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BackgroundHuman brucellosis is a neglected disease transmitted to humans from animals such as cattle, goats, dogs, and swine. The causative agents are bacteria of the genus Brucella, intracellular pathogens usually confined to the reproductive organs of their animal hosts causing sterility and abortions. The objective of the study was to determine the seroprevalence of brucellosis among women with spontaneous abortions (SAW) and compare this seroprevalence with that of healthy pregnant women (HPW).MethodsThe case–control study was designed to determine the seroprevalence and molecular detection of brucellosis in women who suffered from spontaneous abortion and healthy pregnant women of the Haripur District of Pakistan. A total of 770 blood samples (n = 385 for each group) were collected from 9 public and 11 private hospitals in Haripur District from December 2021–March 2023. Data on demographic features, epidemiological variables, and risk factors were collected from each participant by structured questionnaires. Initial screening for brucellosis was performed by Rose Bengal Plate Test followed by qRT-PCR for molecular detection of the genus-specific BCSP-31 gene of Brucella.ResultsThe study showed that anti-Brucella antibodies were more found in SAW 23.63% (91/385) than in HPW 1.29% (5/385). Brucella specific DNA was amplified in 89.01% (81/91) seropositive samples of SAW. Demographic features and risk factors such as age, urbanicity, socioeconomic status, education, occupation, and animal contact were found significantly associated with brucellosis (p ≤ 0.05). Consumption of unpasteurized raw milk (OR = 18.28, 95%CI: 8.16–40.94) was found highly concomitant with seroprevalence.ConclusionThis study reports the first evidence of involvement of brucellosis in spontaneous abortions in women of Pakistan. The study can be used to develop strategies for risk management during pregnancy, to raise awareness for brucellosis, and develop control programs.

This article provides novel data on the microstructure and crystallographic texture of modern giant clam shells (Tridacna squamosa and Hippopus hippopus) from the Coral Triangle region of northeast Borneo. Giant clams have two aragonitic shell layers—the inner and outer shell layer. This dataset focuses on the inner shell layer as this is well preserved and not affected by diagenetic alteration. To prepare samples for analysis, shells were cut longitudinally at the axis of maximum growth and mounted onto thin sections. Data collection involved scanning electron microscopy (SEM) to determine microstructure and SEM based electron backscatter diffraction (EBSD) for quantitative measurement of crystallographic orientation and texture. Post-acquisition reanalysis of saved EBSD patterns to optimize data quality included changing the number of reflectors and band detection mode. We provide EBSD data as band contrast images and colour-coded orientation maps (inverse pole figure maps). Crystallographic co-orientation strength obtained with multiple of uniform density (MUD) values are derived from density distributed pole figures of indexed EBSD points. Raw EBSD data files are also given to ensure repeatability of the steps provided in this article and to allow extraction of further crystallographic properties for future researchers. Overall, this dataset provides 1. a better understanding of shell growth and biomineralization in giant clams and 2. important steps for optimizing data collection with EBSD analyses in biogenic carbonates.

In the original publication, a mistake was observed in Figure 5 as published [...]

Allantoin possesses numerous beneficial properties for the skin, like anti-irritant effects, wound healing, skin hydration, and epithelization. In this paper, we investigated a suitable preparation method for an allantoin hydrogel using the Semi-Solid Control Diagram (SSCD) method and characterized its rheological and consistency behavior. To accomplish this, xanthan gum (XG) was selected as a model gelling agent. Briefly, four hydrogels were prepared, two without allantoin (coded M01 and M02) and two with allantoin (M1 and M2). Similarly, the formulations were either prepared through magnetic stirring (M01 and M1) or homogenization in a mortar (M02 and M2). The prepared hydrogels were evaluated using the SSCD for specific parameters and indexes. The Good Quality Index (GQI) shows a higher value for the formulation, M1 = 6.27, compared to M2 = 5.45. This result is also underlined by the value of M01 = 6.45, which is higher than M02 = 6.38. Considering the consistency, the formulation M01 possessed the highest spreadability, followed by M02 and then the allantoin hydrogels M1 and M2. The rheological behavior had a thixotropic pseudoplastic flow for all the formulations. The use of SSCD pictographs outlined the rheological properties that need improvement, the method that is suitable to prepare the allantoin hydrogels, and the influence of the allantoin suspended in the XG hydrogel.

Staphylococcus aureus is one of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogens among which multidrug resistance has emerged. Resistance to methicillin has resulted in clinicians using the antibiotic of last resort, vancomycin, to treat infections caused by methicillin-resistant S. aureus (MRSA). However, excessive use and misuse of vancomycin are major causes of resistance among S. aureus strains. South Asia encompasses ~25% of the world’s population, and countries in South Asia are often characterized as low- and middle-income with poor healthcare infrastructure that may contribute to the emergence of antibiotic resistance. Here, we briefly highlight the mechanism of vancomycin resistance, its emergence in S. aureus, and the molecular epidemiology of non-susceptible S. aureus to vancomycin in the South Asian region.

Herein, we reported an HPLC method for the simultaneous determination of tibezonium iodide (TBN) and lignocaine hydrochloride (LGN). The method was developed according to the International Conference for Harmonization guidelines (ICH) Q2R1 using Agilent® 1260 with a mobile phase consisting of acetonitrile and phosphate buffer (pH 4.5) in a volumetric ratio of 70:30 and flowing through a C8 Agilent® column at 1 mL/min. The results revealed that TBN and LGN peaks were isolated at 4.20 and 2.33 min, respectively, with a resolution of 2.59. The accuracy of TBN and LGN was calculated to be 100.01 ± 1.72% and 99.05 ± 0.65% at 100% concentration, respectively. Similarly, the respective precision was 100.03 ± 1.61% and 99.05 ± 0.48%. The repeatability for TBN and LGN was found to be 99.05 ± 0.48% and 99.19 ± 1.72%, respectively, indicating that the method was precise. The respective regression co-efficient (r2) for TBN and LGN was found to be 0.9995 and 0.9992. Moreover, the LOD and LOQ values for TBN were 0.012 and 0.037 µg/mL, respectively, while for LGN, they were 0.115 and 0.384 µg/mL, respectively. The calculated greenness of the method for ecological safety was found to be 0.83, depicting a green contour on the AGREE scale. No interfering peaks were found when the analyte was estimated in dosage form and in volunteers’ saliva, depicting the specificity of the method. Conclusively, a robust, fast, accurate, precise and specific method was successfully validated to estimate TBN and LGN.

Both pathogenic as well as nonpathogenic species of staphylococci have been reported in poultry, but these studies have not compared staphylococcal flora of both farmed and household broiler chickens. Staphylococci from farmed (n = 51) and household chicken intestines (n = 43) were isolated and tested for resistance to antimicrobials, presence of resistance genes, and inhibitory activity against other bacteria; correlation of resistance phenotype and genotype was also evaluated. At least 12 staphylococcal species were identified; Staphylococcus carnosus subspecies carnosus was the predominant species from both sources. Most farmed chicken staphylococci were resistant to tigecycline (38/51; 74.8%) while the highest level of resistance among the household chicken staphylococci was to clindamycin (31/43; 72.1%). The mecA gene was only detected in staphylococci from household chickens, whereas ermC and tetK or tetM were found in staphylococci from both groups of birds. Multidrug resistance (resistance ≥ 2 antimicrobial classes) was observed in 88% of resistant staphylococci ranging from 2 to 8 classes and up to 10 antimicrobials. Isolates produced inhibitory activity against 7 clinical bacterial strains primarily Enterococcus faecalis (25/88; 28.4%) and Escherichia coli (22/88; 25%). This study demonstrated that the staphylococcal population among farmed and household chickens varies by species and resistance to antimicrobials. These results may reflect the influence of the environment or habitat of each bird type on the intestinal microflora. As resistance in the staphylococci to antimicrobials used to treat human infections was detected, further study is warranted to determine strategies to prevent transfer of these resistant populations to humans via contamination of the poultry meat.

A background study is important for the conservation and stock management of a species. Terapon jarbua is a coastal Indo-Pacific species, sourced for human consumption. This study examined 134 samples from the central west and east coasts of Peninsular (West) Malaysia and East Malaysia. A 1446-bp concatenated dataset of mtDNA COI and Cyt b sequences was used in this study and 83 haplotypes were identified, of which 79 are unique haplotypes and four are shared haplotypes. Populations of T. jarbua in Malaysia are genetically heterogenous as shown by the high level of haplotype diversity ranging from 0.9167–0.9952, low nucleotide diversity ranging from 0.0288–0.3434, and high FST values (within population genetic variation). Population genetic structuring is not distinct as shown by the shared haplotypes between geographic populations and mixtures of haplotypes from different populations within the same genetic cluster. The gene flow patterns and population structuring observed among these regions are likely attributed to geographical distance, past historical events, allopatric speciation, dispersal ability and water currents. For instance, the mixture of haplotypes revealed an extraordinary migration ability of T. jarbua (>1200 km) via ancient river connectivity. The negative overall value of the neutrality test and a non-significant mismatch distribution are consistent with demographic expansion(s) in the past. The median-joining network concurred with the maximum likelihood haplotype tree with three major clades resolved. The scarcity of information on this species is an obstacle for future management and conservation purposes. Hence, this study aims to contribute information on the population structure, genetic diversity, and historical demography of T. jarbua in Malaysia.

Brucellosis is a neglected zoonotic disease of ruminants. It causes severe health problems in humans and significant economic loss. Only a limited number of studies have been conducted in Pakistan to determine the prevalence of human brucellosis and related risk factors. The objectives of the current cross-sectional study were to determine the prevalence of anti-Brucella antibodies in sera collected from symptomatic patients at three hospitals of Abbottabad using a commercial slide agglutination test (SAT) and to determine risk factors for brucellosis for these patients. Five hundred blood samples were collected. A questionnaire was filled in for each patient to obtain information on age, gender, living area, brucellosis associated symptoms, associated risk factors, pregnancy and abortion history. A total of 13.6% (n = 68) patients were found to be SAT positive and in 83.3% (n = 57) of these samples Brucella DNA was detected by genus specific RT-PCR for BCSP-31 gene. Statistical analysis was performed to determine odd ratios, risk ratios, 95% confidence intervals, and p-values. The prevalence of brucellosis by SAT was reported to be higher in women (14.6%, n = 44) than in men (12.1%, n = 24). The age group 25–50 years was found to be at higher risk for brucellosis (14.5%, n = 50) “animal contact” was reported as the main risk factor followed by “consumption of raw animal products.” Out of 131 pregnant women and 21 patients had abortion, the seropositivity of Brucellosis was 9.9% and 23.8%, respectively. The present study reports a striking prevalence of brucellosis among patients including pregnant women at three hospitals of Abbottabad. These findings may foster strategies for controlling human brucellosis at household level, raising of awareness about brucellosis in hospital and family doctors, and finally in setting up an eradication program in the dairy industry.

Buffalo represent a major source of milk in Pakistan. However, production is impacted by the disease bovine mastitis. Mastitis causes significant economic losses, with Staphylococcus aureus (S. aureus) being one of its major causative agents. While much work has been done understanding the epidemiology of bovine mastitis in Pakistan, detailed molecular characterization of the associated S. aureus is unavailable. In the current study both the epidemiological and molecular characterization of S. aureus from bovine mastitis in the Hazara division of Pakistan are examined. S. aureus was isolated from 18.41% of the animals, and left quarters more prone to infection (69.6%) than right quarters (30.4%). Sub-clinical mastitis (75.31%) was more prevalent than clinical mastitis (24.69%), with infections evenly distributed amongst the eight districts. Molecular characterization revealed that only 19.6% of the isolates were methicillin-resistant, and four strains types identified, including ST9-t7867-MSSA, ST9-MSSA, ST101-t2078-MSSA, and ST22-t8934-MRSA-IVa. Antiseptic resistance genes were not detected in the isolates, and low levels of antibiotic resistance were also noted, however the methicillin-resistant strains had higher overall antibiotic resistance. This study represents the most complete molecular typing data for S. aureus causing bovine mastitis in the Hazara district of Pakistan, and the country as a whole.

In the current study, the reversed-phased high-pressure liquid chromatography (RP-HPLC) method was proposed for the estimation of lignocaine hydrochloride (LIG), hydrocortisone (HYD) and Ketoprofen (KET) according to International Conference for Harmonization (ICH) Q2 R1 guidelines, in a gel formulation. The chromatographic evaluation was executed using Shimadzu RP-HPLC, equipped with a C8 column and detected using UV at 254 nm wavelength, using acetonitrile and buffer (50:50) as a mobile phase and diluent, at flow rate 1 mL/min and n injection volume of 20 μL. The retention time for LIG, HYD, and KET were 1.54, 2.57, and 5.78 min, correspondingly. The resultant values of analytical recovery demonstrate accuracy and precision of the method and was found specific in identification of the drugs from dosage form and marketed products. The limit of detection (LOD) for LIG, HYD, and KET were calculated to be 0.563, 0.611, and 0.669 ppm, while the limit of quantification (LOQ) was estimated almost at 1.690, 1.833, and 0.223 ppm, respectively. The AGREE software was utilized to evaluate the greenness score of the proposed method, and it was found greener in score (0.76). This study concluded that the proposed method was simple, accurate, precise, robust, economical, reproducible, and suitable for the estimation of drugs in transdermal gels.

Agarose (AG) is a naturally occurring biocompatible marine seaweed extract that is converted to hydrocolloid gel in hot water with notable gel strength. Currently, its mucoadhesion properties have not been fully explored. Therefore, the main aim of this study was to evaluate the mucoadhesive potential of AG binary dispersions in combination with Carbopol 934P (CP) as mucoadhesive gel preparations. The gels fabricated via homogenization were evaluated for ex vivo mucoadhesion, swelling index (SI), dissolution and stability studies. The mucoadhesive properties of AG were concentration dependent and it was improved by the addition of CP. Maximum mucoadhesive strength (MS) (27.03 g), mucoadhesive flow time (FT) (192.2 min), mucoadhesive time in volunteers (MT) (203.2 min) and SI (23.6% at 4 h) were observed with formulation F9. The mucoadhesive time investigated in volunteers (MT) was influenced by AG concentration and was greater than corresponding FT values. Formulations containing 0.3%, w/v AG (F3 and F9) were able to sustain the release (~99%) for both drugs till 3 h. The optimized formulation (F9) did not evoke any inflammation, irritation or pain in the buccal cavity of healthy volunteers and was also stable up to 6 months. Therefore, AG could be considered a natural and potential polymer with profound mucoadhesive properties to deliver drugs through the mucosal route.

Agarose (AG) forms hydrocolloid in hot water and possesses a noteworthy gel strength. However, no reasonable scientific work on investigating the mucoadhesive character of AG has been reported. Therefore, the current study was designed to develop AG and carbopol (CP) based buccal gel scaffold for simultaneous release of benzocaine (BZN) and tibezonium iodide (TIB). Gels’ scaffold formulations (F1–F12) were prepared with varied concentrations (0.5–1.25% w/v) of AG and CP alone or their blends (AG-CP) using homogenization technique. The prepared formulations were characterized for solid-state, physicochemical, in vitro, ex vivo, and in vivo mucoadhesive studies in healthy volunteers. The results showed that mucoadhesive property of AG was concentration dependent but improved by incorporating CP in the scaffolds. The ex vivo mucoadhesive time reached >36 h when AG was used alone or blended with CP at 1% w/v concentration or above. The optimized formulation (F10) depicted >98% drugs release within 8 h and was also storage stable up to six months. The salivary concentration of BZN and TIB from formulation F10 yielded a Cmax value of 9.97 and 8.69 µg/mL at 2 and 6 h (tmax), respectively. In addition, the FTIR, PXRD, and DSC results confirmed the presence of no unwanted interaction among the ingredients. Importantly, the mucoadhesive study performed on healthy volunteers did not provoke any signs of inflammation, pain, or swelling. Clearly, it was found from the results that AG-CP scaffold provided better mucoadhesive properties in comparison to pure AG or CP. Conclusively, the developed AG based mucoadhesive drug delivery system could be considered a potential alternative for delivering drugs through the mucoadhesive buccal route.

The aim of the proposed study is to develop a mucoadhesive buccal delivery system for the sustained delivery of metformin (MET) and sitagliptin (SIT) against diabetes mellitus (DM) with improved bioavailability. Polymeric blend of Carbopol® 940 (CP), agarose (AG) or polyvinylpyrrolidone K30 (PVP) as mucoadhesive agents in formulations (R1–R15) were compressed via the direct compression technique. Tablets were characterized for solid state studies, physicochemical and in vivo mucoadhesion studies in healthy volunteers. Outcomes did not reveal any unusual peak or interaction between the drugs and polymers in the physical mixture through Fourier Transform Infrared Spectroscopy (FTIR) and DSC analysis. The mucoadhesive blend of CP and PVP was superior compared to other blends. The formulation R4 revealed exorbitant loading of drugs with complete drug release for 6 h with ex vivo mucoadhesive strength and time of 26.99 g and 8.1 h, respectively. It was further scrutinized to evaluate it as an optimized formulation where it was found to be stable for up to 6 months. The formulation R4 depicted Korsmeyer–Peppas model and first-order mode of release correspondingly for SIT and MET. Moreover, it showed hemocompatibility, biocompatibility and stability with non-significant changes in the dissolution profile. Overall, the CP blend with PVP was found appropriate to yield the desired release coupled with the optimized mucoadhesive properties of the buccal tablets, ensuring sufficient pharmaceutical stability.

The aim of the projected study was to design and develop a novel strategy for evaluating the mucoadhesive potential of polymeric tablets of dexamethasone (DXM) for local delivery against wounds. Therefore, formulations (Q1–Q7) were synthesized via direct compression method by varying the concentrations of polymers, i.e., ethyl cellulose (EC) and agar extract (AG). Moreover, the mucoadhesive polymeric tablets were characterized via physicochemical, in vitro, ex vivo and in vivo experiments. However, physicochemical characteristics such as FTIR showed no interaction with different polymeric combination. Surface pH of all formulations was normal to slightly alkaline. Highest hydration of up to 6.22% and swelling index was comprehended with maximum concentration of AG (50% of total tablet weight). Whereas, ex vivo and in vivo residence time and mucoadhesion were attributed to the increased concentrations of polymers. Moreover, Q7, (optimized formulation), containing 10% of EC and 40% of AG, exhibited maximum release of DXM (100%) over 8 h, along with sufficient mucoadhesive strength up to 11.73 g, following first-order kinetics having r2 value of 0.9778. Hemostatic effects and epithelialization for triggering and promoting wound healing were highly pronounced in cases of Q7. Furthermore, in vivo residence time was 7.84 h followed by salivary drug concentration (4.2 µg/mL). However, mucoadhesive buccal tablets showed stability for 6 months, thus following the standardization (ICH-Iva) stability zone. In summary, DXM mucoadhesive tablets seem to be an ideal candidate for eradication of wound infections via local targeted delivery.

Abstract Objective: To isolate and characterise multidrug resistant strains of Staphylococcus aureus from healthcare workers who are at potential risk of nosocomial infections. Methods: The observational, cross-sectional study was conducted from November 2014 to April 2015 at different hospitals of Haripur and Abbottabad, Pakistan, and comprised ward and operation theatre staff. The isolates were identified on the basis of microbiological and biochemical tests and further confirmed by polymerase chain reaction. Disc diffusion method was used for antibiotic sensitivity testing, and panton valentine leukocidin and methicillin resistance mecA genes were detected using polymerase chain reaction. Results: Of 208 isolates, 108(52%) were from the ward staff and 100(48%) were from the operation theatre staff. Overall, 167(80.3%) isolates were positive for Staphylococcus aureus, and 75(36%) were methicillin-resistant Staphylococcus aureus. The number of antibiotic-resistant isolates was 75(45%) cefoxitin, 60(36%) ofloxacin, 152(91%) erythromycin, 52(31%) doxycycline, 127(76%) lincomycin, 53(32%) amoxicillin-clavulanate, 67(40%) ciprofloxacin, and 89(53%) ceftriaxone. Conclusion: A high number of hospital staff, including those working in operation theatres, were found to be carrying methicillin-resistant Staphylococcus aureus and multidrug resistant strains in their nasal passage that may be a source of infection to patients. Key Words: Methicillin resistance, Nosocomial infections, Vancomycin, MecA gene, Pvl gene. Continuous....

Staphylococcus aureus is an important healthcare-associated bacterium that causes a multitude of infections in humans such as superficial skin and soft tissue infections, necrotizing pneumonia, foodborne illnesses and postsurgical infections. Treatment of S. aureus infections has become more complicated due to the emergence of Methicillin-Resistant Staphylococcus aureus (MRSA), some of which are multidrug resistant. The present study aimed to characterize S. aureus isolates from a tertiary care hospital in the Rawalpindi district of Pakistan. Staphylococci were isolated from 300 clinical samples collected from January 2018 to January 2019 and S. aureus isolates were tested for antimicrobial susceptibility and analyzed using Pulsed-Field Gel Electrophoresis (PFGE), Multi-Locus Sequence Typing (MLST), staphylococcal cassette chromosome mec (SCCmec) and spa typing. Approximately 25.3% (76/300) of the clinical samples were positive for S. aureus; of those, 88.2% (67/76) were mecA+ (MRSA). In addition to the β-lactam antibiotics, high levels of resistance were also found to the fluoroquinolones (ciprofloxacin, gatifloxacin and levofloxacin (73.7% each)). Of the 23 different spa types identified, the majority of isolates belonged to spa type t632 and t657 (9/66; 13.6% each spa type). ST772-t657 (Bengal Bay clone) was the most commonly identified clone in this study although other clones circulating around different regions of the world were also found indicating the diversity in MRSA isolates from this area of Pakistan. This study emphasizes the need to monitor MRSA in the clinical setting for improved infection control and treatment options.

Jellyfish play a vital role in the ocean’s ecosystem, acting as a nursery for young fishes, crabs, and brittle stars, as well as a source of food for certain charismatic megafauna, such as sunfish and sea turtles. They also pose a threat to human activities, with jellyfish blooms negatively impacting fisheries, power generation, and tourism. However, very little information is available on the biodiversity of jellyfish within the waters of Borneo. Here, we present new records of jellyfish found along the coast of Sabah, Malaysia, located in northern Borneo, bordering the megadiverse region of the Coral Triangle. A total of six species belonging to two classes and six families hitherto not recorded to the state are reported, Chironex yamaguchii, Acromitus maculosus, Crambione mastigophora, Linuche aquila, Netrostoma sp., and Phyllorhiza punctata. Of these, two species (C. yamaguchii and L. aquila) are harmful to humans, with C. yamaguchii capable of causing human fatalities. Reports of harmful jellyfish are useful to the medical and tourism industry, as some of these species may inflict stings and adverse reactions to humans ranging from rashes and skin irritation to fatal envenomation. A checklist has also been provided for all collected jellyfish species from Sabah waters of Borneo, Malaysia.

Brucellosis is a neglected zoonotic disease, and occupations with a high risk of infection exist. Limited information is available on brucellosis for these employees at high risk in Pakistan. A total of 459 persons with high-risk occupations, i.e., 211 abattoir workers, 63 milkers, 52 dung cake makers, 44 veterinarians, 44 shepherds, and 45 veterinary students of the districts Kasur and Lahore, Pakistan, were tested in this study. Blood samples and information on place of residence, gender, age, urbanicity, type of occupation, socioeconomic status, contact with animals, consumption of raw milk, contact to women who had a miscarriage, contact to aborted animal fetus or abortion material, pregnancy, miscarriage history, or intrauterine fetal death were collected. Serum samples were examined using Rose Bengal tests for anti-Brucella antibodies and seropositive samples were subjected to genus-specific qPCR for the detection of DNA. Data were analyzed using chi-squared and binary regression. Twenty (4.35%) persons were seropositive for anti-Brucella antibodies. Out of these, 18 (90%) were tested positive by Brucella genus-specific qPCR. Positive sera were more often found in Lahore district (8.3%) than in Kasur district (3.1%). Persons older than 36 years were more often seropositive. Persons involved in the handing of milk and in contact with animals were more often seropositive. Contact with women who had had a miscarriage was also a significant factor for seropositivity for brucellosis. Place of residence, gender, age, urbanicity, and contact with animals were identified as potential risk factors in the present study. The present study confirmed that brucellosis is an occupation hazard for abattoir workers, milkers, dung cake makers, and veterinarians in the districts of Lahore and Kasur of Punjab, Pakistan. The study shows the need for strategies for safety at work to minimize the risk of infection. Raising awareness for the prevention and use of proper personal protection equipment during the slaughtering and treatment of animal is highly needed.

ST239-MRSA-III is probably the oldest truly pandemic MRSA strain, circulating in many countries since the 1970s. It is still frequently isolated in some parts of the world although it has been replaced by other MRSA strains in, e.g., most of Europe. Previous genotyping work (Harris et al., 2010; Castillo-Ramírez et al., 2012) suggested a split in geographically defined clades. In the present study, a collection of 184 ST239-MRSA-III isolates, mainly from countries not covered by the previous studies were characterized using two DNA microarrays (i) targeting an extensive range of typing markers, virulence and resistance genes and (ii) a SCCmec subtyping array. Thirty additional isolates underwent whole-genome sequencing (WGS) and, together with published WGS data for 215 ST239-MRSA-III isolates, were analyzed using in-silico analysis for comparison with the microarray data and with special regard to variation within SCCmec elements. This permitted the assignment of isolates and sequences to 39 different SCCmec III subtypes, and to three major and several minor clades. One clade, characterized by the integration of a transposon into nsaB and by the loss of fnbB and splE was detected among isolates from Turkey, Romania and other Eastern European countries, Russia, Pakistan, and (mainly Northern) China. Another clade, harboring sasX/sesI is widespread in South-East Asia including China/Hong Kong, and surprisingly also in Trinidad & Tobago. A third, related, but sasX/sesI-negative clade occurs not only in Latin America but also in Russia and in the Middle East from where it apparently originated and from where it also was transferred to Ireland. Minor clades exist or existed in Western Europe and Greece, in Portugal, in Australia and New Zealand as well as in the Middle East. Isolates from countries where this strain is not epidemic (such as Germany) frequently are associated with foreign travel and/or hospitalization abroad. The wide dissemination of this strain and the fact that it was able to cause a hospital-borne pandemic that lasted nearly 50 years emphasizes the need for stringent infection prevention and control and admission screening.

Choosing from Whole Cell and Acellular Pertussis Vaccines- Dilemma for the Developing Countries

The coral reefs at the northernmost tip of Sabah, Borneo will be established under a marine protected area: the Tun Mustapha Park (TMP) by the end of 2015. This area is a passage where the Sulu Sea meets the South China Sea and it is situated at the border of the area of maximum marine biodiversity, the Coral Triangle. The TMP includes fringing and patch reefs established on a relatively shallow sea floor. Surveys were carried out to examine features of the coral reefs in terms of scleractinian species richness, and benthic reef assemblages following the Reef Check substrate categories, with emphasis on hard coral cover. Variation in scleractinian diversity was based on the species composition of coral families Fungiidae (n = 39), Agariciidae (n = 30) and Euphylliidae (n = 15). The number of coral species was highest at reefs with a larger depth gradient i.e. at the periphery of the study area and in the deep South Banggi Channel. Average live hard coral cover across the sites was 49%. Only 7% of the examined reefs had > 75% hard coral cover, while the majority of the reef sites were rated fair (51%) and good (38%). Sites with low coral cover and high rubble fragments are evidence of blast fishing, although the observed damage appeared old. Depth was a dominant factor in influencing the coral species composition and benthic reef communities in the TMP. Besides filling in the information gaps regarding species richness and benthic cover for reef areas that were previously without any data, the results of this study together with information that is already available on the coral reefs of TMP will be used to make informed decisions on zoning plans for conservation priorities in the proposed park.