Nocardiosis in Cats - Clinical, Anatomopathological and Morphotintorial Characteristics

Background: Nocardiosis is an infectious bacterial disease that can cause cutaneous/ subcutaneous, pulmonary and systemic lesions in different species of domestic animals. The type of transmission occurs through mechanical lesions on the skin or contamination of wounds, in cases of skin involvement, inhalation of aerosols and ingestion of contaminated materials are involved in the pathogenesis of the respiratory and digestive form of the disease. This paper described 4 cases of nocardiosis in cats, addressing the clinical, anatomopathological and morphotintorial characteristics of Nocardia sp.Cases: Four cases of nocardiosis in cats were reviewed, in which data related to breed, sex, age, origin, clinical signs, macroscopic and histological lesions described in necropsy protocols were evaluated. The histological tissue sections stained with Hematoxylin and Eosin (HE) were evaluated in order to characterize the inflammatory response in each case. In addition, paraffin blocks of fragments from affected organ were selected to perform special histochemical staining techniques of Grocott Methenamine Silver (GMS), modified Ziehl-Neelsen, Gram Brown-Brenn and Giemsa stain which are the most characterized techniques used for histopathological diagnoses and it was also used an immunohistochemical test with polyclonal antibody anti-Nocardia sp. (non-commercial). The animals were adults of both sexes, mixed breed, not castrated and semi-domesticated. Neither immunosuppressive factors nor concomitant diseases were identified in the cases studied. The main clinical signs were apathy, anorexia, dehydration, phlegmon and draining tracts. Macroscopically, skin / subcutaneous tissue (3/4), skeletal muscle (2/4), lymph nodes (2/4), liver (2/4), omentum (1/4), spleen (1/4) were affected. In addition, it could be noted that mandibular bone (1/4), pleural tissue (1/4), left testicle (1/4) and Central Nervous System (CNS) (1/4) were also affected by this disease. Microscopically, regarding all cases, there was a pyogranulomatous inflammation in the affected organs. With respect to cases 1, 3 and 4, filamentous, branched, slightly basophilic structures in loose or individual aggregates in the interior of the pseudo-rosettes and in the necrotic areas were observed in the HE-stained tissue sections. In all cases submitted to special histochemical techniques, filamentous, branched, individual or loose aggregate structures were observed, the samples were impregnated with silver, and bacteria appear as blue using the Brown-Brenn Gram technique, and stained red in the modified Ziehl-Neelsen, and stained faintly pink in Giemsa stain. The bacteria were observed mainly in the border of the pyogranulomas, in the center of the pseudo-rosettes and in the necrotic areas, being compatible with the infection by Nocardia sp. All cases were positive for immunohistochemistry (IHC).Discussion: Nocardiosis was diagnosed in all cats in this study based on the anatomopathological findings associated with the visualization of the agent and its morphotintorial characteristics by using special histochemical stains and being confirmed by IHC. It occurs mainly in the cutaneous and/or subcutaneous tissues, with systemic involvement and death of the affected animals, in addition to affecting bone tissue considered an uncommon site for the disease. The diagnosis can be established based on the anatomopathological findings associated with the morphotintorial characteristics by using special histochemical stains, which are important for evidencing and morphologically characterizing the agent, as well as being confirmed by IHC.Keywords: disease in cat, pyogranulomatous inflammation, Nocardia sp.Título: Nocardiose em gatos - achados clínicos, anatomopatológicos e morfotintoriais Descritores: doença de gato, inflamação piogranulomatosa, Nocardia sp.

ANTIFUNGAL EXTRACT ACTIVITYOF ISOLATE Nocardia sp. ATS-4.1 AGAINST Candida albicans InaCC-Y116

Candida albicans is a microorganism that knows as caused of candidiasis. Nocardia is known to have the ability to produce antifungal bioactive compounds to overcome cases of fungal infections. This research aims to determine the presence of antifungal activity and the good concentration from crude extract of Nocardia sp. ATS-4.1 to inhibit C. albicans InaCC-Y116 and. Antifungal activity test of isolate Nocardia sp. ATS-4.1 was performed using a well diffusion method on Mueller Hinton Agar (MHA) media with a concentration of 92%, 94%, 96%, 98%, 100% and nystatin (positive control) 0,0125% and DMSO 10% (negative control) incubated at 37 ᵒC for 24-48 hours. The results showed that the extract of Nocardia sp. ATS-4.1 can inhibit the growth of C. albicans. Nocardia sp. ATS-4.1 isolate extract concentration of 96% with a resistance diameter of 13.63 ± 0.53 mm with a strong category against the growth of C. albicans InaCC-Y116.

RARE ACTINOMYCETES FROM KUANTAN MANGROVE FOREST SEDIMENT

Actinomycetes are prolific secondary metabolites producer, and they are sought after for their unparalleled capability. Mangrove forests are among the underexplored locations in search of new resources of actinomycetes. Bioprospecting of actinomycetes in Kuantan mangrove forests has revealed highly diverse actinomycetes with high antimicrobial properties. Members of the rare genera successfully isolated include Micromonospora sp., Pseudonocardia sp., Verrucosispora sp., Nocardiopsis sp., Actinophytocola sp., Dietzia sp., Gordonia sp., Micrococcus sp., Mycobacterium sp., Nocardia sp., Saccharopolyspora sp. And Rhodococcus sp. These rare actinomycetes can also be added to the list of genera isolated from this ecological niche, providing solid evidence that considerable diversity of actinomycetes are distributed within Kuantan mangrove forest. PKS-I and NRPS genes are usually related to the ability to produce secondary metabolites in actinomycetes. Interestingly, several of these rare actinomycetes showed the presence of both PKS-I and NRPS genes or either gene, and this exemplifies the potential of these rare actinomycetes may possess. Further studies conducted on these rare actinomycetes may reveal their true potentials that can be exploited for natural product discovery.

Actinomycetes from the Red Sea Sponge Coscinoderma mathewsi: Isolation, Diversity, and Potential for Bioactive Compounds Discovery

The diversity of actinomycetes associated with the marine sponge Coscinoderma mathewsi collected from Hurghada (Egypt) was studied. Twenty-three actinomycetes were separated and identified based on the 16S rDNA gene sequence analysis. Out of them, three isolates were classified as novel species of the genera Micromonospora, Nocardia, and Gordonia. Genome sequencing of actinomycete strains has revealed many silent biosynthetic gene clusters and has shown their exceptional capacity for the production of secondary metabolites, not observed under classical cultivation conditions. Therefore, the effect of mycolic-acid-containing bacteria or mycolic acid on the biosynthesis of cryptic natural products was investigated. Sponge-derived actinomycete Micromonospora sp. UA17 was co-cultured using liquid fermentation with two mycolic acid-containing actinomycetes (Gordonia sp. UA19 and Nocardia sp. UA 23), or supplemented with pure mycolic acid. LC-HRESIMS data were analyzed to compare natural production across all crude extracts. Micromonospora sp. UA17 was rich with isotetracenone, indolocarbazole, and anthracycline analogs. Some co-culture extracts showed metabolites such as a chlorocardicin, neocopiamycin A, and chicamycin B that were not found in the respective monocultures, suggesting a mycolic acid effect on the induction of cryptic natural product biosynthetic pathways. The antibacterial, antifungal, and antiparasitic activities for the different cultures extracts were also tested.

AKTIVITAS ANTIBAKTERI ACTINOMYCETES ASAL DESA CEMPAKA KAPUAS HULU KALIMANTAN BARAT TERHADAP ENTEROPATOGENIK GASTROENTERITIS

AbstrakAntibiotik yang digunakan secara tidak tepat dapat meningkatkan jumlah bakteri penyebab gastroenteritis yang resisten terhadap antibiotik. Beberapa kelompok mikroorganisme tanah diketahui memiliki potensi menghasilkan senyawa aktif untuk menghambat dan membunuh bakteri patogen, seperti Actinomycetes. Tujuan dari penelitian ini adalah untuk mengetahui potensi antibiotik Actinomycetes terhadap bakteri penyebab gastroenteritis yang diisolasi dari sawah tandus di Desa Cempaka Baru, Kabupaten Kapuas Hulu, Kalimantan Barat. Proses penelitian dilakukan dari isolasi Actinomycetes dari sawah tandus, karakterisasi, serta pengujian kemampuan antibiotik terhadap bakteri Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028 dan Staphylococcus aureus ATCC 25923 menggunakan metode sumuran pada media Mueller Hinton Agar. Hasil identifikasi berdasarkan karakteristik morfologi koloni, sel, dan biokimia diperoleh 1 isolat bakteri, yaitu genus Nocardia sp. ATS-4.1 yang mampu menghambat pertumbuhan bakteri-bakteri uji yang dibuktikan adanya rata-rata zona hambat yang terbentuk, yakni 14,51 mm; 16,16 mm; dan 11,10 mm. Hasil uji statistik Friedman diperoleh nilai Asymp. Sig 0,10>0,05, sehingga menunjukkan bahwa pemberian cairan kultur isolat Nocardia ATS-4.1 memberikan pengaruh yang tidak berbeda nyata antara hambatan terhadap bakteri E. coli, S. enterica, dan S. aureus. Hal ini dapat disimpulkan bahwa isolat Nocardia sp. ATS-4.1 berpotensi menghasilkan antibakteri yang setara terhadap bakteri E. coli, S. enterica, dan S. aureus penyebab gastroenteritis.Abstract Using antibiotics incorrectly increases the number of resistant bacteria to gastroenteritis. Soil microorganisms are known to have the potential to produce active compounds to inhibit and kill pathogenic bacteria, for example, Actinomycetes. The purpose of this study was to determine the potential of antibiotics Actinomycetes isolated from rice fields in Cempaka Baru, Kapuas Hulu Regency, West Kalimantan, against gastroenteritis bacteria. The research order was carried out by isolating Actinomycetes from barren rice fields, and then characterizing and testing the ability of antibiotics against Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028, and Staphylococcus aureus ATCC 25923 using the well method on Mueller Hinton Agar. The identification results based on morphological characteristics of colonies, cells and biochemistry showed that one of the bacteria was from genus Nocardia sp. ATS-4.1 which was able to inhibit the three test bacteria by the average inhibition zone 14.51 mm; 16.16 mm; and 11.10 mm, respectively. Friedman’s statistical test resulted in values asymp. sig 0.10>0.05 which showed that the isolate gave inhibition differences insignificantly among the bacteria. In conclusion, isolate Nocardia sp. ATS-4.1 able to produce equal antibacterial activity against bacteria E. coli, S. enterica and S. aureus caused gastroenteritis.