Alternative Traits for Genetic Evaluation of Mastitis Based on Lifetime Merit.

Genetic selection has achieved little progress in reducing mastitis incidence. Mastitis traits are problematic due to the lack of sensitivity of the data and reliance on clinical diagnosis, often missing subclinical cases, and/or on monthly somatic cell count (SCC) measurements. The current measure for mastitis is the lactation average of the somatic cells score (LSCS). We studied two datasets: (1) 148 heifers divided into non-intramammary infected, sub-clinically infected and clinical mastitis groups; (2) data from 89,601 heifers from Israeli Holsteins through the same period divided into "udder healthy" (UH) and "non-healthy" (UNH) by a threshold of SCC 120,000 cells/mL in all nine monthly milk recordings. In study 1, non-infected heifers had significantly (p < 0.05) more partum, production days and overall lifetime milk production compared to clinical and sub-clinically infected. In study 2, UH heifers (20.3%) had significantly higher (p < 0.01) lifetime milk, production days, and lactations. Subdividing datasets by sires, the same analyses detected differences in percentages of UH daughters between the sire groups. Lifetime milk production correlated (r = +0.83, p < 0.001) with udder health status. SCC threshold of less than 120,000 cells/mL during all first lactation measurements indicated healthy udder, providing a valuable insight that this dichotomous trait is advantageous for calculating lifetime net-merit index (NM$) over LSCS.

Complete genome sequence of multi-drug-resistant Salmonella enterica subsp. enterica serovar Typhimurium strain Bnaya isolated from a dairy calf in Israel.

This manuscript reports the complete genome sequence of a Salmonella enterica subsp. enterica serovar Typhimurium strain (designated "Bnaya"), isolated from a dead dairy calf with severe diarrhea in Israel. The isolate exhibited multi-drug resistance, which is highly unusual in bovine Salmonella spp. in Israel, prompting further investigation.

Resistant Bacteria in Broiler Litter Used as Ruminant Feed: Effect of Biotic Treatment.

The use of antimicrobial drugs and coccidiostats in poultry farming is widespread, with a significant proportion of these drugs being excreted and released into the environment. The residues of such drugs in poultry litter (PL) can result in the development of antibiotic-resistant bacteria. The impact of different biotic treatments (aerobic, anaerobic, and stacking) on broiler litter (BL) before its use as animal feed has not been studied extensively, nor have the differences between antimicrobial-dependent and independent broiler farms been investigated. This study aimed to determine the resistant bacteria in BL used as ruminant feed before and after litter treatment. The results show that the most resistant bacteria before BL treatment were the Enterococcus species. This study also found that the quantity of amoxicillin-resistant Enterococcus detected in samples from antimicrobial-dependent farms was significantly higher than in those from antimicrobial-independent farms. Additionally, 14% of bacteria were multi-resistant to tetracycline, sulfafurazole, and erythromycin in antimicrobial-independent farm litters, significantly lower than those measured in antimicrobial-dependent broiler farm litter. This study highlights the importance of better understanding, regulating, managing, and using animal waste appropriately to reduce the number of antibiotic-resistant bacteria and minimize the use of antimicrobials that carry high risks for animals, humans, and the environment.

Upregulation of ica Operon Governs Biofilm Formation by a Coagulase-Negative Staphylococcus caprae.

Staphylococcus caprae is a Gram-positive, coagulase-negative staphylococci (CoNS), which appears as commensal in the skin, as well as a prevalent mastitis pathogen of goats. Occasionally, it is also associated with infections in humans. Biofilm formation has been identified as a putative virulence factor in S. caprae. Biofilms are multicellular communities protected by a self-produced extracellular matrix (ECM), which facilitates the resistance of bacterial cells to antimicrobial treatments. The ECM is constructed by exopolysaccharides, including the major exopolysaccharide-polysaccharide intercellular adhesion (PIA), regulated by the ica operon in Staphylococcus species. The aim of this study was to characterize the expression of the ica operon in relation to biofilm formation in S. caprae. Results showed that within a few hours of growth, S. caprae could adhere to polystyrene surfaces, start to accumulate, and form biofilm. Peak biofilm biomass and maturation were reached after 48 h, followed by a reduction in biomass after 72 h. Confocal laser scanning microscopy showed the expression of matrix-associated proteins and polysaccharides at various time points. The expression dynamics of the ica operon were investigated using real-time reverse transcriptase PCR (RT)-qPCR, which showed elevated expression during the early stages of biofilm formation and subsequent downregulation throughout the biofilm aging process. In conclusion, our results show that the ica operon is essential in regulating biofilm formation in S. caprae, similar to other Staphylococcus species. Furthermore, the robustness of the observed biofilm phenotype could account for the successful intramammary colonization and may explain disease persistence caused by this pathogenic bacterium.

Public and animal health risks associated with spillover of Brucella melitensis into dairy farms.

Brucellosis is a worldwide zoonosis with important public health, animal health and economic implications. Brucella melitensis, commonly associated with small ruminants, is an emerging bovine pathogen in dairy farms. We analysed all B. melitensis outbreaks affecting dairy farms in Israel since 2006, combining traditional and genomic epidemiology to explore the public health implications of this One Health challenge. Whole-genome sequencing was applied to bovine and related human B. melitensis isolates from dairy farm outbreaks. cgMLST-based and SNP-based typing was integrated with epidemiological and investigation data. A secondary analysis combining the bovine-human isolates with endemic human isolates from southern Israel was performed. A total of 92 isolates from dairy cows and related human cases originating from 18 epidemiological clusters were analysed. Most genomic and epi-clusters were congruent, but sequencing showed relatedness between apparently unrelated farm outbreaks. Nine secondary human infections were also genomically confirmed. The bovine-human cohort appeared intermixed with 126 endemic human isolates in southern Israel. We show a persistent and widespread circulation of B. melitensis in dairy farms in Israel with secondary occupational human infection. The genomic epidemiology also uncovered cryptic connections between outbreaks. A regional connection between bovine and endemic human brucellosis cases points to a common reservoir, most probably local small ruminant herds. Control of humans and bovine brucellosis is inseparable. Epidemiological and microbiological surveillance and implementation of control measures across the entire range of farm animals is needed to mitigate this public health challenge.

Evaluation of acoustic pulse technology as a non-antibiotic therapy for bovine intramammary infections: Assessing bacterial cure vs. recovery from inflammation.

Introduction: The spread of antimicrobial resistance (AMR) is a major threat to human and animal health. Therefore, new solutions are needed to prevent returning to a world without effective antibiotics. Mastitis in dairy cows is a major reason for antimicrobial use in food animal production, and mastitis-causing bacteria have the potential to develop AMR. In this study, acoustic pulse technology (APT) was explored as an alternative to antimicrobials for the treatment of mastitis in dairy cows. APT involves the local transmission of mechanical energy through soundwaves which stimulate anti-inflammatory and angiogenic responses in the udder. These responses promote udder recovery and enhance resistance to bacterial infections. Methods: We examined 129 Israeli dairy cows with mastitis in this prospective, controlled study to assess the efficiency of APT treatment on cure and recovery rates. An accurate diagnosis of suspected or confirmed infectious mastitis was made from cows having clinical signs of mastitis and/or somatic cell count (SCC) of above 400,000 cells/mL. The cows were divided into three groups: Group 1 (n = 29), cows with no bacterial findings (NBF); Group 2 (n = 82), cows with clinical signs of mastitis or SCC >400,000 cells/mL in the most recent test; and Group 3 (n = 18), cows with chronic mastitis (two or more tests with SCC >400,000 cells/mL within 3 months). All the cows received APT treatment, which involved 400 pulses on two sides of the infected quarter, delivered in three phases over 3 days. The cure for the mammary gland was indicated by the absence of bacterial growth in post-treatment cultures and recovery by a decrease in SCC to < 250,000 cells/mL in two of three post-treatment tests. Results and discussion: In Group 2, cure and recovery rates were 67.1 and 64.6%, respectively, and were not significantly different between Gram-negative and Gram-positive infections. A similar recovery rate was found in NBF cows. However, in cows with chronic mastitis, both the cure and recovery rates were significantly lower (22.2 and 27.8%, respectively). These results have important implications for dairy farmers, as APT treatment could lead to substantial savings of up to $15,106/year in a 100-cow herd, considering the national estimated prevalence of mastitis and the cost of individual treatment. APT should be further investigated as a viable and sustainable alternative to antimicrobial therapy for mastitis, offering economic benefits to dairy producers and the possibility of preventing AMR.

Infant Botulism, Israel, 2007-2021.

Infant botulism (IB) is an intestinal toxemia that manifests as descending paralysis, constipation, and, in some cases, respiratory failure. Laboratory-confirmed IB cases are rare, and recent data in Israel are lacking. We conducted a national multicenter retrospective study of laboratory-confirmed IB cases reported in Israel during 2007-2021. A total of 8 cases were reported during the study period. During 2019-2021, incidence may have increased because of a cluster of 5 cases. Infant median age for diagnosis was 6.5 months, older than previously reported (3 months). Most cases occurred during March-July. Honey consumption was reported in 1 case, and possible environmental risk factors (living nearby rural or construction areas, dust exposure, and having a father who works as a farmer) were reported in 6 cases. Although IB is rare, its incidence in Israel may have increased over recent years, and its epidemiology and risk factors differ from cases reported previously in Israel.

Photobacterium damselae subspecies damselae Pneumonia in Dead, Stranded Bottlenose Dolphin, Eastern Mediterranean Sea.

Photobacterium damselae subspecies damselae, an abundant, generalist marine pathogen, has been reported in various cetaceans worldwide. We report a bottlenose dolphin in the eastern Mediterranean Sea that was found stranded and dead. The dolphin had a severe case of chronic suppurative pneumonia and splenic lymphoid depletion caused by this pathogen.

Neospora-related abortions in sheep in Israel - A serological diagnostic challenge in an endemic area.

Abortions in sheep flocks is a common multifactorial problem, which affects animal productivity and welfare. Israel is endemic to several infectious abortifacient pathogens, with neosporosis identified as a prominent cause. High seroprevalence of N. caninum is present in the general sheep population, making complicate to associate it as the causative agent of abortion in sheep. Here we describe two investigations of abortion storms in sheep flocks with high seroprevalence of neosporosis. In flock A, higher anti-Neospora antibody titers were demonstrated in aborting versus non-aborting ewes, suggesting that it may be the cause of abortions. In flock B, several infectious abortifacients were identified, but only the seroprevalence of border disease differed statistically between aborting and non-aborting ewes. These reports highlight the difficulty of diagnosing the cause of abortion in a multifactorial situation, and confirm the necessity to assess paired samples from aborting and non-aborting ewes, for reliable interpretation of the results.

What Is Hiding in the Israeli Mediterranean Seawater and Beach Sand.

OBJECTIVE: In the present study, we aimed to investigate the presence of fungi that may affect human health in sand and water on Israeli Mediterranean Sea coast beaches. METHODS: The study included screening of the sand and water of six urban beaches from north to south on the Israeli Mediterranean coast. Sand samples were extracted with water, and the water wash was cultured and quantitated. Water samples were quantitated as well. MALDI-TOF MS analysis and ITS sequencing identified the fungi. RESULTS: The study considered several parameters: 1. Presence of fecal-contamination-related fungi; 2. Presence of dermal-infection-related fungi. 3. Presence of allergy-related fungi; 4. Presence of fungi posing risk for immunocompromised individuals. The screen revealed that about 80% of the isolates were molds and about 20% yeasts. The mold species included opportunistic pathogens and potential allergens: Aspergillus fumigatus and other Aspergillus species, Fusarium, Penicillium, and Mucorales species. Yeast isolates included Candida-including the human commensals Candida albicans and Candida tropicalis-Cryptococcus, and Rhodotorula species. CONCLUSIONS: The results suggest that beaches should be monitored for fungi for safer use, better management, and the benefit of public health.

Fecal Microbiome Features Associated with Extended-Spectrum ß-Lactamase-Producing Enterobacterales Carriage in Dairy Heifers.

Extended-spectrum ß-lactamases (ESBLs) are a growing public health threat, and one key human exposure point is through livestock and the food supply. Understanding microbiome factors associated with fecal ESBL carriage can help detect and ideally assist with controlling and preventing ESBL dissemination among livestock. The objective of this study was to investigate the diversity and composition of the heifer fecal microbiota in ESBL-producing Enterobacterales (ESBL-PE) carriers and noncarriers. A total of 59 fecal samples were collected from replacement heifers between 12 and 18 months old from eight dairy farms in central Israel. Genomic DNA was extracted, and 16S rRNA amplicon sequencing was performed (Illumina short reads), focusing on a comparison between 33 ESBL-PE carriers (55.9%) and 26 (44.1%) noncarriers. Samples were analyzed and compared using QIIME2 (DADA2 pipeline and taxonomic assignment with SILVA database) and associated R packages for alpha and beta diversity and taxonomic abundances. Alpha diversity (Shannon diversity) and beta diversity (unweighted UniFrac) showed no significant difference between ESBL-PE carriers and noncarriers. Heifers from farms feeding calves with pooled colostrum had higher ESBL-PE carriage rates than heifers from farms feeding with individual mother colostrum (p < 0.001). Taxonomical abundance analysis revealed that the most common bacterial phyla were Bacteroidetes (44%) and Firmicutes (38%). There was no significant difference in taxonomic composition between ESBL-PE carriers and noncarriers at the phylum and genus levels. However, LEfSe biomarker discovery analysis identified several genera which were significantly different between carriers and noncarriers. For example, Prevotellacaea, Bacteroides, Rikenellaceae, and uncultured Bacteroidales were more abundant in ESBL carriers than noncarriers. Some aspects of microbiota composition differ between ESBL carriers and noncarriers in dairy heifers, specifically the abundance of certain genera. Feeding with pooled colostrum may play a role in that assembly. These could potentially serve as markers of ESBL-PE carriage. However, further research is needed to determine whether these observed differences have a significant impact on colonization with ESBL-PE.

Pseudomonas aeruginosa isolation from dog grooming products used by private owners or by professional pet grooming salons: prevalence and risk factors.

BACKGROUND: Pseudomonas aeruginosa is the most commonly isolated bacterium from skin lesions of dogs with post-grooming furunculosis (PGF). It is frequently found in human hair and skin care products, and may pose a health risk to consumers. Information regarding the prevalence of P. aeruginosa contamination of dog grooming products is lacking. OBJECTIVES: To investigate the prevalence of P. aeruginosa contamination in nonmedicated dog grooming products after either home or professional use in pet grooming salons, and to identify risk factors that may be associated with contamination. MATERIALS AND METHODS: Of 117 bottles of grooming products sampled for bacterial culture, 97 were used by pet grooming salons and 20 were used by private individuals. The following suspected risk factors were recorded: bottle size, relative remaining volume, content dilution, expiration date and ingredient list. RESULTS: Pseudomonas aeruginosa was isolated from 14 of 117 samples [11.97%, 95% confidence interval (CI) 6.97-19.3%]. Diluted products were contaminated significantly more often compared to undiluted products (odds ratio = 15.5, 95%CI 2.05-117.23; P < 0.01). None of the other variables was significantly associated with P. aeruginosa contamination. CONCLUSIONS AND CLINICAL RELEVANCE: Pseudomonas aeruginosa contamination of dog grooming shampoos and conditioners was significantly associated with product dilution. Contaminated grooming products may predispose dogs to severe bacterial skin infections such as PGF.

Contexte - Pseudomonas aeruginosa est la bactérie la plus couramment isolée des lésions cutanées des chiens atteints de furonculose post-toilettage (PGF). On la trouve fréquemment dans les cheveux humains et les produits de soin de la peau et peut présenter un risque pour la santé des consommateurs. Les informations concernant la prévalence de la contamination par P. aeruginosa des produits de toilettage pour chiens font défaut. Objectifs - Étudier la prévalence de la contamination par P. aeruginosa des produits de toilettage pour chiens non médicamenteux après un usage domestique ou professionnel dans les salons de toilettage pour animaux de compagnie, et identifier les facteurs de risque pouvant être associés à la contamination. Matériels et méthodes - Sur 117 flacons de produits de toilettage prélevés pour culture bactérienne, 97 étaient utilisés par des salons de toilettage et 20 par des particuliers. Les facteurs de risque suspectés suivants ont été enregistrés : taille de la bouteille, volume restant relatif, dilution du contenu, date de péremption et liste des ingrédients. Résultats - Pseudomonas aeruginosa a été isolé dans 14 des 117 échantillons [11,97 %, intervalle de confiance (IC) à 95 % 6,97-19,3 %]. Les produits dilués étaient significativement plus souvent contaminés que les produits non dilués (odds ratio = 15,5, IC à 95 % 2,05-117,23 ; P < 0,01). Aucune des autres variables n'était significativement associée à la contamination par P. aeruginosa. Conclusions et pertinence clinique - La contamination par Pseudomonas aeruginosa des shampoings et après shampoings pour chiens était significativement associée à la dilution du produit. Les produits de toilettage contaminés peuvent prédisposer les chiens à de graves infections cutanées bactériennes telles que la PGF.

Introducción- Pseudomonas aeruginosa es la bacteria más comúnmente aislada de las lesiones cutáneas de perros con furunculosis posterior al aseado (PGF). Se encuentra con frecuencia en productos para el cuidado de la piel y el cabello humano, y puede representar un riesgo para la salud de los consumidores. Falta información sobre la prevalencia de la contaminación por P. aeruginosa en los productos de aseo para perros. Objetivos- investigar la prevalencia de la contaminación por P. aeruginosa en productos de aseo canino no medicados después del uso doméstico o profesional en salones de aseo de mascotas, e identificar los factores de riesgo que pueden estar asociados con la contaminación. Materiales y métodos- de 117 botellas de productos de aseo muestreados para cultivo bacteriano, 97 fueron utilizados por salones de aseo de mascotas y 20 fueron utilizados por particulares. Se registraron los siguientes factores de riesgo sospechosos: tamaño de la botella, volumen restante relativo, dilución del contenido, fecha de caducidad y lista de ingredientes. Resultados- se aisló Pseudomonas aeruginosa en 14 de 117 muestras [11,97 %, intervalo de confianza (IC) del 95 %: 6,97-19,3 %]. Los productos diluidos se contaminaron significativamente más a menudo en comparación con los productos sin diluir (odds ratio = 15,5, IC del 95 %: 2,05-117,23; P < 0,01). Ninguna de las otras variables se asoció significativamente con la contaminación por P. aeruginosa. Conclusiones y relevancia clínica- la contaminación por Pseudomonas aeruginosa de los champús y acondicionadores de aseo para perros se asoció significativamente con la dilución del producto. Los productos de aseo contaminados pueden predisponer a los perros a infecciones cutáneas bacterianas graves como la PGF.

Contexto - Pseudomonas aeruginosa é a bactéria mais frequentemente isolada de lesões cutâneas de cães com furunculose pós-banho (FPB). É frequentemente encontrada em cabelos humanos e cosméticos e pode causar malefícios à saúde dos consumidores. Informações sobre a prevalência de contaminação de produtos de banho caninos por P. aeruginosa são escassas. Objetivos - Investigar a prevalência de contaminação de cosméticos de banho não medicamentosos para cães por P. aeruginosa após uso caseiro ou profissional em pet shops, e identificar os fatores de risco que podem estar associados à contaminação. Materiais e métodos - Dos 117 frascos de cosméticos testados por cultura bacteriana, 97 foram utilizados por pet shops e 20 foram de uso doméstico individual. Os seguintes fatores de risco foram registrados: tamanho do frasco, volume remanescente relativo, diluição do conteúdo, data de validade e lista de ingredientes. Resultados - Pseudomonas aeruginosa foi isolada de 14 das 117 amostras [11,97%, 95% intervalo de confiança (IC) 6,97-19,3%]. Produtos diluídos apresentaram contaminação significativamente mais frequentemente, comparado a produtos não diluídos (odds ratio = 15,5; 95% IC 2,05-117,23; P < 0,01). Nenhuma das outras variáveis foi significativamente associada à contaminação por P.aeruginosa. Conclusões e relevância clínica - A contaminação de shampoos e condicionadores caninos por Pseudomonas aeruginosa foi significativamente associada à diluição do produto. Cosméticos veterinários contaminados podem predispor cães a infecções cutâneas bacterianas como a FPB.

Neck of femur fractures treated with the femoral neck system: outcomes of one hundred and two patients and literature review.

INTRODUCTION: The recently developed femoral neck system (FNS) for treatment of femoral neck fractures (FNF), comprises theoretical biomechanical advantages compared to other implants. The aim of this study was to validate the safety and to report outcomes of patients treated with the FNS. METHOD: A retrospective multicentric analysis of patients treated by FNS with a minimum of three months of follow-up. Details analysed from three medical centres were operative duration, estimated blood loss, initial hospitalisation duration, fixation quality as well as complications and reoperation rate. Patients who had revision surgery were compared to all other patients to identify risk factors for failure. In addition, a literature review was performed to analyse data on FNS clinical implementation and patient's outcomes. The two data sets were combined and analysed. RESULTS: One-hundred and two patients were included in this study cohort with an average follow-up of seven months (range 3-27). Ten papers were included in the literature review, reporting data on 278 patients. Overall, 380 patients were analysed. Average age was 62.6 years, 52% of the fractures were classified as Gardens 1-2. Overall, the revision rate was 9.2% (14 patients diagnosed with cut-out of implant, 10 with AVN, 8 with non-union and 8 with hardware removal). For the 102 patients in the cohort risk factors for reoperation included patients age, surgeon seniority and inadequate placement of the implant. CONCLUSION: This study shows that FNS is a safe treatment option for FNF. Intra-operative parameters and failure rates are comparable to previously reported rates for this implant and other frequently used implants.

Botulinum Neurotoxin C Dual Detection through Immunological Recognition and Endopeptidase Activity Using Porous Silicon Interferometers.

Botulinum neurotoxins (BoNTs) are the most potent toxins known in nature produced by Clostridium botulinum strains, which can cause life-threatening diseases in both humans and animals. The latter is of serious environmental and economic concern, resulting in high mortality, production losses, and rejection of contaminated animal feed. The available in vivo mouse assay is inadequate for real-time and on-site assessment of outbreaks. Herein, we present a reflective-based approach for the detection of BoNT/C while estimating its activity. Two adjacent porous Si Fabry-Pérot interferometers are simultaneously utilized to quantify minute BoNT/C concentrations by a competitive immunoassay and to assess their endopeptidase activity. The reflectivity signals of each interferometer are amplified by biochemical reaction products infiltration into the scaffold or by peptide fragments detachment from the nanostructure. The optical assay is highly sensitive in compliance with the in vivo approach by presenting a detection limit of 4.24 pg mL-1. The specificity and selectivity of the designed platform are cross-validated against BoNT/B and BoNT/D, also relevant to animal health. Finally, the analytical performances of both interferometers for real-life scenarios are confirmed using actual toxins while depicting excellent compliance to complex media analysis. Overall, the presented sensing scheme offers an efficient, rapid, and label-free approach for potential biodiagnostic elucidation of botulism outbreaks.

The association between natural drinking water sources and the emergence of zoonotic leptospirosis among grazing beef cattle herds during a human outbreak.

Leptospirosis is a zoonotic bacterial disease associated with water abundance in tropical and temperate climate zones. Bacterial spread may also occur in dry and warm weather conditions when humans and animals are forced to share depleted water sources. In such settings, farm animals such as beef cattle, which may be present in large numbers in natural water sources, can play a major role in disease spread. However, the risk factors for their infection and the potential control measures to prevent the disease spread have not been adequately studied. In the face of an emerging human leptospirosis outbreak in the dry and warm Israeli 2018 summer, we tested seropositivity to Leptospira serovar Pomona in grazing beef cattle and wild boars located in proximity to the contaminated streams. Additionally, we used the natural setting of the outbreak to identify risk factors for seropositivity in beef cattle. We found high seropositivity to serovar Pomona in grazing beef cattle (233/845), and in wild boars (7/13). Seropositivity was significantly associated with beef cattle drinking from natural water sources compared to beef cattle drinking from water troughs with fresh water supply (Multivariable logistic regression; odds ratio = 18.6, 95% confidence interval = 3-116, p-value<0.01). One Health approach is necessary for mitigating zoonotic Leptospira infections, in which interactions between humans, animals, and the environment play a major role. As the global warming crisis results in severe climate changes, dry and warm weather conditions may become more common worldwide. Under such conditions, reducing inter-species interactions in contaminated natural water sources is essential for protecting public health. Our study demonstrates the role of natural water as a source for beef cattle infection and disease spread. Furthermore, we suggest using water troughs with freshwater supply for preventing future outbreaks in animals and humans in such settings.

Spatial and temporal dynamics of microbiomes and resistomes in broiler litter stockpiles.

Farmers apply broiler chicken litter to soils to enrich organic matter and provide crops with nutrients, following varying periods of stockpiling. However, litter frequently harbors fecal-derived microbial pathogens and associated antibiotic resistance genes (ARGs), and may be a source of microbial contamination of produce. We coupled a cutting-edge Loop Genomics long-read 16S rRNA amplicon-sequencing platform with high-throughput qPCR that targeted a suite of ARGs, to assess temporal (five time points over a 60-day period) and spatial (top, middle and bottom layers) microbiome and resistome dynamics in a broiler litter stockpile. We focused on potentially pathogenic species from the Enterobacteriaceae, Enterococcaceae and Staphylococcaceae families associated with food-borne disease. Bacterial diversity was significantly lower in the middle of the stockpile, where targeted pathogens were lowest and Bacillaceae were abundant. E. coli was the most abundant Enterobacteriaceae species, and high levels of the opportunistic pathogen Enterococcus faecium were detected. Correlation analyses revealed that the latter was significantly associated with aminoglycoside (aac(6')-Ib(aka aacA4), aadA5), tetracycline (tetG), vancomycin (vanC), phenicol (floR) and MLSB (mphB) resistance genes. Staphylococcaceae were primarily non-pathogenic, but extremely low levels of the opportunistic pathogen S. aureus were detected, as was the opportunistic pathogen S. saprophyticus, which was linked to vancomycin (vanSA, vanC1), MLSB (vatE, ermB) and tetracycline (tetK) resistance genes. Collectively, we found that stockpile microbiomes and resistomes are strongly dictated by temporal fluctuations and spatial heterogeneity. Insights from this study can be exploited to improve stockpile management practice to support sustainable antimicrobial resistance mitigation policies in the future.

Protein Biomarker Identification for the Discrimination of Brucella melitensis Field Isolates From the Brucella melitensis Rev.1 Vaccine Strain by MALDI-TOF MS.

Brucella melitensis Rev.1 is a live attenuated vaccine strain that is widely used to control brucellosis in small ruminants. For successful surveillance and control programs, rapid identification and characterization of Brucella isolates and reliable differentiation of vaccinated and naturally infected animals are essential prerequisites. Although MALDI-TOF MS is increasingly applied in clinical microbiology laboratories for the diagnosis of brucellosis, species or even strain differentiation by this method remains a challenge. To detect biomarkers, which enable to distinguish the B. melitensis Rev.1 vaccine strain from B. melitensis field isolates, we initially searched for unique marker proteins by in silico comparison of the B. melitensis Rev.1 and 16M proteomes. We found 113 protein sequences of B. melitensis 16M that revealed a homologous sequence in the B. melitensis Rev.1 annotation and 17 of these sequences yielded potential biomarker pairs. MALDI-TOF MS spectra of 18 B. melitensis Rev.1 vaccine and 183 Israeli B. melitensis field isolates were subsequently analyzed to validate the identified marker candidates. This approach detected two genus-wide unique biomarkers with properties most similar to the ribosomal proteins L24 and S12. These two proteins clearly discriminated B. melitensis Rev.1 from the closely related B. melitensis 16M and the Israeli B. melitensis field isolates. In addition, we verified their discriminatory power using a set of B. melitensis strains from various origins and of different MLVA types. Based on our results, we propose MALDI-TOF MS profiling as a rapid, cost-effective alternative to the traditional, time-consuming approach to differentiate certain B. melitensis isolates on strain level.

Longitudinal study on the effects of growth-promoting and therapeutic antibiotics on the dynamics of chicken cloacal and litter microbiomes and resistomes.

BACKGROUND: Therapeutic and growth-promoting antibiotics are frequently used in broiler production. Indirect evidence indicates that these practices are linked to the proliferation of antimicrobial resistance (AMR), the spread of antibiotic-resistant bacteria from food animals to humans, and the environment, but there is a lack of comprehensive experimental data supporting this. We investigated the effects of growth promotor (bacitracin) and therapeutic (enrofloxacin) antibiotic administration on AMR in broilers for the duration of a production cycle, using a holistic approach that integrated both culture-dependent and culture-independent methods. We specifically focused on pathogen-harboring families (Enterobacteriaceae, Enterococcaceae, and Staphylococcaceae). RESULTS: Antibiotic-resistant bacteria and antibiotic resistance genes were ubiquitous in chicken cloaca and litter regardless of antibiotic administration. Environment (cloaca vs. litter) and growth stage were the primary drivers of variation in the microbiomes and resistomes, with increased bacterial diversity and a general decrease in abundance of the pathogen-harboring families with age. Bacitracin-fed groups had higher levels of bacitracin resistance genes and of vancomycin-resistant Enterococcaceae (total Enterococcaceae counts were not higher). Although metagenomic analyses classified 28-76% of the Enterococcaceae as the commensal human pathogens E. faecalis and E. faecium, culture-based analysis suggested that approximately 98% of the vancomycin-resistant Enterococcaceae were avian and not human-associated, suggesting differences in the taxonomic profiles of the resistant and non-resistant strains. Enrofloxacin treatments had varying effects, but generally facilitated increased relative abundance of multidrug-resistant Enterobacteriaceae strains, which were primarily E. coli. Metagenomic approaches revealed a diverse array of Staphylococcus spp., but the opportunistic pathogen S. aureus and methicillin resistance genes were not detected in culture-based or metagenomic analyses. Camphylobacteriaceae were significantly more abundant in the cloacal samples, especially in enrofloxacin-treated chickens, where a metagenome-assembled C. jejuni genome harboring fluoroquinolone and ß-lactam resistance genes was identified. CONCLUSIONS: Within a "farm-to-fork, one health" perspective, considering the evidence that bacitracin and enrofloxacin used in poultry production can select for resistance, we recommend their use be regulated. Furthermore, we suggest routine surveillance of ESBL E. coli, vancomycin-resistant E. faecalis and E. faecium, and fluoroquinolone-resistant C. jejuni strains considering their pathogenic nature and capacity to disseminate AMR to the environment. Video Abstract.

Seroprevalence of Leptospira spp. in Horses in Israel.

Leptospirosis has been reported in both humans and animals in Israel but has not been reported in horses. In 2018, an outbreak of Leptospira spp. serogroup Pomona was reported in humans and cattle in Israel. In horses, leptospirosis may cause equine recurrent uveitis (ERU). This report describes the first identification of Leptospira serogroup Pomona as the probable cause of ERU in horses in Israel, followed by an epidemiological investigation of equine exposure in the area. Serologic exposure to Leptospira was determined by microscopic agglutination test (MAT) using eight serovars. In 2017, serovar Pomona was identified in a mare with signs of ERU. Seven of thirteen horses from that farm were seropositive for serogroup Pomona, of which three had signs of ERU. During the same time period, 14/70 horses from three other farms were positive for serogroup Pomona. In 2015, two years prior to this diagnosis, 259 horses from 21 farms were sampled and one horse tested seropositive for serovar Icterohaemorrhagiae. In 2018, one year later, 337 horses were sampled on 29 farms, with none testing seropositive. Although horses are not considered a major host of Leptospira spp., it appears that horses may be infected, and clinically affected, in the course of an outbreak in other species. The identification of leptospirosis in stabled horses may impose a significant zoonotic risk to people.

Globetrotting strangles: the unbridled national and international transmission of Streptococcus equi between horses.

The equine disease strangles, which is characterized by the formation of abscesses in the lymph nodes of the head and neck, is one of the most frequently diagnosed infectious diseases of horses around the world. The causal agent, Streptococcus equi subspecies equi, establishes a persistent infection in approximately 10 % of animals that recover from the acute disease. Such 'carrier' animals appear healthy and are rarely identified during routine veterinary examinations pre-purchase or transit, but can transmit S. equi to naïve animals initiating new episodes of disease. Here, we report the analysis and visualization of phylogenomic and epidemiological data for 670 isolates of S. equi recovered from 19 different countries using a new core-genome multilocus sequence typing (cgMLST) web bioresource. Genetic relationships among all 670 S. equi isolates were determined at high resolution, revealing national and international transmission events that drive this endemic disease in horse populations throughout the world. Our data argue for the recognition of the international importance of strangles by the Office International des Épizooties to highlight the health, welfare and economic cost of this disease. The Pathogenwatch cgMLST web bioresource described herein is available for tailored genomic analysis of populations of S. equi and its close relative S. equi subspecies zooepidemicus that are recovered from horses and other animals, including humans, throughout the world. This article contains data hosted by Microreact.