Campylobacter Sp Research Articles

Assessment of the bacteriological qualities of meat and contact surfaces in markets in Abia State, Nigeria

Background: Microbial contamination of meat comes from external sources during cutting, handling, and processing of the meat. This study was carried out to assess the bacteriological qualities of meat and contact surfaces in markets in Abia State, Nigeria. Methods: This research involved the use of a Hazard Analysis Critical Control Point (HACCP) checklist to investigate the sanitation and hygiene practices of meat sellers and a laboratory study of red and white meat, water, and meat-contact surface samples. A total of 425 meat samples collected from 425 meat sellers from some randomly selected markets in Abia State were used for the study. There were also 20 water samples, 22 samples from table surfaces, 22 samples from knife surfaces, and 14 samples from transport vehicles. The multistage simple random sampling technique through balloting was employed to determine communities/markets for the study. Samples for the study were collected and analyzed using standard microbiological techniques such as culturing and the bacteria were enumerated and identified using biochemical and chemical tests. Results: The prevalent bacterial isolates include Staphylococcus sp (78.80%), Bacillus sp (73.17%), Enterococcus sp (64.00%), Escherichia coli (62.11%), Salmonella sp (62.11%), Klebsiella sp (51.29%), Micrococcus sp (44.94%) and Campylobacter sp (43.52%). SPSS analysis using the one-way ANOVA showed no significant difference (p > 0.05) in bacteria isolated from markets in the three Senatorial Zones of the State. Staphylococcus sp was isolated in 61.11% of the tables, 50.00% of vehicles, 41.67% of knives and 46.32% of water; Salmonella sp was isolated in 47.22% of the tables, 36.11% of vehicles, 30.56% of knives and 43.85% of water; Bacillus sp was isolated in 41.67% of the tables, 44.44% of vehicles, 33.33% of knives and 23.70% of water; Campylobacter sp was isolated in 27.78% of the tables, 25.00% of vehicles, 30.56% of knives and none in water. There was no significant difference (p > 0.05) in bacteria isolated from the contact surfaces and water from the markets in the three zones of the State. Conclusion: The bacteriological quality of meat in markets in Abia State could be said to be poor due to the isolation of Indicator bacteria such as E, coli, Salmonella, and Campylobacter from the studied meat samples. The presence of E. coli in the studied meat samples is an indicator of feacal contamination and a red alert for the Public health sector. It is recommended that meat sellers undergo proper training and regularly update their knowledge of meat safety.

Genomic Characterization and Wetland Occurrence of a Novel Campylobacter Isolate from Canada Geese.

Populations of resident, non-migratory Canada geese are rapidly increasing. Canada geese are known to transmit viral and bacterial diseases, posing a possible threat to human health. The most prevalent pathogens vectored by geese are Campylobacter species, yet the current understanding of the identity and virulence of these pathogens is limited. In our previous study, we observed a high prevalence of Campylobacter spp. in the Banklick Creek wetland-a constructed treatment wetland (CTW) located in northern KY (USA) used to understand sources of fecal contamination originating from humans and waterfowl frequenting the area. To identify the types of Campylobacter spp. found contaminating the CTW, we performed genetic analyses of Campylobacter 16s ribosomal RNA amplified from CTW water samples and collected fecal material from birds frequenting those areas. Our results showed a high occurrence of a Campylobacter canadensis-like clade from the sampling sites. Whole-genome sequence analyses of an isolate from Canada goose fecal material, called MG1, were used to confirm the identity of the CTW isolates. Further, we examined the phylogenomic position, virulence gene content, and antimicrobial resistance gene profile of MG1. Lastly, we developed an MG1-specific real-time PCR assay and confirmed the presence of MG1 in Canada goose fecal samples surrounding the CTW. Our findings reveal that the Canada goose-vectored Campylobacter sp. MG1 is a novel isolate compared to C. canadensis that possesses possible zoonotic potential, which may be of human health concern.

The bacteriome of the oral cavity in healthy dogs and dogs with periodontal disease.

To compare the bacteriome of the oral cavity in healthy dogs and dogs with various stages of periodontal disease. Dogs without periodontal disease (n = 12) or with mild (10), moderate (19), or severe (10) periodontal disease. The maxillary arcade of each dog was sampled with a sterile swab, and swabs were submitted for next-generation DNA sequencing targeting the V1-V3 region of the 16S rRNA gene. 714 bacterial species from 177 families were identified. The 3 most frequently found bacterial species were Actinomyces sp (48/51 samples), Porphyromonas cangingivalis (47/51 samples), and a Campylobacter sp (48/51 samples). The most abundant species were P cangingivalis, Porphyromonas gulae, and an undefined Porphyromonas sp. Porphyromonas cangingivalis and Campylobacter sp were part of the core microbiome shared among the 4 groups, and P gulae, which was significantly enriched in dogs with severe periodontal disease, was part of the core microbiome shared between all groups except dogs without periodontal disease. Christensenellaceae sp, Bacteroidales sp, Family XIII sp, Methanobrevibacter oralis, Peptostreptococcus canis, and Tannerella sp formed a unique core microbiome in dogs with severe periodontal disease. Results highlighted that in dogs, potential pathogens can be common members of the oral cavity bacteriome in the absence of disease, and changes in the relative abundance of certain members of the bacteriome can be associated with severity of periodontal disease. Future studies may aim to determine whether these changes are the cause or result of periodontal disease or the host immune response.

Use of TaqMan Array Cards to investigate the aetiological agents of diarrhoea among young infants with severe acute malnutrition

Studies involving less sensitive conventional microscopy and culture-based approaches have identified distinct differences in diarrhoeal aetiology in childhood malnutrition. Our study involved the use of an advanced molecular biology technique, the TaqMan Array Cards (TAC), to elucidate the diarrhoeal aetiology among young infants with severe acute malnutrition (SAM). A total of 113 faecal samples was collected from SAM infants, aged 2-6months, upon admission to the Dhaka Hospital of the International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b) with complications of diarrhoea and related comorbidities. We used TAC for the detection of 29 different diarrhoeal enteropathogens from a single faecal sample. For comparison, we also analysed 25 diarrhoeal samples from well-nourished infants of similar age. Higher odds of detection of all bacterial enteropathogens were associated with diarrhoea among SAM infants. In particular, the detection of Aeromonas sp (aOR: 25.7, p=0.011), Campylobacter sp (aOR: 9.6, p<0.01) and ETEC (aOR: 5.2, p=0.022) was significantly associated with diarrhoea among SAM infants in comparison to well-nourished infants. 80% higher odds of detection of rotavirus and norovirus GII were associated with diarrhoea among well-nourished infants in comparison to SAM infants (aOR: 0.2, p<0.05). Our study findings demonstrate a difference in diarrhoeal aetiology among SAM and well-nourished young infants, which may be useful in providing an evidence-based logic for possible revision of treatment guidelines for treatment of young diarrhoeal infants with SAM in the early management of the menace of antimicrobial resistance.

ASSESSMENT OF THE ANTIBIOTIC RESISTANCE OF BACTERIAL ISOLATES RECOVERED FROM DOG FEED SOLD IN ADO-EKITI METROPOLIS

Animal feed has been incriminated in many animal infectious diseases. This study was carried out to investigate and document the bacteriological safety of dog feed sold in Ado-Ekiti Metropolis. Five feed samples were analyzed, the bacterial load, coliform count, total Salmonella count as well as test for campylobacter was carried out on the feed samples. Pure culture of the isolates were subjected to antibiotic susceptibility test using disc diffusion method. The total bacterial count ranged between 4.41 log10CFU/g and 6.36 log10CFU/g. All feed samples had coliform ranging between 2.09 log10CFU/g and 3.93 log10CFU/g. Salmonella was only recovered from feed sample DED. Only sample DDL harbours Campylobacter sp. Other bacteria isolated from the feed were Escherichia coli, Bacillus sp, Enterobacter sp, Klebsiella sp, Staphylococcus sp and Lactobacillus sp. The antibiogram showed that nitofuratoin and ciproflocacin had the best activity where 93.10 % (54 out of 58) were susceptible to the two antibiotic. Ampicilin was most resisted with only 36.21 % (21) susceptibility. Thirty one (31) of the isolates showed resistance to 3 or more antibiotic constituting 53.44 % of the entire bacterial isolate. Antibiotic resistance pattern mostly encountered were AMP, AMO, TLY and AMP, AMO, FUR with thirteen (13) isolates each. These results showed that dog feed may not be totally safe for the pets. Based on the type of bacteria isolated and the antibiotic resistance, good manufacturing practices should be ensured by manufactures in other to reduce the rate of contamination.

Comparison of media and growth conditions for culturing enterohepatic Helicobacter species.

This research aims to compare the culturing conditions for enterohepatic Helicobacter, evaluating culture media, incubation atmosphere and susceptibility to antimicrobials used to generate selective conditions. Four common media for the closely related genus Campylobacter (Columbia, Bolton, Brucella and CCDA agar), as well as the need for hydrogen in the microaerobic incubation atmosphere, were evaluated. Serial dilutions of 13 strains belonging to six species (H. apodemus, H. bilis, H. canicola, H. canis, H. equorum and Helicobacter sp.) were inoculated in each media and incubated at 37°C for 48 to 96h using CampyGen (OXOID) and gaseous exchange (including hydrogen) in parallel. Columbia or Brucella agars were the most appropriate for culturing EHH (P<0·05). However, there was no significant difference between the atmospheres evaluated (P=0·13). In addition, minimal inhibitory concentration for six antibiotics showed that all isolates were resistant to trimethoprim, whereas for the rest of the antibiotics (cephalothin, cefoperazone, cefsulodin, teicoplanin and vancomycin) the inhibition range was between 8 and 64μgml- 1 . Our findings suggest that Columbia or Brucella media, regardless of the use of hydrogen, can be used for the EHH isolation. In addition, the concentration of antibiotics included in commercial campylobacteria supplements is suitable for EHH species recovery. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterohepatic Helicobacter (EHH) infections have been associated with several diseases in humans such as acute gastroenteritis, inflammatory bowel disease and hepatobiliary diseases. Although they are frequently detected in clinical samples by molecular methods, only occasionally they are isolated using culture conditions described for the taxonomic related pathogen Campylobacter sp. This is because the optimal conditions for the isolation of EHH have not yet been described, which results in an underestimation of the prevalence and clinical importance of these emerging pathogens. Therefore, this study provides insight for culturing EHH species.

Characterization of the nicotinamide adenine dinucleotides (NAD+ and NADP+) binding sites of the monomeric isocitrate dehydrogenases from Campylobacter species

Monomeric isocitrate dehydrogenases (IDHs) have once been proposed to be exclusively NADP+-specific. Intriguingly, we recently have reported an NAD+-specific monomeric IDH from Campylobacter sp. FOBRC14 (CaIDH). Moreover, bioinformatic analysis revealed at least three different coenzyme-binding motifs among Campylobacter IDHs. Besides the NAD+-binding motif in CaIDH (Leu584/Asp595/Ser644), a typical NADP+-binding motif was also identified in Campylobacter corcagiensis IDH (CcoIDH, His582/Arg593/Arg638). Meanwhile, a third putative NAD+-binding motif was found in Campylobacter concisus IDH (CcIDH, Leu580/Leu591/Ala640). In this study, CcIDH was overexpressed in Escherichia coli and purified to homogeneity. Gel filtration chromatography demonstrated that the recombinant CcIDH exists as a monomer in solution. Kinetic analysis showed that the Km value of CcIDH for NADP+ was over 49-fold higher than that for NAD+ and the catalytic efficiency (kcat/Km) of CcIDH is 115-fold greater for NAD+ than NADP+. Thus, CcIDH is indeed an NAD+-specific enzyme. However, the catalytic efficiency (kcat/Km = 0.886 μM−1 s−1) of CcIDH for NAD+ is much lower (<5%) when compared to that of the typical monomeric NADP-IDHs for NADP+. Then, the three core NAD+-binding sites were evaluated by site-directed mutagenesis. The mutant CcIDH (H580R591R640) showed a 51-fold higher Km value for NAD+ and 21-fold lower Km value for NADP+ as compared to that of the wild type enzyme, respectively. The overall specificity of the mutant CcIDH was 12-fold greater for NADP+ than that for NAD+. Thus, the coenzyme specificity of CcIDH was converted from NAD+ to NADP+. Isocitrate dependence of enzyme kinetics showed that although the mutant H580R591R640 preferred NADP+ as its coenzyme, its catalytic efficiency for isocitrate reduced to half of that for the wild-type CcIDH as using NAD+. The finding of both NAD+ and NADP+-binding sites in monomeric IDHs from Campylobacter species will provide us a chance to explore the evolution of the coenzyme specificity in monomeric IDH subfamily.

Ocurrence of Campylobacter spp. in chilled chicken carcasses slaughtered in the west region of Santa Catarina, Brazil

Background: The thermophilic bacteria of the genus Campylobacter are important agents of alimentary gastroenteritis, called campylobacteriosis. These microorganisms multiply in temperatures ranging from 25ºC to 46ºC, however, low temperatures are incompatible with their multiplication. For this reason, the seasons of the year may interfere with the level of contamination by Campylobacter sp. The main sources of transmission are contaminated meat and giblets from poultry during poorly conducted slaughter operations. The disease may present itself with a different range of forms of disease: from mild signs of gastrointestinal infection to more severe cases, such as the Guillain-Barré syndrome.Material, Methods &amp; Results: Due to the great importance of western Santa Catarina to the poultry industry, it was necessary to verify the occurrence of the pathogen in cold carcasses of broilers slaughtered in this region, and its variation through the seasons of the year. From January 2013 to February 2015 broiler carcasses were collected weekly, after the water cooling process, in slaughterhouses under Federal Inspection of the three largest microregions of western Santa Catarina in terms of number of broilers slaughtered, totaling 808 samples. The assessment of thermophilic Campylobacter was performed according to the methodology recommended by ISO 10272-1: 2006. Of the 808 samples analyzed, the frequency of isolation of thermophilic Campylobacter was 1.82% (8/440) in microregion 1, 4.95% (10/202) in microregion 2 and 13.86% (23/166) in the microregion 3, totaling 5.07% of positive samples (41/808). Comparing the microregions, it was verified that there was no significant difference (P &gt; 0.05) between the isolation frequencies of microregions 1 and 2. However, there was a significant difference (P &lt; 0.05) between the isolation rates of Microregion 3 in relation to microregions 1 and 2. Microregion 1 presented the lowest percentage of Campylobacter thermophilic isolates.Discussion: Although the rates found are lower than expected when compared to those already published, consumption of these products may provide risks to consumers, mainly through cross-contamination of foods that will be consumed raw, requiring greater controls in the entire poultry industry in order to ensure a safe product for the consumer. Analyzing each season of the year individually (winter and summer), 5.88% (14/238) of the samples processed during summer (October to March) were positive for the assessed microorganism and in 4,74% (27/570) of the samples analyzed in the winter (April to September), the presence of thermophilic Campylobacter was detected. Nevertheless, there was no significant difference (P &gt; 0.05) between the seasons. However, it is evident the need for a longer study to evaluate the actual influence of the seasons (winter and summer) on the isolation rate of thermophilic Campylobacter in broiler carcasses shortly after the cooling process. Even though the study demonstrated low levels of thermophilic Campylobacter isolation rates in the evaluated regions, just the fact of isolating the agent already shows that there is an imminent risk to consumers, mainly due to cross-contamination of foods that will be consumed raw; therefore, higher standards of microbiological controls should be practiced in the production chain and slaughter of poultry, respecting good manufacturing practices, hygiene and handling practices within the industry.

Exploring the microbiome of healthy and diseased peri‐implant sites using Illumina sequencing

To compare the microbiome of healthy (H) and diseased (P) peri-implant sites and determine the core peri-implant microbiome. Submucosal biofilms from 32 H and 35 P sites were analysed using 16S rRNA sequencing (MiSeq, Illumina), QIIME and HOMINGS. Differences between groups were determined using principal coordinate analysis (PCoA), t tests and Wilcoxon rank sum test and FDR-adjusted. The peri-implant core microbiome was determined. PCoA showed partitioning between H and P at all taxonomic levels. Bacteroidetes, Spirochetes and Synergistetes were higher in P, while Actinobacteria prevailed in H (p<.05). Porphyromonas and Treponema were more abundant in P while Rothia and Neisseria were higher in H (p<.05). The core peri-implant microbiome contained Fusobacterium, Parvimonas and Campylobacter sp. T.denticola, and P.gingivalis levels were higher in P, as well as F.alocis, F.fastidiosum and T.maltophilum (p<.05). The peri-implantitis microbiome is commensal-depleted and pathogen-enriched, harbouring traditional and new pathogens. The core peri-implant microbiome harbours taxa from genera often associated with periodontal inflammation.

The microbiome of oral leukoplakia shows enrichment in Fusobacteria and Rothia species

ABSTRACTThe current study was carried out to determine if changes in the oral microbiome were associated with oral leukoplakia. Swabs of oral leukoplakias were taken from 36 patients. Contralateral normal tissue was also swabbed. Swabs from 35 control patients without symptoms of leukoplakia were also taken. DNA was extracted and the V1V2 region of the 16s rRNA gene was sequenced using the Illumina MiSeq and analysed using the Mothur software package.The structure of oral mucosal communities was most affected by smoking and the location of the site (AMOVA p < 0.01). Analysis of the constituents of these communities using LEfSe showed that Fusobacterium sp. and Leptotrichia sp. were enriched on leukoplakia sites. Patients with leukoplakia also showed enrichment for Rothia mucilaginosa and Campylobacter sp. Quantitative RT-PCR also showed that leukoplakias from lingual sites were more likely to be colonised by Candida sp. Analysis of these enrichments identified specific co-localisation patterns (Pearson correlation P <0.01) including Leptotrichia sp., Prevotella sp. and Campylobacter concisus; F. nucleatum, Alloprevotella tannerae and C. gracilis, amongst others.Fusobacteria have been implicated in the progression of colorectal carcinoma and further studies are now required to determine if these microorganisms are linked to the development of OSCC.

Inflammatory bacteriome featuring Fusobacterium nucleatum and Pseudomonas aeruginosa identified in association with oral squamous cell carcinoma

Studies on the possible association between bacteria and oral squamous cell carcinoma (OSCC) remain inconclusive, largely due to methodological variations/limitations. The objective of this study was to characterize the species composition as well as functional potential of the bacteriome associated with OSCC. DNA obtained from 20 fresh OSCC biopsies (cases) and 20 deep-epithelium swabs (matched control subjects) was sequenced for the V1-V3 region using Illumina’s 2 × 300 bp chemistry. High quality, non-chimeric merged reads were classified to species level using a prioritized BLASTN-algorithm. Downstream analyses were performed using QIIME, PICRUSt, and LEfSe. Fusobacterium nucleatum subsp. polymorphum was the most significantly overrepresented species in the tumors followed by Pseudomonas aeruginosa and Campylobacter sp. Oral taxon 44, while Streptococcus mitis, Rothia mucilaginosa and Haemophilus parainfluenzae were the most significantly abundant in the controls. Functional prediction showed that genes involved in bacterial mobility, flagellar assembly, bacterial chemotaxis and LPS synthesis were enriched in the tumors while those responsible for DNA repair and combination, purine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, ribosome biogenesis and glycolysis/gluconeogenesis were significantly associated with the controls. This is the first epidemiological evidence for association of F. nucleatum and P. aeruginosa with OSCC. Functionally, an “inflammatory bacteriome” is enriched in OSSC.

Antimicrobial activity of Zatacin against bacterial diarrheal pathogens

Background: Calf diarrhea is an important disease that is caused by different pathogens including bacteria, virus and parasites and is associated with economic losses. Objectives: In this study, we evaluated the antibacterial activities of Zatacin (Z. multiflora aerial part ethanol extract), colistin, co-Trimoxazole and enrofloxacin against clinical isolates of Salmonella sp, E. coli and Campylobacter sp isolated from diarrheic calves. Methods: Disc diffusion method and microbroth dilution assay were used for antimicrobial evaluation. Results: In disc diffusion method, the antibacterial activity of Zatacin increased dose dependently. The sensitivity of different isolates of E. coli, Salmonella sp and Campylobacter sp to Zatacin was almost the same. The antibacterial activity of Zatacin was lower than that of enrofloxacin but it was higher than co-Trimoxazole and colistin. The means of MIC values of Zatacin for E.coli were higher than that of Campylobacter sp and Salmonella sp but its means of MBC values for E. coli were lower than that of two other bacteria. Conclusions: Zatacin can be used as an antimicrobial agent in treatment of infectious causes of calf scours instead of antibiotics with undesired adverse effects on animal and humans.

English

The present study aimed to evaluate the efficiency of the irradiation process in the control of Campylobacter spp. in chilled chicken heart samples, since this microorganism is related to the contamination of meat and chicken giblets and is responsible for enteritis in humans. The methodology and standards recommended by RDC no. 12 (Brazil, 2001) were applied for the bacteriological analyses. The chilled chicken heart samples were acquired in an industry that undergoes sanitary inspection, located in the West Zone of Rio de Janeiro. Samples were divided into two groups, non-contaminated (NC - originally from the industrial plant) and contaminated (CAMPY– contaminated with C. jejuni ATCC 33291/CCAMP/FIOCRUZ 00262 strains by CCAMP/LABZOO/IOC/FIOCRUZ), subsequently separated into four groups: NC and CAMPY control groups and samples irradiated at 1.5, 3.0 and 4.5 kGy. The eight subgroups were analyzed for the presence of Campylobacter sp. No statistically significant difference was observed between the four groups namely the non-irradiated controls and the 1.5, 3.0 and 4.5 kGy irradiated samples (p> 0.05). Elimination of Campylobacter sp. was observed, with no bacterial growth in any of the irradiated, non-contaminated (CN) and infected (CAMPY) samples. Thus, the efficiency of the Co60 irradiation process of chilled chicken heart for the elimination of the surveyed microorganisms was proven. The lowest dose applied was sufficient to eliminate the enteric pathogen which is of great significance in a public health point of view. However, it should be noted that the Brazil legislation determining the microbiological standards for food does not include a microbiological standard for Campylobacter sp. This means that any amount of this enteric pathogen may bring public health risks. Key words: Food irradiation; chicken heart; Campylobacter sp.; Foodborne illness; public health.

Un cas d’encéphalomyélite aiguë disséminée post-infectieuse découvert devant une rétention vésicale fébrile

IntroductionMRI should be performed in the presence of an acute febrile urinary retention, when septic and obstructive causes are eliminated. We report a case of post-infectious probable acute disseminated encephalomyelitis (ADEM) with a mostly spinal cord tropism of involving Campylobacter. Case reportA 32-year-old man with no medical history was admitted for an acute febrile urinary retention. He reported severe diarrhea 3 days before. Clinical course was then complicated by a progressive tetraparesis predominating in the lower limbs. Medullar MRI showed thoracic myelitis. A five-day course of intravenous corticosteroids allowed a full recovery of both the motor and urinary symptoms. Fecal culture isolated Campylobacter sp. Final diagnosis was post-bacterial ADEM. ConclusionClinical findings and MRI allow clinicians to suspect acute disseminated encephalomyelitis. This hypothesis implies to actively look for recent infections or vaccinations preceding the clinical presentation.

Isolation of a novel thermophilic Campylobacter from cases of spotty liver disease in laying hens and experimental reproduction of infection and microscopic pathology

The condition known as spotty liver disease or spotty liver syndrome can cause significant mortality in free range laying hen flocks. It has been described in Europe and Australia but the aetiology has not been established. There are similarities between spotty liver disease and avian vibrionic hepatitis, a condition which was reported in the 1950s. A Vibrio-like organism was suspected to be the cause of avian vibrionic hepatitis, although this organism was never fully characterised.We report the isolation of a novel Campylobacter from five separate outbreaks of spotty liver disease. The conditions required for culture, the growth characteristics, electron microscopical morphology and results of the phenotypic tests used in the identification of this novel Campylobacter sp. are described. The novel Campylobacter is slow growing and fastidious and does not grow on media routinely used for isolating Campylobacter sp. The morphology is typical for a Campylobacter sp. and phenotypic tests and a duplex real time PCR test differentiate the novel Campylobacter from other members of the genus. 16S rRNA analysis of 19 isolates showed an identical sequence which appears to represent a hitherto unknown sub lineage within the genus Campylobacter. Experimental intraperitoneal infection of four week old SPF chickens produced microscopic liver pathology indistinguishable from natural disease and the novel Campylobacter was recovered from the experimentally infected chicks. The isolates described appear to be a possible causal organism for spotty liver disease.

Novel type II and monomeric NAD+ specific isocitrate dehydrogenases: phylogenetic affinity, enzymatic characterization, and evolutionary implication.

NAD+ use is an ancestral trait of isocitrate dehydrogenase (IDH), and the NADP+ phenotype arose through evolution as an ancient adaptation event. However, no NAD+-specific IDHs have been found among type II IDHs and monomeric IDHs. In this study, novel type II homodimeric NAD-IDHs from Ostreococcus lucimarinus CCE9901 IDH (OlIDH) and Micromonas sp. RCC299 (MiIDH), and novel monomeric NAD-IDHs from Campylobacter sp. FOBRC14 IDH (CaIDH) and Campylobacter curvus (CcIDH) were reported for the first time. The homodimeric OlIDH and monomeric CaIDH were determined by size exclusion chromatography and MALDI-TOF/TOF mass spectrometry. All the four IDHs were demonstrated to be NAD+-specific, since OlIDH, MiIDH, CaIDH and CcIDH displayed 99-fold, 224-fold, 61-fold and 37-fold preferences for NAD+ over NADP+, respectively. The putative coenzyme discriminating amino acids (Asp326/Met327 in OlIDH, Leu584/Asp595 in CaIDH) were evaluated, and the coenzyme specificities of the two mutants, OlIDH R326H327 and CaIDH H584R595, were completely reversed from NAD+ to NADP+. The detailed biochemical properties, including optimal reaction pH and temperature, thermostability, and metal ion effects, of OlIDH and CaIDH were further investigated. The evolutionary connections among OlIDH, CaIDH, and all the other forms of IDHs were described and discussed thoroughly.

Co-infection of the Siberian hamster (Phodopus sungorus) with a novel Helicobacter sp. and Campylobacter sp.

We report the isolation of a novel helicobacter isolated from the caecum of the Siberian hamster (Phodopus sungorus). Sequence analysis showed 97% sequence similarity to Helicobacter ganmani. In addition, we report the co-infection of these Siberian hamsters with a Campylobacter sp. and a second Helicobacter sp. with 99% sequence similarity to Helicobacter sp. flexispira taxon 8 (Helicobacter bilis), a species isolated previously from patients with bacteraemia. Gross necropsy and histopathology did not reveal any overt pathological lesions of the liver and gastrointestinal tract that could be attributed to the Helicobacter or Campylobacter spp. infections. This is the first helicobacter to be identified in the Siberian hamster and the first report of co-infection of Helicobacter spp. and Campylobacter sp. in asymptomatic Siberian hamsters.

Efek Dekontaminasi Karkas Ayam Pedaging Menggunakan Asam Asetat, Asam Sitrat dan Kombinasinya Terhadap Angka Lempeng Total Campylobacter sp.

(Effect of poultry carcas decontamination by acetic acid, citric acid and its combination to total plate count of Campylobacter sp).ABSTRACT. This research aimed to detect the total number of Campylobacter sp. on poultry carcass after decontamination by acetic acid, citric acid and combination of both. This research was conducted in the Laboratory of Veterinary Public Health Syiah Kuala University, Banda Aceh. The research was factorial completely randomized designed. Samples of poultry carcass were obtained from the Lamnyong market, Banda Aceh. Sixty poultry carcasses were divided into three groups of treatment and one group without treatment. Observation of each treatment was five time replicated at 0, 2, 4, 6 and 8 hours after immersion for ± 30 seconds in each of the decontamination material. Observation of Campylobacter sp. was done by inoculating every sample on CM0739 selective Campylobacter Blood free selective agar base and CCDA Selective supplement SR0155E. Measurements of Total Plate Count (TPC) Campylobacter sp according to Standard Plate Count (SPC). Data was variance analyzed (ANOVA) using SPSS. The results of research showed that the growing colonies of Campylobacter sp. were indicated by white colony with surrounded by black zone. Immersion of poultry carcass with acetic acid, citric acid and combination of the both and the observation time had no influence (P0,05) to ALT Campylobacter sp. The research was concluded that carcass to Campylobacter sp. The immersion by acetic acid, citric acid and combination of both and the observation time were not reduce Campylobacter sp. in poultry carcass.

Antimicrobial resistance of bacterial enteropathogens isolated from stools in Madagascar

BackgroundDiarrheal diseases are a major public health problem in developing countries, and are one of the main causes of hospital admissions in Madagascar. The Pasteur Institute of Madagascar undertook a study to determine the prevalence and the pathogenicity of bacterial, viral and protozoal enteropathogens in diarrheal and non-diarrheal stools of children aged less than 5 years in Madagascar. We present here the results of the analysis of antimicrobial susceptibility of the bacteria isolated during this study.MethodsThe study was conducted in the community setting in 14 districts of Madagascar from October 2008 to May 2009. Conventional methods and PCR were used to identify the bacteria; antimicrobial susceptibility was determined using an agar diffusion method for enterobacteriaceae and MICs were measured by an agar dilution method for Campylobacter sp. In addition to the strains isolated during this study, Salmonella sp and Shigella sp isolated at the Pasteur Institute of Madagascar from 2005 to 2009 were included in the analysis to increase the power of the study.ResultsTwenty-nine strains of Salmonella sp, 35 strains of Shigella sp, 195 strains of diarrheagenic E. coli, 203 strains of C. jejuni and 71 strains of C. coli isolated in the community setting were tested for antibiotic resistance. Fifty-five strains of Salmonella sp and 129 strains of Shigella sp isolated from patients referred to the Pasteur Institute of Madagascar were also included in the study. Many E. coli and Shigella isolates (around 80%) but fewer Salmonella isolates were resistant to ampicillin and trimethoprim/sulfamethoxazole. A small proportion of strains of each species were resistant to ciprofloxacin and only 3% of E. coli strains presented a resistance to third generation cephalosporins due to the production of extended-spectrum beta-lactamases. The resistance of Campylobacter sp to ampicillin was the most prevalent, whereas less than 5% of isolates were resistant to each of the other antibiotics.ConclusionThe highest prevalence of antimicrobial resistance was to ampicillin and trimethoprim/sulfamethoxazole. Antibiotic treatment is not recommended for children with diarrhea in Madagascar and the emphasis should be placed on oral rehydration.

Persistence and Biofilm Assessment of Campylobacter Jujeni in Poultry Abattoir

Persistence of Campylobacter sp and its biofilm forming ability was assessed in two poultry abattoirs at two weeks intervals. Average prevalence (63.75%) of Campylobacter spp. was observed on assessing a total of 160 samples collected from the surfaces of packaging table (80%), dressing table (75%), floor source (70%) and washing table (30%). Biofilm assessment formed by Campylobacter jejuni within 5-days at 37°C were in decreasing order of washing table > packaging table > dressing table > floor. An average rate (19.6%) of isolates to develop biofilm observed in both sites was considered relatively low. Absorbance value (Optical Density-OD 590nm ) of formed biofilms ranged from 0.483 – 0.952. Wastewater from the facilities showed higher TDS (643 – 820 mgl −1 ), TSS (1200 – 1775 mgl −1 ), COD (152 – 141 mgl −1 ) and BOD (30.3 – 32.5mgl −1 ) than the WHO standards of 500 mgl −1 , 100 mgl −1 , 10 mgl −1 and 6 mgl −1 respectively. This is a clear indication of heavy microbial presence in the wastewater. Total bacterial count (TBC) was slightly higher in site A (4.4 × 10 5 CFU/ml) than site B (3.5 × 10 5 CFU/ml). Efficiency index ratio (≈/> 1) observed in all tested drugs suggests their effectiveness in campylobacteriosis management. Decreasing drug sensitivity pedigree was observed with streptomycin > erythromycin & gentamincin > tetracycline & neomycin > penicillin > riphapicin & ampicillin > norflaxicin & cephalexin. These results of frequency and biofilm forming tendencies of Campylobacter spp. observed in this study can be of value in checkmating campybacteriosis outbreak from poultry abattoir facility.