with spread via the teat canal. The organisms were largely confined to the ductular/secretary lumen and there was little invasion of the parenchyma. The severity of the disease was considered due to the absence of the inflammatory response seen in mid lactation.

Postnatal care.

PIP: Early contact between mother and baby and early breastfeeding are essential elements of postnatal care. They promote bonding and a better breastfeeding performance, and cannot be overemphasized. Thereafter, in hospital practice, the next essential is a separation of abnormal from normal cases so that those in greatest need may receive the care they require. Commonly occurring postnatal problems should be managed according to standard protocols, so that effective management can be instituted by midwives or junior medical staff as necessary. Special attention must be paid to those with severe puerperal sepsis. There should be a readiness to recognize and treat such rare but curable conditions as acute tubular necrosis. Proper advice to the mother and the recording of significant events of the pregnancy in a document kept by the mother is the doctor's final responsibility.^ieng

Treatment of puerperal infection.

Ninety-two patients with puerperal infection admitted to Siriraj Hospital from April 1, 1980 to March 1, 1983 were studied. The treatment in this study was both medical and surgical, blood transfusion was given in some cases with low hematocrit level. The medical treatment alone was based on the causative organisms which were detected by cervical & intrauterine swab, smear & gram stain and cultures. PGS & Kanamycin were the most frequently used antibiotics which were intended to treat both gram-positive and gram-negative bacteria and adjunctive chloramphenicol for anaerobes. The surgical procedures which were performed in combination with medical treatment included total abdominal hysterectomy, uterine curettage, appendectomy and drainage of subdiaphragmatic abscess. The result of the treatment was satisfactory, 96.7 per cent improved after therapy with slight morbidity in some patients.

Diagnosis of postpartum infections: clinical criteria are better than laboratory parameter.

BACKGROUND: Physiologic increases in the leukocyte count and segmental neutrophil percentage during pregnancy alter the ability of these indices to predict infection. METHODS: Women with cesarean deliveries were assessed by leukocyte counts and segmented neutrophil percentage as well as clinical signs to detect postoperative infection. RESULTS: In 157 consecutive patients undergoing cesarean deliveries there was a significant physiologic increase between the preoperative and postoperative leukocyte count (12.4 +/- 3.9/uL vs. 14.8 +/- 3.4/uL, P < 0.001). The leukocyte count after delivery did not discriminate whether or not the patient would develop a postpartum infection or have significant hyperpyrexia. CONCLUSION: Due to physiologic changes associated with pregnancy, the leukocyte count and segmented neutrophil percentage do not predict infection. Therefore clinical findings are most important in diagnosing postpartum infections.

Puerperal retroperitoneal abscess caused by Clostridium difficile: case report and review of the literature.

BACKGROUND: Retroperitoneal infection can be lethal. Optimal management is still elusive to describe because of the small number of case reports. We presented here a case of retroperitoneal abscess caused by Clostridium difficile arising in the puerperal period. METHODS: Case report and review of recent English-language literature. RESULTS: The patient presented with surgical incision dehiscence. A gas-forming fluid collection was discovered in the pelvic retroperitoneal fascia by computed tomography, but the patient did not show marked symptoms of sepsis. Emergency laparotomy drainage and debridement were performed. Clostridium difficile was isolated, and she was treated with a three-week course of vancomycin. The patient recovered without major morbidity. Recent case reports describe variation in the course of the disease and management options for puerperal retroperitoneal infection. CONCLUSION: Puerperal retroperitoneal abscess caused by C. difficile can present with minimal symptoms. Prompt recognition, early surgical intervention, and optimal use of antibiotics can reduce morbidity and prevent death.