Disseminated paracoccidioidomycosis in a black lion tamarin (Leontopithecus chrysopygus) confirmed by histopathology, molecular diagnostics and phylogenetic analysis of DNA from FFPE tissues

First identification of Cryptosporidium meleagridis in wild white-eyed parakeet (Psittacara leucophthalmus).

In this research, we investigated the genetic diversity of Cryptosporidium spp. in 65 free-living white-eyed parakeets (Psittacara leucophthalmus) from the central-eastern region of São Paulo state, Brazil. Using real-time nested PCR of the 18S rRNA gene and conventional nested PCR of the actin and heat shock protein (HSP-70) genes, two samples were positive for Cryptosporidium spp., representing an prevalence of 3.08% (95% CI: 0.85-10.54%). However, oocysts of this protozoan were not observed through coproparasitological examination, nor were clinical signs characteristic of cryptosporidiosis noted in any bird. The amplicons obtained in this study were subjected to bidirectional Sanger sequencing. Among the positive samples, one presented sequences of sufficient quality for analysis of the 18S rRNA and HSP70 genes, exhibiting 100% and 99.64% genetic similarity, respectively, with Cryptosporidium meleagridis sequences deposited in GenBank. Phylogenetic analysis using HSP-70 gene sequences from Cryptosporidium spp. confirmed that the amplified Cryptosporidium sequence is closely related to C. meleagridis sequences, grouping within the same clade. Nested PCR targeting the parasite surface glycoprotein gene (GP60) was performed; however, a sufficient number of amplicons were not obtained, making subgenotyping impossible. We believe this to be the first report of C. meleagridis, a protozoan with zoonotic potential in the synanthropic white-eyed parakeets (P. leucophthalmus).

Molecular detection of Rickettsia rhipicephali and hemotropic Mycoplasma sp. in Dermacentor panamensis from a Mexican Hairy Dwarf Porcupine (Coendou mexicanus) in Veracruz, Mexico

Histopathological and molecular evidence of systemic Sarcocystis spp. infection in a free-living Guira Cuckoo (Guira guira)

Background: Sarcocystis species are protozoans that require intermediate and definitive hosts to complete their life cycle. Several species of Sarcocystis can infect birds and cause severe lesions. This study aimed to report the anatomopathological and molecular findings compatible with Sarcocystis spp. in guira cuckoos (Guira guira). Case Description: Two guira cuckoos were found dead and submitted for necropsy. Tissues were collected, fixed in 10% buffered formalin, and processed according to routine procedures. The sections were stained with hematoxylin and eosin. Nested polymerase chain reaction (PCR) targeting the 18s ribosomal ribonucleico acid region was performed to detect Sarcocystis spp., followed by digestion with restriction enzymes. Macroscopically, the lungs were edematous and dark red. One animal had partially digested cockroaches in the gizzard. Microscopically, diffuse pulmonary hemorrhage was associated with schizonts compatible with Sarcocystis. Schizonts associated with lymphocytic infiltrates were detected in the liver, skeletal muscle, and muscle tissues of the proventriculus. Encephalitis and gliosis were observed in the brain of one animal. Deoxyribonucleic acid amplification detected products compatible with Sarcocysts spp. in the tongue, heart, liver, pectoral muscle, brain, and esophagus. Conclusion: Together, the histopathological and molecular findings confirm systemic sarcocystosis in guira cuckoos and support their involvement in the local transmission cycle, with the presence of cockroaches suggesting a potential epidemiological link in parasite dissemination.

Mycobacterium shimoidei causing granulomatous inflammation in a captive bird species, the white-cheeked turaco

Mycobacteriosis in avian species is generally caused by Mycobacterium avium subsp. avium and M. genavense, although sporadic reports also exist of other nontuberculous mycobacterium species in birds. Here, we describe disseminated mycobacteriosis caused by M. shimoidei (a rare and infrequently reported pathogen in humans and animals) in a bird species. A captive, adult white-cheeked turaco (Tauraco leucotis) was presented because of persistent dyspnea unresponsive to treatment. The animal was treated unsuccessfully with antimicrobials and later euthanized on welfare grounds. At autopsy, granulomas were identified in the lungs and spleen. Histologically, the coalescing granulomas with central necrosis contained numerous chains of slender, slightly curved, 2-µm acid-fast bacilli, suggesting mycobacterial infection. Bacterial isolation confirmed Mycobacterium spp., which was identified as M. shimoidei by partial sequencing of the 16S rRNA and rpoB genes. Our report expands the known host range of M. shimoidei and highlights the importance of molecular testing methods in identifying atypical nontuberculous mycobacteria (NTM) in zoologic collections. Our case also underscores the need for enhanced epidemiologic surveillance of NTM infections in avian species, given their potential implications for both animal and public health.

A novel Candidatus Rickettsia chiangmaiensis and occurrence of Candidatus Rickettsia laoensis in hard ticks from Chiang Mai Province, Thailand.

Hard ticks are important vectors of zoonotic pathogens, including bacteria that cause rickettsiosis, Q fever, and anaplasmosis. In this study, 167 ticks were collected from vegetation in Chiang Mai Province, northern Thailand, and examined using morphological and molecular methods. Five tick species were identified: Dermacentor tamokensis (n = 81), D. bellulus (n = 49), D. pasteuri (n = 9), Ixodes acutitarsus (n = 26), and Haemaphysalis hystricis (n = 2), with the first three species newly characterized at the molecular level. Moreover, D. tamokensis and D. bellulus are newly recorded in Thailand. Phylogenetic analysis using the multilocus sequence analysis (MLSA) approach revealed two distinct genotypic groups of spotted fever group rickettsiae (SFGRs): a novel Candidatus Rickettsia chiangmaiensis in I. acutitarsus (20/26) and Candidatus Rickettsia laoensis in D. tamokensis (12/81) and D. bellulus (10/49). The latter SFGR was closely related to strains found only in western Thailand and Laos. Additionally, Coxiella-like endosymbionts (CLEs) were detected in D. tamokensis (61/81), D. bellulus (13/49), and H. hystricis (1/2), while Anaplasma sp. was found in D. tamokensis (2/81). Notably, double infections: Ca. R. laoensis-CLEs, Anaplasma-CLEs, and a triple infection: Ca. R. laoensis-CLE-Anaplasma were reported herein. These findings demonstrate the diverse bacterial associations in ticks, suggesting the ecological complexity of forested areas in northern Thailand.

First report of Angiostrongylus vasorum parasitizing lesser grison (Galictis cuja) (Mustelidae) in Brazil

Angiostrongylus vasorum is a metastrongylid nematode that parasitizes the pulmonary arteries and right heart of canids, being the etiological agent of canine angiostrongylosis, a disease of growing veterinary concern due to its expanding geographic distribution and the involvement of domestic and wild hosts. In Brazil, the parasite has been reported mainly in domestic dogs and some wild canids, and its occurrence in mustelids has been documented only once previously. This study reports the first occurrence of A. vasorum infecting a lesser grison (Galictis cuja) in Brazil. The mustelid was found after being accidentally run over by a vehicle on a highway in the south of the country. Clinical, physical, and complementary examinations were performed for clinical screening. In the parasitological examination of feces, nematode larvae were recovered and identified morphometrically and molecularly through conventional PCR targeting to the COX1 gene, followed by sequencing and phylogenetic analysis. The results confirmed the presence of A. vasorum, showing 99–100% identity with sequences of the same species deposited in GenBank. Therefore, we document here the first report of A. vasorum parasitizing G. cuja. Thus, the number of known host species parasitized by this helminth is expanded. Furthermore, we provide a new molecular sequence, contributing to the knowledge of this nematode in South America.