Short communication: Outbreak of methicillin-resistant Staphylococcus aureus (MRSA)-associated mastitis in a closed dairy herd.

Cows are probably the main source of contamination of raw milk with Staphylococcus aureus. Mammary glands with subclinical mastitis can shed large numbers of Staph. aureus in milk. Because of the risk of this pathogen to human health as well as animal health, the aim of this paper was to describe an outbreak of mastitis caused by methicillin-resistant Staph. aureus (MRSA), oxacillin-susceptible mecA-positive Staph. aureus (OS-MRSA), and methicillin-susceptible Staph. aureus (MSSA) on a dairy farm. Milk samples were obtained from all quarters, showing an elevated somatic cell count by the California Mastitis Test. The isolates were identified by phenotypic and genotypic methods. Staphylococcus spp. were isolated from 53% (61/115) of the milk samples, with 60 isolates identified as Staph. aureus (98.4%) and 1 isolate identified as Staphylococcus epidermidis (1.6%). The presence of the mecA gene was verified in 48.3% of Staph. aureus isolates. Of the Staph. aureus isolates, 23.3% were MRSA and 25.0% were OS-MRSA. The total of mastitis cases infected with MRSA was 12.2%. The detection of this large percentage of mastitis cases caused by MRSA and OS-MRSA is of great concern for the animals' health, because ß-lactams are still the most important antimicrobials used to treat mastitis. In addition, Staph. aureus isolates causing bovine mastitis represent a public health risk.

Detection of icaA, icaD, and bap genes and biofilm production in Staphylococcus aureus and non-aureus staphylococci isolated from subclinical and clinical bovine mastitis / Detecção dos genes icaA, icaD e bap e produção de biofilme por Staphylococcus aureus e estafilococos não-aureus isolados de mastite clínica e subclínica bovina

Algumas espécies de Staphylococcus causam infecções crônicas intramamárias e podem levar à formação de biofilme. No presente estudo, levantou-se a hipótese de que as espécies de Staphylococcus isolados da mastite bovina são capazes de formar biofilme in vitro associado à presença dos genes icaA, icaD ou bap. Um total de 200 isolados de Staphylococcus, sendo 100 Staphylococcus aureus de casos de mastite subclínica e 100 estafilococos não aureus (ENA) de casos de mastite subclínica e clínica, obtidos em duas fazendas leiteiras, no estado de São Paulo, foram avaliados quanto à capacidade de produzir biofilmes in vitro. A presença de icaA, icaD e bap foi confirmada por PCR, e a produção de biofilme em ágar vermelho congo (Congo Red Agar - CRA) e em teste de microplaca (Microtiter Plate - MtP) foi avaliada nos isolados de S. aureus e ENA. Os resultados mostraram a presença dos genes icaA, icaD e bap em S. aureus, mas não em ENA. A produção de biofilme pode estar associada à presença de outros fatores ou genes que estimulam a produção de biofilme in vitro. O ensaio de MtP serve como um modelo quantitativo para o estudo da aderência de espécies de estafilococos associados à mastite bovina.(AU)

Detection of Toxoplasma gondii DNA in the milk of naturally infected ewes.

Toxoplasmosis is the major parasitic disease affecting sheep. It is important for veterinary medicine, animal science and public health since it causes reproductive and economic losses in the herd, as well as damaging human health due to consumption of contaminated meat and milk, which can facilitate zoonotic transmission. Detection of Toxoplasma gondii in ovine milk and lack of data in the literature describing differentiation between acute and chronic disease for this species stimulated the elaboration of the present research project. To achieve the aim of this study, the animals were allocated to two groups of 20 ewes each, of which group 1 was composed of animals with positive serology and group 2 with negative serology. Acute and chronic stages of the disease were differentiated by modified direct agglutination test (MAT), in which antigens were fixed with formalin (MAT-AF) and methanol (MAT-AM). The parasite was detected in milk by polymerase chain reaction (PCR), and the molecular identity of the amplified products was confirmed by sequencing. The serological results indicated that sheep had a chronic infection profile. T. gondii DNA was detected in seven milk samples from five seropositive sheep, and twice in milk of two sheep. Sequences of species shared 97-100% identity with T. gondii. These findings allowed the hypothesis that the peripartum period may also lead to the resurgence of tissue T. gondii tachyzoites cysts which can circulate again and be excreted in the milk. This study used sheep naturally infected with T. gondii as a prerequisite for further investigations on the possible participation of this species in toxoplasmosis epidemiology and as a potential transmission route related to consumption of milk from infected sheep.

Road-killed wild animals: a preservation problem useful for eco-epidemiological studies of pathogens

Road-killed wild animals have been for years used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies. In the current study, fungal infection was evaluated by PCR and nested-PCR in tissue samples collected from 19 road-killed wild animals. The necropsies were carried out and samples were collected for DNA extraction. Results, using PCR with a panfungal primer and nested PCR with specific primers, indicated that some animals are naturally infected with Amauroascus aureus, Metarhizium anisopliae, Aspergillus flavus, Aspergillus oryzae, Emmonsia parva, Paracoccidioides brasiliensis or Pichia stipitis. The approach employed herein proved useful for detecting the environmental occurrence of several fungi, as well as determining natural reservoirs in wild animals and facilitating the understanding of host-pathogen relationships.