Evidence of Powassan/deer tick virus in adult black‐legged ticks (Ixodes scapularis) recovered from hunter‐harvested white‐tailed deer (Odocoileus virginianus) in Pennsylvania: A public health perspective

Studies reporting tick infection rates for Powassan virus (POWV), an emerging zoonotic arthropod‐borne pathogen responsible for POWV disease in the Commonwealth of Pennsylvania, are limited. To determine the presence and ascertain a statewide prevalence of POWV, ticks were collected from 9,912 hunter‐harvested white‐tailed deer (Odocoileus virginianus) heads presented to six regional Pennsylvania Game Commission Chronic Wasting Disease sampling stations in early December of 2013, 2014 and 2015. Of the 2,973 ticks recovered, 1,990 (66.9%) were identified as adult Ixodes scapularis (black‐legged tick). The 1,990 I. scapularis ticks were PCR‐tested for the presence of POWV. The ticks had a statewide Powassan/deer tick virus infection rate of 0.05%, providing evidence of this pathogen in Pennsylvania's adult I. scapularis ticks and supporting the need for more comprehensive pathogen prevalence assessment strategies, as well as increased public health awareness for this emerging zoonotic arthropod‐borne pathogen of public health concern.     

White‐Tailed Deer (Odocoileus virginianus) as a Potential Sentinel for Human Lyme Disease in Indiana

We assessed the potential of white‐tailed deer (WTD) (Odocoileus virginianus) to be a sentinel for human cases of Lyme disease (LD) in Indiana using location data from a 3‐year survey of approximately 3400 hunted deer with associated tick Ixodes scapularis and Borrelia burgdorferi (Bb) data. Data on human LD cases at the county level were obtained from the Indiana Department of Health. All data were assigned to county centroids to match the resolution of the LD data before creating optimized trend surfaces for LD incidence, hunted deer count, Ixodes scapularis and Bb prevalence. To determine whether LD was spatially associated with the areas of high densities of deer, deer with Ixodes scapularis and deer with ticks infected with Bb, we used spatial analysis with distance indices (SADIE). The SADIE analysis found significant spatial association between LD and the distribution of three organismal predictor variables, that is, WTD, Ixodes ticks and Bb. Lyme disease incident rate varied between 0.08 cases per 10 000 habitants (Johnson county) and 5.9 cases per 10 000 habitants (Warren county). In conclusion, WTD can be used as an accurate and cost‐effective sentinel for human LD. This method will permit public health workers to identify potentially endemic areas independently of human case reports.     

Review of Equine Piroplasmosis

Equine piroplasmosis is caused by one of 2 erythrocytic parasites Babesia caballi or Theileria equi. Although the genus of the latter remains controversial, the most recent designation, Theileria, is utilized in this review. Shared pathogenesis includes tick‐borne transmission and erythrolysis leading to anemia as the primary clinical outcome. Although both parasites are able to persist indefinitely in their equid hosts, thus far, only B. caballi transmits across tick generations. Pathogenesis further diverges after transmission to equids in that B. caballi immediately infects erythrocytes, whereas T. equi infects peripheral blood mononuclear cells. The recent re‐emergence of T. equi in the United States has increased awareness of these tick‐borne pathogens, especially in terms of diagnosis and control. This review focuses in part on factors leading to the re‐emergence of infection and disease of these globally important pathogens.     

Detection of Invasive Borrelia burgdorferi Strains in North‐Eastern Piedmont,Italy

Following reports of human cases of Lyme borreliosis from the Ossola Valley, a mountainous area of Piemonte, north‐western Italy, the abundance and altitudinal distribution of ticks, and infection of these vectors with Borrelia burgdorferi sensu lato were evaluated. A total of 1662 host‐seeking Ixodes ricinus were collected by dragging from April to September 2011 at locations between 400 and 1450 m above sea level. Additional 104 I. ricinus were collected from 35 hunted wild animals (4 chamois, 8 roe deer, 23 red deer). Tick density, expressed as the number of ticks per 100 m², resulted highly variable among different areas, ranging from 0 to 105 larvae and from 0 to 22 nymphs. A sample of 352 ticks (327 from dragging and 25 from wild animals) was screened by a PCR assay targeting a fragment of the 16S rRNA gene of B. burgdorferi s.l. Positive samples were confirmed with a PCR assay specific for the 5S‐23S rRNA intergenic spacer region and sequenced. Four genospecies were found: B. afzelii (prevalence 4.0%), B. lusitaniae (4.0%), B. garinii (1.5%) and B. valaisiana (0.3%). Phylogenetic analysis based on the ospC gene showed that most of the Borrelia strains from pathogenic genospecies had the potential for human infection and for invasion of secondary body sites.     

Seasonality and anatomical location of human tick bites in the United Kingdom

Tick bites on humans can occur in a variety of habitats and may result in the transmission of tick‐borne pathogens, such as the causative agent of Lyme borreliosis (LB), Borrelia burgdorferi sensu lato. As the risk of transmission of this pathogen to the host increases with the duration of tick feeding, the recognition and removal of ticks as soon as possible following attachment is important for reducing the risk of infection. Performing a thorough body examination for ticks following potential exposure is recommended by tick awareness campaigns. Knowledge of where on the body feeding ticks are frequently found, and at which times of year peak tick exposure occurs, provides important information for public health messaging and may aid those bitten by ticks to engage more effectively with tick‐checking behaviour. This paper summarizes human tick bites in the United Kingdom (UK) during 2013–2018 reported to Public Health England's passive Tick Surveillance Scheme and further examines the anatomical location and seasonality of bites from the most commonly encountered tick and LB vector Ixodes ricinus. A total of 1,328 tick records from humans were received of which 93% were I. ricinus. Humans were most commonly bitten by I. ricinus nymphs (70% bites). Tick bites were recorded on all parts of the body, but there were significant differences in their anatomical location on adults and children. Most tick bites on adults occurred on the legs (50%), whereas on children tick bites were mostly on the head and neck (43%). Bites from I. ricinus were recorded throughout the year but were most numerous during May to August. This study adds to the body of research on the seasonality and anatomical location of human tick bites in temperate Europe and highlights the importance of data collected through passive surveillance in addition to research and epidemiological studies.     

Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.     

Humoral response of Borrelia burgdorferi outer surface protein A (OspA) vaccination in equids

The objective of this study was to assess the safety and humoral response of outer surface protein A (OspA) Borrelia burgdorferi vaccine in equids via the transdermal or subcutaneous route over 355 days. Prior to vaccination, serological testing confirmed the vaccination and exposure status to B. burgdorferi. Vaccine was administered on Days 0, 22 and 226. Equids were examined for vaccine reactions at 24 h post‐vaccination. Antibodies to outer surface proteins were quantified over 355 days. A total of 42 healthy adult equids of various ages and breeds were used. These equids were selected based on unlikely exposure to B. burgdorferi. The equids were grouped according to full size, miniature, age and sex to create two relatively heterogeneous mirrored groups of 20 equids. Group TD (20 equids) was administered 1 mL (1/2 mL/site) vaccine transdermally over the pectoral region. Group SQ (19 equids) was administered 2 mL in a single injection subcutaneously at the left cervical region. Group C (3 equids) was unvaccinated. Vaccine was administered on Days 0, 22 and 226. Antibodies to outer surface proteins were quantified over 355 days. Both vaccinated groups responded with a significant increase in OspA antibodies as compared with the control group (P<0.0001) and to themselves prevaccination. The mean response was greater in vaccinated equids (Group SQ, P<0.0080; Group TD, P<0.0016) as compared prevaccine and control. Equids responded to the OspA vaccine via either route. This data may aid in strategic vaccination protocols and the development of a USDA approved vaccine for equids in the prevention of Lyme disease using the OspA vaccine.     

The Presence of Borrelia miyamotoi,A Relapsing Fever Spirochaete,in Questing Ixodes ricinus in Belgium and in The Netherlands

Borrelia miyamotoi is a tick‐borne bacterium that may cause relapsing fever in humans. As this pathogen has been discovered in Europe only recently, only little is known about its local impact on human health and its spatial distribution. In this study, we show the results of PCR screenings for B. miyamotoi in flagged Ixodes ricinus from Belgium and the Netherlands. B. miyamotoi was detected in nine of thirteen, and three of five locations from the Netherlands and Belgium, respectively. These outcomes indicate that B. miyamotoi is more spread than previously thought. The mean infection rate B. miyamotoi was 1.14% for Belgium and 3.84% for the Netherlands.     

Prevalence of Salmonella spp. in Cane Toads (Bufo marinus) from Grenada,West Indies,and their Antimicrobial Susceptibility

Cloacal swabs and caecal contents sampled from 58 cane toads (Bufo marinus) in St George’s parish, Grenada, during a 7‐month period in 2011 were examined by an enrichment and selective culture method for presence of Salmonella spp. Twenty‐four (41%) toads were positive for Salmonella spp. of which eight were Salmonella enterica serovar Javiana, and eight were S. enterica serovar Rubislaw. The other serovars were as follows: Montevideo, 6; Arechavaleta, 1; and serovar: IV:43:‐:‐, 1. The high frequency of isolation of serovar Javiana, an emerging human pathogen associated with several outbreaks in the recent years in the eastern United States, suggests a possible role for cane toads in transmission of this serovar. Although S. Rubislaw has been isolated from lizards, bats and cases of some human infections, there is no report of its carriage by cane toads, and in such high frequency. The rate of carriage of S. Montevideo, a cause for human foodborne outbreaks around the world was also over 10% in the 58 toads sampled in this study. The antimicrobial drug susceptibility tests against amoxicillin‐clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, imipenem, nalidixic acid, streptomycin, tetracycline and trimethoprim‐sulfamethoxazole showed that drug resistance is minimal and is of little concern. Antimicrobial resistance was limited to ampicillin and amoxicillin‐clavulanic acid in one isolate of S. Javiana and one isolate of S. Rubislaw. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars not identified previously in cane toads in Grenada, West Indies.     

Modelling the Phenological Relationships of Questing Immature Ixodes Ricinus (Ixodidae) Using Temperature and NDVI Data

All active stages of the tick Ixodes ricinus were collected monthly at two sites in northern Spain between the years 2000 and 2007. We used percentile accumulation of the active stage in the environment to evaluate simple and coherent correlations between accumulation of the active stages of larvae and nymphs and medium‐resolution MODIS satellite‐derived information on the climate, including monthly and accumulated temperature and the Normalized Difference Vegetation Index (NDVI). This framework is not intended to predict the actual abundance of ticks in the field as a measure of the hazard to humans, but to provide a basic structure for addressing the phenology of the tick in its geographic range. We demonstrated that the accumulation of larval ticks in the active stage is a sigmoid function of the accumulated temperature from the beginning of the calendar year. We also demonstrated that the accumulated temperature necessary to recruit nymphs from the questing larval stage is a function of the changes in accumulated larvae and nymphs and the accumulated temperature and NDVI recorded by the Aqua sensor. The low p‐values obtained in the regressions confirmed that such recruitment can be calculated using time intervals to estimate, for example, the beginning of the questing period or the time of the year when a population peak can be expected. The comparison among predicted and actual accumulated temperatures between larvae and nymph recruitment had an averaged error of ±20 days in one complete year. The use of accumulated temperature and NDVI proposed in this study opens up the re‐evaluation of reports on the phenology of the tick in Europe. This framework is intended to evaluate the same correlations along the tick's range and predict its phenological patterns in areas of pathogen transmission risk for humans.     

The effects of the supplementation of Bacillus subtilis RX7 and B2A strains on the performance,blood profiles,intestinal Salmonella concentration,noxious gas emission,organ weight and breast meat quality of broiler challenged with Salmonella typhimurium

An experiment was conducted to evaluate the effects of B. subtilis RX7 and B. subtilis B2A on growth performance, blood profiles, intestinal Salmonella population, noxious gas emission, organ weight and breast meat quality of broilers under S. typhimurium challenge. A total of 120, one‐day‐old Ross 308 male broiler chicks were assigned to four dietary treatments, composed of six replications, with five birds per replication, for 10 day. The dietary treatment groups were negative control (NC; no antibiotic, no B. subtilis), positive control (PC; NC + 0.1% virginiamycin), B. subtilis RX7 (NC + 0.1% B. subtilis RX7 1.0 × 10⁹ cfu/g) and B. subtilis B2A (NC + 0.1% B. subtilis 1.0 × 10⁹ cfu/g). All birds were orally challenged with 2 ml suspension, containing 10⁴ cfu/ml of S. typhimurium KCCM 40253. Results indicated that the body weight gain, feed intake and feed conversion did not differ, among all comparative treatments. Serum haptoglobin concentration was lower in Bacillus treatments (RX7 + B2A) than the NC treatment (p < 0.05). Intestinal and excreta Salmonella number, and excreta ammonia gas emission in the PC treatment or Bacillus treatments, was lower than the NC treatment (p < 0.05). Breast pH, colour and water‐holding capacity were not affected by supplementation of B. subtilis RX7 and B2A. However, drip loss at 1 day post‐slaughter from birds fed with B. subtilis RX7 and B2A decreased, compared with the positive control birds (p < 0.05). Relative gizzard weights of birds fed B. subtilis RX7 and B2A were significantly higher than the NC birds under S. typhimurium challenge. It is concluded from the results that B. subtilis RX7 and B2A increased the gizzard weight and decreased the intestinal and excreta Salmonella population and excreta ammonia gas, and drip loss of breast meat after being stored for 1 day, under stress caused by the S. typhimurium challenge.     

Distribution of Neoehrlichia mikurensis in Ixodes ricinus ticks along the coast of Norway: The western seaboard is a low‐prevalence region

Neoehrlichia mikurensis is a tick‐borne pathogen widespread among ticks and rodents in Europe and Asia. A previous study on Ixodes ricinus ticks in Norway suggested that N. mikurensis was scarce or absent on the south‐west coast of Norway, but abundant elsewhere. The aim of this study was to further investigate the prevalence and distribution of N. mikurensis along the western seaboard of Norway in comparison with more eastern and northern areas. The second aim of the study was to examine seasonal variation of the bacterium in one specific location in the south‐eastern part of Norway. Questing I. ricinus were collected from 13 locations along the coast of Norway, from Brønnøysund in Nordland County to Spjærøy in Østfold County. In total, 11,113 nymphs in 1,113 pools and 718 individual adult ticks were analysed for N. mikurensis by real‐time PCR. The mean prevalence of N. mikurensis in adult ticks was 7.9% while the estimated pooled prevalence in nymphs was 3.5%. The prevalence ranged from 0% to 25.5%, with the highest prevalence in the southernmost and the northernmost locations. The pathogen was absent, or present only at low prevalence (<5%), at eight locations, all located in the west, from 58.9°N to 64.9°N. The prevalence of N. mikurensis was significantly different between counties (p < .0001). No significant seasonal variation of N. mikurensis prevalence was observed in the period May to October 2015. Our results confirm earlier findings of a low prevalence of N. mikurensis in the western seaboard of Norway.     

Optimization of surfactin production from Bacillus subtilis in fermentation and its effects on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers

Bacillus species are commonly used as probiotics in the poultry feed industry for preventing infectious diseases and improving productivity by altering gastrointestinal microbiota. The growth parameters of Bacillus subtilis for surfactin production in fermentation and the benefits of surfactin on broiler chickens remain unclear. In this study, we examined the growth parameters of B. subtilis in fermentation and evaluated the effects of surfactin from B. subtilis‐fermented products on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers. Results showed that the highest viable biomass of B. subtilis was observed at 10% molasses and 2% yeast supplementation during fermentation. The 4‐ and 6‐day fermented B. subtilis products were heat‐, acid‐ and bile‐resistant. Furthermore, the 4‐day fermented B. subtilis products with the highest surfactin concentration showed the maximal antimicrobial activity against pathogens, including Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and C. perfringens. Dietary B. subtilis‐fermented product supplementation in broilers significantly improved intestinal morphology and necrotic lesions under C. perfringens challenge. Bacillus subtilis treatments could enhance broiler productivity, as well as promote bone quality and intestinal morphology. These results together indicate that B. subtilis‐fermented products containing surfactin have potential for the development as feed additives and use as possible substitutes for antibiotics to treat C. perfringens in the poultry industry.     

Multidrug‐Resistant Outbreak‐Associated Salmonella Strains in Irrigation Water from the Metropolitan Region,Chile

Salmonella enterica (S. enterica) is the main cause of foodborne diseases in the Chilean population. With the aim of characterizing the presence of S. enterica in bodies of water, samples from 40 sources were obtained, including rivers and irrigation canals used by agricultural farms in the most populated regions of Chile. As result, 35 S. enterica isolates belonging to several serotypes were detected, with the highest frequency represented by Typhimurium and Enteritidis. All strains showed phenotypic antimicrobial resistance, and most of them were multiresistant to critically important antimicrobials. In addition, the pulse‐field gel electrophoresis analysis using XbaI and BlnI endonucleases showed that seven Salmonella isolates belonging to serotypes Typhimurium, Enteritidis and Infantis had identical pulsotypes to outbreak‐associated clinical isolates detected in the Chilean population, suggesting a public health risk of water pollution in this region. Among sampling sites, the higher detection rates were observed in rural than urban and peri‐urban areas, suggesting that the animal husbandry might contribute for environmental dispersion of this pathogen. Future efforts should address the characterization of cause‐and‐effect relationship between water contamination and foodborne disease, including the implementation of surveillance programmes to tackle potential risks for both human and animal populations.     

Quantitative expression of mRNA encoding BMP/SMAD signalling genes in the ovaries of Booroola carrier and non‐carrier GMM sheep

The GMM sheep is a carrier of Booroola fecundity (FecB) gene, which produces the twins and triplets in one lambing. The homozygous carrier GMM (FecB^BB), non‐carrier GMM and Malpura (FecB⁺⁺) ewes were synchronized by progesterone sponges, and the plasma progesterone concentration was measured by RIA. The results showed that the progesterone concentration did not differ significantly (p > .05) in homozygous carrier GMM (5.74 ± 1.2 ng/ml), non‐carrier GMM (5.42 ± 1.4 ng/ml) and non‐carrier Malpura ewes (5.67 ± 1.5 ng/ml). Further, quantitative expression of BMP factors/receptors and SMAD signalling genes were analysed in the ovaries of sheep by qRT‐PCR. The study showed that the expression of BMP2 was slightly higher (p > .05) in carrier GMM than that of non‐carrier GMM, but it was almost similar to Malpura ewes. Expression of BMP4 and BMP7 was significantly higher (p < .001; p < .05) in carrier GMM than that of non‐carrier GMM and Malpura ewes. Although BMP6 expression was higher (p > .05) in carrier GMM than that of non‐carrier GMM, but lower (p > .05) than the Malpura ewes. Expression of BMP15 (p < .05), GDF5 (p < .01) and GDF9 (p < .05) was significantly higher in carrier GMM than non‐carrier GMM ewes. Surprisingly, BMPR1B expression was significantly higher (p < .001) in non‐carrier GMM and Malpura than the carrier GMM ewes, while TGFβRI did not differ significantly (p > .05) among both GMM genotypes. On the other hand, expression of BMPR1A (p > .05) and BMPRII (p < .05) was higher in carrier GMM than the non‐carrier GMM, but significantly lower (p < .001) than the Malpura ewes. It was interesting to note that the expression of SMAD1 (p > .05), SMAD2 (p < .001), SMAD3 (p < .05), SMAD4 (p < .001), SMAD5 (p < .001) and SMAD8 (p < .001) was lower in the carrier GMM than that of non‐carrier GMM ewes. It is concluded that the FecB mutation alters the expression of BMPR1B and SMAD signalling genes in the ovaries of homozygous carrier GMM ewes.     

Maternal dietary linoleic acid supplementation promotes muscle fibre type transformation in suckling piglets

As meat quality is basically dependent on muscle fibre characteristics, it is important to know how muscle fibres are regulated and transformed. This study aimed to investigate the effect of maternal dietary supplementation on muscle fibre types using 3% saturated fatty acid (palmitic acid, PA) or 3% unsaturated fatty acid (linoleic acid, LA) from 80 days of gestation to the weaning of offspring (25 days post‐natal). The results indicated that higher mRNA levels of MyHCI type genes were found in the soleus muscles of piglets that suckled from LA‐supplemented sows than from PA‐supplemented sows. In addition, LA treatment increased the gene expression of the type I muscle fibre marker troponin I (p < 0.01), suggesting that LA promoted muscle fibre type transformation to type I fibres. Moreover, PGC‐1α (p < 0.01) and MEF2c (p < 0.05) mRNA levels were higher in the piglets from the LA treatment group than in those from the PA treatment group. Furthermore, LA supplementation also significantly increased AMP‐activated protein kinase (AMPK) mRNA levels (p < 0.05), which is an upstream regulator of PGC‐1α. Collectively, these findings demonstrated that maternal dietary LA supplementation promoted muscle fibre transformation to type I fibre and that this process may be mediated through an AMPK‐dependent pathway.     

Urban foci of murine typhus involving cat fleas (Ctenocephalides felis felis) collected from opossums in Mexico City

Murine typhus, a neglected rickettsiosis caused by Rickettsia typhi, is a common disease in several Latin‐American countries. The sylvatic life cycle of R. typhi encompasses the presence of several wild mammals, particularly opossums of the genus Didelphis and their associated fleas. Due to the colonization of wild environments by human populations, the increase in contact with opossum fleas has generated the presence of urban outbreaks of typhus. For this reason, the aim of our study was to identify the presence and diversity of Rickettsia sp. in fleas collected from opossums of an urban reserve in Mexico City. Opossums were captured from February to September 2017. For the detection of Rickettsia DNA, fragments of 800 bp of the citrate synthase (gltA) and the outer membrane protein B (ompB) were amplified. A total of 141 fleas (111 ♀, 30 ♂) of a single species (Ctenocephalides felis felis) were recovered from 31 Didelphis virginiana. Rickettsia DNA was detected in 17.7% (25/141) of the analysed fleas, recovered from seven infested opossums. The Maximum likelihood of sequences exhibited an identity of 99%–100% with sequences of R. typhi from southern United States. This work represents the first record of R. typhi in fleas from opossums in Mexico.     

Rickettsial Infection in Animals,Humans and Ticks in Paulicéia,Brazil

A previous study in Paulicéia Municipality, south‐eastern Brazil, reported 9.7% of the Amblyomma triste ticks to be infected by Rickettsia parkeri, a bacterial pathogen that causes spotted fever in humans. These A. triste ticks were shown to be associated with marsh areas, where the marsh deer Blastocerus dichotomus is a primary host for this tick species. During 2008–2009, blood serum samples were collected from 140 horses, 41 dogs, 5 opossums (Didelphis albiventris) and 26 humans in farms from Pauliceia Municipality. Ticks were collected from these animals, from vegetation and from additional wildlife in these farms. Overall, 25% (35/140) of the horses, 7.3% (3/41) of the dogs, 3.8% (1/26) of the humans and 100% (5/5) of the opossums were seroreactive (titre ≥64) to spotted fever group (SFG) Rickettsia spp. Multivariate statistical analysis indicated that horses that were allowed to forage in the marsh were 4.8 times more likely to be seroreactive to spotted fever group (SFG) Rickettsia spp than horses that did not forage in the marsh. In addition, horses that had been living in the farm for more than 8.5 years were 2.8 times more likely to be seroreactive to SFG Rickettsia spp than horses that were living for ≤8.5 years. Ticks collected from domestic animals or from vegetation included Amblyomma cajennense, Amblyomma coelebs, Amblyomma dubitatum, Dermacentor nitens and Rhipicephalus microplus. By PCR analyses, only one pool of A. coelebs ticks from the vegetation was shown to be infected by rickettsiae, for which DNA sequencing revealed to be Rickettsia amblyommii. Ticks (not tested by PCR) collected from wildlife encompassed A. cajennense and Amblyomma rotundatum on lizards (Tupinambis sp), and A. cajennense and A. triste on the bird Laterallus viridis. Our results indicate that the marsh area of Paulicéia offers risks of infection by SFG rickettsiae.     

The occurrence of Salmonella,extended‐spectrum β‐lactamase producing Escherichia coli and carbapenem resistant non‐fermenting Gram‐negative bacteria in a backyard poultry flock environment

Increase in the number of small‐scale backyard poultry flocks in the USA has substantially increased human‐to‐live poultry contact, leading to increased public health risks of the transmission of multi‐drug resistant (MDR) zoonotic and food‐borne bacteria. The objective of this study was to detect the occurrence of Salmonella and MDR Gram‐negative bacteria (GNB) in the backyard poultry flock environment. A total of 34 backyard poultry flocks in Washington State (WA) were sampled. From each flock, one composite coop sample and three drag swabs from nest floor, waterer‐feeder, and a random site with visible faecal smearing, respectively, were collected. The samples were processed for isolation of Salmonella and other fermenting and non‐fermenting GNB under ceftiofur selection. Each isolate was identified to species level using MALDI‐TOFF and tested for resistance against 16 antibiotics belonging to eight antibiotic classes. Salmonella serovar 1,4,[5],12:i:‐ was isolated from one (3%) out of 34 flocks. Additionally, a total of 133 ceftiofur resistant (Cef^R) GNB including Escherichia coli (53), Acinetobacter spp. (45), Pseudomonas spp. (22), Achromobacter spp. (8), Bordetella trematum (1), Hafnia alvei (1), Ochrobactrum intermedium (1), Raoultella ornithinolytica (1), and Stenotrophomonas maltophilia (1) were isolated. Of these, 110 (82%) isolates displayed MDR. Each flock was found positive for the presence of one or more Cef^R GNB. Several MDR E. coli (n = 15) were identified as extended‐spectrum β‐lactamase (ESBL) positive. Carbapenem resistance was detected in non‐fermenting GNB including Acinetobacter spp. (n = 20), Pseudomonas spp. (n = 11) and Stenotrophomonas maltophila (n = 1). ESBL positive E. coli and carbapenem resistant non‐fermenting GNB are widespread in the backyard poultry flock environment in WA State. These GNB are known to cause opportunistic infections, especially in immunocompromised hosts. Better understanding of the ecology and epidemiology of these GNB in the backyard poultry flock settings is needed to identify potential risks of transmission to people in proximity.     

Detection of Zoonotic Protozoa Toxoplasma gondii and Sarcocystis suihominis in Wild Boars from Spain

Food safety regulations require the control of the presence of protozoa in meats destined for human consumption. Wild boar (Sus scrofa) meat may constitute a source of zoonoses. A 23.8% (688/2881) seroprevalence of anti‐Toxoplasma gondii antibodies and 72.2% (662/910) Sarcocystis sarcocysts prevalence were detected among wild boars hunted in Southwestern areas of Spain. Identity of Sarcocystis spp. was performed by RFLP‐PCR and sequencing, detecting S. miescheriana (7/8) and the zoonotic S. suihominis (1/8). Risk assessment studies of these coccidian in meats destined to human consumption are needed.