At-Home Administration of Gantenerumab by Care Partners to People with Early Alzheimer's Disease: Feasibility, Safety and Pharmacodynamic Impact.

BACKGROUND: Monoclonal antibodies that target amyloid-beta and remove amyloid plaques can slow cognitive and functional decline in early Alzheimer's disease. Gantenerumab is a subcutaneously administered fully-human anti-amyloid-beta monoclonal antibody with highest affinity for aggregated amyloid-beta. Since the phase 3 GRADUATE trials did not meet the primary endpoint (change from baseline to Week 116 in Clinical Dementia Rating scale - Sum of Boxes), development of gantenerumab in sporadic Alzheimer's disease was stopped and all ongoing trials were terminated early due to sponsor decision. Subcutaneous administration at the clinic or at home by care partner would be an important option for other therapies in this class in order to increase flexibility and reduce overall burden. The insights obtained from the experience with gantenerumab home administration by care partner in the phase 2 GRADUATION trial will serve to guide the ongoing efforts with other anti-amyloid-beta antibodies. OBJECTIVES: To evaluate the pharmacodynamic effects on brain amyloid load of once weekly subcutaneous administration of gantenerumab and the safety and feasibility of home administration by care partners. DESIGN: Phase 2, open-label, single arm study. SETTING: Multicenter trial conducted in 33 sites in 8 countries from November 2020 to March 2023. PARTICIPANTS: Participants aged 50 to 90 with early symptomatic Alzheimer's disease (mild cognitive impairment/mild dementia due to Alzheimer's disease), and evidence of amyloid positron emission tomography positivity. INTERVENTION: Participants could receive up to 255 mg gantenerumab once-weekly, administered subcutaneously at site or at home by healthcare professionals or non-healthcare-professional care partners. MEASUREMENTS: The primary endpoint was the change from baseline to Week 52 and to Week 104 in brain amyloid load as measured by PET centiloid levels. The secondary endpoints were responses to the home administration questionnaire, plasma concentrations and safety. RESULTS: The overall number of participants enrolled was 192, with a mean (standard deviation) amyloid PET load at baseline of 101.80 (29.80) centiloids. At the time of early study termination by sponsor, 149 participants had valid Week 52 amyloid PET data (primary endpoint), and 12 participants had an early termination PET within the pre-defined time range of Week 104. The mean change in amyloid PET from baseline to Week 52 and Week 104 was -26.19 centiloids (range: -75.6-15.8; n=149) and -35.48 centiloids (range: -63.2--7.0; n=12), respectively. Responses to the home administration questionnaire at Week 52 (n=148) indicated that the majority of care partners (88-97%) considered administration of study drug at home easy (30.4%) or very easy (57.4%), and convenient (25.7%) or very convenient (70.9%). Care partners felt confident (31.1%) or very confident (62.2%) and satisfied (29.7%) or very satisfied (64.9%) with giving the injection at home. Responses by care partners at Week 36 (n=72), Week 76 (n=126) and Week 104 (n=29) and participant (patient) assessment of convenience and satisfaction at these time points were similar. There were no new safety findings associated with gantenerumab administered subcutaneously once weekly at 255 mg or safety issues associated with at-home injections by non-healthcare professional care partners. CONCLUSIONS: Once-weekly subcutaneous home administration of the anti-amyloid-beta antibody gantenerumab by non-healthcare-professional care partners to participants with early Alzheimer's disease was feasible, safe, well tolerated, and considered as a convenient option by both the care partners and participants with Alzheimer's disease. Although gantenerumab's development has been stopped due to lack of efficacy, this approach has the potential to reduce the frequency of hospital/outpatient clinic visits required for treatment with other anti-amyloid-ß antibodies and can increase flexibility of drug administration for people living with Alzheimer's disease and their families.

Generating data to facilitate more equitable distribution of health resources: an illustration of how local health surveys can identify probable need in mixed socio-economic regions.

OBJECTIVES: This study aimed to (1) encourage allocation of governmental and grant funds to the administration of local area health surveys and (2) illustrate the predictive impact of socio-economic resources on adult health status at the local area level to provide an example of how health surveys can identify residents with the greatest health needs. STUDY DESIGN: Randomly sampled and weight-adjusted regional household health survey (7501 respondents) analyzed with categorical bivariate and multivariate statistics, combined with Census data. Survey sample consists of the lowest, highest, and near highest ranked counties in the County Health Rankings and Roadmaps for Pennsylvania. METHODS: Socio-economic status (SES) is measured regionally with Census data consisting of seven indicators and individually with Health Survey data consisting of five indicators based on poverty level, overall household income, and education. Both of these composite measures are examined jointly for their predictive effects on a validated health status measure using binary logistic regression. RESULTS: Once county-level measures of SES and health status are broken down into smaller areas, better identification of pockets of health need is possible. This was most strongly revealed in an urban county, Philadelphia, which is ranked lowest of 67 counties on health measures in the state of Pennsylvania, yet when broken down into 'neighborhood clusters' contained both the highest- and lowest-ranked local area in a five-county region. Overall, regardless of the SES level of the County subdivision one lives in, a low-SES adult has close to six times greater odds of reporting 'fair or poor health status' than does a high-SES adult. CONCLUSION: Local health survey analysis can lead to a more precise identification of health needs than surveys attempting to cover broad areas. Low-SES communities within counties, and low-SES individuals, regardless of the community they live in, are substantially more likely to experience fair to poor health. This adds urgency to the need to implement and investigate socio-economic interventions, which can hopefully improve health and save healthcare costs. Novel local area research can identify the impact of intervening variables such as race in addition to SES to add more specificity in identifying populations with the greatest health needs.

Post-operative Day 1 Serum Transaminase Levels in Relation to Morbidity After Liver Resection.

BACKGROUND: Post-operative serum transaminases have been proposed as possible early predictors of morbidity after liver resection. This study aimed to verify the clinical value of post-operative serum transaminases. METHODS: Clinical data from 2001 to 2016 in a single non-academic referral HPB center were collected from a prospectively held database. Post-operative day 1 serum aspartate transaminase (AST) and alanine transaminase (ALT) were tested for their relationship with post-operative major morbidity, defined by a Clavien-Dindo score 3 or higher, and mortality. RESULTS: For this analysis, 371 patients were included, including 149 (40%) undergoing major liver resections. In total, 17% of the patients developed major morbidity. Stepwise logistic regression demonstrated that AST, and not ALT, is an independent predictor for major morbidity (p = 0.017). The probability of major morbidity significantly increased with increasing AST values. A threshold value of 242 U/L was found to be predictive for one or more major complications. CONCLUSIONS: In this study, post-operative serum AST on day 1 was a predictive factor for major morbidity after liver resection. For patients with low AST value, early discharge could be considered. However, because of the substantial inter-individual variability of AST values, more studies are needed to translate these results into clinical practice.

Thirty-Six-Month Amyloid Positron Emission Tomography Results Show Continued Reduction in Amyloid Burden with Subcutaneous Gantenerumab.

Previous findings from the positron emission tomography (PET) substudy of the SCarlet RoAD and Marguerite RoAD open-label extension (OLE) showed gantenerumab doses up to 1200 mg every 4 weeks administered subcutaneously resulted in robust beta-amyloid (Aß) plaque removal over 24 months in people with prodromal-to-moderate Alzheimer's disease (AD). In this 36-month update, we demonstrate continued reduction, with mean (standard error) centiloid values at 36 months of -4.3 (7.5), 0.8 (6.7), and 4.7 (8.0) in the SCarlet RoAD (double-blind pooled placebo and active groups), Marguerite RoAD double-blind placebo, and Marguerite RoAD double-blind active groups respectively, representing a change of -57.0 (10.3), -90.3 (9.0), and -74.9 (10.5) centiloids respectively. These results demonstrate that prolonged gantenerumab treatment, at doses up to 1200 mg, reduces amyloid plaque levels below the amyloid positivity threshold. The ongoing GRADUATE Phase III trials will evaluate potential clinical benefits associated with gantenerumab-induced amyloid-lowering in people with early (prodromal-to-mild) AD.

Transoral thyroid surgery vestibular approach: does size matter anymore?

BACKGROUND: Various approaches for endoscopic and minimally invasive thyroid operations have been reported, with some becoming popular to date. The aim of these procedures is to reduce and prevent visible scars in the neck. This led to the transoral endoscopic thyroidectomy vestibular approach (TOETVA) technique applicable in small thyroid specimen. To amend TOETVA and to broaden the indication for transoral thyroid surgery, our aim was to combine the TOETVA with the retroauricular endoscopic cephalic access thyroid surgery (EndoCATS) facelift approach. METHODS: After successful implementation of the TOETVA technique in 2017, we evaluated the transoral technique regarding feasibility and safety in well-selected patients in Germany and Austria. With the combination of TOETVA and EndoCATS, we were able to optimize the extraction process of the specimen by avoiding damage to the thyroid capsule and to broaden the indication of transoral surgery by operating on thyroid glands with higher volume. Patients' characteristics, surgical outcome and complications were determined. RESULTS: Seventy-one transoral procedures were performed in 70 patients. The TOETVA procedure was performed in 60 patients, and 10 female patients underwent TOVARA with transoral thyroid lobe mobilization and removal of the specimen via the retroauricular access. Overall, median operation time was 205 min (range 96-370 min) and permanent RLNP rate was 1% accordant to 99 nerves at risk (39 hemithyroidectomies, 29 total thyroidectomies, 2 parathyroid resections, and 1 thyroglossal cyst resection). No conversion to conventional open surgery was necessary. Long-term mental nerve injury occurred in two patients. No infection was identified. CONCLUSIONS: Transoral thyroid and parathyroid surgery via the vestibular approach is both feasible and safe in Western Europe. The combination of the TOETVA with the retroauricular endoscopic cephalic access thyroid surgery (EndoCATS) facelift approach, called TOVARA, is a promising opportunity to allow for thyroid surgery without a visible scar in specimen with volume of more than 40 ml also.

Possible Preventable Causes of Unplanned Readmission After Elective Liver Resection, Results from a Non-academic Referral HPB Center.

INTRODUCTION: Unplanned readmission is a common event after liver resection, and it is a burden for both patients and healthcare policy makers. This study evaluates the incidence of and reasons for unplanned readmission after liver resection, in order to identify possible preventable causes. METHODS: In this single-center cohort study, data from patients who underwent liver resection for both malignant and benign indications from 2001 to 2016 at our institute were collected from a database with prospective data. Readmissions were analyzed for their reasons and risk factors. Patients with general complaints with no specific complications were categorized as failure to thrive. RESULTS: In 406 patients, the readmission rate was 11.6%. Most patients were readmitted because of failure to thrive (35%), deep and superficial surgical site infection (28%), or cardiopulmonary complications (15%). A multivariate analysis revealed that unplanned readmission was associated with the occurrence of complications during index admission-with an odds ratio of 4.69 (CI 2.41-9.12, p < 0.001). CONCLUSION: Readmission occurs in more than 1 in 10 patients after liver resection, and it is associated with a complicated course during index admission. One-third of readmissions occur because of failure to thrive and might be preventable. Future research in strategies to reduce readmission rates should focus on both the prevention of complications during index admission and programs at the interface between primary and secondary care.

Characterisation of mecA gene negative Staphylococcus aureus isolated from bovine mastitis milk from Northern Germany.

Staphylococcus aureus (S. aureus) is an important causative agent of contagious intermammary infections in dairy cattle. S. aureus is also considered as an important foodborne pathogen and cause of food poisoning cases and outbreaks worldwide. In order to understand the molecular ecology of S. aureus, the present study compared phenotypic and genotypic characteristics of 70 S. aureus isolates from bovine mastitis milk samples collected during the period from August 2001 to March 2014 in different regions of Northern Germany. The S. aureus isolates were characterised phenotypically, as well as genotypically for their genetic diversity using multi-locus sequence typing (MLST), spa typing and the presence of virulence genes encoding 16 staphylococcal enterotoxins (sea-selu), toxic shock syndrome toxin (tst), thermonuclease (nuc), clumping factor (clfA and clfB), coagulase (coa) and the methicillin resistance gene mecA. A total of 16 sequence types were grouped into eight clonal complexes (CCs), and 17 spa types were identified. These included six novel sequence types and one novel spa type. The majority of bovine mastitis milk-associated sequence types belonged to the clonal complex CC5, CC97, CC133, and CC151 and showed closely related genotypes or lineages with sequence types of human origin. The genotype CC133 (ST133-t1403) was predominant, constituting 27.1% of the isolates. In addition, the S. aureus isolates displayed nine different enterotoxigenic profiles. All S. aureus were methicillin-susceptible (MSSA). The current study provides new information on phenotypic and genotypic traits of S. aureus isolates from bovine mastitis. The comparison of characteristics of isolates from the present study originating from mastitis milk showed similarities with human isolates. This might help to better understand the distribution of S. aureus in the one health context.

[Transoral thyroid and parathyroid surgery : Implementation and evaluation of the transoral endoscopic technique via the vestibular approach (TOETVA)]. / Transorale Chirurgie der Schilddrüse und Nebenschilddrüsen : Implementierung und Evaluation der transoralen endoskopischen Technik über den Vestibularzugang (TOETVA).

BACKGROUND: More than 10 years ago various research groups in Germany first reported the possibility of transoral surgery of the thyroid and parathyroid glands. These were developed and evaluated within the framework of natural orifice transluminal endoscopic surgery (NOTES). While development of these innovative and new techniques that enabled surgery without visible scars did not become well accepted in Europe and America, it led to an optimized transoral endoscopic thyroidectomy vestibular approach (TOETVA), which was implemented particularly in Asia. We report the preparation, step-by-step implementation, and first promising results for TOETVA of an international surgical cooperation. MATERIAL AND METHODS: A Thai-Austrian-German cooperation was started in June 2017. Between June and November 2017 the first 10 TOETVA procedures were performed in female and male patient(s) presenting with single thyroid nodules, sporadic primary hyperparathyroidism and solitary parathyroid adenoma or thyroglossal duct cysts. The TOETVA technique was performed using 3 laparoscopic ports inserted at the oral vestibule and a CO2 insufflation pressure at 6-8 mm Hg. Each surgery was performed using laparoscopic instruments and ultrasonic or bipolar devices. Surgical outcome, complications and conversions to an open technique were recorded. RESULTS: Of the patients six presented with single thyroid nodules, two patients had multinodular goitre with scintigraphically cold nodules and multifocal hyperfunctioning thyroid tissue, one patient suffered from sporadic primary hyperparathyroidism with a left sided parathyroid adenoma and one patient suffered from a thyroglossal duct cyst. No conversion to conventional open surgery was necessary. Average tumor size was 3.1 cm (range 1-4 cm). There was no relevant loss of blood or subsequent bleeding. No temporary or permanent hoarseness or mental nerve injury occurred, while transient hypoparathyroidism was evident after successful parathyroid resection. Of the patients five developed a slight postoperative hematoma in the submandibular region. No infections or disorders of wound healing occurred. CONCLUSION: The TOETVA technique is feasible and safe in selected patients; however, further prospective studies are necessary to evaluate the value of the procedure, risks and long-term results.

GPs' assessment of cooperation with other health care providers involved in cancer care-a cross-sectional study.

Cancer is a complex disease requiring the involvement of several health care providers. A possible constant in the cancer care process is the general practitioner (GP). The aim of this project was to evaluate GPs' satisfaction with cooperation with other health care providers in the cancer care process of their patients and to explore which variables are associated with higher satisfaction with cooperation with other health care providers. We considered the following health care providers: outpatient oncology specialists, physicians in relatively small hospitals (≤400 beds), physicians in relatively large hospitals (>400 beds), home care services, outpatient psycho(onco)logists/psychotherapists, hospice/palliative care units and specialised palliative home care. The cross-sectional study was carried out as a postal survey all over Germany. Data were analysed descriptively and by means of logistic regression. Overall satisfaction with cooperation with other health care providers involved in cancer care was rather high. Only cooperation with outpatient psycho(onco)logists/psychotherapists was rated as not assessable/irrelevant by a majority of GPs. For all other health care providers under review, both communication and the transfer of sufficient information in good time were associated with overall satisfaction with cooperation. Little association was found between GP and practice variables and overall satisfaction with cooperation with the considered health care providers.

Comparative phenotypic analysis of "Clostridium neonatale" and Clostridium butyricum isolates from neonates.

"Clostridium neonatale" was recently described as a new species within the Cluster I of the Clostridium genus sensu stricto. In this study, we characterized "C. neonatale" isolates (n = 42) and compared their phenotypic properties with those of Clostridium butyricum (n = 26), a close related species. Strains isolated from fecal samples of healthy neonates were tested for different phenotypic characteristics. Compared to C. butyricum, "C. neonatale" showed a significant higher surface hydrophobicity (p = 0.0047), exopolysaccharide production (p = 0.0069), aero-tolerance (p = 0.0222) and viability at 30 °C (p = 0.0006). A lower swimming ability (p = 0.0146) and tolerance against bile (0.3%) (p = 0.0494), acid (pH 4.5) (p < 0.0001), osmolarity (NaCl 5%, p = 0.0188) and temperature at 50 °C (p = 0.0013) characterized "C. neonatale" strains. Our results showed that "C. neonatale" behaves very differently from C. butyricum and suggests specific responses to environmental changes. Besides it is the first study on clinical isolates for these two anaerobic members of the newborns' gut microbiota and broadens our knowledge about their phenotypic traits.

Biomechanical evaluation of different osteosynthesis methods after mandibular sagittal split osteotomy in major advancements.

The aim of this study was to assess the biomechanical stability of six different osteosynthesis methods after sagittal split osteotomy. Sixty polyurethane hemimandibles were divided into two groups, with six subgroups in each. After 10-mm advancement of the distal segment (group 1) and 10-mm advancement combined with 20° counterclockwise rotation (group 2), the bone segments were fixed using 2.0-mm plates/screws as follows: subgroup A, one conventional straight plate; subgroup B, two conventional straight plates; subgroup C, one conventional sagittal plate; subgroup D, one locking straight plate; subgroup E, two locking straight plates; subgroup F, one locking sagittal plate. The hemimandibles were tested for compressive strength by three-point biomechanical test, until there was 3mm of displacement between the segments. The fixations showed better performance in group 1 than in group 2 in all cases, with statistical significance for subgroups A, C, and D. In both groups, the use of two straight miniplates showed the most resistance, followed by the sagittal miniplates. However, in counterclockwise rotations, no statistically significant difference was found between two conventional straight plates and the sagittal locking plate. This study shows that the use of two plates is the form of fixation with the minimum displacement. If the clinician opts to use one plate, a sagittal plate is the best alternative.

Interferon γ is a STAT1-dependent direct inducer of BCL6 expression in imatinib-treated chronic myeloid leukemia cells.

B-cell CLL/lymphoma 6 (BCL6) exerts oncogenic effects in several human hematopoietic malignancies including chronic myeloid leukemia (CML), where BCL6 expression was shown to be essential for CML stem cell survival and self-renewal during imatinib mesylate (IM) treatment. As several lines of evidence suggest that interferon γ (IFNγ) production in CML patients might have a central role in the response to tyrosine kinase inhibitor (TKI) therapy, we analyzed if IFNγ modulates BCL6 expression in CML cells. Although separate IFNγ or IM treatment only slightly upregulated BCL6 expression, combined treatment induced remarkable BCL6 upregulation in CML lines and primary human CD34+ CML stem cells. We proved that during combined treatment, inhibition of constitutive signal transducer and activator of transcription (STAT) 5 activation by IM allowed the specific enhancement of the STAT1 dependent, direct upregulation of BCL6 by IFNγ in CML cells. By using colony-forming assay, we found that IFNγ enhanced the ex vivo colony or cluster-forming capacity of human CML stem cells in the absence or presence of IM, respectively. Furthermore, inhibition of the transcriptional repressor function of BCL6 in the presence of IM and IFNγ almost completely blocked the cluster formation of human CML stem cells. On the other hand, by using small interfering RNA knockdown of BCL6, we demonstrated that in an IM-treated CML line the antiapoptotic effect of IFNγ was independent of BCL6 upregulation. We found that IFNγ also upregulated several antiapoptotic members of the BCL2 and BIRC gene families in CML cells, including the long isoform of MCL1, which proved to be essential for the antiapoptotic effect of IFNγ in an IM-treated CML line. Our results suggest that combination of TKIs with BCL6 and MCL1 inhibitors may potentially lead to the complete eradication of CML stem cells.

Maxilla reconstruction with autogenous bone block grafts: computed tomography evaluation and implant survival in a 5-year retrospective study.

This retrospective study was performed to evaluate the bone thickness of the anterior maxillary region after reconstruction with autogenous bone blocks at 6 months and 5 years after surgery using computed tomography (CT) and to determine the implant survival rate. Eleven patients with a horizontal bone deficiency were treated with reconstructive procedures and implant placement. CT measurements were obtained before surgery (T0) and at 6 months (T1) and 5 years (T2) after surgery. The values were analysed statistically (analysis of variance and Tukey's test; P<0.05). Implant survival was evaluated at follow-up. The mean width of the lower region of the ridge (±standard deviation, in millimetres) was 3.8±1.6 at T0, 7.0±1.6 at T1, and 6.5±1.0 at T2; the mean width of the upper region of the ridge was 5.7±2.3 at T0, 8.3±2.2 at T1, and 7.3±1.6 at T2. The mean total thickness of the ridge was 4.7mm at T0, 7.6mm at T1, and 6.9mm at T2; the average increase in horizontal thickness was 2.9mm at T1 and 2.2mm at T2. A statistically significant difference was observed in the mean width of the lower portion at T1 and T2 compared to the width at T0. The implant survival rate was 94.1%. This technique demonstrated high predictability for implant survival, with a reduction in the graft bone during the follow-up period.

Effect of a nano-silver coating on the quality of fresh turkey meat during storage after modified atmosphere or vacuum packaging.

Nano-silver is used in consumer products due to its antibacterial properties. The aim of this study was to evaluate the effect of a nano-silver-coated film on the quality of turkey meat during vacuum-sealed and modified atmosphere packaging up to 12 days of storage. In the first part of the experiment, turkey breasts were packaged using either vacuum packaging or modified atmosphere packages (MAPs) and contained films with or without a nano-silver coating (control film). Parameters such as pH, electrical conductivity, color (lightness L*, redness a*), myoglobin redox forms, thiobarbituric acid-reactive substances (TBARS), biogenic amines (BAs), total viable bacterial counts, Pseudomonas species counts, and Enterobacteriaceae species counts were evaluated on storage days 4, 8, and 12. In the second part of the study, the antimicrobial effect of a nano-silver-coated film on turkey breast was evaluated after inoculation with Escherichia coli (E. coli). Turkey meat packaged with the nano-silver film exhibited lower a* values on days 1 (3.15 ± 0.62), 4 (3.90 ± 0.68), and 8 (4.27 ± 0.76) compared to the packaged meat with the control film (3.41 ± 0.73, 4.35 ± 0.94, 4.85 ± 0.89, respectively), indicating special optical properties of nanoparticles. Concerning the BAs, silver packaged meat showed higher values of tyramine on day 12 (1274 ± 392 ng/g meat) and cadaverine on day 4 (1224 ± 435 ng/g meat) compared to the normal packaged products (647 ± 576 and 508 ± 314 ng/g meat, respectively). MAP meat revealed higher L* and TBARS values and lower microbial counts than the vacuum packaged products on all days. The MAP meat also showed lower a* results on days 4 and 8 and higher metmyoglobin (metMb) values on days 8 and 12 compared to th E: vacuum products. In the inoculation study, the microbial counts of the turkey meat were comparable between the two film types. The study showed that the nano-silver coating did not exhibit any advantageous effects on the quality and microbiological parameters of the turkey meat.

Microbiology and foodborne pathogens in honey.

Honey has been considered a relatively safe foodstuff due to its compositional properties, with infant botulism caused by Clostridium botulinum being the most prominent health risk associated with it. Our review is focused on the honey microflora along the food chain and evaluates the pathogenic potential of those microorganisms found in honey. This product may contain a great variety of bacteria and, particularly, fungi that eventually entered the food chain at an early stage (e.g., via pollen). For many of these microorganisms, opportunistic infections in humans have been recorded (e.g., infections by Staphylococcus spp., Citrobacter spp., Escherichia coli, Hafnia alvei, Aspergillus spp., Fusarium spp., Trichoderma spp., Chaetomium spp.), although direct infections via honey were not registered.

Genome Sequences of Malonate-Positive Cronobacter sakazakii Serogroup O:2, Sequence Type 64 Strains CDC 1121-73 and GK1025, Isolated from Human Bronchial Wash and a Powdered Infant Formula Manufacturing Plant.

We introduce draft genome sequences of strains CDC1121-73 (human bronchial wash isolate) and GK1025 (powdered infant formula manufacturing facility isolate), which are both malonate-positive Cronobacter sakazakii serogroup O:2, sequence type 64. Assemblies for these strains have sizes of 4,442,307 and 4,599,266 bp and % G+C contents of 56.9 and 56.7, respectively.

Development and validation of a loop mediated isothermal amplification (LAMP) assay for the detection of Staphylococcus aureus in bovine mastitis milk samples.

Staphylococcus (S.) aureus is one of the most important animal pathogens causing bovine mastitis. Also, it is a major human pathogen that may produce a variety of toxins which cause staphylococcal food poisoning. In the present study a LAMP assay based on gene nuc to identify S. aureus was developed and validated. The specificity of the LAMP assay was confirmed by using 70 S. aureus isolates and 21 non-S. aureus strains. The optimal temperature-time combination to amplify gene nuc successfully was 65 °C and 30 min. The analytical sensitivity of the developed LAMP assay was 0.26 pg of S. aureus DNA per reaction. The limit of detection evaluated with milk spiked with S. aureus was 9 × 102 CFU mL-1. The final results of this assay were available within less than 2 h. The present study showed that the LAMP assay based on gene nuc appeared to be rapid and simple, and could also be used to identify S. aureus isolates from mastitis milk of dairy cows.

Application of a loop-mediated isothermal amplification (LAMP) assay for molecular identification of Trueperella pyogenes isolated from various origins.

In the present study 28 Trueperella pyogenes strains isolated from various origins could successfully be identified with a newly designed loop-mediated isothermal amplification (LAMP) assay based on gene cpn60 encoding chaperonin. No cross reaction could be observed with control strains representing four species of genus Trueperella and seven species of closely related genus Arcanobacterium. The present cpn60 LAMP assay might allow a reliable and low cost identification of T. pyogenes also in laboratories with less specified equipment.

Multilocus sequence typing of Campylobacter jejuni and Campylobacter coli isolates from poultry, cattle and humans in Nigeria.

AIMS: To determine the genetic diversity of Campylobacter jejuni and Campylobacter coli isolates from Nigeria and to identify the association between multilocus sequence types and hosts (poultry, cattle and humans). METHODS AND RESULTS: Isolates were identified using multiplex PCR assays. Multilocus sequence typing (MLST) was used to determine the genetic diversity of 36 Camp. jejuni and 24 Camp. coli strains isolated from poultry, cattle and humans. Of the 36 Camp. jejuni genotyped, 21 sequence types (ST) were found, 9 (43%) were new while of the 24 Camp. coli isolates genotyped, 22 STs were identified with 14 (64%) being new. The most prevalent sequence type was ST1932 followed by ST1036 and ST607 while the prevalent clonal complexes were CC-828, CC-460 and CC-353. CONCLUSIONS: Campylobacter isolates from Nigeria were found to be diverse with novel genotypes. There was overlap of CC-828, CC-460 and CC-353 between the poultry, cattle and human isolates. Genetic exchange was also detected in two of the Camp. coli isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the genetic diversity of Campylobacter strains in Nigeria, demonstrating that Camp. jejuni and Camp. coli isolates are diverse and have both local and global strains. The predominant sequence types and clonal complexes found in this study differ from other countries; this exemplifies that different predominant Campylobacter populations exist between countries.