Source Of Human Infection Research Articles

Pilot Study of Risk Group Human Seroprevalence to Coxiella burnetii (Q Fever) in Latvia

Abstract Q fever is an important zoonotic disease worldwide. The main sources of human infection are inhalation of aerosols containing Coxiella burnetii bacteria and exposition to infected materials during parturition or slaughtering. The high-risk group includes people who work directly with infected livestock, such as farmers, veterinarians, veterinary medicine students, slaughterhouse and laboratory staff. Here we present a pilot study of risk-group human seroprevalence to C. burnetii in Latvia. The study included 240 sera samples — 190 from the risk groups and 50 from the control group. Samples were tested with Coxiella burnetii (Q-fever) Phase 1 and 2 IgG ELISA kits. All sera from the control group were negative. The seroprevalence among risk group persons was 8.04–11.54%. No statistically significant differences were observed between genders. The highest percentage of seropositive and equivocal sera samples (25%) were detected in age categories 39–48 years and 49–58 years. Working as a practicing veterinarian or former veterinarian was the only risk factor identified as statistically significant, and belonging to the risk group in general. The geographical distribution of seropositive risk group participants indicated that they tend to located more in the northern, central, and eastern part of the country.

Protective effect of Lactiplantibacillus plantarum LP5 in a murine model of colonisation by Campylobacter coli DSPV458.

Thermotolerant Campylobacter species are the leading cause of foodborne bacterial diarrheal disease worldwide. Campylobacter coli, abundant in pigs and pork products, have been identified as a source of human infection. In this study, we propose the use of Lactiplantibacillus plantarum LP5 as a probiotic to reduce colonisation of this intestinal pathogen in a murine colonisation model of C. coli DSPV458. Six-week-old adult female Balb/cCmedc mice were housed in groups: Control, Campy and Pro-Campy. Control and Pro-Campy groups received antibiotics for 5 days and the Campy group for 12 days. Pro-Campy group was inoculated for 7 days with 8.78 log10 cfu total of L. plantarum LP5 suspended in De Man, Rogosa and Sharpe broth. All groups were inoculated with 6.72 log10 cfu of C. coli DSPV458 suspended in brain heart infusion broth. L. plantarum LP5 was recovered only in the Pro- Campy group. C. coli DSPV458 was recovered at higher levels in the Control and Campy groups. The differences with the Pro-Campy group were significant. As regards faeces, Control and Campy groups reached 7.41 and 7.84 log10 cfu/g, respectively, and the Pro-Campy group only 4.62 log10 cfu/g. In the caecum, Control and Campy groups reached 8.01 and 9.26 log10cfu/g, respectively, and the Pro-Campy group only 4.51 log10 cfu/g. In the ileum, Control and Campy groups reached 3.43 and 3.26 log10 cfu/g, respectively, and the Pro-Campy group did not show detectable levels. The reduction of C. coli DSPV458 in the Pro-Campy group compared to the Control group in faeces, caecum and ileum was 99.55, 99.98 and 100%, respectively. Animals were maintained under normal health conditions, and haematological parameters were within the standard values for Balb/cCmedc. The incorporation of a probiotic generated a protective effect in the mice colonisation model. The protective effect would also apply to intestinal colonisation by indigenous enterobacteria. Therefore, the strategy used in this study is of great importance to understand the protection mechanisms in a murine model, as well as its application in food-producing animals.

Enrichment Free qPCR for Rapid Identification and Quantification of Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis in Chicken Meat Samples by a New Couple of Primers.

Campylobacter is the main cause of bacterial foodborne disease and poultry meat is the principal source of human infections. Rapid methods for Campylobacter detection are urgently needed to decrease high bacterial prevalence in poultry products. In this study, we developed new primers, CampyPFw and CampyPRv, that target the 16S-23S rRNA genes of Campylobacter jejuni, C. coli, C. lari and C. upsaliensis. The primers were tested on positive and negative reference strains in pure cultures and in inoculated poultry meat samples before their application in real-time PCR (qPCR) protocol for analyzing chicken meat samples. In parallel, the samples were tested by using the ISO 10272-1:2006 method. The qPCR protocol based on CampyPFw and CampyPRv showed good sensitivity, with the limit of detection of 4.6 × 102 cells/mL in chicken samples without enrichment steps.

A Highlight on Avian Toxoplasmosis: One Health Disease with a Special Reference to the Current Egyptian Situation

This review article was developed to the infection of avian species with Toxoplasma gondii (T. gondii), diagnosis, pet bird and human infection, and control methods with a special reference to the current status of infection among the Egyptian poultry farms and population. Toxoplasmosis is a zoonotic disease caused by a unicellular, protozoan parasite T. gondii. Different domesticated and wild animals, as well as birds can harbor T. gondii and may be a potential source of infection to humans. Avian species could be infected with T. gondii through the ingestion of contaminated food, soil, and water with oocysts shed in the excreta of infected animals, especially cats. Poor sanitation and hygienic conditions increase the risk of infection. Consumption of food or water, as well as undercooked poultry meat or meat products containing the oocysts of the parasite, are the main sources of human infection with T. gondi. Diagnosis of T. gondii in the infected host depends on the serological detection of specific antibodies and molecular detection of the parasite. Microscopic demonstration of the oocysts and other developmental stages of the parasite in the intestine, liver, brain, and skeletal muscles tissues is another means for rapid diagnosis. Generally, a high prevalence of the disease is also reported in pet birds. Toxoplasmosis in humans is associated with abortion, congenital disorders, stillbirth, and other complications, especially in immunocompromised patients. Application of hygienic measures, as well as public awareness, are essential for the prevention and control of toxoplasmosis. In different Egyptian governorates, a high prevalence of T. gondii has been detected in animals, birds, and humans. High incidence of infection was recorded due to the contact with Toxoplasma oocysts shed mainly from infected cats or other carriers. Egyptian chicken and turkey flocks and backyard birds revealed the presence of different developmental stages of the parasite and even its antibodies. In addition, human populations showed signs of toxoplasmosis with severe complications.

Influence of Season, Population and Individual Characteristics on the Prevalence of Leptospira spp. in Bank Voles in North-West Germany

Simple SummaryLeptospirosis is a worldwide emerging zoonotic disease. Clinical symptoms in humans range from mild flu-like symptoms to severe clinical disease with kidney failure and multiple organ dysfunction. Infections occur after contact with infected animals or through water and soil contaminated by urine of infected animals. Cases are mostly linked to occupational risk groups such as field workers or farmers, but contact with pets and recreational activities like fresh water sports also lead to a higher exposure risk. This study was conducted to evaluate the prevalence and species distribution of Leptospira in bank voles in Germany. We analyzed the DNA of 1817 kidney samples and detected a mean prevalence of 7.5% with the following pathogenic Leptospira species: L. interrogans, L. kirschneri, and L. borgpetersenii. The individual infection risk in bank voles depended on season, body weight and abundance of bank voles. Even if leptospirosis case numbers in Germany are low, our study shows that pathogenic Leptospira spp. are present and a potential source for human infection, which should be recognized by clinicians and veterinarians.Leptospirosis is a worldwide zoonotic disease with more than 1 million human cases annually. Infections are associated with direct contact to infected animals or indirect contact to contaminated water or soil. As not much is known about the prevalence and host specificity of Leptospira spp. in bank voles (Clethrionomys glareolus), our study aimed to evaluate Leptospira spp. prevalence and genomospecies distribution as well as the influence of season, host abundance and individual characteristics on the Leptospira prevalence. Bank voles, which are abundant and widely distributed in forest habitats, were collected in the years 2018 to 2020 in North-West Germany, covering parts of North Rhine-Westphalia and Lower Saxony. The DNA of 1817 kidney samples was analyzed by real-time PCR targeting the lipl32 gene. Positive samples were further analyzed by targeting the secY gene to determine Leptospira genomospecies and multilocus sequence typing (MLST) to determine the sequence type (ST). The overall prevalence was 7.5% (95% confidence interval: 6.4–8.9). Leptospira interrogans (83.3%), L. kirschneri (11.5%) and L. borgpetersenii (5.2%) were detected in bank voles. Increasing body weight as a proxy for age increased the individual infection probability. Only in years with high bank vole abundance was this probability significantly higher in males than in females. Even if case numbers of human leptospirosis in Germany are low, our study shows that pathogenic Leptospira spp. are present and thus a persisting potential source for human infection.

Risk factor analysis for Salmonella contamination of broiler chicken (Gallus gallus) hatcheries in Great Britain

Salmonellosis is the second most commonly reported zoonosis in the European Union and contaminated meat from broiler chickens (Gallus gallus) is an important source of human infection. In Great Britain (GB), prevalence of Salmonella enterica in broiler flocks is low, having declined considerably since the introduction of the Salmonella National Control Programme in 2010. However, this decreasing trend has stabilised in recent years and serovars with known ability to persistently colonise hatcheries have been isolated from broiler flocks with increasing frequency, indicating that further controls on hatchery contamination are required. The broiler industry in GB has changed dramatically over the last 15 years, with greater intensification and dominance by a small number of very large companies which rely on relatively few hatcheries. An investigation of risk factors for Salmonella contamination in GB broiler hatcheries was therefore carried out so that relevant up-to-date advice on Salmonella control can be provided. Twenty-two hatcheries, representing most commercial scale GB broiler hatcheries, were visited between 2015 and 2018. Salmonella contamination was comprehensively investigated at each hatchery by collecting between 108 and 421 environmental swab samples per hatchery (6990 samples in total from all hatcheries). An in-depth questionnaire on hatchery operations was completed for each hatchery, and results were incorporated into a risk factor analysis (univariable followed by multivariable mixed effects logistic regression) to identify factors associated with Salmonella occurrence. Overall, 6.0 % (416/6990) of environmental samples were Salmonella-positive and Salmonella was isolated from 17/22 hatcheries. Ten different serovars were isolated, the most common being S. Senftenberg and S. Mbandaka which are known hatchery colonisers. Sixty-four risk factor variables were investigated. Twenty-two of these were initially retained based on univariable analyses (p ≤ 0.25) and six were ultimately left in the final multivariable model (p ≤ 0.05). Salmonella detection was positively associated with having ≥30 hatchers in regular use compared to fewer (Odds ratio [OR] 23.7, 95 % confidence interval [CI] 6.7–84.2), storing trays in process rooms (OR 28.8, CI 7.8–106.3), drying set-up trolleys in corridors (OR 15.6, CI 5.9–41.4) and having skips located in enclosed areas (OR 8.99, CI 5.89–41.35). Using a closed waste disposal system was negatively associated with Salmonella detection (OR 0.08, CI 0.04−0.18) and the odds of detecting Salmonella in hatcheries with 31−60 total workers was lower compared to hatcheries with ≤30 staff (OR 0.16, CI 0.06−0.40). Despite the complexities of hatchery enterprises, changes to a relatively small number of features may significantly reduce the occurrence of hatchery contamination.

A Validation System for Selection of Bacteriophages against Shiga Toxin-Producing Escherichia coli Contamination

Shiga toxin-producing Escherichia coli (STEC) can cause severe infections in humans, leading to serious diseases and dangerous complications, such as hemolytic-uremic syndrome. Although cattle are a major reservoir of STEC, the most commonly occurring source of human infections are food products (e.g., vegetables) contaminated with cow feces (often due to the use of natural fertilizers in agriculture). Since the use of antibiotics against STEC is controversial, other methods for protection of food against contaminations by these bacteria are required. Here, we propose a validation system for selection of bacteriophages against STEC contamination. As a model system, we have employed a STEC-specific bacteriophage vB_Eco4M-7 and the E. coli O157:H7 strain no. 86-24, bearing Shiga toxin-converting prophage ST2-8624 (Δstx2::cat gfp). When these bacteria were administered on the surface of sliced cucumber (as a model vegetable), significant decrease in number viable E. coli cells was observed after 6 h of incubation. No toxicity of vB_Eco4M-7 against mammalian cells (using the Balb/3T3 cell line as a model) was detected. A rapid decrease of optical density of STEC culture was demonstrated following addition of a vB_Eco4M-7 lysate. However, longer incubation of susceptible bacteria with this bacteriophage resulted in the appearance of phage-resistant cells which predominated in the culture after 24 h incubation. Interestingly, efficiency of selection of bacteria resistant to vB_Eco4M-7 was higher at higher multiplicity of infection (MOI); the highest efficiency was evident at MOI 10, while the lowest occurred at MOI 0.001. A similar phenomenon of selection of the phage-resistant bacteria was also observed in the experiment with the STEC-contaminated cucumber after 24 h incubation with phage lysate. On the other hand, bacteriophage vB_Eco4M-7 could efficiently develop in host bacterial cells, giving plaques at similar efficiency of plating at 37, 25 and 12 °C, indicating that it can destroy STEC cells at the range of temperatures commonly used for vegetable short-term storage. These results indicate that bacteriophage vB_Eco4M-7 may be considered for its use in food protection against STEC contamination; however, caution should be taken due to the phenomenon of the appearance of phage-resistant bacteria.

MERS-CoV in sheep, goats, and cattle, United Arab Emirates, 2019: Virological and serological investigations reveal an accidental spillover from dromedaries.

The recent COVID-19 pandemic has demonstrated again the global threat posed by emerging zoonotic coronaviruses. During the past two decades alone, humans have experienced the emergence of several coronaviruses, such as SARS-CoV in 2003, MERS-CoV in 2012, and SARS-CoV-2 in 2019. To date, MERS-CoV has been detected in 27 countries, with a case fatality ratio of approximately 34.5%. Similar to other coronaviruses, MERS-CoV presumably originated from bats; however, the main reservoir and primary source of human infections are dromedary camels. Other species within the Camelidae family, such as Bactrian camels, alpacas, and llamas, seem to be susceptible to the infection as well, although to a lesser extent. In contrast, susceptibility studies on sheep, goats, cattle, pigs, chickens, and horses obtained divergent results. In the present study, we tested nasal swabs and/or sera from 55 sheep, 45 goats, and 52 cattle, collected at the largest livestock market in the United Arab Emirates, where dromedaries are also traded, for the presence of MERS-CoV nucleic acid by RT-qPCR, and for specific antibodies by immunofluorescence assay. All sera were negative for MERS-CoV-reactive antibodies, but the nasal swab of one sheep (1.8%) repeatedly tested positive for MERS-CoV nucleic acid. Next generation sequencing (NGS) of the complete N gene of the sheep-derived MERS-CoV revealed>99% nucleotide identity to MERS-CoV sequences of five dromedaries in nearby pens and to three reference sequences. The NGS sequence of the sheep-derived MERS-CoV was confirmed by conventional RT-PCR of a part of the N gene and subsequent Sanger sequencing. All MERS-CoV sequences clustered within clade B, lineage 5. In conclusion, our study shows that noncamelid livestock, such as sheep, goats, and cattle do not play a major role in MERS-CoV epidemiology. The one sheep that tested positive most likely reflects an accidental viral spillover event from infected dromedaries in nearby pens.

Rhodococcus equi-Occurrence in Goats and Clinical Case Report.

Background: Rhodococcus equi infection is commonly known in equine medicine to cause frequently fatal rhodococcosis. Infections in other species and people are also reported. Clinical manifestation in goats is relatively similar to horses and humans, but data regarding bacterium prevalence are scarce. Thus, the study aimed to estimate the occurrence of R. equi in goats. Methods: During post mortem examination, submandibular, mediastinal, and mesenteric lymph nodes were collected. Standard methods were used for bacteria isolation and identification. Results: A total of 134 goats were examined, and 272 lymph node samples were collected. R. equi was isolated from four animals. All four isolates carried the choE gene, and one also had traA and pVAPN plasmid genes. Conclusions: To the authors’ best knowledge, this is the first report of R. equi occurrence and genetic diversity in goats. The results may help create a model for treating rhodococcosis in other animal species and assessing the role of meat contamination as a potential source of human infection. This research should be considered a pilot study for further application of the goat as a model of R. equi infection in horses and humans.

Evaluation of the Correlation between the mRNA Expression Levels of ystA and ymoA Genes in Y. enterocolitica Strains with Different Enterotoxic Properties.

Yersinia enterocolitica is one of the main causative agents of human diarrhea. Pigs are a reservoir and the most common source of infection for humans. The aim of this study was to analyze the expression of ystA and ymoA genes in Y. enterocolitica strains with different enterotoxic properties, isolated from humans and pigs. The experiment involved two groups of Y. enterocolitica strains producing and not producing enterotoxin YstA, which were isolated from humans and pigs. All strains were ystA- and ymoA-positive. The expression of ystA and ymoA genes was analyzed by quantitative real-time PCR (qPCR). The relative expression level of the ystA gene was significantly higher than the expression level of the ymoA gene in Y. enterocolitica strains isolated from humans with clinical symptoms of yersiniosis. In other strains, a significant decrease in ystA gene transcription was observed, and the relative expression level of the ymoA gene was significantly higher than the expression level of the ystA gene. Statistically significant differences were not observed in either group of strains isolated from pigs. The results of our study revealed a correlation between mRNA expression levels of ystA and ymoA genes in Y. enterocolitica strains isolated from humans.

Modelling the introduction and transmission of Campylobacter in a North American chicken flock.

Campylobacter is the second leading cause of foodborne illness in the United States. Although many food production animals carry Campylobacter as commensal bacteria, consumption of poultry is the main source of human infection. Previous research suggests that the biology of Campylobacter results in complete flock colonization within days. However, a recent systematic review found that the on-farm prevalence of Campylobacter varies widely, with some flocks reporting low prevalence. We hypothesized that the low prevalence of Campylobacter in some flocks may be driven by a delayed introduction of the pathogen. The objectives of this study were to (a) develop a deterministic compartmental model that represents the biology of Campylobacter, (b) identify the parameter values that best represent the natural history of the pathogen in poultry flocks and (c) examine the possibility that a delayed introduction of the pathogen is sufficient to replicate the observed low prevalence examples documented in the literature. A deterministic compartmental model was developed to examine the dynamics of Campylobacter in chicken flocks over a 56-day time period prior to movement to the abattoir. The model outcome of interest was the final population prevalence of Campylobacter at day 56. The resulting model that incorporated a high transmission rate (β=1.04) was able to reproduce the wide range of prevalence estimates observed in the literature when pathogen introduction time is varied. Overall, we established that the on-farm transmission rate of Campylobacter in chickens is likely high and can result in complete colonization of a flock when introduced early. However, delaying the time at which the pathogen enters the flock can reduce the prevalence observed at 56days. These results highlight the importance of enforcing strict biosecurity measures to prevent or delay the introduction of the bacteria to a flock.

Toxocara Species Infection in Pet Dogs and Cats Ensured by Loop-Mediated Isothermal Amplification Assay

Background: Toxocariasis is a parasitic disease with global distribution. Many paratenic or transport hosts have been identified as the sources of infection, including vertebrates such as birds and humans. The definitive hosts of Toxocara are dogs and cats whose parasites live in their guts. Hence, the diagnosis of the disease in these animals is important. Objective: The present study was performed using the loop-mediated isothermal amplification (LAMP) method to diagnose toxocariasis. Materials and Methods: In the present study, 60 samples of pet feces were collected and subsequently evaluated by direct smear, formalin-ether, and LAMP. Results: Toxocara infection in dogs by microscopic and formal ether methods and LAMP method was 10% and 13.3%, respectively, while feline infection with this parasite was not observed either microscopically or molecularly. Conclusion: The present findings revealed that pet dogs can be considered as a source of infection for humans. Therefore, it is recommended that animal owners receive the necessary training to prevent the risks of transmitting zoonotic diseases.

Detection of Lassa virus in wild rodent feces: Implications for Lassa fever burden within households in the endemic region of Faranah, Guinea

Lassa arenavirus (LASV) is the cause of Lassa Fever in humans in West Africa. The multimammate mouse (Mastomys natalensis) is a reservoir host of LASV and the primary source of human infections. Humans are assumed to become infected due to contact with this animal or its excretions. Thus far, the available literature does not describe the sampling of feces as a means to detect LASV in M. natalensis populations. More evidence is needed to know if feces of naturally infected M. natalensis can be LASV-positive and an exposure risk to humans. This study sampled feces deposits in households from three villages in the LASV-endemic region of Faranah, Guinea. PCR analysis found 10 out of 88 samples to be positive for LASV, and sequencing showed clustering to previously identified Yarawelia and Dalafilani strains. We conclude that feces sampling is a viable, non-invasive method for the determination and sequencing of LASV strains.

Genomic Characterization of Multidrug-Resistant Salmonella Serovars Derby and Rissen From the Pig Value Chain in Vietnam.

Nontyphoidal Salmonella (NTS) is the most reported cause of bacterial foodborne zoonoses in Vietnam, and contaminated pork is one of the main sources of human infection. In recent years, the prevalence of NTS carrying multiple antimicrobial resistance genes (ARGs) have been increased. The genomic characterization along the pig value chain and the identification of ARGs and plasmids have the potential to improve food safety by understanding the dissemination of ARGs from the farm to the table. We report an analysis of 13 S. Derby and 10 S. Rissen isolates, collected in 2013 at different stages in Vietnamese slaughterhouses and markets. VITEK 2 Compact System was used to characterize the phenotypical antimicrobial resistance of the isolates. In addition, whole-genome sequencing (WGS) was used to detect ARGs and plasmids conferring multidrug resistance. Whole genome single nucleotide polymorphism typing was used to determine the genetic diversity of the strains and the spread of ARGs along the pig value chain. Altogether, 86.9% (20/23) of the samples were resistant to at least one antibiotic. Resistance to ampicillin was most frequently detected (73.9%), followed by piperacillin and moxifloxacin (both 69.6%). At least one ARG was found in all strains, and 69.6% (16/23) were multidrug-resistant (MDR). The observed phenotype and genotype of antimicrobial resistance were not always concordant. Plasmid replicons were found in almost all strains [95.6% (22/23)], and the phylogenetic analysis detected nine clusters (S. Derby, n = 5; S. Rissen, n = 4). ARGs and plasmid content were almost identical within clusters. We found six MDR IncHI1s with identical plasmid sequence type in strains of different genetic clusters at the slaughterhouse and the market. In conclusion, high rates of multidrug resistance were observed in Salmonella strains from Vietnam in 2013. Genomic analysis revealed many resistance genes and plasmids, which have the potential to spread along the pig value chain from the slaughterhouse to the market. This study pointed out that bioinformatics analyses of WGS data are essential to detect, trace back, and control the MDR strains along the pig value chain. Further studies are necessary to assess the more recent MDR Salmonella strains spreading in Vietnam.

Wildlife Is a Potential Source of Human Infections of Enterocytozoon bieneusi and Giardia duodenalis in Southeastern China.

Wildlife is known to be a source of high-impact pathogens affecting people. However, the distribution, genetic diversity, and zoonotic potential of Cryptosporidium, Enterocytozoon bieneusi, and Giardia duodenalis in wildlife are poorly understood. Here, we conducted the first molecular epidemiological investigation of these three pathogens in wildlife in Zhejiang and Shanghai, China. Genomic DNAs were derived from 182 individual fecal samples from wildlife and then subjected to a nested polymerase chain reaction–based sequencing approach for detection and characterization. Altogether, 3 (1.6%), 21 (11.5%), and 48 (26.4%) specimens tested positive for Cryptosporidium species, E. bieneusi, and G. duodenalis, respectively. Sequence analyses revealed five known (BEB6, D, MJ13, SC02, and type IV) and two novel (designated SH_ch1 and SH_deer1) genotypes of E. bieneusi. Phylogenetically, novel E. bieneusi genotype SH_deer1 fell into group 6, and the other genotypes were assigned to group 1 with zoonotic potential. Three novel Cryptosporidium genotypes (Cryptosporidium avian genotype V-like and C. galli-like 1 and 2) were identified, C. galli-like 1 and 2 formed a clade that was distinct from Cryptosporidium species. The genetic distinctiveness of these two novel genotypes suggests that they represent a new species of Cryptosporidium. Zoonotic assemblage A (n = 36) and host-adapted assemblages C (n = 1) and E (n = 7) of G. duodenalis were characterized. The overall results suggest that wildlife act as host reservoirs carrying zoonotic E. bieneusi and G. duodenalis, potentially enabling transmission from wildlife to humans and other animals.

Serological survey of Coxiella burnetii infections in dairy cattle, sheep, goats and zoo animals in Hungary - Short communication.

Q fever is a disease of high zoonotic potential, but interest in its causative agent is rather low although it causes some public health problems in Hungary. The prevalence of Q fever is highly variable by country. The main reservoirs of the disease are the same domestic ruminant species everywhere, but the epidemiological profile depends on the features of the specific reservoir. The aim of this large-scale study was to demonstrate the importance of Q fever in different species as a possible source for human infection in most regions of Hungary. A total of 851 serum samples from 44 dairy farms, 16 sheep flocks, 4 goat farms and 3 zoos located in different parts of Hungary were tested. The presence of antibodies to Coxiella burnetii was surveyed in dairy cattle (n = 547), goats (n = 71), sheep (n = 200) and zoo animals (n = 33). The animal species tested in Hungary showed different seroprevalence values of C. burnetii infection. Seropositivity by the enzyme-linked immunosorbent assay was found in 258 out of 547 (47.2%) cows and in 69 out of 271 (25.5%) small ruminants, among them in 47 out of 200 (23.5%) sheep and in 22 out of 71 (31.0%) goats. Antibodies to C. burnetii were not detected in zoo animals. Seropositivity was demonstrated in 44 out of 44 (100%) dairy cattle farms, with at least one serum sample found to be positive on each farm. The seropositivity rate of small ruminant farms was 55.0% (11 positive out of 20 tested), with 9 out of 16 (56.3%) sheep flocks and 2 out of 4 (50.0%) goat herds showing seropositivity.

Retail chicken meats as potential sources of Clostridioides difficile in Al-Jouf, Saudi Arabia

Presence of Clostridioides difficile in stool of food birds and animals is a risk for contamination of their meats to become potential sources of human infection. The main virulence factors of C. difficile are its resistance to antibiotics, production of toxins and spores. As far as I know, this is the first study to evaluate C. difficile prevalence in chicken meats, its toxigenic activities and antibiotics sensitivity patterns in Al-Jouf, Saudi Arabia. Totally, 250 raw chicken meat samples were examined. Standard microbiological and biochemical procedures were used for C. difficile isolation and identification. The suspected colonies were tested by L-proline and C. difficile test kits then confirmed by Vitek 2 compact system. Xpect C. difficile toxin A/B test was used to detect A/B toxins production. Antibiotics susceptibility patterns were detected by Epsilon tests. C. difficile was isolated from 11/250 (4.40%) chicken meat samples; 5/65 (7.69%) legs, 3/65 (4.61%) thighs, 2/60 (3.33%) wings and 1/60 (1.67%) breasts (p = 0.4). All isolates were non-toxigenic. Although all isolates were vancomycin sensitive, some isolates were intermediate/resistant to metronidazole, tetracycline, clindamycin or moxifloxacin antibiotics with variable degrees. C. difficile might contaminate retail chicken meats. Although low level of contamination by non-toxigenic strains was detected, chicken meats should be investigated as C. difficile infection sources for humans especially elders, immune-compromised and long terms wide spectrum antibiotics-used persons. Decreased sensitivity of C. difficile to antibiotics is emerging.

Detection of Multidrug Resistance Salmonella spp. from Chicken Meat by Multiplex PCR and VITEK 2 system

Salmonellae are highly pathogenic foodborne bacteria able to cause infection even at low doses. Infection by Salmonella from contaminated foods leads to gastrointestinal disease known as salmonellosis. Raw chicken can be a source of human infection if the meat products are not properly handled, stored or cooked. The aim of the present study was to investigate the prevalence of Salmonella Enteritidis and Salmonella Typhymurium serovars in retail raw chicken meat from retail market in Erbil, as well as, to assess their antimicrobial resistance. A total of 100 raw retail chicken meat samples were collected and plating on Xylose Lysine Deoxycholate and Salmonella-Shigella agar media. In addition, the typical black colonies were identified and stored for further analysis. In addition, Salmonella spp. isolates recovered from the samples were identified and tested for antibiotics susceptibility by using VITEK 2 automated system. The multiplex polymerase chain reaction was employed to detect Salmonella Enteritidis and Salmonella Typhymurium in the meat samples. Out of 100 raw chicken meat analysed samples, 5 (5%) were positive for Salmonella Typhimurium, while none of the samples were contaminated with Salmonella Enteritidis. The resistance levels of the isolates against 12 different antimicrobial agent were tested: all the strains were suscebtible to Imipenem, Meropenem, and Ciprofloxacin. However, the resistance rates in the bacterial isolates were 20% each for Amikacin, Gentamicin, and Tobramycin, also 40% and 30% resistance for Minocycline, and Trimethoprim/Sulfamethoxazole. All isolates of the Salmonella Typhymurium were multiresistant to three or more antimicrobial agents. Preventive measure such as proper temperature control as well as proper handling of raw chicken meat in the market place are crucial to the minimization of any potential health hazard by this foodborne pathogen.

IS26 Is Responsible for the Evolution and Transmission of blaNDM-Harboring Plasmids in Escherichia coli of Poultry Origin in China.

ABSTRACTCarbapenem-resistant Enterobacteriaceae are some of the most important pathogens responsible for nosocomial infections, which can be challenging to treat. The blaNDM carbapenemase genes, which are expressed by New Delhi metallo-β-lactamase (NDM)-producing Escherichia coli isolates, have been found in humans, environmental samples, and multiple other sources worldwide. Importantly, these genes have also been found in farm animals, which are considered an NDM reservoir and an important source of human infections. However, the dynamic evolution of blaNDM genetic contexts and blaNDM-harboring plasmids has not been directly observed, making it difficult to assess the extent of horizontal dissemination of the blaNDM gene. In this study, we detected NDM-1 (n = 1), NDM-5 (n = 24), and NDM-9 (n = 8) variants expressed by E. coli strains isolated from poultry in China from 2016 to 2017. By analyzing the immediate genetic environment of the blaNDM genes, we found that IS26 was associated with multiple types of blaNDM multidrug resistance regions, and we identified various IS26-derived circular intermediates. Importantly, in E. coli strain GD33, we propose that IncHI2 and IncI1 plasmids can fuse when IS26 is present. Our analysis of the IS26 elements flanking blaNDM allowed us to propose an important role for IS26 elements in the evolution of multidrug-resistant regions (MRRs) and in the dissemination of blaNDM. To the best of our knowledge, this is the first description of the dynamic evolution of blaNDM genetic contexts and blaNDM-harboring plasmids. These findings could help proactively limit the transmission of these NDM-producing isolates from food animals to humans.IMPORTANCE Carbapenem resistance in members of the order Enterobacterales is a growing public health problem that is associated with high mortality in developing and industrialized countries. Moreover, in the field of veterinary medicine, the occurrence of New Delhi metallo-β-lactamase-producing Escherichia coli isolates in animals, especially food-producing animals, has become a growing concern in recent years. The wide dissemination of blaNDM is closely related to mobile genetic elements (MGEs) and plasmids. Although previous analyses have explored the association of many different MGEs with mobilization of blaNDM, little is known about the evolution of various genetic contexts of blaNDM in E. coli. Here, we report the important role of IS26 in forming multiple types of blaNDM multidrug resistance cassettes and the dynamic recombination of plasmids bearing blaNDM. These results suggest that significant attention should be paid to monitoring the transmission and further evolution of blaNDM-harboring plasmids among E. coli strains of food animal origin.

Sero-Monitoring of Horses Demonstrates the Equivac® HeV Hendra Virus Vaccine to Be Highly Effective in Inducing Neutralising Antibody Titres

Hendra virus (HeV) is a high consequence zoonotic pathogen found in Australia. The HeV vaccine was developed for use in horses and provides a One Health solution to the prevention of human disease. By protecting horses from infection, the vaccine indirectly protects humans as well, as horses are the only known source of infection for humans. The sub-unit-based vaccine, containing recombinant HeV soluble G (sG) glycoprotein, was released by Pfizer Animal Health (now Zoetis) for use in Australia at the end of 2012. The purpose of this study was to collate post-vaccination serum neutralising antibody titres as a way of assessing how the vaccine has been performing in the field. Serum neutralization tests (SNTs) were performed on serum samples from vaccinated horses submitted to the laboratory by veterinarians. The SNT results have been analysed, together with age, dates of vaccinations, date of sampling and location. Results from 332 horses formed the data set. Provided horses received at least three vaccinations (consisting of two doses 3–6 weeks apart, and a third dose six months later), horses had high neutralising titres (median titre for three or more vaccinations was 2048), and none tested negative.