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In this study, a total of 228 raw meat samples (pork: 76, beef: 76, chicken: 76) and 301 raw seafood samples (fish: 199, shrimp: 67, squid: 35) were collected randomly at traditional markets in Ho Chi Minh City (Vietnam). In meat, the ratio of Salmonella spp. was 70.61% (161/228). Among the contaminated meat samples, pork was infected with a ratio of 90.79 % (69/76) while the contamination ratios in beef and chicken were 43.42% (33/76) and 77.63% (59/76), respectively. Salmonella contamination was detected in fish (40.20%), shrimp (7.46%) and squid (17.14%). Because of sulfonamide group is used in Salmonella treatment, the study focused on sulfonamide resistance. In fresh seafood comparison, there were 32.56%, 40% and 10% Salmonella showing resistance to sulfamethoxazol in fish, squid and shrimp, respectively. In fresh meat comparison, there were 31.58%, 16.67% and 55.56% Salmonella showing resistance to sulfamethoxazol in pork, beef and chicken, respectively. Interestingly, there were 21 serovars including 19 identified serovars including S. Kentucky (8), S. Agona (2), S. Infanis (4), S. Saintpaul (1), S. Indiana (1), S. Braenderup (1), S. Potsman (2) and 2 unidentified serovars showing different phenotype to this antibiotic. Among the 21 serovars, only 23.81% strains carried both genes (sul1, sul2). For the sul1 gene, 61.9% strains were presented while sul2 occupied at a lower rate than sul1 with the rate of 52.38%. The study is very interesting and useful to go more functional analysis in sulfonamide resistance.

Background and Aim: For understanding the epidemiology of leptospirosis, the confined abundance of several species of pathogenic leptospires and knowledge on the serovar(s) prevalent in the reservoir and carrier hosts may be a useful indicator of transmission to incidental/accidental hosts in a geographical niche. The present study was carried out to ascertain the frequency distribution of Leptospira serovars and the prevalence of anti-leptospiral antibodies in small ruminants (sheep and goats) in the epidemiological units (villages) in the coastal districts of enzootic regions in South Peninsular India. Materials and Methods: A total of 1167 serum samples (sheep n=299 and goats n=868) from apparently healthy animals, randomly collected from various epidemiological units were tested in microscopic agglutination test (MAT) using 18 reference Leptospira serovars antigens. Results: The overall seroprevalence of 40% (at 95% confidence intervals [CI]: 36.82-42.43) in small ruminants (44% [95% CI: 40.49-52.26] in sheep and 38% [95% CI: 34.96-41.41] in goats) was observed with the predominance of Icterohaemorrhagiae, Javanica, Australis, Hurstbridge, and Pyrogenes serogroup anti-leptospiral antibodies in the studied region. The Chi-squared test revealed that the presence of anti-leptospiral antibodies is significantly not independent (associated) across the administrative division (Chi-square=105.80, p

Introduction Leptospirosis is a bacterial disease that affects both humans and animals, the occurrence of which increases markedly during and after heavy rainfall and flooding. The aim of this study was to determine the serological prevalence of leptospiral infection in livestock after a voluminous flood in 10 districts of the Malaysian state of Kelantan. Material and Methods In December 2014, Kelantan was hit by an extensive flood. A total of 1,728 serum samples were collected from livestock from the state, comprised of 1,024 from cattle, 366 from goats and 338 from sheep, and they were tested using the microscopic agglutination test (MAT). Results Altogether, 203 (11.75%; 203/1728; 95% CI: 10.20%–13.30%) of the tested sera were found to be positive serologically. Cattle had the highest prevalence of 14.16% (145/1024), while goats and sheep had 11.20% (41/366) and 5.03% (17/338) respectively. The most frequent serovars detected were Hardjo-bovis (3.70%; 64/1728), Hebdomadis (2.08%; 36/1728) and Pomona (1.04%; 18/1728). There was a statistically significant association (P < 0.05) between livestock that were exposed to the flood and seropositivity. Conclusion This study showed that flood is a risk factor that can play a role in the epidemiology of leptospiral infection in livestock.

Leptospirosis is an acute, subacute and chronical contagious disease of animals and humans. Causative agents of this disease belong to the genus Leptospira, family Leptospiraca. As a disease of wild animals, leptospirosis is widespread through Europe. Certain wild animals (rodents, fox and wild boars) are important reservoirs and highly probable vectors for the spread of infection into domestic animals and humans. During the hunting season, hunting dogs are often in direct or indirect contact with wild animals that could be carriers of this disease, and the possibility of appearance and spreading within this cohort of dogs is very high. The main reasons for this study on the prevalence of Leptospirosis in Bosnia and Herzegovina are the regular contact between hunting dogs and wild animals (carriers), and the lack of data about Leptospirosis in hunting dogs. In total, 175 serum samples from 15 towns of Bosnia and Herzegovina were tested. Twelve serovars of L. interrogans were used in the microscopic agglutination test. Presence of antibodies of four serovars was confirmed. Prevalence of seropositive dogs was 15.4% (27/175). Most positive dogs had a reaction to the Pomona serovar 51.8% (n=14), while the prevalence of the Sejroe serovar was 33.3%, Icterohaemorrhagiae serovar 11.1% and Bratislava serovar 3.7%. The highest number of positive reactions 55.5% (n=15) was with serum dilution of 1:200. This study showed that most infections in dogs were caused by serovars that are currently not included in commercial vaccines. One of the most efficient preventive measure could be vaccination with the serovars most often found in wild animals, as they appear to be the most common source of the infection., Leptospiroza je akutna, subakutna i kronična zarazna bolest životinja i ljudi. Uzročnici ove bolesti pripadaju rodu Leptospira, porodici Leptospiraca. Leptospiroza kao bolest divljih životinja široko je rasprostranjena u Europi. Određene divlje životinje (glodavci, lisice i divlje svinje) kao važan rezervoar i vrlo vjerojatan vektor zaraze proširile su bolest na domaće životinje i ljude. Lovački psi tijekom sezone lova često su u direktnom ili indirektnom kontaktu s divljim životinjama koje bi mogle biti nositelji ove bolesti, a mogućnost pojave i širenja unutar kategorije takvih pasa vrlo je velika. Glavni razlozi ove studije o učestalosti leptospiroze na području Bosne i Hercegovine su redoviti kontakti lovačkih pasa i divljih životinja (nositelji) i nedovoljni podatci o prisutnosti leptospiroze u kategoriji lovačkih pasa. Ukupno je istraženo 175 uzoraka seruma iz 15 gradova Bosne i Hercegovine. U mikroskopskom testu aglutinacije korišteno je dvanaest serovara L. Interrogans. Potvrđena je prisutnost antitijela na četiri serovara. Prevalencija seropozitivnih pasa bila je 15,4 % (27/175). Većina pozitivnih pasa imala je reakciju na serovar Pomona 51,8 % (n=14), dok je u serovaru Sejroe prevalencija 33,3 %, serovar Icterohaemorrhagiae 11,1 % i serovar Bratislava 3,7 %. Najveći broj pozitivnih reakcija bio je s razrjeđivanjem seruma od 1:200 ili 55,5 % (n=15). Studije pokazuju da su većinu infekcija u pasa prouzročili serovari koji nisu uključeni u komercijalna cjepiva. Jedna od najučinkovitijih preventivnih mjera mogla bi biti cijepljenje sa serovarovima koji se najčešće nalaze u divljih životinja jer su one najčešći izvor zaraze.

Salmonella enterica is a major causative pathogen of human and animal gastroenteritis. Antibiotic resistant strains have emerged due to the production of extended-spectrum β-lactamases (ESBLs) posing a major health concern. With the increasing reports on ESBL-producing Enterobacterales that colonize companion animals, we aimed to investigate ESBL dissemination among ESBL-producing Salmonella enterica (ESBL-S) in hospitalized horses. We prospectively collected ESBL-S isolates from hospitalized horses in a Veterinary-Teaching Hospital during Dec 2015–Dec 2017. Selection criteria for ESBL-S were white colonies on CHROMagarESBL plates and an ESBL phenotypic confirmation. Salmonella enterica serovars were determined using the Kaufmann-White-Le-Minor serological scheme. ESBL-encoding plasmids were purified, transformed and compared using restriction fragment length polymorphism (RFLP). Whole genome sequencing (Illumina and MinION platforms) were performed for detailed phylogenetic and plasmid analyses. Twelve ESBL-S were included in this study. Molecular investigation and Sequence Read Archive (SRA) meta-analysis revealed the presence of three unique Salmonella enterica serovars, Cerro, Havana and Liverpool, all reported for the first time in horses. PFGE revealed the clonal spread of S. Cerro between seven horses. All twelve isolates carried blaCTX–M–3 and showed an identical multidrug resistance profile with co-resistance to trimethoprim/sulfamethoxazole and to aminoglycosides. Plasmid RFLP proved the inter-serovar horizontal spread of a single blaCTX–M–3-encoding plasmid. Complete sequence of a representative plasmid (S. Havana strain 373.3.1), designated pSEIL-3 was a -86.4 Kb IncM2 plasmid, that encoded nine antibiotic resistance genes. pSEIL-3 was virtually identical to pCTX-M3 from Citrobacter freundii, and showed high identity (>95%) to six other blaCTX–M–3 or blaNDM–1 IncM2 broad host range plasmids from various Enterobacterales of human origin. Using a specific six gene-based multiplex PCR, we detected pSEIL-3 in various Enterobacterales species that co-colonized the horses’ gut. Together, our findings show the alarming emergence of ESBL-S in hospitalized horses associated with gut shedding and foal morbidity and mortality. We demonstrated the dissemination of CTX-M-3 ESBL among different Salmonella enterica serovars due to transmission of a broad host range plasmid. This report highlights horses as a zoonotic reservoir for ESBL-S, including highly transmissible plasmids that may represent a ‘One-Health’ hazard. This risk calls for the implementation of infection control measures to monitor and control the spread of ESBL-S in hospitalized horses.

This study was conducted to evaluate the degree of biosecurity level with especial reference toE. coliSppas an example to explain the expected causes and risk factors that leads to spread them in poultry flocks in Egypt and to evaluate its sensitivity to most common disinfectants used in Egypt. About 300 samples (100 cloacal swabs, 100 liver and intestinal samples,100 litter samples) were collected from 10 broiler farms with different age (at 0 old day, one week,2,4 and 6 weeks of age). The samples were investigated forE. coli Sppand subsequently identified based on biochemical and serological tests. The obtained results showed that 44 isolates were isolated (27±1.99%) ;(11±0.42%) and (7±0.72%) from cloacal swab; liver and litter, respectively. Mean prevalence ofE. coli spp.was 15±1.22%.E. coliserotypes were: O78 (31.81%), O2: H6 (18.18), O1: H7 (15.9), O91: H21(11.36), O128: H2 (9.09), O26: H11(4.54) O146: H21, O124, O44: H18 and O153: H2. The most common serovars were O124, O44: H18 and O153: H2 (2.27%). In absence of organic matter; there was great statistical significant difference in the sensitivity ofE. colito the most common disinfectants(Pas Verkon- S®achieved 3 log reduction,after 5-minute,Formalin and Phenique were achieved 4 log and 3log reductions respectively,Aldekol Des- Gda®achieved 4 log reduction, TH4®,Biosentry®904 and Iodophore achieved 2 log reductions after 5 minutes.

Multiplex Polymerase Chain Reaction for amplification of virulent genes was carried out onSalmonella serovarsisolated. All theSalmonella serovarsisolated and identified contained plasmids which were identified using replicon typing.The plasmids contained genes coding for virulence factors and multiple antibiotic resistance which include: ParcC (480bp), GyrA (251bp), CatA (198bp) were present the latter coded for chloramphenicol resistance. InvA plasmid genes (284bp and 389bp) typical of virulentSalmonella serovarswere identified. Most of the serovars possessed plasmids that contained TetB gene in the 16sRNA region (571bp) coding for tetracycline resistance, SitC gene (578bp) and spvA gene (604bp) were also identified. Sex distribution dynamics of multidrug resistance of differentSalmonella serovarscompared the overall percentage susceptibility of the eight antibiotics used. GEN had the highest % susceptibility of 100% followed by NIT (72.2%) and COT (66.7%) before and after plasmid curing. Differences observed in curing status was insignificant (T = 0.33, P>0.05). The sex distribution ofSalmonellainfected Patients in relation to antibiotic resistance before plasmid curing was determined. The highest number of patients that resisted antibiotic was 18 as observed in AUG where 11 patients were male and 7 were female.More males compared to females showed significant antibiotic resistance before plasmid curing using the Mann whitney test (W = 84.0, P≤0.05 and after plasmid curing.There was no difference between both sexes after plasmid curing using the Mann Whitney test (W = 73.0, P>0.05).

Background & objectives: Salmonellosis due to the consumption of contaminated poultry products is a well-known public health concern, and assessing the distribution of Salmonella serovars among poultry becomes important for better prevention and control. The objective of the present study was to assess the distribution of Salmonella serovars among poultry. Methods: The isolates received at National Salmonella and Escherichia Centre during 2011-2016 were subjected to biochemical identification, followed by serological characterization to identify the Salmonella serovars, and the data were presented to exhibit the distribution of Salmonella serovars among poultry. Results: Salmonella was found to be present in poultry in all the regions included in the study. Salmonella Typhimurium, S. Gallinarum and S. Enteritidis were the most prevalent serovars accounting for 96.2 per cent of isolates. Salmonella was identified in poultry from all major egg-producing and egg-consuming States. Other serovars which were scantly identified included S. Infantis (2.7%), S. Montevideo (0.64%), S. Newport (0.26%) and S. Pullorum (0.13%). Interpretation & conclusions: Diverse distribution of Salmonella serovars in poultry in India, with known potential to infect human population and/or other poultry flocks, requires urgent nationwide stringent control measures.

Salmonella has been known as the most important foodborne pathogen, which can infect humans via consuming contaminated food. Chicken meat has been known as an important vehicle to transmit Salmonella by the food supply chain. This study determined the prevalence, antimicrobial resistance, and genetic characteristics of Salmonella at different chicken slaughtering stages in East China. In total, 114 out of 200 (57%) samples were Salmonella positive, while Salmonella contamination was gradually increasing from the scalding and unhairing stage (17.5%) to the subdividing stage (70%) throughout the slaughtering. Whole-genome sequencing (WGS) was then performed to analyze the serotype, antimicrobial resistance gene profiles, and genetic relationship of all Salmonella isolates. The most common serotypes were S. Kentucky (51/114, 44.7%) and S. Enteritidis (37/114, 32.5%), which were distributed throughout the four slaughtering stages, and were also identified in the corresponding environments. The multilocus sequence typing (MLST) analysis revealed that seven sequence types (STs) were occupied by six different serotypes, respectively. Only S. Kentucky had two STs, ST314 was the predominant ST shared by 50 isolates, while the ST198 has 1 isolate. The antimicrobial resistance gene analysis demonstrated that most of the strains belonging to S. Kentucky (39/51, 76.5%) and S. Indiana (15, 100%) contained over five groups of antimicrobial resistance genes. Based on the core genome analysis, 50 S. Kentucky isolates were genetically identical, indicating that one S. Kentucky strain with the same genetic background was prevalent in the chicken slaughtering line. Although 37 S. Enteritidis isolates only had three different antimicrobial resistance gene profiles, the core genome sequence analysis subtyped these S. Enteritidis isolates into five different clusters, which revealed the diverse genetic background of S. Enteritidis in the slaughterhouse. The antimicrobial resistance phenotypes were consistent with the presence of the corresponding resistance genes of S. Kentucky and S. Enteritidis, including tetA, floR, blaTEM-1B, strA/B, sul1/sul2, and gyrA (D87Y). Our study observed a high prevalence of Salmonella in the chicken slaughter line and identified the slaughtering environment as a main source of causing Salmonella cross-contamination during chicken slaughtering. Further studies will be needed to limit the transmission of Salmonella in the slaughterhouse.

Non‐typhoidal salmonellosis remains an important public health problem worldwide. Dogs may harbour Salmonella in their intestines and can easily shed Salmonella in the environment with the possibility of transmission to humans. Thus, monitoring is essential to understand the role of dogs in zoonotic transmission. The objectives of this study were to determine the shedding of Salmonella by owned, apparently healthy dogs in Grenada, West Indies, to identify the serovars, and to examine their antimicrobial susceptibility profiles. Faecal samples collected during August to October, 2016 from 144 non‐diarrhoeic owned dogs were examined by enrichment and selective culture for the presence of Salmonella spp. Eight (5.6%) of the tested animals were culture positive, yielding 35 Salmonella isolates that belonged to six serovars of Salmonella enterica subspecies enterica. These were serovars Arechavaleta from two dogs, Arechavaleta and Montevideo from one dog, and Javiana, Rubislaw, Braenderup and Kiambu from one dog each. All these serovars have been reported as causes of human salmonellosis globally. Antimicrobial susceptibility tests on 35 isolates showed absence of resistance to the currently used drugs for cases of human salmonellosis, including ciprofloxacin and cefotaxime. One isolate (2.9%) was resistant to neomycin, two isolates (5.7%) showed intermediate susceptibility to neomycin, and another (2.9%) had intermediate susceptibility to tetracycline. This is the first report of isolation and antimicrobial susceptibilities of non‐typhoidal Salmonella serovars from dogs in Grenada. This study shows that dogs in Grenada may be involved in the epidemiology of salmonellosis., Faecal samples from 144 non‐diarrhoeic owned dogs collected during August to October, 2016 were examined by enrichment and selective culture for the presence of Salmonella spp.

Aim. An attempt to use MALDI-TOF mass-spectrometry method for identification of leptospiral isolates on the serovar level. Materials and methods. 8 reference Leptospira spp. and 11 leptospira strains isolated from leptospiral patients and infected animals in the North-VNfestem region of Russia were included into the study. Mass-spectra of all the studied strains were obtained by direct profiling of cell extracts. The created main spectral profiles (MSP) of reference strains were used for identification of isolates. Evaluation of identification was carried out by calculating coefficients of matching rate of separate spectra of each isolate with MSP of all the reference strains. Results. Results of identification have shown the similarity of spectra of isolates belonging to Pomona, Icterohaemorrhagiae and Canicola serogroups, with MSP of saprophyte strain L. biflexa Patoc I. It is assumed that spectra of the studied strains contained peaks of polysaccharide O-antigens. Wherein maximum mean values of matching rate coefficients between spectra of isolates and MSP of pathogenic reference strains of leptospira correctly matched serovar type of the isolate. Conclusion. Further extended studies may form the base of development of a simple and rapid method of typing of leptospirosis causative agents on the level of serovars using MALDI-TOF mass-spectrometry.

Background Ureaplasma spp. have been implicated in a variety of clinical conditions and certain serovars are likely to be disease-associated. Hence, the ascending trend of Ureaplasma spp. resistance to antimicrobials should deserve more attention. Here we assessed the extent of antimicrobial resistance of Ureaplasma serovars in Tunisia, and investigated the underlying molecular basis. Methods This study included 101 molecularly typed Ureaplasma spp. clinical strains isolated over a 12-year time period (2005–2017). The antimicrobial susceptibility was tested against nine antibacterial agents using the broth microdilution method. Neighbor-joining tree was constructed to establish the phylogenetic relationships among isolates. Results We found that all ureaplasma isolates were resistant to ciprofloxacin and erythromycin, intermediately resistant to azithromycin, and susceptible to doxycycline, moxifloxacin and josamycin. Ofloxacin and levofloxacin resistance was found in 73.27 and 17.82%, respectively, while 37.62% of isolates proved resistant to tetracycline. Consequently, we detected an elevated multidrug resistance rate among ureaplasma isolates (37.62%), particularly among serovars 2, 5, 8, and 9 (77.77% overall), as well as serovars 4, 10, 12, and 13 (52.63% overall). In most cases, drug resistance was found to be associated with known molecular mechanisms, yet we have identified two novel mutations in the L22 protein, which might be associated with macrolide-resistance. Conclusion To our knowledge, this is the first study that reports the widespread expansion of multidrug resistance among Ureaplasma serovars, a finding of importance in terms of both surveillance and antimicrobial usage.

Cloacal swabs from 45 Grenada bank tree boas (Corallus grenadensis) were sampled during a 12‐month period (2011–2012) from the rain forests and scrublands of Grenada. Cloacal swabs were examined by enrichment and selective culture for the presence of Salmonella spp. In all, 16 (35.6%) of the snakes were positive for Salmonella, and six serovars of Salmonella were isolated. The most common serovar was Rubislaw (31.3%), the most frequent serovar recently isolated from green iguanas in Grenada, followed by serovar Braenderup (18.8%), and serovar IV:48:g,z51:‐ (formerly, S. Marina) (18.8%), also found in green iguanas in this country. The remaining three less frequent serovars were, IV:53:g,z51:‐, I:6,7:e,h:‐ and IIIb:38:i:z. Antimicrobial susceptibility tests conducted by a disc diffusion method against amoxicillin–clavulanic acid, ampicillin, cefotaxime, ciprofloxacin, enrofloxacin, gentamicin, imipenem, nalidixic acid, streptomycin, tetracycline and trimethoprim–sulfamethoxazole showed that drug resistance is minimal, with intermediate susceptibility, only to streptomycin. This is the first report of isolation and antimicrobial susceptibilities of Salmonella serovars from wild Grenadian tree boas., To understand the epidemiology of salmonellosis, it is important to know about the actual serovar prevalence in animals including wildlife.

International audience; Salmonella is a genus of Gram-negative bacteria in the Enterobacteriaceae family causing various illnesses. The ability of the different serovars of Salmonella enterica subsp. enterica to infect a host and to induce pathology relies in part on their cellular and molecular interactions with the intestinal epithelium. In the current study, an in vitro approach using non-polarized or polarized IPEC-1 porcine intestinal epithelial cells were used in order to assess the relation between adhesion, invasion, and induction of the immune response as a function of the serotype of Salmonella. Five serovars, Choleraesuis (host-adapted), Typhimurium (ubiquitous), Typhisuis (host-restricted), which are relevant for pig infection, and Dublin and Gallinarum, which are host-restricted or host-adapted, were studied. A strong variation was observed in the percentages of adhesion and invasion amongst the S. enterica serovars used to interact with the non-polarized and polarized cells. Subsequently, differences were identified between serovars in terms of immune response induced. Serovars Typhimurium and Typhisuis induced a strong innate immune response four and half hours after the beginning of cell stimulation while Choleraesuis, Gallinarum, and Dublin did not. A strong inflammatory response could limit the spread of the porcine serovars to the gut while, with a weak response, bacteria may not be constrained by the immune response enabling severe systemic diseases. Different repertoires of adhesion factors and of secreted protein effectors between Salmonella serovars interacting with IPEC-1 cells probably explains the differences in their early pathogenic behaviours.

Not all Salmonella enterica serovars cause the same disease. S. enterica represents an incredibly diverse species comprising >2,600 unique serovars. While some S. enterica serovars are host-restricted, others infect a wide range of hosts. The diseases that nontyphoidal Salmonella (NTS) serovars cause vary considerably, with some serovars being significantly more likely to cause invasive disease in humans than others. Furthermore, while genomic analyses have advanced our understanding of the genetic diversity of these serovars, they have not been able to fully account for the observed clinical differences. One overarching challenge is that much of what is known about Salmonella’s general biology and virulence strategies is concluded from studies examining a select few serovars, especially serovar Typhimurium. As targeted control strategies have been implemented to control select serovars, an increasing number of foodborne outbreaks involving serovars that are less frequently associated with human clinical illness are being detected. Harnessing what is known about the diversity of NTS serovars represents an important factor in achieving the ultimate goal of reducing salmonellosis-associated morbidity and mortality worldwide. In this review we summarize the current understanding of the differences and similarities among NTS serovars, highlighting the virulence mechanisms, genetic differences, and sources that characterize S. enterica diversity and contribute to its success as a foodborne pathogen.

Glässer’s disease (GD) is an important infectious disease of swine caused byHaemophilus (Glaesserella) parasuis. Vaccination with inactivated whole cell vaccines is the major approach for prevention ofH. parasuisinfection worldwide, but the immunity induced is predominantly against the specific polysaccharide capsule. As a consequence, the available vaccines may not induce adequate protection against the field strains, when the capsules present in the vaccine strains are different from those in strains isolated from the farms. Therefore, it is crucial to mapH. parasuisserovars associated with regional outbreaks so that appropriate bacterin vaccines can be developed and distributed for prevention of infection. In this study, 459H. parasuisfield strains isolated from different Glässer’s disease outbreaks that occurred in 10 different Brazilian States were analyzed for serotype using PCR-based approaches. Surprisingly, non-typeable (NT) strains were the second most prevalent group of field strains and along with serovars 4, 5 and 1 comprised more than 70% of the isolates. A PCR-based approach designed to amplify the entire polysaccharide capsule locus revealed 9 different band patterns in the NT strains, and 75% of the NT strains belonged to three clusters, suggesting that a number of new serovars are responsible for a substantial proportion of disease. These results indicate that commercially available vaccines in Brazil do not cover the most prevalentH. parasuisserovars associated with GD.

Background The colonization of Ureaplasma species in genital tract is related with male infertility. However, it has been postulated based upon limited study that virulence is related to serotype specificity. The aim of this study was to determine the distribution of Ureaplasma serovars in genital tract of infertile males and analyze their role in male infertility. Methods A total of 358 urethral swabs samples were obtained from infertile males. The culture of Ureaplasma species were performed using a commercially available Mycoplasma IST 2 kit. Serovars were determined by real-time polymerase chain reaction (real-time PCR). Results A total of 92 (25.7%) infertile males were positive for Ureaplasma spp; among them, Ureaplasma parvum (UPA) was detected in 73 (79.3%) isolates, and Ureaplasma urealyticum (UUR) was detected in 19 (20.7%) isolates. Serovars 1, 6, or in combination accounted for 63.0% (46/73) of UPA isolates. Serovar 9 (alone and in combination of other serovars) was the most common serovar in UUR (47.4%, 9/19). Multiple serovars were detected in 21 (22.8%) isolates, and serovars 4, 5, 7, and 12 were not detected in any sample. Conclusion The distribution of 14 Ureaplasma serovars in genital tract of infertile males was identified for the first time by real-time PCR assay. UPA serovars 1 and 6, and UUR serovar 9 are the most common serovars colonization in urogenital tract of infertile males.

s Objective This is the first study was conducted to identify the Leptospirosis seroprevalence and serovar distribution among healthy people in different regions in Vietnam. Methods This cross-sectional study was conducted with 600 asymptomatic people in three provinces, representing for the North, the northern Central, and the South of Vietnam from November to December 2019. Each participant was collected a 3 ml venous blood sample by a certified phlebotomist. The microscopic agglutination test (MAT) was applied to determine the leptospiral seroprevalence and serogroups with a cutoff titre for seropositivity of ≥1:100. Results The overall seroprevalence of leptospiral antibody among healthy Vietnamese people was 9.5%. In total eleven serovars of Leptospira were isolated. The most frequent serovars identified were strains of Hebdomadis (14.0%), Pomona (14.0%), Saxkoebing (12.3%), and Panama (8.8%). Among seropositive cases, farmer was majority, accounting for 63.2%, other jobs accounted from 3 to 8%. Conclusions Overall seropositivity Leptospirosis among healthy people was relatively high in Vietnam. Diverse serovars of Leptospira have been confirmed to circulate in different regions in the country. Further research need to be conducted to determine factors for high seroprevalence in the population of Vietnam. In addition, health education programs should be considered to prevent leptospirosis among Vietnamese people, especially for farmers, in the future.

Leptospirosis in New Zealand has been strongly associated with animal-contact occupations and with serovars Hardjo and Pomona. However, recent data suggest changes in these patterns, hence, serovar-specific epidemiology of leptospirosis from 1999 to 2017 was investigated. The 19-year average annual incidence is 2.01/100,000. Early (1999–2007) and late (2008–2017) study period comparisons showed a significant increase in notifications with serovar Ballum (IRR: 1.59, 95% CI: 1.22–2.09) in all cases and serovar Tarassovi (IRR: 1.75, 95% CI: 1.13–2.78) in Europeans and a decrease in notifications with serovars Hardjo and Pomona in all cases. Incidences of Ballum peaked in winter, Hardjo peaked in spring and Tarassovi peaked in summer. Incidence was highest in Māori (2.24/100,000) with dominant serovars being Hardjo and Pomona. Stratification by occupation showed meat workers had the highest incidence of Hardjo (57.29/100,000) and Pomona (45.32/100,000), farmers had the highest incidence of Ballum (11.09/100,000) and dairy farmers had the highest incidence of Tarassovi (12.59/100,000). Spatial analysis showed predominance of Hardjo and Pomona in Hawke’s Bay, Ballum in West Coast and Northland and Tarassovi in Waikato, Taranaki and Northland. This study highlights the serovar-specific heterogeneity of human leptospirosis in New Zealand that should be considered when developing control and prevention strategies.

Salmonella spp. remain among the most important agents of foodborne diseases worldwide. The importance of Salmonella spp. in public health is linked to their wide range of antimicrobial resistance and to their pathogenicity and virulence in both human and animal hosts. The aim of this study was to determine the antimicrobial resistance patterns for Salmonella serotypes isolated from poultry sources in Brazil and to detect virulence-associated genes and verify their association with specific serotypes. A total of 163 strains of Salmonella enterica isolated from poultry sources in Southern Brazil were selected, and each belonged to one of 11 different serotypes. They were tested against ten antibiotics and examined for the presence of 26 virulence-associated genes by PCR. S. Typhimurium, S. Bredeney, S. Schwarzengrund and S. Tennessee showed the highest overall resistance rates. Approximately 18% of Salmonella strains were classified as multidrug-resistant strains. Our results indicate associations between antimicrobial resistance and specific serotypes. Most of the investigated genes presented a high frequency and a regular distribution, regardless of the serotype. Eight genes are positively or negatively associated with at least one serotype. The observed associations between antimicrobial resistance and specific serotypes are useful in developing specific control and treatment measures for each serotype. Despite the virulence genes being evenly distributed among the serotypes, some of these genes are associated with specific serotypes, and sefA, sopEand lpfA were selected as possible markers of Salmonella serotypes.

Purpose. Chlamydiatrachomatis is responsible for trachoma-associated blindness as well as the most common sexually transmitted bacterial infection worldwide, although the genovars for the former are typically A-C, whilst for the latter they are D-K and for the uncommon infection lymphogranuloma venereum they are L1-3. Nucleotide variations within the ompA gene facilitate the identification of C. trachomatis genovars. This study describes a colorimetric multiplex PCR/RLB typing assay (mPCR-RLB) directed to the VD2 region of the ompA gene for general C. trachomatis positivity and the identification of 14 individual C. trachomatis genovars.Methodology. The assay was validated by analysing 40 blinded samples that included reference strains of C. trachomatis genovars and other non-chlamydial micro-organisms that had been analysed previously using quantitative PCR (qPCR). Ninety clinical samples that had previously been found to be C. trachomatis-positive by qPCR were also evaluated using the mPCR-RLB assay.Results. The mPCR-RLB assay showed 100% agreement with the qPCR in the detection of C. trachomatis reference strains and no cross-reaction of non-chlamydial micro-organisms was observed. In the analysis of the chlamydial clinical samples, 97.8% were C. trachomatis-positive by mPCR/RLB assay and there was a 96.6% concordance with the qPCR at the group identification level and a 92.2% concordance at the genovar level.Conclusion. The mPCR-RLB assay is a rapid and sensitive methodology for the identification of C. trachomatis genovars associated with urogenital infections, trachoma or lymphogranuloma venereum diseases that can be implemented in clinical settings, helping to identify reinfections and treatment failures and establish the appropriate treatment course.

Highlights • Analysis of complete capsule loci in all 18 serovars of A. pleuropneumoniae. • Novel insights into evolution of capsule loci in A. pleuropneumoniae. • Development of two mPCRs for comprehensive capsule typing., Problems with serological cross-reactivity have led to development of a number of PCRs (individual and multiplex) for molecular typing of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. Most of these assays were developed for detection of specific amplicons within capsule biosynthetic genes before the availability of complete sequences for the different serovars. Here we describe comparative analysis of the complete capsular loci for all 18 serovars of A. pleuropneumoniae, and development of two multiplex PCRs for comprehensive capsule typing of this important pig pathogen.

Salmonella serovars sampled from meat products in Southern Spain (Andalucía) during the period 2002-2007 were analyzed in this study. The serovars most frequently detected (in order) were Typhimurium, Enteritidis, Derby, Anatum and Rissen. Isolates (n = 43) were tested for sensitivity to biocides, including the quaternary ammonium compounds benzalkonium chloride (BC), cetrimide (CT) and hexadecylpyridinium chloride (HDP), and the bisphenols triclosan (TC) and hexachlorophene (CF). The minimum inhibitory concentration (MIC) for the quaternary ammonium compounds was in the range of 25 to 50 mg/L for most isolates, although a few isolates required much higher concentrations, up to 250 mg/L. Bisphenols showed higher inhibitory activity, with a MIC of 2.5 to 25 mg/L. A few isolates showed a “non-wildtype” MIC for TC of up to 250 mg/L. These results indicate a low incidence of tolerance towards quaternary ammonium compounds and triclosan among Salmonella from meats and meat products.

Salmonella serotyping remains the gold-standard tool for the classification of Salmonella isolates and forms the basis of Canada’s national surveillance program for this priority foodborne pathogen. Public health officials have been increasingly looking towards whole genome sequencing (WGS) to provide a large set of data from which all the relevant information about an isolate can be mined. However, rigorous validation and careful consideration of potential implications in the replacement of traditional surveillance methodologies with WGS data analysis tools is needed. Two in silico tools for Salmonella serotyping have been developed, the Salmonella in silico Typing Resource (SISTR) and SeqSero, while seven gene MLST for serovar prediction can be adapted for in silico analysis. All three analysis methods were assessed and compared to traditional serotyping techniques using a set of 813 verified clinical and laboratory isolates, including 492 Canadian clinical isolates and 321 isolates of human and non-human sources. Successful results were obtained for 94.8%, 88.2% and 88.3% of the isolates tested using SISTR, SeqSero, and MLST, respectively, indicating all would be suitable for maintaining historical records, surveillance systems, and communication structures currently in place and the choice of the platform used will ultimately depend on the users need. Results also pointed to the need to reframe serotyping in the genomic era as a test to understand the genes that are carried by an isolate, one which is not necessarily congruent with what is antigenically expressed. The adoption of WGS for serotyping will provide the simultaneous collection of information that can be used by multiple programs within the current surveillance paradigm; however, this does not negate the importance of the various programs or the role of serotyping going forward.

Recent studies have reported that the virulence factors (VFs) were detected more frequently in amoxicillin-clavulanate (AMC) susceptible clinical isolates of Escherichia coli. Here, we have evaluated the relationship between VFs and AMC-resistance phenotype in clinical isolates of Y. enterocolitica biovar 1A. The presence/absence of VFs was compared with their minimum inhibitory concentrations for AMC in strains of two serovars. We observed that the strains of the serovar O: 6, 30-6, 31 showed a similar relationship between the number of VFs and resistance to clavulanic acid as in E. coli but not of serovar O: 6, 30. Variations in the promoters/complete coding sequences (CCDSs) of β-lactamase gene (bla A) or the serological characteristics could not account for unusual susceptibility to AMC displayed by the strains of the serovar O: 6, 30. Therefore, we speculate that since the clinical strains of serovar O: 6, 30-6, 31 originated from the environment they were less exposed to antibiotics compared to clinical strains of serovar O: 6, 30. Thus, AMC susceptibility seems to be influenced by factors other than serotypes or promoters/CCDS of β-lactamase genes.

This study aims to determine the prevalence of fluoroquinolone resistance of Ureaplasma biovars and serovars isolated from urogenital clinical samples and determine the underlying molecular mechanism for quinolone resistance for all resistant isolates. Of 105 samples confirmed as positive for U. urealyticum/U. parvum, 85 were resistant to quinolones by the Mycoplasma–IST2 kit. However, only 43 out of 85 quinolone resistant isolates had amino acid substitutions in GyrA, GyrB, ParC and ParE proteins underlining that this assay have mis-identified as fluoroquinolone resistant 42 isolates. The known ParC E87K and ParC S83L mutations were found in 1 and 10 isolates, respectively. An original mutation of ureaplasmal ParC (E87Q, 1 isolate) was found. Furthermore, we found a ParE R448K mutation in one isolate, already described. Among the additional alterations detected, the most prevalent mutation found was L176F in GyrA protein in 18 isolates with single infection and in 3 isolates with mixed ureaplasma infections. Mutations in GyrB (E502Q, 4 isolates), ParE (Q412K, Q412P, Q412T, 3 independent isolates), whose role is unknown, were also found. Other sporadic mutations in the four genes were identified. This investigation is the result of monitoring the data for molecular fluoroquinone resistance in Ureaplasma spp. in Italy. Resulting that this acquired resistance is high and that continued local epidemiological studies are essential to monitor and document their antimicrobial resistance trends.

This study aims to determine the antibiotic resistance phenotypes of Salmonella isolates of broiler meat and chicken samples against a total of 23 antimicrobial agents. For this, 64 meat and 79 chicken Salmonella isolates, which were obtained from retail meat and farm samples analyzed in our laboratory from February 2006 to January 2009, were used. All Salmonella isolates were resistant to at least 4 antimicrobials tested, with the common highest resistance rates to erythromycin (100.0%), bacitracin (99.3%), and rifampicin (98.6%), regardless of the sample type. Additionally, meat isolates were 100.0% resistant to bacitracin, clindamycin, erythromycin, rifampicin, and chicken isolates were 100.0% resistant to erythromycin, spiramycin, penicillin G, doxycycline hydrochloride, and tetracycline. Four to 7 Multi Drug Resistant (MDR) Salmonella conformed 82.83% of all meat isolates, and 10, 12, 15, and 17 MDR Salmonella conformed 55.68% of all chicken isolates analyzed. The B/DA/E/RD and N/B/CT/E/SP/P/RD/DO/OT/TE resistances were the most common patterns as 35.94% and 13.92% within meat and chicken isolates, respectively. The considerably high resistance rates and MDR types to commonly used antimicrobial agents among the Salmonella isolates tested pose a significant risk factor for the treatment of foodborne Salmonella infections in humans, Bu çalışmada tavuk eti ve tavuk orijinli Salmonella izolatlarında 23 antimikrobiyal ajana karşı dirençlilik fenotiplerinin belirlenmesi amacı ile perakende satış yerleri ve çiftlik örneklerinden elde edilen 64 adet et ve 79 adet tavuk Salmonella izolatı, Şubat 2006 ile Ocak 2009 arasında laboratuvarımızda analiz edildi. Tüm Salmonella izolatlarının, örnek tipi göz önünde bulundurulmaksızın, test edilen antimikrobiyalleden en yüksek direnç oranı ile sırası ile eritromisin’e (%100.0), basitrasin’e (%99.3) ve rifampisin’e (%98.6) olmak üzere en az 4 antimikrobiyale dirençli olduğu belirlendi. Ayrıca, et izolatlarının basitrasin, klindamisin, eritromisin, rifampisin’e; tavuk izolatlarının ise eritromisin, spiramisin, penisilin G, doksisiklin hidroklorür ve tetrasiklin’e %100.0 dirençli olduğu bulundu. Tüm et izolatlarının %82.83’ü 4 ile 7 Çoklu İlaç Direnci (ÇİD) gösterirken, tüm tavuk izolatlarının %55.68’i 10, 12, 15 ve 17 antimikrobiyale ÇİD gösterdiği belirlendi. Et izolatlarında %35.94 oranında ve tavuk izolatlarında ise %13.92 oranında olmak üzere en sık rastlanan ÇİD paternleri sırasıyla B/DA/E/RD ve N/B/CT/E/SP/P/RD/DO/OT/TE olarak bulundu. Salmonella izolatlarında karşılaşılan bu yüksek dirençlilik oranlarının ve ÇİD paternlerinin insanlardaki Salmonella infeksiyonlarının tedavisinde risk oluşturabileceği belirtildi

Background & objectives: Limited data are available on the typing of Chlamydia trachomatis in India. Serovars D to K of C. trachomatis are chiefly responsible for urogenital infections. Thus, this study was conducted to determine the distribution of C. trachomatis serovars in patients with urogenital infections and to characterize omp A gene of the detected C. trachomatis isolates by sequence analysis. Presence of other co-infections was also evaluated. Methods: Endocervical swabs were collected from 324 women and urethral swabs/urine were collected from 193 men attending the sexually transmitted diseases outpatient clinic. The samples were screened for C. trachomatis by cryptic plasmid PCR and omp A gene PCR. Genotyping was performed by PCR-restriction fragment length polymorphism (RFLP) and sequencing of the omp A gene. Samples were screened for genital mycoplasmas, Neisseria gonorrhoeae, Treponema pallidum and human immunodeficiency virus (HIV). Results: C. trachomatis was found in 15.0 per cent men and 10.8 per cent women. Serovar D was the most prevalent followed by serovars E, F, I and G. Twenty two C. trachomatis isolates were selected for omp A gene sequencing. No mixed infection was found. Variability in omp A sequences was seen in 31.8 per cent cases. Both PCR-RFLP and omp A gene sequencing showed concordant results. The presence of Ureaplasma spp. and Mycoplasma hominis was observed in 18.7 and 9.5 per cent patients, respectively. Co-infection of C. trachomatis was significantly associated with Ureaplasma urealyticum and HIV. Interpretation & conclusions: The high occurence of C. trachomatis infections warrants its screening in addition to other sexually transmitted infections namely U. urealyticum and HIV. Genotyping of the omp A gene may provide additional information for vaccine development.

To determine whether rapid emergence of Salmonella enterica serovar Infantis in Israel resulted from an increase in different biotypes or spread of 1 clone, we characterized 87 serovar Infantis isolates on the genotypic and phenotypic levels. The emerging strain comprised 1 genetic clone with a distinct pulsed-field gel electrophoresis profile and a common antimicrobial drug resistance pattern.

Aim. B. thuringiensis (Bt) are gram-positive spore-forming aerobic or facultative anaerobic bacteria able to form during sporulation species specific crystal-like inclusions of protein nature, consisting of particular thermolabile d-endotoxins. Serological Bt variants produce different entomotoxins; their synthesis in many respects depends on the conditions of cultivation. There was accumulated a vast information on the entomotoxins, their origin, synthesis, structure, toxic properties and mechanisms of action on insects. These bacteria are dominating in the microbiomethods of pest control in plants and animals. There are more than 70 serovariants of Bt selectively specific to the definite groups of host insects. However, the description of new variants not always looks justified considering the phylogenetic systematization based on phenotype signs. Methods. A comparative phylogenetic analysis of the Bt intraspecific interrelations was performed on the basis of the cloned 16S rRNA genes of entomopathogenic bacteria BtH1, BtH10, BtH14. Results. The phylogenetically homogeneous lines were investigated – a homology of 16S rRNA of the strains 1 and 10 ranged from 90,0 to 94,0 %; no distinct genetic isolation among the strains of 14th and 10th serovars was revealed. Conclusions. The comparison of nucleotides sequences of 16S rRNA has shown the existence of strains polymorphism within the group of entomopathogens BtH1, BtH10, BtH14, connected with their entomocide activity

Single nucleotide polymorphisms can be used for epidemiologic and evolutionary studies worldwide., Chlamydia trachomatis is a global cause of blinding trachoma and sexually transmitted infections (STIs). We used comparative genomics of the family Chlamydiaceae to select conserved housekeeping genes for C. trachomatis multilocus sequencing, characterizing 19 reference and 68 clinical isolates from 6 continental/subcontinental regions. There were 44 sequence types (ST). Identical STs for STI isolates were recovered from different regions, whereas STs for trachoma isolates were restricted by continent. Twenty-nine of 52 alleles had nonuniform distributions of frequencies across regions (p

Salmonellosis is a foodborne disease caused by bacteria of the genus Salmonella, being pigs and pork-products potentially important for its occurrence. In recent decades, some serovars of Salmonella have shown increase of resistance to conventional antimicrobials used in human and animal therapy, with serious risks for public health. The aim of this study was to evaluate feces (n=50), mediastinal (n=50), mesenteric (n=50) and mandibular (n=50) lymph nodes obtained from slaughter houses for Salmonella spp. Positive samples were serotyped and subjected to an in vitro antimicrobial susceptibility test, including the extended-spectrum beta-lactamase (ESBL) production. Salmonella species were identified in 10% (20/200) of total samples. From these, 20% (10/50) were identified in the submandibular lymph nodes, 18% (9/50) in the mesenteric lymph nodes, 2% (1/50) in feces and 0% (0/50) in the mediastinal lymph nodes. The serotypes found were Salonella Typhimurium (55%), S. enterica subsp. enterica 4,5,12: i: - (35%), S. Brandenburg and S. Derby with 5% (5% each). All strains showed resistance to at least one antimicrobial; 90% were resistant to four or more antimicrobials, and 15% were multidrug-resistant. Resistance to ciprofloxacin, tetracycline and nalidixic acid was particularly prevalent amongst the tested serovars. Here, we highlighted the impact of pigs in the epidemiological chain of salmonellosis in domestic animals and humans, as well as the high antimicrobial resistance rates of Salmonella strains, reinforcing the necessity for responsible use of antimicrobials for animals as an emergent One Health issue, and to keep these drugs for human therapy approaches. RESUMO: Nas últimas décadas, o aumento de cepas circulante de Salmonella concomitantemente a resistência microbiana tem despertado a preocupação dos órgãos de Saúde Pública. Deste modo, o objetivo do presente trabalho foi pesquisar a presença de Salmonella a partir de fezes (n=50), linfonodos mediastinos (n=50), mesentéricos (n=50) e submandibular (n=50) oriundos de um abatedouro suíno. As cepas isoladas foram sorotipadas e testadas quanto a resistência antimicrobiana. A presença de Salmonella isolada foram em 10% (20/200) do total de amostras, sendo 20% dos linfonodos submandibulares, 18% dos linfonodos mesentéricos e 2% das fezes. Os sorotipos encontrados foram S. Typhimurium (55%), S. enterica subsp. enterica 4,5,12: i: - (35%), S. Brandenburg (5%) e S. Derby (5%). Todas a cepas apresentaram resistência a pelo menos um antimicrobiano testado, sendo 90% resistente pelo menos quatro antimicrobianos. Destes, 15% foram classificadas como multidrogas resistentes. Os antimicrobianos mais resistentes entre os sorovares isolados foram a ciprofloxacina, tetraciclina e o ácido nalidixico. A presença de cepas de Salmonella resistente a antimicrobianos na espécie suína tem gerado um grande impacto epidemiológico entre homem e animal, reforçando cada vez mais a necessidade do uso adequado de drogas principalmente relacionado com o tema “One Health”.

The aim of the present study was to contribute to the knowledge on extended-spectrum beta-lactamases (ESBL:s), AmpC beta-lactamases and integrons in Enterobacteriaceae isolated from horses, which is still limited. The susceptibility of 1581 clinical isolates from animals to ceftiofur was tested. Most of these isolates (n = 1347) originated from horses. Seven ceftiofur-resistant equine isolates (four Escherichia coli and three Klebsiella pneumoniae) were identified and all seven were multidrug-resistant. These isolates were further studied for the presence of ESBL's, AmpC beta-lactamases and class 1 integrons. The potential for the horizontal transfer of resistance genes among these clinical isolates was also studied. ESBL-type resistance genes were found in five isolates, AmpC-type genes in one isolates and integrons in six isolates. Nucleotide sequence analysis revealed that the isolates carried the bla(CTX-M-1), bla(CMY-2), bla(TEM-1) and/or bla(SHV-1) genes. This is the first report describing the in vitro conjugal transfer of the blaCTX-M-1 genes from a clinical E. coli isolate to Salmonella isolates. Gene cassettes encoding resistance to aminoglycosides (aadA1, aadA2 and aadA5), and trimethoprim (dfrA1, drfA12 and dfrA17) were found on the integrons present in the isolates. The cassette arrays of the dfrA17-aadA5 and dfrA1-aadA1 genes in the two integrons of a single E coli isolate have not yet been described before. To our knowledge this is the first report on ESBL:s and AmpC beta-lactamases in equine E. coli and Klebsiella isolates. (c) 2007 Elsevier B.V. All rights reserved.

Os produtos de origem avícola podem ser importantes veículos de transmissão de Salmonella spp. para humanos e, dentre os vários parâmetros que determinam a qualidade de um alimento, destacam-se os que definem suas características microbiológicas. Objetivou-se detectar e quantificar Salmonella spp. na tecnologia de abate de frangos de corte por microbiologia convencional (MC) e número mais provável miniaturizado (mNMP). As coletas foram realizadas em duas visitas a três abatedouros sob Inspeção Federal e em seis pontos de coleta em triplicata, definidos como: recepção das aves (swabs de cloaca e esponjas de gaiolas de transporte antes e após a higienização) e carcaças (após pré resfriamento em chiller, após o gotejamento e antes da embalagem primária e congeladas a -12oC por 24 horas), totalizando 108 amostras. Identificou-se Salmonella spp. em três dos seis pontos do fluxograma de abate e em dois dos três estabelecimentos amostrados, independentemente do método utilizado, perfazendo 5,5% de positividade, onde destaca-se a contaminação nas gaiolas de transporte das aves após a higienização. Não foi possível correlacionar os resultados da microbiologia convencional e do mNMP ou mesmo quantificar a contaminação ao longo da tecnologia de abate, o que indica a necessidade de se utilizar um método qualitativo aliado ao método de quantificação quando Salmonella estiver presente em quantidades inferiores ao limite de detecção do mNMP proposto (0,13 NMP/mL). Os sorovares identificados foram Typhimurium, Panama, Lexington e Rissen, consideradas paratíficos e, portanto, potencialmente capazes de causar infecções em humanos, embora estes sorovares não tenham sido isolados em produtos finais e sim na chegada dos frangos aos abatedouros (swabs de cloaca e gaiolas de transporte). A identificação de Salmonella spp. nas gaiolas de transporte após a higienização é um indicativo da necessidade de revisão e adequação dos métodos automatizados de lavagem atualmente utilizados nos abatedouros. Poultry products can be important modes of transmission of Salmonella spp. to humans and, among several parameters used to determine food quality, microbiological characteristics play an essential role. The aim of this study was to determine and quantify Salmonella spp. at broiler slaughtering facilities. This was done by conventional microbiology and by the miniaturized most probable number (mMPN) methods. Three federally-inspected slaughterhouses were visited, where samples were collected in triplicate from six sites: reception of live birds (cloacal swabs and sponge samples from transport cages before and after sanitation) and carcass processing (after pre-chiller, after dripping, and before primary packaging and refrigeration at -12oC for 24h), totaling 108 samples. Three of the six surveyed sites and two of the three slaughterhouses were contaminated with Salmonella spp., showing an infection rate of 5.5% independently of the method used, and revealing that transport cages were contaminated after sanitation. No correlation could be established between the results of conventional microbiology and mMPN methods, and contamination along the slaughtering line could not quantified. This indicates the importance of combining qualitative and quantitative methods for the enumeration of Salmonella when detection rates are lower than the proposed mMPN limit (0.13 MPN/mL). Typhimurium, Panama, Lexington and Rissen, which are paratyphoid organisms and are potentially infectious to humans, were identified. However, these serovars were isolated at the reception of live birds (from cloacal swabs and from transport cages) rather than from the end products. Given that Salmonella spp. was detected in transport cages after sanitation, it is paramount that automated washing procedures currently used in slaughterhouses be reassessed and adjusted.

Leptospirosis is a zoonosis of worldwide distribution. The aim of this study was to examine the presence of antibodies against 21 Leptospira reactive serovars in Chaetophractus villosus in La Pampa province, Argentina, using the microscopic agglutination test (MAT). Pathologic changes compatible with leptospirosis and in situ detection of the agent by immunohistochemistry were studied in 24 and 3 individuals respectively. Only 35/150 (23.3%) serum samples had antibodies against Leptospira sp. Six percent of the samples reacted with serovar Canicola, 4.7% with serovar Castellonis, 1.3% with serovar Icterohemorrhagieae and 0.7% with serovar Hardjo. Sixteen (10.6%) serum samples agglutinated with Castellonis-Icterohemorrhagiae and Canicola-Castellonis serovars, both with 4.7%, and Canicola-Hardjo and Castellonis-Canicola-Icterohemorrhagiae both with 0.6%. Fourteen animals had variable degrees of lesions, which were more severe in animals with higher serological titers (3200), and Leptospira sp. was detected in 3 animals by immunohistochemistry. These results represent the first record of the presence of Leptospira in C. villosus in La Pampa. Fil: Kin, Marta S.. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales; Argentina Fil: Brihuega, Bibiana. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Fort, Marcelo. Instituto Nacional de Tecnologia Agropecuaria. Centro Regional La Pampa-San Luis; Argentina Fil: Delgado, Fernando. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Bedotti, Daniel. Instituto Nacional de Tecnología Agropecuaria. Centro Regional la Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Casanave, Emma Beatriz. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina

A total of 296 Shigella spp. were received from State Public Health Laboratories, during the period from 1999 to 2004, by National Reference Laboratory for Cholera and Enteric Diseases (NRLCED) - IOC/Fiocruz, Rio de Janeiro, Brazil. The frequency of Shigella spp. was: S. flexneri (52.7%), S. sonnei (44.2%), S. boydii (2.3%), and S. dysenteriae (0.6%). The most frequent S. flexneri serovars were 2a and 1b. The highest incidence rates of Shigella isolation were observed in the Southeast (39%) and Northeast (34%) regions and the lowest rate in the South (3%) of Brazil. Strains were further analyzed for antimicrobial susceptibility by disk diffusion method as part of a surveillance program on antimicrobial resistance. The highest rates of antimicrobial resistance were to trimethoprim-sulfamethozaxole (90%), tetracycline (88%), ampicillin (56%), and chloramphenicol (35%). The patterns of antimicrobial resistance among Shigella isolates pose a major difficulty in the determination of an appropriate drug for shigellosis treatment. Continuous monitoring of antimicrobial susceptibilities of Shigella spp. through a surveillance system is thus essential for effective therapy and control measures against shigellosis.

Avian salmonellosis is a worldwide problem to the poultry industry, from the point of view of animal health and public health as well. The aim of the present study was to survey the most common Salmonella serovars in commercial breeders or broiler flocks from several regions in Brazil. The results of the present study indicated a high incidence of S enterica subspecies enterica serovar Enteritidis in breeders (57.5%) and broilers flocks (84.0%). The importance of these findings lies in the fact that S. Enteritidis has become the most frequent serovar responsible for foodborne outbreaks and sporadic cases of salmonellosis in humans.

In Colombia, little information is available concerning the epidemiology of leptospirosis in urban environments. Furthermore, the role of dogs in the transmission cycle of leptospirosis in the urban setting is unclear. To explore the potential role of canines in the transmission of leptospirosis in Cali, a serological study was conducted with 197 serum samples collected from stray dogs during 2001 and 2003. Serum specimens were screened with the Microscopic Agglutination Test (MAT) and 7 serovars--Icterohaemorrhagiae, Canicola, Gryppotyphosa, Hardjo strain Hardjobovis, Pomona, Hardjo strain Hardjoprajitno and Bratislava. All serovars were provided by the Instituto Colombiano Agropecuario (ICA), Tulua, Colombia. The MAT was considered positive when 50% or more leptospiras were agglutinated with one or more serovars in a serum dilution of 1:100. At least one serovar showed evidence of infection in 41.1% of the dogs. The most prevalent serovar was Icterohaemorrhagiae, found in 55.6% of the seropositive dogs. 48.1% were co-agglutinations. No reactions against the serovars Pomona, Hardjo strain Hardjoprajitno and Bratislava were observed. These findings suggested that stray dogs are potential reservoirs of Leptospira in Cali and underscored the need to study the epidemiology of this disease in Colombia.

Detection of Salmonella by bacteriologic methods is known to be time consuming. Therefore, we have developed a real-time probe-specific polymerase chain reaction (PCR) to rapidly detect Salmonella invA gene-based PCR products from chicken feces and carcasses by a fluorescence resonance energy transfer assay. The sensitivity and the specificity of this system were determined as 3 colony-forming units ml(-1) and 100%, respectively. Overnight tetrathionate broth enrichment cultures of chicken feces and carcass samples were used in template preparation for PCR. Also, a standard bacteriology was performed (National Poultry Improvement Plan-U.S. Department of Agriculture, Bacteriological Analytical Manual-Food and Drug Administration Center for Food Safety and Applied Nutrition) for confirmation. Seventy-two cloacal swab, 147 intestine, and 50 carcass (neck) samples were examined. Thirteen (8.8%) and 25 (17%) of the intestinal samples were found to harbor Salmonella by bacteriology and PCR, respectively. Forty-five of 50 (90%) carcass samples were Salmonella positive by both methods. Salmonella was not detected from cloacal swab samples. Results indicate that this assay has the potential for use in routine monitoring and detection of Salmonella in infected flocks and carcasses.

Este relato de caso descreve a manifestacao clinica de diferentes sorovares de Salmonella em bezerros leiteiros. A S . typhimurium foi identificado em amostras fecais de um bezerro com prolapso rectal e em amostras fecais e linfonodos mesentericos de um bezerro com necrose na ponta da cauda. Ja a S . agona foi identificada em amostras de fezes de um bezerro com diarreia, mas sem nenhuma outra manifestacao clinica, e a S . dublin foi observada em amostras de fezes e de orgaos a partir de um bezerro com sinais clinicos neurologicos. Diagnostico diferencial para os principais patogenos entericos ( E. coli enteropatogenica, rotavirus e coronavirus) foram realizados e foram negativos. Devido ao impacto negativo da salmonelose e a ocorrencia de diferentes sorotipos e manifestacoes clinicas em bezerros, o diagnostico correto e importante para identificar formas de controle e medidas profilaticas no rebanho leiteiro.

Background: Leptospirosis and brucellosis are zoonosis worldwide distributed that have great economic importance, especially in goats. This study aimed to carry out a cross-sectional study to determine the prevalence of anti-Leptospira spp. and anti-Brucella abortus antibodies and to determine epidemiological variables associated to infection by these ethiological agents in goats.Materials, Methods & Results: Serum samples were collected from 1055 goats of 95 properties distributed in 18 regional centers of Paraná State from April to August of 2010. Collected samples were submitted to microscopic agglutination test (MAT) for the detection of anti-Leptospira spp. antibodies with 22 reference serovars: Australis, Bratislava, Autumnalis, Butembo, Castellonis, Bataviae, Canicola, Fortbragg, Whitcombi, Cynopteri, Grippotyphosa, Hebdomadis, Copenhageni, Icterohaemorrhagiae, Panama, Pomona, Pyrogenes, Hardjo, Wolffi, Shermani, Tarassovi and Londrina 1 (L1). The samples were also submitted to buffered acidified plate antigen test (BAPA) and 2-mercaptoethanol test (2-ME) to detect anti-Brucella abortus antibodies. The results obtained from epidemiological and serological study were analysed by Epi Info 3.5.3. We observed 9.9% (104/1055) of goats reagents in at least one serovar of Leptospira spp. and no reagent animal to Brucella abortus. The variables that were statistically significant with leptospirosis were: production type meat, sewer destination in dry sump, frequent abortion, shared use of pastures, slaughter of animals on the property, the presence of pigs and wild animals such as deer and capybaras on the property.Discussion: This was the first study about antibodies prevalence against Leptospira spp. and Brucella abortus in goat herds from Paraná State. The presence of both animal species were statiscally associated to risk of leptospirosis. It is necessary avoid the acess of these animals in herds, decreasing the dissemination of the agent. The production type meat is a form of creation in which the animals have no care of zookeepers and continuous veterinary care. The final destination of sewage was statistically associated to risk of leptospirosis in goats and this is a big problem in rural properties, requiring greater attention of authorities for proper disposal of this wastewater, reducing water and food contamination. Among the variables related to management, the herds that used rented pasture or shared pastures also were statistically associated to risk of leptospirosis in goats; with the sharing of pastures a greater flow of animals transiting in the pickets occurs, allowing not only flocks of goats, but other species also, to use the same pastures and become more exposed to certain diseases. The slaughter of pigs, sheep, cattle and goats on the property is a problem for animal and human health due to lack of sanitary inspection. Abortion is one of the main clinical signs of leptospirosis in farm animals, particularly goats. The knowledge of these results will be useful to develop control programs for the disease in Paraná State herds. The absence of circulating anti-Brucella abortus antibodies in goat herds has also been reported in other Brazilian States.

A . E Y I G O R , K . T A Y F U N C A R L I A N D C . B . U N A L . 2002. Aims: The present study describes the implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry. Methods and Results: Real-time PCR with Salmonella invA-specific primers and a standard bacteriological method was applied to detect Salmonella in tetrathionate enrichment cultures of 492 intestinal homogenates and 27 drag swabs from 47 poultry flocks. The number of positive individual samples by real-time PCR and culture method was 65 (12AE5%) and 35 (6AE8%), respectively. The number of Salmonella-positive flocks was 13 (27AE7%) by both methods. PCR detection required 25 min for up to 32 samples. Melting curve analysis revealed the Tm for Salmonella-specific PCR product as 87 a 1∞C. Conclusions: Implementation of real-time PCR to tetrathionate broth enrichment step reduces the Salmonella detection time to 18 h and 25 min. Isolation of Salmonella should be carried out with PCR to determine the serovar. Significance and Impact of the Study: Real-time PCR is a powerful tool in rapid and accurate Salmonella monitoring in poultry companies, together with standard bacteriology.

Objective To analyze antigenic and genetic variations of Chlamydia trachomatis among the serovars obtained from Japanese infants. Methods The polymerase chain reaction (PCR) was used to amplify a large part of the major outer-membrane protein gene, and restriction fragment length polymorphism (RFLP) was used to identify the serovars of C. trachomatis from nasopharyngeal and conjunctival swabs from Japanese infants and neonates. Results The typing of 10 nasopharyngeal isolates gave the following results: seven E, one H, and two unclassified serovars. The typing of seven conjunctival isolates gave the following results: five D, one F, and one unclassified serovar. Reactive patterns of these unclassified strains, determined by PCR-RFLP, to monoclonal antibodies were different from those of 15 reference serovars. Conclusions Characterization of unclassified variants will allow more detailed epidemiologic studies of perinatal C. trachomatis infections in Japan.

The proportion of positive Leptospira microscopic agglutination tests for 23,005 dogs significantly increased from 2002 to 2004 (p < 0.002) regardless of the positive cutoff titer used and was highest (p < 0.05) for serovars Autumnalis and Grippotyphosa. The strongest positive serologic correlation (r = 0.72) was between serovars Autumnalis and Pomona.

Salmonella enterica subsp. enterica comprises a number of serovars, many of which pose an epidemiological threat to humans and are a worldwide cause of morbidity and mortality. Most reported food infection outbreaks involve the serovars Salmonella Enteritidis and Salmonella Typhimurium. Rapid identification to determine the primary sources of the bacterial contamination is important to the improvement of public health. In recent years, many DNA-based techniques have been applied to genotype Salmonella. Herein, we report the use of a manual TRS-PCR approach for the differentiation of the Salmonella enterica subspecies enterica serovars in a single-tube assay. One hundred seventy Salmonella strains were examined in this work. These consisted of serovars S. Enteritidis, S. Typhimurium, S. Infantis, S. Virchow, S. Hadar, S. Newport and S. Anatum. Five of the TRS-primers, N6(GTG)4, N6(CAC)4, N6(CGG)4, N6(CCG)4 and N6(CTG)4, perfectly distinguished the S. Enteritidis and S. Typhimurium serovars, and the N6(GTG)4 primer additionally grouped the other five frequently isolated serovars. In our opinion, the TRS-PCR methodology could be recommended for a quick and simple DNA-based test for inter-serovar discrimination of Salmonella strains.

Listeria monocytogenes is an important foodborne human pathogen. Human infection is associated with high mortality rates. Epidemiological investigation and molecular subtyping can be useful in linking human illness with specific sources of infection. This retrospective study describes the use of PFGE to examine relationships of 222 isolates from human and non-human sources in Ireland. Human clinical isolates from other countries were also examined. Eight small clusters of human and non-human isolates (mostly serotype 4b) that were indistinguishable from one another were detected, suggesting potential sources for human infection. For non-human isolates, some PFGE types appeared to be exclusively associated with a single source, whereas other PFGE-types appeared to be more widely disseminated. Indistinguishable, or highly related clusters of isolates of Irish and non-Irish origin suggest that some PFGE patterns may be globally distributed.

Prevention of Salmonella contamination of poultry products requires detailed knowledge of the main sources associated with its presence in the production system. The aims of this study were to determine the main sources of Salmonella contamination in broiler production during growing, to assess the risk factors for Salmonella contamination at the end of the rearing period and to determine the main serovars involved in broiler production systems in Eastern Spain. A total of 65 different broiler houses from different farms were sampled. Each house was sampled at different times during the rearing period. First, when the previous flock was taken to the slaughterhouse, samples of dust, surfaces and previous flock faeces were collected. After cleaning and disinfection (C&D), samples of dust and surfaces were also taken. On the first day of rearing, samples of water, bedding, farming boots, meconiums, delivery-box liners and feed were collected. During rearing, feed samples were taken directly from the truck and from feeders. On slaughter day, samples of dust, surfaces, water, feed and faeces were also collected. Finally, two days after slaughter, carriers (rodents, flies and beetles) were trapped. All samples collected were analysed according to ISO 6579:2002 (Annex D) and positive samples were serotyped in accordance with Kauffman-White-Le-Minor technique. Our results showed that all different types of samples collected were contaminated with Salmonella (prevalence ranged between 1.5% and 38.6%). The most contaminated samples related with poultry production were: delivery-box liners (32.0%), faeces samples (31.2%), dust samples (25.0%), farming boots (19.7%) and feed from feeders (16.0%). However, the most important risk factors for Salmonella contamination of the flocks at the end of the rearing period were Salmonella status of the house after cleaning and disinfection, Salmonella status of day-old chick flocks and feed from feeders. Twenty-one different serovars were isolated from the samples analysed. The most prevalent were in decreasing order: Salmonella Enteritidis (52.9%), S. Hadar (17.8%), S. Virchow (8.9%) and S. Ohio (5.4%). The study suggested that there are many sources for Salmonella contamination and persistence in broiler production. Hence, the whole production chain needs to be controlled to eradicate the bacteria from primary production. © 2010 Elsevier B.V.

A proof of principle of a multi-target assay for genotyping Salmonella has been developed targeting 62 genomic marker sequences of Salmonella related to pathogenicity. The assay is based on multiplex ligation detection reaction (LDR) followed by customized ArrayTube (R) microarray detection. The feasibility of the developed assay was verified in a method comparison study with conventional PCR using 16 Salmonella 'test' strains comprising eight serovars. Subsequently, the feasibility of the LDR microarray assay was also tested by analyzing 41 strains belonging to 23 serovars. With the exception of four serovars each serovar was characterized by a unique virulence associated gene repertoire. The LDR microarray platform proved to be a convenient, rapid and easy to use tool with potential in tracing a Salmonella contamination in the food chain, for outbreak studies, and to provide data for risk assessors that support bio-traceability models. (C) 2010 Elsevier B.V. All rights reserved

This study presents Salmonella Enteritidis incidence in chicken layer flocks in Turkey determined by real-time PCR (rPCR) and by International Organization for Standardization (ISO) method 6579:2002/Amd 1:2007. A total of 259 samples, composed of 1,036 individual samples each pooled into 4, including 175 cloacal swab, 14 intestine, 35 gizzard swab, and 35 cecal swab samples, belonging to 6 major companies, were collected from 50 layer flocks and tested by rPCR and ISO culture methods. Overall incidence of Salmonella in layer flocks by rPCR and culture was 61.0 and 55.6%, respectively, where 70.1% of these Salmonella isolates were determined as Salmonella Enteritidis. Incidences of Salmonella Enteritidis in culture-positive samples were 65.3% in cloacal swabs, 50.0% in intestines, 73.9% in gizzard swabs, and 87.5% in cecal swabs. The rPCR results were in 100% agreement (100% sensitivity and specificity) with culture results when cecal swabs were selected as the sample type. The relative accuracy of rPCR was 92.4, 91.4, and 84% for intestine, gizzard, and cloacal swab samples, respectively. As a result, by using rPCR and ISO culture, we determined that the Salmonella Enteritidis incidence in layer flocks in Turkey was high and that the use of cecal swab and intestine samples in Salmonella detection would yield reliable results. To reduce this high Salmonella Enteritidis incidence in layer flocks, Salmonella Enteritidis-specific vaccination should be implemented properly in conjunction with a well-designed biosecurity plan, including verifiable corrective actions.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:24:42Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:46:48Z : No. of bitstreams: 1 2-s2.0-78649409673.pdf: 126420 bytes, checksum: d21fc7b8647e972ba81dca2321877c6a (MD5) Made available in DSpace on 2014-05-27T11:24:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-06-01 Leptospirosis may affect all domestic and wild animals as well as human beings. Some serological studies have shown the involvement of wild species in the epidemiology of the disease. Once captive wild animals are not much studied, especially in Brazil, the present study aimed to detect anti- Leptospira spp. antibodies in animals from Ribeirão Preto city zoo, in São Paulo state, Brazil. Blood samples were collected from captive birds, fish, reptiles and mammals, sinanthropics and free-living animals and also from employees between March and October, 2006. Four hundred and three blood samples were obtained, 388 animals' samples (110 reptiles, 143 birds, 110 mammals and 25 fish) and 15 humans'. The sera were analysed by Microscopic Agglutination Test using 22 serovars from pathological leptospiras and two from non-pathological serovars. Among the animal samples, 339 were from captive animals, and 49 from free-living ones, captured with traps inside the zoo. One hundred and three (103/388 = 26.5%) samples reacted to leptospirosis, ninety-two (92/339 = 27.1%) samples were from captive animals and eleven (11/49 = 22.4%) from free-living ones. All humans' samples were negative. Serological titles varied from 40 to 5.120, with predominance of titles between 40 and 80 and the most frequent serovars were Patoc, Andamana, Canicola, Icterohaemorrhagiae and Panama. Departamento de Medicina Veterinária Preventiva e Reprodução Animal Faculdade de Ciências Agrárias e Veterinária Universidade Estadual Paulista, Jaboticabal-SP Instituto Mamíferos Aquáticos, Salvador-BA Médica Veterinária Autônoma Bosque Zoológico Municipal 'Dr. Fábio de Sá Barreto', Ribeirão Preto-SP Departamento de Medicina Veterinária Preventiva e Reprodução Animal Faculdade de Ciências Agrárias e Veterinária Universidade Estadual Paulista, Jaboticabal-SP