CAMP Test Research Articles

Expanding capabilities to evaluate readiness for return to duty after mTBI: The CAMP study protocol.

The purpose of this observational longitudinal study is to: 1- establish typical performance parameters for ADSM on the CAMP test battery 2- determine the element of the CAMP battery that demonstrate the greatest differences from standard performance and serve as predictors for successful return to duty and 3- develop clinician-facing feedback algorithms and displays and 4-develop materials for clinical dissemination. This ongoing multi-site study is currently funded through the CDMRP and has been approved by the Naval Medical Center Portsmouth IRB. ADSM undergoing post-concussion rehabilitation at the Intrepid Spirit Centers will be tested within one week of their initial Physical Therapy evaluation and after completing Physical Therapy. Control participants will include males, females, and ADSM from the Special Operations community. Participants will complete an intake form that includes questions about demographics, military service, deployment and concussion history, and profile and duty status. Other measures include those that explore concussion symptoms, sleep quality, post-traumatic stress, and perceptions of resilience. The CAMP includes three separate 10-15 minute tasks. Movement is recorded by wearable inertial sensors and heart rate variability is recorded with a POLAR10 monitor. The "Run-Roll" task requires rapid position changes, combat rolls and quick running forwards and backwards while carrying a simulated weapon. Visual stability before and after the task is also performed. The "Dual-Task Agility" task includes rapid running with and without a weighted vest and a working memory task. The "Patrol Exertion" task requires repeated stepping onto an exercise step while watching a virtual patrol video. Additional tasks include monitoring direction of travel, observing for signs of enemy presence, and reacting to multiple auditory signals embedded in the video. Measures that evaluate relevant skills are vital to support safe return to duty for ADSM who may be exposed to imminent danger as part of training or mission demands. The CAMP is designed to be an ecologically valid and clinically feasible assessment that may be more sensitive to capturing subtle impairments that impact duty performance as test skills are integrated into dual and multi-tasks that reflect occupational demands. Assessment results may serve as a more robust indicator of readiness for full return to duty after concussion.

Screening of Group B Streptococcal and Candida Infections in Antenatal Women Visiting a Tertiary Care Hospital and the Neonatal Outcome

Reproductive tract infections in antenatal women are frequently occurring public health concern affecting the quality of life in the infected women and further affects the neonatal outcome negatively. Vaginal infections are predominantly caused due to the disturbance in the normal vaginal microflora. Group B Streptococcus (GBS) is a major peritoneal pathogen leading to morbidity and mortality in both mother and neonates leading to complications like preterm labour, chorioamnionitis and Preterm rupture of membrane (PROM). Further, Vulvovaginal candidiasis (VVC) is a frequently encountered gynaecological disease causing morbidity in 3/4th of women at least once in their life span. It leads to infertility in non-pregnant women. It results in adverse pregnancy outcomes such as chorioamnionitis and congenital abnormalities in the neonates. The aim of this study is to screen for potential bacterial and yeast infections with focus on GBS and Candida infections and its neonatal outcome. Candida species and GBS were identified by routine culture-based tests. HiChrome agar was used for speciation of Candida species. CAMP test was performed for GBS; further identification was done using Streptococcal grouping kit. GBS was found in 15% of the antenatal women and Candida Species was found to be most common with an infection rate of 50%. Lower segment Cesarean section (LSCS) rate in GBS positive women was 60% and in Candida Non albicans positive women was found to be 65.38%. 33% of the neonates born to GBS positive mothers had respiratory distress. Infection in antenatal women negatively influenced the mode of delivery and the mean body weight of the neonates born to infected mothers were comparatively less.

Serotype distribution, antimicrobial resistance, and molecular characterization of group B Streptococcus isolates from Chinese pregnant woman

Objective This study was aimed to investigate the serotypes, antibiotic susceptibilities, and multi-locus sequence type (MLST) profiles of group B Streptococcus (GBS) in the Beijing area. Methods Lower vaginal and rectal swabs were obtained from pregnant women of 35–37 gestational weeks (GWs) who attended the Beijing Obstetrics and Gynecology Hospital. All GBS isolates were identified with Gram staining, catalase reaction assays, and CAMP tests, followed by antibiotic susceptibility testing, serotype identification, multilocus sequence typing and erythromycin resistance gene analysis (ermB and mefE). Results From July 2020 to June 2022, 311 (5.17%) of 6012 pregnant women that were screened for GBS colonization were detected positive. Of the eight serotypes identified (III, Ia, Ib, IV, II, VIII, V, and NT), serotypes III (43.09%), Ia (34.08%) and Ib (17.04%) were the predominant species. In the antimicrobial susceptibility experiments, the resistant rates measured for erythromycin, clindamycin, levofloxacin, and tetracycline were 76.21%, 63.99%, 50.80%, and 81.03%, respectively, and 7.6% of GBS isolates showed inducible clindamycin in resistance (D-test phenotype). Meanwhile, the multilocus sequence typing analysis showed that sequence type 19 (ST19) (30.34%) and ST10 (18.62%) were the dominant sequence types. Among the 237 erythromycin-resistant isolates, 176 harbored ermB (128, 54.00%) or mefE (48, 20.30%) gene alone. Conclusion The infection rates, serotypes or MSLT distribution, and antimicrobial resistance of GBS in Beijing area were investigated, which may be applied in analyses of the epidemiological characteristics of GBS. This contributes to the basic knowledge required for successful GBS vaccine development suited for disease prevention and treatment in China, as well as the implementation of effective clinical antimicrobials.

<i>Streptococcus agalactiae</i> in asymptomatic bacteriuria in pregnant women

BACKGROUND: Asymptomatic bacteriuria caused by group B gram-positive streptococcus, Streptococcus agalactiae, in pregnant women is a leading cause of sepsis in newborns and young children, transmitted from their mothers. In the present study, the frequency of S. agalactiae inoculation from the urine of pregnant women with asymptomatic bacteriuria was studied with the determination of the sensitivity of isolated strains to antibiotics.
 AIMS: This study aimed to investigate the inoculation of group B streptococcus in asymptomatic bacteriuria of pregnant women and to determine the sensitivity of the isolated cultures of the pathogen to the main antibiotics.
 MATERIALS AND METHODS: The analysis involved bacteriological studies of 2,559 urine samples collected from pregnant women. Urine was inoculated on UriSelect 4 Medium chromogenic medium with subsequent identification of S. agalactiae using a strepto-latex test or a CAMP test with a Staphylococcus aureus culture. The sensitivity of microorganisms to antibacterial drugs was determined using the disk diffusion method. Data analysis was conducted using the Statistica 12.0 software package.
 RESULTS: Out of 2,559 urine samples studied, asymptomatic bacteriuria was detected in 317 samples (12.4%), and 330 strains of microorganisms were isolated. S. agalactiae was isolated by culture from 108 pregnant women (4.2% of the total subjects), with 50 cases (16%) showing asymptomatic bacteriuria caused by S. agalactiae. The high significance of Escherichia coli in asymptomatic bacteriuria in pregnant women was shown, with a lesser extent of involvement observed with Enterococcus spp. Furthermore, associations of S. agalactiae with other microorganisms were identified. The high sensitivity of S. agalactiae strains to clindamycin, to a lesser extent to levofloxacin, chloramphenicol, and, especially, to erythromycin, has been demonstrated.
 CONCLUSIONS: The study revealed a high culturability of group B streptococcus in asymptomatic bacteriuria in pregnant women. Low sensitivity of isolated streptococci to erythromycin, moderate sensitivity to levofloxacin and chloramphenicol, and high sensitivity to clindamycin were also revealed.

Group B Streptococci recto-vaginal colonization, antimicrobial susceptibility pattern, and associated factors among pregnant women at selected health facilities of Wolaita Sodo Town, Southern Ethiopia.

Streptococcus agalactiae or Group B Streptococcal colonization of the gastrointestinal and genital tracts of pregnant women usually remains asymptomatic, even though it is the critical determinant of infection in neonates and young infants. It causes early and late onset of invasive Group B Streptococcus (GBS) disease manifesting as septicemia, meningitis, and pneumonia. Now it is recognized as an important cause of maternal and neonatal morbidity and mortality in many parts of the world including Ethiopia, where the magnitude of the problem has been little studied. The aim of this study was to assess the prevalence of GBS colonization and to identify associated risk factors and antimicrobial susceptibility patterns among pregnant women at selected health facilities of Wolaita Sodo Town, Southern Ethiopia. A health-facility-based cross-sectional study design was conducted at WSUCSH & Wolaita Sodo Health Center from June to August, 2022. A total of 279 pregnant women who were in ANC follow-up at 35-37 weeks of gestation were included. For GBS isolation, recto-vaginal swabs were inoculated in 1 mL Todd-Hewitt broth medium supplemented with 10 μg/mL colistin and 15 μg/mL nalidixic acid, followed by identification of isolates based on colonial morphology, gram stains, catalase reaction, and CAMP tests. Antimicrobial susceptibility testing was performed using a modified Kirby-Bauer disc diffusion method. All collected data were entered in Epi info 4.6.0.2, then transferred and tabulated using SPSS version 20. Logistic regression analysis was used to see the association between variables. Finally, a p-value <0.05 was considered statistically significant. In the present study, 279 pregnant mothers, aged between 15 to 38 years with a mean of 26.5 ± 4.5 years, were included. Of all participants, the highest proportion (120) (43.01%) were housewives. The overall carriage rate of GBS was 67 (24.0%). GBS colonization showed a statistically significant association with college and above levels of maternal education [AOR = 6.610, 95% CI (1.724-25.349), p = 0.01]. High susceptibility of GBS isolate was seen with Penicillin G & Chloramphenicol (92.5%), Ampicillin, Ceftriaxone (89.6%), Vancomycin (74.62%), and Erythromycin (77%). Relatively, GBS showed high resistance to Tetracycline (88%). In this study, the overall prevalence of GBS colonization was 24.0%. College and above educational level was statistically significant with GBS colonization. This study aimed to draw attention to the management of Group B Streptococci in pregnant women by making GBS culture one of the routine diagnoses during ANC follow-up and to prevent infection with early detection.

Antimicrobial susceptibility of Rhodococcus equi strains isolated from foals in Chile

Rhodococcus equi is responsible for foal pneumonia worldwide, with a significant economic impact on the production and breeding of horses. In Chile, the first case was reported in 2000, and since then, its incidence has been increasing. Distinctive characteristics of R. equi as an intracellular pathogen in macrophages, emergence of virulence plasmids encoding surface lipoprotein antigens, and appearance of antibiotic resistance against macrolides and rifampicin have significantly complicated the treatment of R. equi pneumonia in foals. Therefore, in vitro susceptibility studies of first-line and newer antibiotics against R. equi are the first step to establishing effective treatments and optimizing new therapeutic options. The aim of the present study is to determine the susceptibility profile of fourteen strains of R. equi isolated from foals in Chile to several antibiotics of the macrolide group including azithromycin, amikacin, tildipirosin and gamithromycin as well as others such as rifampicin, doxycycline and ceftiofur. Identification of R. equi in collected isolates from foals in Chile has been performed by CAMP test and PCR based on detecting of the gene encoding the 16 S rRNA. The presence of genes encoding virulence plasmids was also determined using PCR. Results obtained have demonstrated presence of virulent R. equi strains in Chile. In vitro susceptibility pattern to different antibiotics has shown better results for doxycycline and rifampicin similar to previous studies performed. Current macrolides have been evaluated in order to consider alternative treatment options in a context of emerging resistance to classic macrolides and rifampicin, obtaining better results with gamithromycin (MIC range of 0.125 to 128 mg/ml) than with tildipirosin (MIC range of 16 to 128 mg/ml). An adequate diagnosis of bacterial susceptibility based on antibiograms is necessary to treat the Rhodococcus equi infection in foals.

Analysis of molecular characteristics of CAMP-negative Streptococcus agalactiae strains.

Streptococcus agalactiae can produce CAMP factor, which can promote the β-hemolysin activity of Staphylococcus aureus, forming an arrow-shaped hemolysis enhancement zone at the intersection of the two bacterial species on a blood agar plate. This characteristic feature of Streptococcus agalactiae has led to the widespread use of the CAMP test as an identification method. Vaginal/rectal swabs, collected from women at 35-37 weeks of pregnancy, were first inoculated into a selective enrichment broth media, then subcultured onto GBS chromogenic agar and 5% sheep blood agar sequentially. The VITEK-2 automatic identification system and MALDI-TOF MS were initially employed for identification, followed by the CAMP test. CAMP-negative strains underwent 16S rDNA and cfb gene sequence analysis, as well as bacterial multilocus sequence typing. A total of 190 strains were isolated, with 15 identified as CAMP-negative. Further 16S rDNA gene sequence analysis confirmed that all 15 strains were Streptococcus agalactiae. The MLST typing assay revealed that these 15 strains were of the ST862 type. The cfb gene was amplified and electrophoresed, but no specific fragments were found, indicating that these strains lack the CAMP factor due to cfb gene deletion. Antibiotic susceptibility tests demonstrated no resistance to penicillin, ampicillin, vancomycin and linezolid among the GBS strains. However, there are significant differences in resistance rates to tetracycline. This study found that 7.9% of GBS strains isolated from the vagina/rectum of pregnant women were CAMP-negative, suggesting that the CAMP test method or primers targeting the cfb gene should not be used as the sole presumptive test for GBS identification.

Microbiological monitoring of antibiotic resistance of strains of Streptococcus agalactiae among pregnant women

Group B Streptococcus (GBS) is the causative agent in 2–7% of all urinary tract infections (UTI), including asymptomatic bacteriuria (AB), cystitis, and pyelonephritis. We used the bacteriological quantitative method of sowing urine samples of pregnant women on Columbia agar with 5% lamb blood), Strepto B chromogenic agar and Todd Hewitt broth, identification of GBS strains with determination of sensitivity to bacitracin, hippurate hydrolysis test and CAMP test. Susceptibility to antimicrobial drugs was determined by the disk-diffusion method according to recommendations of the European Committee on Antimicrobial Susceptibility Testing. For the period of 2021, out of 3,356 urine samples of pregnant women, there were 149 samples of the investigated biomaterial with a positive result for GBS (resultancy – 4.4%). It was established that among the studied contingent, 41 strains of GBS were isolated in the first trimester of pregnancy (27.5%), in the second trimester – 59 strains (39.5%), in the third trimester – 49 strains (33.0%). It is established that out of the 149 strains of GBS, 38 strains (25.0%) were resistant to norfloxacin, 45 strains (30.0%) to erythromycin, 41 strains (28.0%) to clindamycin,125 strains (84.0%) to tetracycline, 20 strains (13.0%) to levofloxacin, 3 strains (2.0%) to nitrofurantoin. All strains of Streptococcus agalactiae were found to be sensitive to benzylpenicillin. The number of resistant strains of SGB identified from the urine of pregnant women was 47 strains. All resistant strains were resistant to at least three groups of antibacterial drugs, except beta-lactams. The results of the scientific research will allow us to obtain dynamic data on the antibiotic resistance of GBS strains in order to study the rate of development of antibiotic sensitivity of this microorganism. In the future, the research data can be used by scientists when reviewing protocols for the treatment of urinary tract infections in pregnant women.

Listeria monocytogenes isolated from raw milk: phenotypic and molecularcharacterization, pathogenicity testing, and multidrug resistance profiling

Foodborne infections are a worldwide public health emergency that actually impact a variety of disorders. In the current research, a total of 300 samples comprising raw milk (100),milk products (100), and chicken (100) were screened for detection of pathogenic Listeria species by using the USDA method. On conventional biochemical characterization, three isolates were identified as Listeria monocytogenes (from raw milk samples)indicating an overall prevalence of Listeria monocytogenes to the tune of 1%. All other samples comprising milk products and chicken samples (100) showed negativity for the presence of any of the Listeria species. Further, all three isolates were subjected to polymerase chain reaction (PCR) targeting genus-specific (prsA and iap) and species-specific gene (isp) in which all three were turn out positive for both the genes, endorsing their identification as Listeria monocytogenes. All three confirmed Listeria monocytogenes isolates were phenotypically assessed for in-vitro pathogenicity tests like hemolysis on 7% sheep blood agar, CAMP test, and PI-PLC assay. The results revealed their highly pathogenic nature. Subsequently, all these isolates were also assessed for their virulent nature by PCR, targeting the array ofmarkers including virulence-associated genes viz. hlyA, actA, plcA. The results endorsed pathogenic nature of all isolates showing amplification of all targeted virulence genes. The antibiotic resistance profiling revealed occurrence of multidrug-resistant pathogenic L. monocytogenes in foods of animal origin with maximum multiple antibiotic resistance (MAR) index of 0.6and a minimum MAR index of 0.48 in MDR, which is a matter of concern from public health point of view.

Vertical Transmission, Risk Factors, and Antimicrobial Resistance Patterns of Group B Streptococcus among Mothers and Their Neonates in Southern Ethiopia.

Background Group B Streptococcus (GBS) contributes to maternal and neonatal morbidity and mortality by increasing intrauterine infection or vertical transmission at the time of birth. Despite many efforts to reduce the potential risk of vertical transmission, GBS remains the main cause of serious disease (neonatal sepsis, meningitis, and/or pneumonia) in vulnerable newborns during the first week of life. This study aimed to assess vertical transmission, risk factors, and antimicrobial resistance patterns of GBS among pregnant women and their neonates. Methods A facility-based cross-sectional study was conducted among mothers and their neonates from February to May 2021. A total of 201 pregnant women with their neonates participated in this study. A well-designed questionnaire was used to collect sociodemographic and clinical data. A vaginal swab from mother before delivery and neonatal nasal and ear canal swab samples were taken as soon as after delivery within 30 minutes. Vaginal swabs, neonatal ear canal, and nasal swabs were placed into Todd–Hewitt broth and incubated at 37°C for 18–24 hours at 35–37°C in 5% CO2 conditions and then subcultured on 5% sheep blood agar for 18–48 hours. Presumptive identification of GBS was made by morphology, Gram stain, catalase, and hemolytic activity on sheep blood agar plates. CAMP and bacitracin susceptibility tests were used as confirmatory tests for GBS. Data were analyzed using SPSS version 21. P value ≤0.05 was considered statistically significant. Results Vertical transmission rates of GBS (mother to neonates) were 11.9%. The prevalence of GBS among pregnant women and newborns was 24/201 (11.9%) (95% CI = 7.5–16.9) and 11/201 (5.5%) (95% CI = 2.5–9.0), respectively. The history of prolonged rupture of membranes (AOR = 3.5, CI = 2.2–18.8) and urinary tract infection (AOR = 2.9, CI = 1.7–16.3) were associated factors for maternal GBS colonization. Gestational age of <37 weeks (p=0.008), low birth weight of <2.5 kg (p=0.001), and maternal history of vaginal discharge (p=0.048) were associated factors for neonatal GBS colonization. Low antibiotic resistance was observed for erythromycin 8.6%, clindamycin 5.7%, and chloramphenicol 2.9%. Conclusion In this study, high vertical transmission (mother to neonates) rate was observed. The prevalence of vaginal GBS colonization of women at delivery was 11.9% and significantly associated with the history of prolonged rupture of membranes and urinary tract infections. Gestational age of <37 weeks, low birth weight of <2.5 kg, and maternal history of vaginal discharge were associated with neonatal GBS colonization. Hence, there is a need for antenatal culture-based GBS screening, risk factor-based interventions, and regular follow-up of drug resistance patterns for proper treatment and management of GBS.

Genomic analysis of virulence factors and antimicrobial resistance of group B Streptococcus isolated from pregnant women in northeastern Mexico.

Group B Streptococcus (GBS) causes infections in women during pregnancy and puerperium and invasive infections in newborns. The genes lmb, cylE, scpB, and hvgA are involved with increased virulence of GBS, and hypervirulent clones have been identified in different regions. In addition, increasing resistance of GBS to macrolides and lincosamides has been reported, so knowing the patterns of antibiotic resistance may be necessary to prevent and treat GBS infections. This study aimed to identify virulence genes and antibiotic resistance associated with GBS colonization in pregnant women from northeastern Mexico. Pregnant women with 35-37 weeks of gestation underwent recto-vaginal swabbing. One swab was inoculated into Todd-Hewitt broth supplemented with gentamicin and nalidixic acid, a second swab was inoculated into LIM enrichment broth, and a third swab was submerged into a transport medium. All samples were subcultured onto blood agar. After overnight incubation, suggestive colonies with or without hemolysis were analyzed to confirm GBS identification by Gram staining, catalase test, hippurate hydrolysis, CAMP test, and incubation in a chromogenic medium. We used latex agglutination to confirm and serotype GBS isolates. Antibiotic resistance patterns were assessed by Vitek 2 and disk diffusion. Periumbilical, rectal and nasopharyngeal swabs were collected from some newborns of colonized mothers. All colonized women and their newborns were followed up for three months to assess the development of disease attributable to GBS. Draft genomes of all GBS isolates were obtained by whole-genome sequencing. In addition, bioinformatic analysis to identify genes encoding capsular polysaccharides and virulence factors was performed using BRIG, while antibiotic resistance genes were identified using the CARD database. We found 17 GBS colonized women out of 1154 pregnant women (1.47%). None of the six newborns sampled were colonized, and no complications due to GBS were detected in pregnant women or newborns. Three isolates were serotype I, 5 serotype II, 3 serotype III, 4 serotype IV, and 2 serotype V. Ten distinct virulence gene profiles were identified, being scpB, lmb, fbsA, acp, PI-1, PI-2a, cylE the most common (3/14, 21%). The virulence genes identified were scpB, lmb, cylE, PI-1, fbsA, PI-2a, acp, fbsB, PI-2b, and hvgA. We identified resistance to tetracycline in 65% (11/17) of the isolates, intermediate susceptibility to clindamycin in 41% (7/17), and reduced susceptibility to ampicillin in 23.5% (4/17). The tetM gene associated to tetracyclines resistance was found in 79% (11/14) and the mel and mefA genes associated to macrolides resistance in 7% (1/14). The low prevalence of colonization and the non-occurrence of mother-to-child transmission suggest that the intentional search for GBS colonization in this population is not justified. Our results also suggest that risk factors should guide the use of intrapartum antibiotic prophylaxis. The detection of strains with genes coding virulence factors means that clones with pathogenic potential circulates in this region. On the other hand, the identification of decreased susceptibility to antibiotics from different antimicrobial categories shows the importance of adequately knowing the resistance patterns to prevent and to treat GBS perinatal infection.

Experimental Study on Streptococcus agalactiae Genotype and Erythromycin Resistance in Neonatal Sepsis.

This study aimed to evaluate Streptococcus agalactiae genotype and erythromycin resistance in neonatal sepsis. After obtaining the mothers' informed consent, trained nurses sampled 430 neonatal specimens of sepsis from the ear canal, oral cavity and umbilical cord immediately after childbirth and implemented a cross-sectional study. By Gram staining, morphology, hemolysis mode, catalase and CAMP tests, the isolate was identified as S. agalactiae. All 455 isolates were tested for antimicrobial susceptibility by the disc diffusion method. Multilocus sequence typing was used to serotype S. agalactiae involving sequencing of 7 housekeeping genes. The erythromycin resistance genes-erm (B), erm (A) and mef (A) were detected by PCR. Results showed that there were 286 cases (66.51%) of neonates delivered naturally, and 144 cases (33.49%) of neonates delivered by cesarean section. A total of 455 strains were tested, including 253 strains (55.60%) of Gram-positive bacteria with 100 strains (21.98%) of S. agalactiae and 52 strains (11.43%) of Staphylococcus epidermidis, 178 strains (39.12%) of Gram-negative bacteria with 45 strains of Klebsiella pneumoniae (9.89%), 36 strains of Escherichia coli (7.91%), 36 strains of Pseudomonas aeruginosa (7.91%), and 323 strains of Citrobacter freundii (7.03%). S. agalactiae had the highest resistance of 87 (87.00%) to erythromycin, followed by resistance to azithromycin 83 (83.00%) and clindamycin 78 (78.00%). In children with neonatal sepsis, S. agalactiae serotypes were mainly Ia, Ib, and III, accounting for 29.00%, 35.00%, and 19.00% respectively. The main genotypes were ST651, ST103 and ST176, which account for 19.00%, 17.00% and 15.00% respectively. The ST19 type 13.00%, ST27 type 8.00%, ST17 Type 11.00%, ST10 type 12.00%, ST485 type 5.00%. The ST103 and ST485 isolates were classified as serotype Ia, the ST10 and ST176 isolates were classified as serotype Ib, and ST17 and ST19 isolates were classified as serotype III. Among the strains of S. agalactiae, 40.23% (35/87) carry erm (A) gene, 35.63% (31/87) carry erm (B) gene, and 24.14% (21/87) carry mef (A) gene. erm (A) gene was the most common gene in ST19 strain (7/11, 63.64%), and erm (B) gene was the most common gene in ST176 and ST651 strains (6/12, 50.00%; 8/18, 44.44%), while mef (A) gene was the most common gene in ST17 strain (5/11, 45.45%). In general, S. agalactiae genotypes in neonatal sepsis were mainly ST651, ST103 and ST176, and the main serotypes are Ia, Ib, and III. There was good consistency between ST and serotype, and a significant difference was shown in erythromycin resistance and ST distribution, which highlights the value of new epidemiological trend detection by monitoring multiple characteristics and provides inspiration for the development of multivalent S. agalactiae vaccines.

Incidence of Vaginal Colonization of Group B Streptococci among Pregnant Women Attending a Tertiary Care Hospital – Chennai, India

Streptococcus agalactiae or Group B Streptococcus (GBS) is an important pathogen, which can cause serious illness and sometimes death especially in newborns, pregnant women and in people with compromised immune system. The major risk factor for neonatal disease is vaginal colonization of GBS during pregnancy and delivery. 1) To study the incidence of vaginal colonization of Group B Streptococcus (GBS) among pregnant women. 2) To compare the efficacy of sheep blood agar vs selective medium like HiCHROME Strep B Selective agar (HIMEDIA) in the isolation of GBS. Vaginal swabs were collected from 150 antenatal women at 35-37 weeks of gestation or at any gestational age if additional risk factors were present. The swabs were cultured on non-selective medium like Blood agar and a chromogenic medium (HiCHROME Strep B Agar, HiMedia). GBS was identified by standard microbiological techniques. Bluish, purple-colored colonies were confirmed as GBS by CAMP test and latex agglutination test. Only 3 out of 150 women (2%) of the screened antenatal women were colonized with GBS. Incidence was quite low, compared to the literature from the West. All the 3 babies born to the three women colonized with GBS were healthy with no indication of invasive GBS disease. The chromogenic medium supported the growth of Group B Streptococcus and Enterococcus spp. only and was a useful selective medium for isolation of GBS.

Mucoid bovine mastitis isolates of Streptococcus uberis - difficulties in identification

Highly mucoid Streptococcus uberis were isolated on blood agar from five milk samples originating from two dairy cow farms. All the isolates were CAMP test and esculin hydrolysis negative. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) following prior extraction with 70% formic acid was used for identification to the species level. Given that it is plausible that such strains could be isolated in the future from dairy cows by laboratories in Serbia and the Balkans, we consider this case report to be a useful instruction for veterinary bacteriology laboratories.

Prevalence and Risk Factors of Group B Streptococcus Colonization in Pregnant Women: A Pilot Study in Palestine.

Background Maternal Streptococcus agalactiae (Group B Streptococcus (GBS)) colonization is an important cause of complications in mothers and neonates during gestation and after delivery. The data regarding GBS colonization among pregnant women in Palestine is scarce. The aim of this study is to determine the prevalence of GBS colonization, its associated risk factors, and the antibiotic sensitivity patterns in Nablus, West Bank, Palestine. Methods A cross-sectional, single center study conducted at Rafidia Governmental Hospital in Nablus, West Bank, Palestine. Samples were collected between November 2019 and January 2020. Vaginal swabs from 200 pregnant women (≥35 weeks of gestation) attending the labor and delivery department were plated directly on CHROMagarTM StrepB (CHROM agar, France) and placed in an incubator at 35–37°C. After 24 and 48 hours, the plates were checked for growth and classified into three categories: growth of GBS with mauve colonies on chromogenic media, no growth, or other growth. The identification of the mauve colonies was confirmed by the CAMP test. Identified GBS isolates were tested for susceptibility to vancomycin, ampicillin, clindamycin, cefotaxime, erythromycin, and levofloxacin using the disc diffusion method. Clinical and demographic information were collected using a questionnaire. Result The overall prevalence of GBS colonization was 12%. The median age of the study population was 27 years. GBS colonization was significantly associated with age (p=0.013), history of previous preterm delivery (p=0.013), and parity (p=0.015). No association was noted with smoking, previous abortion, previous history of fetal demise, vaginitis, or urinary tract infection. Resistance to ampicillin, vancomycin, cefotaxime, erythromycin, clindamycin, and levofloxacin was found to be 91.7%, 54.2%, 45.8%, 29.2%, 25%, and 8.3%, respectively. Conclusion The prevalence of vaginal GBS in this study was 12% from Nablus, West Bank. Further research is needed to determine the GBS serotypes common in West Bank and the burden they cause on the health system. Moreover, this study also highlights the need to establish a screening program suited to a developing country with low control on the antibiotic's prescription protocols.

Investigation of antimicrobial susceptibility and virulence factor genes in Trueperella pyogenes isolated from clinical mastitis cases of dairy cows.

Trueperella pyogenes is an opportunistic pathogen causing important diseases including mastitis and metritis in domestic animals such as dairy cows leading to prominent economic losses in food production industry. The aim of this study was to investigate bacterial species, antimicrobial susceptibility, and presence of virulence factor genes and genotyping of T. pyogenes isolates associated with summer mastitis cases from 22 different farms around Tehran, Iran. Fifty‐five percent of dairy cows with clinical mastitis symptoms was infected by T. pyogenesis indicated that this pathogen is the most important contributor to clinical mastitis in dairy cows in the present study. A significant correlation was illustrated between presence of virulence factor genes of isolated pathogen, biochemical patterns, and the utter infected types. Multidrug resistance susceptibility observed between isolates indicated the important need for prudent use of antimicrobials in treatment of mastitis caused by T. pyogenes and increased concerning of consumer health associated with recent problems of antimicrobial resistance. The categorization of isolates was implemented into seven different clonal related types by COX‐PCR at 80% of similarity cutoff with significance relationship to clonal types, CAMP test result and sampling time and biochemical profile. Regarding to the results obtained at the present study, T. pyogenes can be considered as an important typically cause of purulent and acute form of clinical bovine mastitis and loss of dairy productivity. Further studies with more sample size and high‐throughput omic methods in various sampling time and areas are suggested for study of this pathogen precisely.

Study of listeriosis in spontaneous abortions during preganacy at tertiary care centre

Listeriosis is an emerging zoonotic disease. Listeria monocytogenes is an uncommon cause of illness in general population, however, in some high risk groups including neonates, pregnant women, elderly persons, immunosuppressed transplant recipients and others with impaired cell mediated immunity, it is important cause of life threatening bacteremia meningoencephalitis. Listeriosis is 18 times more common in pregnancy (12/100,000) than in the non-pregnant population (0.7/100,000) and 16-27% of all infections with Listeria occur in pregnant women. Isolation of bacteria by conventional methods of microscopy and culture on selective agar PALCUM agar after enrichment of samples by using UVM1, UVM2 .Speciation of listeria done by carbohydrate fermentation test followed by study of pathogenicity by hemolysis on sheep blood agar and CAMP test followed by antimicrobial susceptibility test done.Out of total 131 patients having history of Spontaneous abortion were screened for Listeria, in that 10(7.6%) were carrying Listeria out of which 3(2.29%) of them found to be positive for L. monocytogenes and 07(5.34%) were other Listeria spp. The prevalence of L. monocytogenes in present study was 03(2.29%) from spontaneous abortion cases.Based on the results of present study it is concluded that Listeria monocytogenes is responsible for spontaneous abortions in spontaneous abortion during pregnancy in humans.

Role of PCR in Rapid Detection Group B Streptococcus in Pregnant Females in Al-Quwayiyah General Hospital, Riyadh, Saudi Arabia

Background: Group B streptococcus is one of the most common causes of severe neonatal infections.&#x0D; Aim: To detect the prevalence of group B. Streptococcus and their antimicrobial susceptibility and to assess the role of PCR as a rapid method of its detection.&#x0D; Place and Duration of the Study: A cross sectional and prospective cohort study was carried out from September 2019 to February 2020 in Gynaecology and Obstetrics OPD and inpatient units in Al- Quwayiyah General hospital, Riyadh, Saudi Arabia&#x0D; Methodology: Paired rectal/vaginal specimens were collected from 540 pregnant females with gestational age 35 or more, Each swab was inoculated into selective medium, Todd Hewitt, One swab is streaked onto blood agar plates incubated in 5% CO2 for 24h at 36ºC. β-hemolytic colonies growth is identified by Gram’s stain, colony morphology and CAMP test. The confirmation and antimicrobial susceptibility were done by Vitek II machine, The second swab was used for PCR to identify atr gene.&#x0D; Results: Out of 540 pregnant women 87 (16.1%) were colonized with GBS isolates. The positive GBS women aged 25 (22-34) and negative GBS women aged 23 (24-35) with no statistical difference. Patients aged more than 35 years old has the higher rate of positive GBS, 46.2%. No significant association detected between GBS and gestational age at delivery, antenatal visits, BMI and gravidity. The GBS strains isolated from pregnant women was 100% susceptible to linezolid and vancomycin followed by ampicillin (93.1%) and tobramicin (83.9%) then gentamicin (81.6%) and levofloxacin(78.2%) and showed least antibiotic susceptibility to erythromycin (26.4%). The PCR was positive in 145 (26.9%). Using culture as gold method, PCR sensitivity was 100% (95% CI: 91.62-100), while specificity was 83.2% (95% CI: 82.61-91.02). Negative and positive predictive values were 100% and 61% respectively. Kappa between the two methods was 0.71, which indicate major agreement.&#x0D; Conclusion: The GBS prevalence among the pregnant females in Al Quwayiyah General Hospital was 16.1%. Detection of GBS using new PCR technique was found to have high sensitivity and faster results, allowing efficient management of GBS and reduction in newborn morbidity and mortality however the cost is high for some laboratories. Further studies should be assessed to be both low cost and accurate rapid screening.

Continuous medical education activities; Answers to Case No. 3

The considerations were Listeria, Corynebactrium, Erysipelothrix and Arcanobacterium species. The isolate was catalase-negative, which rules out Corynebacteria and Listeria species. The Gram-positive card of the Vitek 2 system can identify Erysipelothrix species, which eliminated that genus. At this point, a reverse CAMP test was performed to determine if the microorganism might be Arcanobacterium hemolyticum (A. hemolyticum). Arcanobacterium hemolyticum typically inhibits the beta hemolysis of Staphylococcus aureus on blood agar, hence resulting in a positive reverse CAMP test. The patient's isolate neither inhibited nor enhanced the β-hemolysis of S. aureus and was not A. hemolyticum. Additional biochemical tests were performed. The mystery microorganism is a tiny, Gram positive rod that is strongly β-hemolytic, catalase negative, ferments xylose and demonstrates no reaction on the reverse CAMP test. What is your diagnosis? (To be continued…).

Usefulness Of Microbial Cytotoxins In The Diagnosis Of Selected Bacterial Infections

Abstract The reaction of synergistic hemolysis, which is the basis of the CAMP test, is used in the preliminary identification of selected bacterial pathogens, including L. monocytogenes, C. perfringens, S. agalactiae. The experimental part was to determine the influence blood cells of used, incubation time, distance between growth lines and the type of strains on the intensity of CAMP. Synergistic hemolysis was observed in the classic CAMP test, when 10% human blood cell is used in the medium. It has been shown that the optimal substrate for the CAMP test to identify Listeria sp. is an agar with the 5% addition of human blood cells. While sheep’s erythrocytes were the best substrate for hemolysis of C. perfringens. The optimum distance and incubation time for the tested bacterial species was 1–4 mm and 24 h. When performing preliminary identification of pathogenic microorganisms using the CAMP test, different types of erythrocytes should be used, due to strain and species-specific variabilieties.