Group B Streptococcus Sequence Type 103 as Human and Bovine Pathogen, Brazil.

Group B Streptococcus sequence type 103 is known primarily as a bovine mastitis pathogen. In Brazil, it has circulated in cattle and humans since the 1990s. It lacks scpB and, in humans, was found only among carriage isolates. Bovine-human interspecies transmission may have contributed to its evolution and spread.

Validation of a Questionnaire for Equine Personality Assessement.

We propose a practical method of assessing the personality of horses using five personality axes. Four evaluators empirically judged 19 horse individuals on specific adjectives for each axis. To validate the questionnaire, four behavioral tests were conducted with these same animals (social interactions, novel object test, bridge test,and arena test). In this tests, the frequency of specific behaviors were evaluated to create a scale related to the same personality adjectives and judge the animals based on their reactions.The questionnaire was reliable in determining the personality of horses, since the results were consistent with those obtained through behavioral tests. Additionally, in this group of horses attention reactions were more frequent than fear reactions, but significant differences occurred among tests. This study proposes a practical questionnaire for owners and trainers to assess the personality of their horses. The application of this tool can improve the relationship between humans and horses, directing a more empathic approach in the everyday routine with these animals.

Long-Term Co-Circulation of Host-Specialist and Host-Generalist Lineages of Group B Streptococcus in Brazilian Dairy Cattle with Heterogeneous Antimicrobial Resistance Profiles.

Group B Streptococcus (GBS) is a major cause of contagious bovine mastitis (CBM) in Brazil. The GBS population is composed of host-generalist and host-specialist lineages, which may differ in antimicrobial resistance (AMR) and zoonotic potential, and the surveillance of bovine GBS is crucial to developing effective CBM control and prevention measures. Here, we investigated bovine GBS isolates (n = 156) collected in Brazil between 1987 and 2021 using phenotypic testing and whole-genome sequencing to uncover the molecular epidemiology of bovine GBS. Clonal complex (CC) 61/67 was the predominant clade in the 20th century; however, it was replaced by CC91, with which it shares a most common recent ancestor, in the 21st century, despite the higher prevalence of AMR in CC61/67 than in CC91, and high selection pressure for AMR from indiscriminate antimicrobial use in the Brazilian dairy industry. CC103 also emerged as a dominant CC in the 21st century, and a considerable proportion of herds had two or more GBS strains, suggesting poor biosecurity and within-herd evolution due to the chronic nature of CBM problems. The majority of bovine GBS belonged to serotype Ia or III, which was strongly correlated with CCs. Ninety-three isolates were resistant to tetracycline (≥8 µg/mL; tetO = 57, tetM = 34 or both = 2) and forty-four were resistant to erythromycin (2.0 to >4 µg/mL; ermA = 1, ermB = 38, mechanism unidentified n = 5). Only three isolates were non-susceptible to penicillin (≥8.0 µg/mL), providing opportunities for improved antimicrobial stewardship through the use of narrow-spectrum antimicrobials for the treatment of dairy cattle. The common bovine GBS clades detected in this study have rarely been reported in humans, suggesting limited risk of interspecies transmission of GBS in Brazil. This study provides new data to support improvements to CBM and AMR control, bovine GBS vaccine design, and the management of public health risks posed by bovine GBS in Brazil.

Identification of Escherichia coli isolated from flies (Insecta: Diptera) that inhabit the environment of dairy farms harboring extraintestinal virulence markers.

AIMS: We investigate extraintestinal pathogenic genes (ExPEC) related to virulence of Escherichia coli in flies from the dairy environment. METHODS AND RESULTS: We collected 217 flies from nine dairy farms, which were submitted to microbiological culture. Fifty-one E. coli were identified using mass spectrometry. Eleven dipteran families were identified, with a predominance of Muscidae, and a minor frequency of Tachinidae, Drosophilidae, Sphaeroceridae, Ulidiidae, Syrphidae, Chloropidae, Calliphoridae, Sarcophagidae, and Piophilidae. A panel of 16 virulence-encoding genes related to ExPEC infections were investigated, which revealed predominance of serum resistance (traT, 31/51 = 60.8%; ompT, 29/51 = 56.9%), iron uptake (irp2, 17/51 = 33.3%, iucD 11/51 = 21.6%), and adhesins (papC, 6/51 = 11.8%; papA, 5/51 = 9.8%). CONCLUSIONS: Our findings reveal Dipterans from milking environment carrying ExPEC virulence-encoding genes also identified in clinical bovine E. coli-induced infections.

In Vitro Activity of Quaternary Ammonium in Prototheca Isolated from Clinical Bovine Mastitis Identified by Mass Spectrometry and PCR Sequencing of the cytb Gene Marker.

The in vitro algaecide activity of quaternary ammonium (QA) against Prototheca isolated from bovine clinical mastitis was investigated, in which the clinical severity was scored, milk samples were subjected to microbiological culture, and algal species were identified by molecular typing. A total of 4275 milk clinical samples of different cows from ten large dairy farms were used. Forty-four (1%) samples of cows from three dairy farms yielded growth of Prototheca, of which 88.6% (39/44) were identified as Prototheca bovis and 11.3% (5/44) as Prototheca sp. by MALDI-TOF MS, whereas 100% of the isolates were identified as P. bovis using PCR sequencing of the cytb gene. Among cows for which clinical severity scoring was available, 78.8% (26/33) and 21.2% (7/33) had mild and moderate infections, respectively, whereas no animal showed severe clinical signs. The algaecide activity of QA in Prototheca was observed in low concentrations among all isolates, in 20.4% (9/44) at 35 ppm, 36.4% (16/44) at 17 ppm, and 43.2% (19/44) at an 8 ppm, in addition to activity on three reference Prototheca strains. Overall, the study highlights the predominance of P. bovis as the causative agent of algal mastitis in bovines. Prototheca induced abnormalities preponderantly in the milk and mammary gland tissue of cows, and to our knowledge, our study is the first to apply clinical severity scoring in protothecal mastitis. In addition, the study underlines the activity of QA in low concentrations against Prototheca, indicating its potential use as an antiseptic/disinfectant in milking facilities and dairy environments.

Flies (Insecta, Diptera) collected in the environment of dairy farms as carriers of Rotavirus A and betacoronavirus.

AIMS: We aimed to investigate the prevalence of rotavirus and coronavirus in dipterans that commonly inhabit the environment of dairy farms. METHODS AND RESULTS: We collected 217 insect specimens from nine dairy farms, which were examined through hemi-nested RT-PCR followed by Sanger sequencing in search of VP1 and N genes for rotavirus and bovine coronavirus-BCoV, respectively. With a predominance of Muscidae (152/217 = 70%) 11 families of Diptera were identified. Rotavirus A (RVA) and betacoronavirus (BCoV) were detected in 14.7% (32/217) and 4.6% (10/217) of the dipterans, respectively. Sequencing of the amplicons was possible for 11.5% (25/217) of RVA and 0.5% (1/217) of BCoV, confirming the presence of these pathogens. CONCLUSIONS: Our findings highlight the role of dipterans as carriers of RVA and BCoV of great relevance for public and animal health.

Relationship between Biofilm Production and High Somatic Cell Count in Streptococcus agalactiae Isolated from Milk of Cows with Subclinical Mastitis.

Streptococcus agalactiae (S. agalactiae) is one of the main agents that causes mastitis in dairy cows, mainly inducing the subclinical form, which is characterized by a high somatic cell count (SCC). The aim of this study was to correlate the increase in SCC caused by S. agalactiae in cows with subclinical mastitis to the presence of genes related to adhesion and invasion in bovine mammary epithelial cells (BMEC) and biofilm formation. Considering the 145 isolates tested, 57.2% presented the capsular type Ia and 42.8% presented type III. We identified the virulence genes among the isolates and determined nine genetic profiles. The most common profile was identified in 69 isolates (47.5%): Ia, fbsA+, fbsB-, pI1-, pI2a-, pI2b+, and hylb+. All isolates produced biofilm, with 58.6% classified as strong producers, 29% as moderate producers and 12.4% as weak producers. No statistical correlation was found between the presence of virulence genes and increased SCC or biofilm production. However, biological evidence was observed between increased SCC and biofilm production. One isolate from each profile was randomly subjected to adhesion and invasion assays, and all of them adhered to BEMC, but none were able to invade. Our results showed that different genetic profiles do not provide advantages for bacteria to invade BMEC in vitro. In addition, biofilm production appears to be related to high SCC.

Genome-Based Characterization of Multidrug-Resistant Escherichia coli Isolated from Clinical Bovine Mastitis.

Mastitis occurrence in dairy cows is a broad topic that involves several sectors, from antimicrobial resistance and virulence of strains to economic implications and cattle management practices. Here, we assessed the molecular characterization (antimicrobial resistance determinants, virulence genes, sequences type, serotypes, and plasmid types) of 178 Escherichia coli strains isolated from milk samples from cows with clinical mastitis using a genome-based k-mers approach. Of these, 53 (29.8%) showed multidrug resistance by disc diffusion. We selected eight multidrug-resistant mastitis-associated E. coli for whole-genome sequencing and molecular characterization based on raw data using k-mers. We assessed antimicrobial resistance genes, virulence factors, serotypes, Multilocus Sequence Typing (MLST), and plasmid types. The most antimicrobial resistance gene found were blaTEM-1B (7/8), tetA (6/8), strA (6/8), strB (6/8), and qnrB19 (5/8). A total of 25 virulence factors were detected encoding adhesins, capsule, enzymes/proteins, increased serum survival, hemolysin, colicins, and iron uptake. These virulence factors were associated with Extraintestinal Pathogenic E. coli. Three pandemic clones were found: ST10, ST101, and ST69. Two E. coli were assigned in the O117 serogroup and one in the O8:H25 serotype. The most common plasmid groups were IncFII (7/8) and IncFIB (6/8). Our findings contribute to the knowledge of virulence mechanisms, epidemiological aspects, and antimicrobial resistance determinants of E. coli strains obtained from clinical mammary infections of cows.

Characterization of mammary pathogenic Escherichia coli reveals the diversity of Escherichia coli isolates associated with bovine clinical mastitis in Brazil.

Mammary pathogenic Escherichia coli (MPEC) is one of the most common pathogens associated with clinical mastitis. We analyzed isolates obtained from milk samples of cows with clinical mastitis, collected from 10 farms in Brazil, to verify molecular and phenotypic characteristics. A total of 192 (4.5%) mammary pathogenic E. coli isolates were obtained from 4,275 milk samples analyzed, but we tested 161. We assigned most of these isolates to E. coli phylogroups B1 (52.8%) and A (36.6%), although phylogroups B2, C, D, E, and unknown also occurred. All isolates were assessed for the presence of several genes encoding virulence factors, such as adhesins (sfaDE, papC, afaBC III, ecpA, fimH, papA, and iha), toxins (hlyA, cnf1, sat, vat, and cdt), siderophores (iroN, irp2, iucD, ireA, and sitA), an invasion protein (ibeA), and serum resistance proteins (traT, KpsMTII, and ompT), and isolates from phylogroups B1, B2, and E showed up to 8 genes. Two isolates harbored the locus of enterocyte effacement (escN+) and lack the bundle-forming pilus (bfpB-) operon, which corresponds to a molecular profile of a subgroup of diarrheagenic E. coli (aEPEC), thus being classified as hybrid MPEC/aEPEC isolates. These isolates displayed a localized adherence-like pattern of adherence in HeLa cells and were able to promote F-actin polymerization underneath adherent bacteria. Based on the pulsed-field gel electrophoresis analyses, considerable genetic variability was observed. A low index of antimicrobial resistance was observed and 2 extended-spectrum ß-lactamase-producing E. coli were identified, both harboring blaCTX-M15 gene, and were classified as ST10 and ST993 using multilocus sequence typing. A total of 148 (91.2%) isolates were weak biofilm producers or formed no biofilm. Because raw milk is still frequently consumed in Brazil, the occurrence of virulence factor-encoding genes from extraintestinal or diarrheagenic E. coli added to the presence of extended-spectrum ß-lactamase-producing isolates can turn this veterinary medicine problem into a public health concern.

Occurrence of Salmonella spp. in fecal samples from foals with and without diarrhea in the state of São Paulo: microbiological diagnosis, antimicrobial susceptibility profile, and molecular detection / Ocorrência de Salmonella spp. em amostras fecais de potros com e sem diarreia no estado de São Paulo: diagnóstico microbiológico, perfil de susceptibilidade aos antimicrobianos e detecção molecular

The present study investigated Salmonella spp. in the feces of 200 foals up to one year of age (100 with clinical signs of diarrhea and 100 without clinical signs of diarrhea). Bacteriological culture, serotyping, antimicrobial susceptibility, and real-time PCR (qPCR SYBR® Green or a TaqMan®) for detecting the invA gene (with and without a selective pre-enrichment step in tetrathionate broth) were performed. Bacterial culture revealed 15% (n=30) of positive animals (21 animals with diarrhea and nine without diarrhea). Among the 30 isolates, 13 different serovars were identified: S. Infantis, S. Minnesota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. Braenderup, S. Give, S. Newport, S. IIIb 61:c:z35, S. 109:-:1.5, S. I.4.12:d:-, S. I.6.8:-:-. Multidrug resistance was found in 43.33% (n=13) of the isolates, with one isolate obtained from animals without diarrhea and 12 isolates from animals with diarrhea. All qPCR techniques used in the study classified more samples as positive for Salmonella spp. than the bacterial culture of feces. In addition, all qPCR techniques detected more positive animals in the diarrhea group than in the diarrhea-free group. The results confirm the utility of the qPCR method without the pre-enrichment step in tetrathionate as a rapid test for Salmonella spp. in carrier animals. In animals with clinical signs of diarrhea, it can be combined with bacterial culture (antimicrobial susceptibility testing and serotyping). The isolation of Salmonella spp. in nine animals without diarrhea confirms the importance of asymptomatic carrier animals in the epidemiology of the disease. The multidrug resistance observed highlights the importance of rational antimicrobial use in horses and adopting biosecurity protocols that are efficacious in controlling the spread of infections between animals and zoonotic transmission in farms.

O presente estudo investigou a ocorrência de Salmonella spp. em fezes de 200 potros com até um ano de idade (100 com sinais clínicos de diarreia e 100 sem sinais clínicos de diarreia), utilizando as técnicas de cultivo bacteriológico e PCR em tempo real (qPCR) pelos métodos de corante fluorescente (SYBR® Green) e sonda específica (Taqman®) para a detecção do gene invA com e sem etapa de pré-enriquecimento seletivo em caldo de tetrationato. O cultivo bacteriológico revelou 15% (n=30) de animais positivos (21 animais com diarreia e nove animais sem diarreia). Dentre esses 30 isolados, 13 sorovares diferentes foram identificados: S. Infantis, S. Minnesota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. Braenderup, S. Give, S. Newport, S. IIIb 61:c:z35, S. 109:-:1.5, S. I.4.12:d:-, S. I.6.8:-:-. Multirresistência foi constatada em 43,33% (n=13) dos isolados, sendo um isolado obtido de animal sem diarreia e 12 isolados de animais com diarreia. Todas as técnicas de qPCR empregadas no estudo apresentaram maior número de amostras classificadas como positivas para Salmonella spp. comparadas ao cultivo bacteriológico de fezes. Adicionalmente, em todas as técnicas de qPCR houve maior número de animais detectados como positivos no grupo de animais com diarreia em relação aos animais sem diarreia. Os resultados confirmaram a utilidade do método qPCR sem a etapa de pré-enriquecimento em tetrationato, como um teste rápido para detecção de Salmonella spp. em animais portadores ou em animais com sinais clínicos de diarreia. O cultivo bacteriológico deve ser associado para a realização do teste de sensibilidade aos antimicrobianos e sorotipificação. O isolamento de Salmonella spp. em nove animais sem diarreia, confirma a importância dos animais portadores assintomáticos na epidemiologia da doença. A multirresistência observada evidencia a importância do uso racional de antimicrobianos em equinos e a importância da adoção de protocolos de biossegurança que sejam eficazes para controlar a disseminação de infecções entre animais e a transmissão zoonótica nas fazendas.

Genetic and Antimicrobial Resistance Profiles of Mammary Pathogenic E. coli (MPEC) Isolates from Bovine Clinical Mastitis.

Mammary pathogenic E. coli (MPEC) is one of the main pathogens of environmental origin responsible for causing clinical mastitis worldwide. Even though E. coli are strongly associated with transient or persistent mastitis and the economic impacts of this disease, the virulence factors involved in the pathogenesis of MPEC remain unknown. Our aim was to characterize 110 MPEC isolates obtained from the milk of cows with clinical mastitis, regarding the virulence factor-encoding genes present, adherence patterns on HeLa cells, and antimicrobial resistance profile. The MPEC isolates were classified mainly in phylogroups A (50.9%) and B1 (38.2%). None of the isolates harbored genes used for diarrheagenic E. coli classification, but 26 (23.6%) and 4 (3.6%) isolates produced the aggregative or diffuse adherence pattern, respectively. Among the 22 genes investigated, encoding virulence factors associated with extraintestinal pathogenic E. coli pathogenesis, fimH (93.6%) was the most frequent, followed by traT (77.3%) and ompT (68.2%). Pulsed-field gel electrophoresis analysis revealed six pulse-types with isolates obtained over time, thus indicating persistent intramammary infections. The genes encoding beta-lactamases detected were as follows: blaTEM (35/31.8%); blaCTX-M-2/blaCTX-M-8 (2/1.8%); blaCTX-M-15 and blaCMY-2 (1/0.9%); five isolates were classified as extended spectrum beta-lactamase (ESBL) producers. As far as we know, papA, shf, ireA, sat and blaCTX-M-8 were detected for the first time in MPEC. In summary, the genetic profile of the MPEC studied was highly heterogeneous, making it impossible to establish a common genetic profile useful for molecular MPEC classification. Moreover, the detection of ESBL-producing isolates is a serious public health concern.

Effects of Acupuncture in the Treatment of Dogs with Neurological Sequels of Distemper Virus.

Background: Acupuncture (AP) has been empirically used to relieve post-canine distemper virus (CDV) infection neurological signs in veterinary clinics. Objectives: This clinical study aimed to investigate the effects of AP combined with electroacupuncture (EA) on neurological function in dogs infected by CDV. Methods: Twenty-four CDV-infected dogs with neurological sequelae were recruited to receive weekly AP/EA sections for 24 weeks. Neurological improvements were assessed before each AP/EA session using a modified scoring system. Data were analyzed using the McNemar test, Friedman test, Fisher's exact test, and Kaplan-Meier curves (p < 0.05). Results: Neurological scores improved from seven to 24 weeks after AP/EA treatment compared with pretreatment scores (p < 0.001). Significant improvements were recorded over time for functional limb recovery, cranial nerve deficits, mental status (p = 0.025 - 0.014), and urinary function (p < 0.001). Myoclonus was improved and entirely reversed in 75% and 25% of the dogs, respectively. At the end of treatment, the proportion of dogs with normal proprioception, posture, hopping (p < 0.001), and superficial pain sensation responses (p = 0.004) was greater than pretreatment values. Conclusion: AP/EA therapy promoted significant neurological recovery in CDV-infected dogs and may be considered within the chronic phase of the disease to improve motor and sensory rehabilitation. However, these results are preliminary and must be confirmed by further investigations.

Overview of antimicrobial resistance and virulence factors in Salmonella spp. isolated in the last two decades from chicken in Brazil.

Foodborne infections caused by Salmonella have been linked to a variety of poultry products. The aim of this study was to compare the molecular profile of virulence genes considering different serotypes of Salmonella, isolates were from chicken breast sampled during the last two decades (1999 to 2010 and 2011 to 2018). The resistance to antimicrobials was also evaluated, establishing a comparative epidemiological parameter on the pathogenic potential on this bacterium over time. We tested 238 Salmonella isolates, and 18 different serotypes were observed. These being S. Enteritidis (42.3%, 58/137) and S. Ohio (28.3%, 36/137), the most frequent in the first decade; and S. Heidelberg (25.7%, 26/101) and S. Typhimurium (21.8%, 22/101), in the second. We found four (1.68%) multidrug resistant isolates from the first decade and 28 (11.76%) in the second. All extended spectrum beta-lactamase (ESBL) positive isolates belonged to the S. Heidelberg serotype, and were also detected in the second decade. Considering the nine different antimicrobial classes tested, an increase in the number of resistant isolates was observed over time: from five classes with resistant isolates in the first decade to eight classes in the second, with cefotaxime being the antimicrobial with the highest number of resistant isolates in both decades. All isolates (100%) presented the invA, sitC and tolC genes. In sequence, the most frequent genes were flgL (99.6%), sopB (98.3%), flgK (97.9%), fljB (96.6%), sipA (94.9%), sipB (88.6%), sifA (86.4%), sipD (66.1%), ssaR (51.3%), sopD (37.3%) and spvB (34.3%) was the least frequent; and 13 isolates showing all 14 virulence genes investigated. The ability of these isolates to resist certain antimicrobials, and to express genes encoding virulence factors, reinforce their marked pathogenic potential; while the possibility to trigger diseases in humans through the food chain is a serious public health threat through.

Síndrome de Felty / Felty's syndrome

RESUMEN Introducción: El síndrome de Felty es una rara complicación extraarticular de la artritis reumatoide, descrito por primera vez en 1924. Felty describió 5 pacientes que se caracterizaban por la presencia de una tríada clínica de artritis reumatoide, esplenomegalia y neutropenia, que aparece fundamentalmente en los pacientes con largos años de evolución de su enfermedad. Es una entidad clínica con alto grado de mortalidad, dado a que es poco diagnosticada y por tanto, no tratada. Objetivo: Presentar un paciente con síndrome de Felty, sus características clínicas y el procedimiento terapéutico empleado. Caso clínico: Paciente femenina de 64 años de edad, con antecedentes de artritis reumatoide e hipotiroidismo; ambas enfermedades sin tratamiento regular. Asistió al servicio de urgencias con fiebre y presencia de lesiones ulcerativas en miembro inferior izquierdo, con fondo necrótico. Debido a leucopenia acompañada de esplenomegalia se le diagnosticó neutropenia febril (sepsis), secundaria a síndrome de Felty. Se instauró tratamiento oportuno y se logró paulatinamente la cicatrización de las lesiones ulcerativas, la erradicación de la infección con recuperación de los valores normales de neutrófilos y la posterior recuperación de la paciente. Conclusiones: Al realizar el diagnóstico de síndrome de Felty, lo esencial es alcanzar un control inmediato de la enfermedad de base, de las comorbilidades y a través del tratamiento obtener un incremento de la supervivencia de los pacientes.

ABSTRACT Introduction: Felty's syndrome is a rare extra-articular complication of rheumatoid arthritis, first described in 1924. Felty described 5 patients who were characterized by the presence of a clinical triad of rheumatoid arthritis, splenomegaly, and neutropenia that appears fundamentally in patients with long years of evolution of their disease. It is a clinical entity with a high degree of mortality, given that it is a poorly diagnosed disease and therefore not treated. Objective: Present a clinical case of Felty´s syndrome, its clinical characteristics and the therapeutic procedure used. Clinical case: 64-year-old female patient with a history of rheumatoid arthritis and hypothyroidism; both diseases without regular treatment. She attended to emergency with fever and the presence of ulcerative lesions on the left lower limb with a necrotic background. Due to leukopenia accompanied by splenomegaly, she was diagnosed with febrile neutropenia (sepsis) secondary to Felty's syndrome. Treatment was established and healing of the ulcerative lesions was gradually achieved, the eradication of the infection with recovery of normal neutrophil values and the subsequent recovery of the patient. Conclusions: When diagnosing Felty's syndrome, the essential thing is to achieve immediate control of the underlying disease and the comorbidities; and through treatment obtain an increase in patient survival.

Short communication: Association between the accessory gene regulator (agr) group and the severity of bovine mastitis caused by Staphylococcus aureus.

Staphylococcus aureus can elicit mild to more severe degrees of mastitis in cattle, depending on the response of the host's immune system and the virulence factors of the specific isolate. Several virulence factors are controlled by a global regulatory system, designated accessory gene regulator (agr). Thus, the objective was to examine associations between different capsular and agr types and the severity of bovine mastitis caused by S. aureus. All isolates were obtained from bovine subclinical (n = 50), mild clinical (n = 73), and moderate clinical mastitis cases (n = 28). Isolates containing the agrI gene and lacking the agr locus (agr-) were more prevalent among subclinical than clinical mastitis cases, whereas isolates containing the agrII and agrIII genes were more prevalent among clinical mastitis cases. The capsular types 5 (cap5) and 8 (cap8) were found in 42 and 44%, respectively, of the isolates obtained from subclinical cases and in 38.6 and 58.4%, respectively, of those isolated from clinical mastitis cases. Capsular type was not associated with type of mastitis (subclinical, mild clinical, or moderate clinical). We found a strong association between agr type and type of mastitis, suggesting that knowledge of S. aureus genetic profiles could be an additional tool to control this disease.

Epidemiological aspects of Ancylostoma spp. infection in naturally infected dogs from São Paulo state, Brazil.

Ancylostoma spp. are the most prevalent canine nematode parasites in Brazil. Despite their widespread parasitism in juvenile dogs, infections may occur regardless of host age. Although eosinophilia is a frequent finding in experimental infections, it is uncommon in naturally infected dogs. This study aimed to evaluate the prevalence of Ancylostoma spp. and the eosinophil blood counts (EBC) in naturally infected dogs, with or without comorbidities, admitted to the Veterinary Teaching Hospital of the São Paulo State University (UNESP), School of Veterinary Medicine and Animal Science, Botucatu campus, São Paulo state, Brazil, from 2009 to 2018. All retrospective data were gathered from veterinary medical records (VMR); diagnosis of Ancylostoma infection required the identification of eggs in fecal samples by the simple flotation test (SFT). Ancylostoma-infected animals were evaluated for other intestinal parasitic coinfections (IPC) by either the SFT or the centrifugal-flotation test. Dogs free of any gastro-intestinal parasites were prospectively included in control group (Group C). Ancylostoma-infected animals were defined: Ancylostoma spp. only intestinal parasite infection (Group A), Ancyslostoma spp. with concurrent IPC (Group B), Ancylostoma spp. only intestinal parasite infection with concurrent systemic disorders (Group D), and Ancylostoma spp. with both IPC and concurrent systemic disorders (Group E). The overall prevalence of Ancylostoma spp. was 12.1% (207/1715), that was decreased from 2014 to 2018 (9.7%) relative to the 2009-to-2013 period (13.9%). Prevalence was not significantly different between dogs <1-year-old (10.7% [51/478]) and ≥ 1 year-old (11.7% [130/1109]). IPC was observed in 45.4% (93/205) of the animals positive for Ancylostoma spp., while dogs <1 year old experienced IPC more often (58.8% [30/51]) than dogs ≥1 year old (38.5% [50/130]) (P = 0.02). Group A (n = 35) exhibited median EBC of 1.05 × 109/L, and an eosinophilia ratio of 34.3% that was significantly higher (P < 0.05) than Group C (0.45 × 109/L and 4.1%, respectively). Both variables did not differ in Group B (n = 20), D (n = 39) or E (n = 36) in comparison to Group C (P > 0.05). By ROC curve analysis, only Group A generated a significant area under the curve (0.72). With EBC cutoff of 0.85 × 109 eosinophils/L, sensitivity and specificity were 65.7% and 70.8%, respectively. Eosinophil counts alone may be helpful in raising suspicion of an Ancylostoma spp. infection if further intestinal parasites and concurrent disorders are absent. However, local prevalence data and epidemiological findings should also be evaluated, since eosinophilia is less frequently observed with Ancylostoma spp. infections in the presence of comorbidities.

First investigation of Staphylococcus argenteus in a Brazilian collections of S. aureus isolated from bovine mastitis.

BACKGROUND: Staphylococcus argenteus is a new specie positive coagulase staphylococci. We investigate the presence of S. argenteus in isolates previously classified as S. aureus, obtained from the milk of cows with mastitis in Brazil. RESULTS: Among 856 S. aureus tested in chocolate agar, tryptone soya agar and salt egg yolk agar, white or colorless colonies were observed in 185 (21.6%) isolates. Regarding the ctrOPQMN operon, 111 (60%) presented the complete cluster. Despite some missing genes in this cluster, the remaining strains (74) were confirmed as S. aureus using the nrps gene. CONCLUSIONS: As far as we know, this is the first review of S. aureus collection in Brazil and S. argenteus does not appear to be a significant problem in Brazilian herds.

Environmental persistence and virulence of Salmonella spp. Isolated from a poultry slaughterhouse.

Salmonella spp. is responsible for severe foodborne disease, and is one of the main agents involved in foodborne outbreaks worldwide. Contamination occurs mainly as a result of poultry and egg consumption since they can carry some serotypes pathogenic to humans. The aim of the study was to evaluate the persistence and pathogenic potential of Salmonella spp. (n = 40) isolated from poultry slaughterhouse mats, using adhesion and invasion assays, antimicrobial susceptibility by disc diffusion, and biofilm production as phenotypic tests and genotypic analyses. Polystyrene mats presented 3.2 times greater chance of isolating Salmonella than canvas mats. Besides, we observed resistance to tetracycline (17.5%), ampicillin (10%), cefotaxime (7.5%), trimethoprim-sulfamethoxazole (5%), and chloramphenicol (2.5%). All strains possessed the invA, sipB, sipD, ssaR, sifA, sitC, iroN, tolC, flgK, fljB, and flgL genes. The genes sopB and sipA were both present in 92.5% of the isolates, while sopD and spvB were observed in 90% and 32.5% of strains, respectively. All strains adhered to and invaded HeLa cells. Regarding biofilm production, 31 (77.5%) strains were able to produce biofilm on polystyrene microplates. Using PFGE, we detected the persistence of clones in the environment for up to 18 fromthe 20 weeks. The ability of these strains to produce a biofilm and thus persist in the environment and disperse through contact surfaces in the processing plant favors the contamination of food, aggravated by the pathogenic potential of these isolates demonstrated by their adhesion capacity, invasion and resistance to various antibiotic agents.

Short communication: Investigation of extra-intestinal pathogenic Escherichia coli virulence genes, bacterial motility, and multidrug resistance pattern of strains isolated from dairy cows with different severity scores of clinical mastitis.

Escherichia coli is a major pathogen involved in the etiology of environmentally derived bovine mastitis and is characterized by a variety of virulence factors (VF). Mammary infections with E. coli have shown a wide range of clinical signs, causing changes in milk (score 1, or mild), abnormal appearance of milk and udder inflammation (score 2, or moderate), and abnormalities in milk, udder inflammation, and systemic signs of illness (score 3, or severe). Nevertheless, to date, the profile of the genes related to the virulence of the pathogen in mammary infections and the severity scores of cases have not been thoroughly elucidated. Therefore, a panel of 18 virulence-encoding genes associated with extra-enteric pathogenicity of E. coli (ExPEC) were investigated in addition to in vitro swimming and swarming motility profiles and antimicrobial susceptibility/resistance patterns among 114 E. coli strains isolated from cows with clinical mastitis and different severity scores. Of 114 clinical cases, 39.5, 54.4, and 6.1% were mild, moderate, and severe, respectively. The main genes related to VF harbored by isolates were adhesins (fimH 100%; ecpA 64.0%, fimA 31.6%), serum resistance (traT 81.6%; ompT 35.1%), siderophores (irp2 9.6%), and hemolysin (hlyA 7%). Among the isolates studied, 99.1% showed in vitro resistance to bacitracin and cloxacillin, and 98.2% to lincosamin. Of the total isolates, 98.2% were considered multidrug resistant based on the multiple antimicrobial resistance index. No significant difference was observed between mean swimming (13.8 mm) and swarming (13.5 mm) motility, as well as severity scores of clinical mastitis and the ExPEC genes studied. The isolation of strains resistant to various antimicrobials, even though tested only in vitro, highlights the importance of rational use of antimicrobials for mastitis treatment. The high prevalence of the genes related to serum resistance (traT and ompT) and adhesion (ecpA) of the pathogen, in addition to main associations between the genes fimH, ecpA, and traT among cows with severity scores of 1 (15%) and 2 (22.6%), indicates that the genes traT, ecpA, and ompT could be further studied as biomarkers of ExPEC for clinical intramammary infections. In addition, the ExPEC genes ompT (protectin), ibe10 (invasin), and ecpA (adhesin) were investigated for the first time among cows with mastitis, where scores of clinical severity were assessed. Results of this study contribute to the characterization of virulence mechanisms and antimicrobial resistance profile of ExPEC variants that affect dairy cows with different scores of clinical mastitis.